Horticulture

The explanation that I find most likely is a reduction in water intake. Weight loss can indicate a reduction in fat mass or lean mass, but it can also indicate a reduction in water mass, or dehydration. The food in the experimental condition was purposely designed to provide sufficient calories while being lower in mass and volume than everyday food. One of the best ways to create energy-dense food is to reduce water content . Consequently, the food in the experimental condition had lower water content than the food in the control condition. It is therefore likely that participants consumed less water from food during the experimental condition, and that they did not sufficiently make up for this difference by drinking more fluids. This hypothesis is supported by the results of the post hoc analyses, described below. In the post hoc analyses, it was found that participants in the experimental condition reported consuming 2,800.82 total grams of food + fluid each day, whereas participants in the control condition reported consuming 3,180.42 total grams of food + fluid each day. Therefore, participants in the experimental condition reported consuming an average of 379.6 grams less over the course of a day, even though they did not consume fewer macro-nutrients .

Aside from macro-nutrients,container vertical farming the weight of food usually results from a substantial amount of water and a small amount of fiber and alcohol, along with a relatively small amount of ash . Consequently, it appears likely that participants in the experimental condition consumed less water. This would not be surprising, given that the experimental food was designed to be energy dense. In support of this theory, participants in the experimental condition lost an average of 2.157 pounds, or 1.25% of their body weight, which can easily occur from dehydration. Some researchers have defined mild dehydration as a 1-2% decrease in body weight, moderate dehydration as a 2-5% decrease in body weight, and severe dehydration as a 5% or greater decrease in body weight . It is possible that during the experimental condition, participants consumed less water from food and fluids, resulting in mild dehydration that contributed a 1.25% decrease in body weight. For comparison, when participants were in the control condition they lost 0.04% of their body weight. If participants were, indeed, mildly dehydrated during the experimental condition, is this a cause for concern? During the experimental condition, participants consumed approximately 2,410.82 grams from non-energy-yielding sources . Even if the ash and fiber comprised 15% of this mass, participants would have consumed over 2,000 grams of water – the minimum intake required by NASA, based on participants’ average caloric intake in this study .

Additionally, decrements in cognitive or physical performance usually do not occur until individuals have lost 2% or more of their body weight . Indeed, during the experimental condition, participants showed no significant changes in psychological or physical outcomes other than decreased weight. These factors would indicate that any mild dehydration experienced by participants is little cause for concern. However, although NASA’s ISS requirements stipulate astronauts should receive at least 2 liters of water a day, the Institute of Medicine recommends more than that . Furthermore, numerous factors make it impossible to determine whether sufficient water was consumed in the experimental condition: participants’ fluid reports were questionable, participants were allowed to consume caffeine, and participants did not record their exercise. Nevertheless, at present, the data do not indicate participants were worrisomely dehydrated while eating the experimental diet. In future studies of commercial space food, incorporating measures that can differentiate between fat mass and lean mass and measures that can assess changes in dehydration would help to determine the cause of weight loss.NASA’s ISS requirement is that astronauts be provided with 12-15% of their calories from protein, 30-35% of their calories from fat, and 50-55% of their calories from carbohydrates . The Institute of Medicine sets forth adequate macro-nutrient proportions of 10-35% of calories from protein, 25-35% of calories from fat, and 45-65% of calories from carbohydrate. During the experimental condition, participants met both NASA and the Institute of Medicine’s recommended macro-nutrient proportions.

During the control condition, participants exceeded NASA’s protein and fat requirements, and failed to meet the lower limit of NASA’s carbohydrate requirement. During the control condition, participants also consumed fat in excess of the Institute of Medicine’s recommendations. In summary, during the experimental condition, participants consumed macro-nutrients in the recommended proportions, whereas during the control condition they did not.Participants expressed a desire for more animal protein, indicating that satisfaction with the diet may increase if more animal protein were added. Increasing the amount of animal protein in the diet could also assist with assembling a low fiber diet, which I aimed to achieve but which was difficult to attain, as many “healthy” prepackaged foods are high in fiber. Increasing the amount of protein in the experimental diet would not necessarily be unhealthy, as 14.5% of calories in the diet came from protein, and the Institute of Medicine’s upper limit for protein is 35% . However, NASA has an upper limit of 15% because a high protein diet, especially one from animal sources, could lead to hypercalcemia and increased risk of developing renal stones . Renal stones are especially a concern for astronauts as physiological changes during spaceflight may already lead to increased risk of renal stone formation . Consequently, there is rationale for keeping protein intake relatively low. As discussed in Table 1, given that many of the foods in the experimental diet contained a mix of animal and vegetable proteins, and that I did not have access to a calorimeter, it was not possible for me to calculate the experimental diet’s percentage of protein from animal vs. vegetable sources. However, based on a visual analysis of the types of food in the experimental diet, it is possible that diet did not meet NASA’s recommendation to have 2/3 of protein come from animal sources . It therefore seems advisable to at least shift away from plant-based protein bars and toward more substantial animal sources, such as peppered beef jerky. Various brands and varieties should be investigated to identify a meat source that is tender, salty, spicy, and substantial enough to provide a “meal-like” sensation,hydroponic vertical garden without exceeding the sodium limit and without having small pieces that could flake off and float away in a microgravity environment.NASA only includes items that score, on average, 6.0 or higher on the 9-point Hedonic Scale. In this study, participants rated 50% of the experimental diet items as 6.0 or higher. Because these items were so well liked, I recommend keeping these items in the diet. See all green-colored items in Table 10 for a list of these foods. Nearly one-third of the items in the experimental diet were rated between 5.0 and 5.9. These foods were rated below NASA’s acceptable score of 6.0 and their elimination should therefore be considered. However, it’s not clear that participants’ ratings in this study were equivalent to NASA’s food ratings because in this study, participants ate and rated the food in a naturalistic setting, rather than in a laboratory. Research has shown that the environment can greatly impact food ratings; individuals rate food differently depending upon whether they are in a laboratory, a restaurant, or a cafeteria . In this study, participants may have eaten in the cafeteria with colleagues, alone at their desk, or at home.

Sometimes, when they ate, they saw just their own food, and atother times they saw a plethora of freshly prepared food that their colleagues were eating, yet which they could not eat. Consequently, it is not clear how the multiple environments in this study should be categorized and how participants’ ratings would have been different, had they been doing a taste test in a laboratory. Furthermore, some items that received a mean rating between 5.0 and 5.9 were controversial, meaning that some participants loved them and some hated them. Consequently, when designing future versions of the experimental diet, I recommend that these 5.0- to 5.9- scored items be put back into the pool of options for consideration. Instead of eliminating all these items, perhaps reducing their frequency would be wiser. For instance, all three noodle/potato entrées received mean ratings between 5.0 and 5.9. Yet one participant specifically requested more of these items, indicating a strong preference for them. Keeping just one noodle entrée in the menu may be a better choice than eliminating them completely. However, eliminating all 10 items that were rated lower than 5.0 is advised. Because participants were provided with over 3,000 calories a day, these 10 items could be completely removed from the diet while still providing over 2,400 calories per day – well over the 1,810.71 calories that participants, on average, consumed each day of the experimental condition. Removing these 10 items from the diet would still provide crew with 38 different food items for their 4-day trip and would, if the ratings for the remaining food items did not change, raise the mean food rating of the diet to 6.18 on the Hedonic Scale. All experimental food had to be eaten at room temperature. Room temperature food is less odiferous than heated food, and I had therefore anticipated the smell of food would not be a strong factor in food choice. Consequently, participants were not asked to rate the smell of foods. However, the strong odor of certain foods became apparent to me while evaluating leftovers. This experience highlighted a major difference between the study environment and the Crew Transportation Vehicle: in the study, the open-air setting likely muted the smell of foods, whereas in the enclosed Crew Transportation Vehicle, the smell of foods would be easily noticeable. This is a concern because smells could exacerbate the symptoms of space motion sickness, which is commonly experienced by astronauts during early flight. Former astronauts have recalled that, “Even if the sick people wisely stuck to broth and crackers, odors from the meals being enjoyed by those fortunate enough to feel better could be overwhelming. Some commanders made the law of ‘no stinky food on day 1 or 2.’ That precluded such odiferous items as tuna, salmon, or beef with barbeque sauce” . It would therefore be wise to remove strongly odiferous foods from the diet. The odor of foods could be evaluated in the same way that materials used in spaceflight are evaluated, by having panelists rate the smell from 0 = “undetectable” to 4 = “revolting” . Foods that are rated 2.5 or higher could be eliminated from the diet . Another factor that became quickly apparent was the large variation in participants’ food preferences. Some participants craved the noodle entrées whereas others said the noodles were inedible. Food choice is impacted by not only the qualities of the food itself, but also demographic , sociocultural, and economic variables . When NASA initially designed the Space Shuttle menu, a standard menu was developed for all missions . After four missions, the menu was changed to be crew-selected, so that crew could provide input but all crew members were ultimately provided with the same food. However, crew members expressed a desire for even more personalized menus, and consequently starting with the seventh Space Shuttle mission , individualized menus were provided for each crew member. Crew members were allowed to select the food they wanted from a menu of over 350 food items. Crew members’ choices were then analyzed by a dietitian, who would recommend substitutions to obtain a balanced diet. Based on the large variation in food preferences observed in this study, it seems the individualized menu would be better for achieving high food satisfaction than the standard menu. However, most commercial aerospace companies hope to fly not just eight astronauts a year, but hundreds. Personalizing the diet for each astronaut could become onerous. Instead, I recommend letting crew members pick the food they want from a collection of 100+ food items. Each crew member could be provided with a container and be allowed to select whatever food they wanted as long as it fit in that container and met certain mass restrictions.

The sclerotia of Polyporus umbellatus Fries is widely used to promote urination and prevent dampness.Our previous studies systematically demonstrated that ergosta‐4,6,8,22‐tetraen‐3‐one isolated from Polyporus umbellatus Fries showed significant antitumor, diuretic, and renoprotective effects.Our recent study demonstrated that ergone inhibited NF‐κB signaling and α‐SMA expression in 5/6 nephrectomised and unilateral ureteral obstruction rats.The Wnt signaling pathway is an evolutionarily conserved developmental signaling cascade that plays a critical role in regulating organ development and tissue homeostasis. Leucine‐rich repeat‐containing G protein‐coupled receptor 5 is a novel functional marker in glioma stem cells that promotes EMT by activating the Wnt/β‐catenin signaling pathway.Sry‐like high‐mobility group box 8 regulated cancer stem‐like properties and cisplatin‐induced EMT by the Wnt/β‐catenin signaling pathway in tongue squamous cell carcinoma.In addition,stackable planters bone marrow mesenchymal stromal cells suppressed EMT by inhibiting the Wnt/β‐catenin signaling pathway in silica‐induced pulmonary fibrosis.

Our previous study showed the activation of the canonical Wnt/β‐catenin signaling pathway accompanied by the upregulation of proinflammatory and pro‐oxidative protein expression in the NF‐κB signaling pathway and downregulation of the anti‐inflammatory Nrf2 signaling pathway in patients with chronic kidney disease compared with the Nrf2 signaling pathway in healthy controls.Many compounds inhibit the Wnt signaling pathway, and these compounds may suppress EMT in cancer and fibrosis .FH535, a β‐catenin/Tcf inhibitor, not only increased radio‐sensitivity but also suppressed EMT in the radio resistant KYSE‐150R esophageal cancer cell line, which indicated that inhibitors of the Wnt signaling pathway might be effective anticancer agents with the potential to be anticancer drugs.In addition, it was reported that FH535 alleviated multiple types of cancer, including colorectal cancer, gastric cancer, and hepatocellular carcinoma.Isoquercitrin, a bio-active flavonoid from Bidens bipinnata L., inhibited the Wnt/β‐catenin signaling pathway and hepatocyte growth factor/scatter factor‐induced EMT in NBT‐II cells.In addition, salinomycin was previously used as an antibiotic and also showed significant anticancer activity by suppressing EMT.Salinomycin‐inhibited EMT by suppressing the Wnt/β‐catenin signaling pathway in epithelial ovarian cancer cells, indicating that small molecules targeting the Wnt/β‐catenin signaling pathway might have anticancer properties by reversing EMT.Poria cocos is a well‐known fungus that exhibits an effective therapeutic effect to improve kidney function in clinic.Our previous studies have confirmed that extracts of the surface layer of Poria cocos show remarkable antihyperlipidemic, diuretic, and renoprotective effects.

Recently, our group isolated more than 90 triterpenoid compounds from the surface layer of Poria cocos, some of which exhibited significant anti-fibrotic properties.New triterpenoids, including poricoic acid ZC, poricoic acid ZD, and poricoic acid ZE, significantly down regulated the expression of Wnt1, active β‐catenin, Snail, Twist, MMP‐7, PAI‐1, and FSP1 in HK‐2 cells induced by TGF‐β1 and angiotensin II and mice with unilateral ureteral occlusion. Moreover, new triterpenoids, including poricoic acid ZG and poricoic acid ZH, improved renal fibrosis by targeting the phosphorylation of Smad3 signaling and the Wnt/β‐catenin signaling pathwa . Alismatis rhizome , the dried rhizome of Alisma orientale Juzep, exhibited diuretic, antihyperlipidemic, and renoprotective effects that were also confirmed by our previous studies.Our recent study demonstrated that triterpenoid was the main component of AR,and further study showed that the novel tetracyclic triterpenoid 25‐O‐methylalisol F inhibited EMT by suppressing the Wnt/β‐catenin signaling pathway as well as the phosphorylation of Smad3 signaling in both NRK‐52E and NRK‐49F cells . Additionally, it was also observed that ergone inhibited extracellular matrix accumulation in HK‐2 cells and attenuated podocyte injury through inhibiting the activation of the Wnt/β‐catenin signaling pathway induced byangiotensin II.Taken together, these data indicate that tetracyclic triterpenoid and steroid compounds show significant anti-fibrotic properties in renal fibrosis.Akt is a serine/threonine‐specific protein kinase that plays a key role in various cellular activities. Many cellular signals are transduced through the Akt signaling pathway, and Akt is involved in EMT in cooperation with other proteins. For example, tripartite motif‐containing 14 is an oncogene that regulated EMT and the metastasis of human gastric cancer by activating Akt signaling.M3 muscarinic acetylcholine receptors regulated EMT, perineural invasion, and metastasis in cholangiocarcinoma via the Akt pathway.

Canopy homologue 2 promoted EMT via activating the Akt/GSK3 pathway in non–small‐cell lung cancer.In addition, Akt also promoted EMT cooperation with miRNAs. MiR‐944 inhibited EMT and the metastasis of gastric cancer by the metastasis‐associated in colon cancer 1 /Met/Akt signaling pathway.MiR‐1296 inhibited EMT and the metastasis of hepatocellular carcinoma via the serine/threonine‐protein kinase 1/PI3K/Akt signaling pathway.Taken together, these data suggest that Akt plays a key role in EMT cellular signal transduction and that targeting Akt might be an effective approach to treat fibrosis and tumors . Simvastatin is a 3‐hydroxy‐3‐methylglutaryl coenzyme A inhibitor that was originally used to treat cardiovascular diseases. Recently, it was reported that simvastatin had a therapeutic effect in several cancers. The administration of simvastatin suppressed EMT through the phosphatase and tensin homologue /PI3K/Akt pathway in EC9706‐R cells, suggesting a new therapeutic function for simvastatin in cancers.Trichostatin A, a histone deacetylase inhibitor, is an antifungal antibiotic. In addition to its antibiotic properties, trichostatin A alleviated EMT in bleomycin‐ induced lung injury in mice via inhibiting the Akt signaling pathway.Ubenimex inhibits multiple proteases, including arginyl aminopeptidase, leukotriene A4 hydrolase, alanyl aminopeptidase, and leucyl/cysteinyl aminopeptidase, a membrane dipeptidase used to treat acute myelocytic leukemia and lymphedema. Ubenimex alleviated acquired sorafenib resistance in renal cell carcinoma via suppressing the Akt pathway.Luteolin is widely distributed in plants and has shown anti inflammatory, antioxidant, antimicrobial, and antitumor properties. Luteolin attenuated TGF‐β1‐induced EMT by mediating the PI3K/Akt/NF‐κB‐Snail pathway in lung cancer cells. In addition, luteolin attenuated the progression of gastric cancer by reversing EMT via the Notch signaling pathway.Moreover, luteolin also suppressed the metastasis of triple‐negative breast cancer via blocking EMT by the down regulation of β‐catenin.Furthermore, luteolin suppressed EMT by down regulating the expression of cyclic AMP‐responsive element binding protein 1175 in colorectal cancer cells.Collectively, luteolin showed therapeutic effects in both tissue fibrosis and tumors through various signaling pathways and might be a promising candidate to treat fibrosis and tumors. α‐Mangostin derived from the pericarp of the mangosteen fruit has been shown to have various cellular functions, such as arresting the cell cycle, inhibiting cell viability, inducing apoptosis, and differentiation, reducing inflammation and decreasing adhesion.

α‐Mangostin suppressed viability and EMT by downregulating the PI3K/Akt pathway in pancreatic cancer.Icaritin, a hydrolytic product of icariin that is isolated from members of the Epimedium genus, induced the trans‐differentiation of embryonic stem cells into cardiomyocytes, prevented steroid‐ associated osteonecrosis and stimulated neuronal differentiation.Icaritin inhibited invasion and EMT via targeting the PTEN/Akt/HIF‐1α signaling pathway.As a highly conserved cell signaling system, the Notch signaling pathway is widely involved in cell proliferation and differentiation during embryonic and adult development. Extensive studies have demonstrated that the Notch signaling pathway is also critical for EMT in tumorigenesis and fibrogenesis. The over expression of Notch1‐induced EMT in PC‐3 cells, and claudin‐1 contributed to EMT by the Notch signaling pathway in human bronchial epithelial cells.In addition, Notch and TGF‐β1 generated a reciprocal positive regulatory loop and cooperatively regulated EMT in epithelial ovarian cancer cells,stackable flower pots which provided new insight into the mechanism of EMT.Moreover, miR‐34a downregulation induced by hypoxia enhanced EMT via the Notch signaling pathway in tubular epithelial cells, which indicated that the Notch signaling pathway is critical for EMT during fibrosis.These cases suggest that a targeted intervention in the Notch signaling pathway might be an effective strategy to treat cancer and fibrosis. DAPT, a γ‐secretase inhibitor, decreased the expression of Snail and vimentin and increased E‐cadherin expression in two oral squamous cell carcinoma cell lines, Tca8113 and CAL27, indicating that the targeted inhibition of the Notch signaling pathway might be a new therapeutic strategy to treat cancer. Another γ‐secretase inhibitor, RO4929097, not only inhibited EMT, invasion, and metastasis in cervical cancer HeLa and CaSki cells but also exerted significant therapeutic effects in patients with recurrent malignant glioma, cervical and colon cancer and advanced solid tumors in clinical trials.3,6‐Dihydroxyflavone is ubiquitous in vegetables and fruits and blocks EMT in breast cancer cells through suppressing the Notch signaling pathway.Moreover, luteolin inhibited EMT in gastric cancer by the Notch signaling pathway.In addition, emodin is a main bioactive component of Polygonum cuspidatum that suppressed EMT in alveolar epithelial cells via the Notch signaling pathway. Therefore, it is a promising prospect in treating pulmonary fibrosis.Furthermore, berberine was isolated from Berberis vulgaris and reversed EMT by blocking the Notch/Snail signaling pathway in mice with diabetic nephropathy.The RAS signaling pathway not only regulates blood pressure and fluid balance but also is involved in many kinds of diseases, including cancer and fibrosis. Recently, emerging evidence has suggested that the RAS signaling pathway plays a key role in EMT during fibrosis and tumorigenesis. Angiotensin II promoted EMT by the interaction between hematopoietic stem cells and the stromal cell‐derived factor‐1/CXR4 axis in intrahepatic cholangiocarcinoma.In addition, the overexpression of angiotensin II type 1 receptor induced EMT and promoted tumorigenesis in human breast cancer cells, and the silencing of angiotensin II type 1 receptor suppressed EMT that was induced by high glucose through inactivating the mTOR/p70s6k signaling pathway in the human proximal tubular epithelial HK‐2cell line.

Based on these results, it was suggested that suppressing RAS might be an effective therapy to treat cancer and fibrosis. Losartan is an AT1R antagonist that improved renal fibrosis by suppressing EMT in rats with hyperglycemia.Although many RAS inhibitors showed beneficial effects in tumors and fibrosis, few were reported to inhibit EMT in the treatment of cancer and fibrosis.MiRNAs are endogenous small noncoding RNAs that bind the 3′‐untranslated region of messenger RNAs to regulate gene expression. Recently, many miRNAs have been found to promote or suppress EMT in fibrosis and tumors. For example, miRNA‐497/Wnt3a/c‐Jun regulated growth and EMT, and miR‐497 served as a tumor suppressor in glioma cells.Moreover, miR‐205 inhibited tumor growth, invasion, and EMT via targeting semaphorin 4C in hepatocellular carcinoma.Furthermore, the upregulation of miR‐183‐5p induced apoptosis and inhibited EMT, proliferation, invasion, and migration by the downregulation of ezrin in human endometrial cancer cells.Collectively, EMT is inhibited by many miRNAs, including miR‐145, miR‐497, miR‐145‐5p, miR‐138, miR‐200a, miR‐200b, miR‐655, miR‐30‐5p, and miR‐32.In addition, EMT is also promoted by many miRNAs, including miR‐221, miR‐222, miR‐214‐3p, and miR‐181a.The flavonoids rhamnetin from cloves, berries and cirsiliol from Cirsium lineare Sch.‐Bip. showed anti inflammatory and antitumor properties. Both rhamnetin and cirsiliol induced radio‐senitization and inhibited EMT by miR‐34a/Notch1 signaling in non–small‐cell lung cancer cells.Quercetin alleviated TGF‐β1‐induced fibrosis in HK‐2 cells via downregulating miR‐21 expression and upregulating PTEN and TIMP metallopeptidase inhibitor 3 expression.Sophocarpine from Sophora alopecuroides L. inhibited tumor progression and reversed EMT by targeting miR‐21 in head and neck cancer.In addition, sophocarpine exerted a profound antitumor effect through inhibiting EMT induced by TGF‐β. Zerumbone from Zingiber zerumbet Smith exhibited anti‐inflammatory and anticancer properties. Recently, it was reported that zerumbone inhibited the β‐catenin pathway via miR‐200c to block EMT and cancer stem cells.In addition, nicotine upregulated FGFR3 and RB1 and promoted EMT by the downregulation of miR‐99b and miR‐192 in non–small‐cell lung cancer cells.50 Moreover, osthole alleviated EMT‐mediated metastasis by inhibiting miR‐23a‐3p.Furthermore, resveratrol inhibited proliferation, invasion, and EMT via the upregulation of miR‐200c in HCT‐116 colorectal cancer cells.RNA‐binding proteins such as RNA‐binding Fox protein 2 , epithelial splicing regulatory protein 1 , and ESRP2 control the splicing of many gene transcripts and splice nascent RNAs to functionally and structurally different miRNAs to regulate the process of EMT. For example, bleomycin inhibited ESRP1 expression, leading to the increased alternative splicing of FGFR2 to its mesenchymal isoform IIIc, which induced EMT in lung fibrosis.In addition, over expressed ESRP1 contributed to EMT in ovarian cancer, inducing a cell‐specific variant of CD44 and a protein‐enabled homologue.Moreover, Rbfox2 was upregulated during the EMT, and the depletion of Rbfox2 suppressed the expression of mesenchymal marker genes.Several studies have revealed that small molecules regulate the expression of RNA‐binding proteins to mediate EMT. For example, caffeine, an alkaloid in tea and coffee, reduced p53α expression and upregulated p53β expression through altering the expression of serine/arginine‐rich splicing factor 3 to regulate EMT.Both tissue fibrosis and tumors lead to high morbidity and mortality worldwide; thus, effective therapeutic strategies are urgently needed. Mounting studies have demonstrated that EMT plays a critical role in fibrosis and tumors, suggesting that drugs targeting EMT may be an effective therapy against fibrosis and tumors. Although TGF‐β1 is a potent inducer of EMT, new targets are needed due to the controversial role of TGF‐β1, which has been shown to have multiple beneficial roles in various bio-activities. As summarized above, myriad mediators, including many transcription factors , signaling pathways , RNA‐binding proteins and miRNAs, regulate EMT.

To determine acute mortality via oral exposure, A. pisum were treated by being individually dipped in a particular pesticide solution. They were removed from the solution with a fine paint brush, and placed on a paper towel in a fume hood until excess pesticide had dripped off and the aphids had dried sufficiently and were able to crawl. 10 topically-treated adult A. pisum were placed in a test arena, and a single untreated H. convergenswas added and allowed to feed for a period of 48 h. To test the effects of residual exposure, individual arenas were treated by adding 4 ml of a given pesticide solution and turning 360 degrees in order to coat the entire inside surface before the solution was poured out. Vials were inverted, suspended on a rack, and allowed to dry in a fume hood for a period of 24 h. Approximately 10 untreated adult A. pisum were then added as a food source to the treated arenas. A single untreated H. convergenswas added per arena and allowed to crawl on the treated surface for a period of 48 h. For topical exposure,hydroponic fodder system individual H. convergens were placed in an 8.5 cm petri dish with filter paper lining and treated in a Potter spray tower set to 68.9 kPa. For each pesticide solution, the spray volume per application was 1.4 ml, resulting in a spray deposit of 2.50 mg cm-2 .

This deposit is similar to that used in other studies on non- target insects as well as by the IOBC Working Group “Pesticides and Beneficial Organisms” . The treated insects were allowed to dry, and were placed individually into an untreated arena with 10 untreated A. pisum adults as a food source for a period of 48 h. The statistical program R was used for all analyses. Generalized linear models with binomial errors, or quasibinomial errors in cases where there was over dispersion, were used to analyze the effects of pesticide, pesticide concentration , life stage , and exposure route on the acute mortality of H. convergens. For all GLMs, model reduction and log likelihood ratio tests were used to assess the statistical significance of each factor and their interactions on acute mortality at α = 0.05. In a first analysis, the uncorrected acute mortality data were analyzed. Starting with a full model that included all four factors and interactions, the data set was finally subdivided into eight separate models with no significant interaction between factors to compare the relative toxicity of the different pesticides tested. From these models, the pesticides and pesticide concentrations with similar effects on acute mortality were grouped , and their toxicity was characterized according to the criteria developed by the International Organisation for Biological Control for classifying pesticide selectivity on beneficial insects in a laboratory bio-assay: <30% mortality is harmless, 30-79% mortality is slightly harmful, 80-99% mortality is moderately harmful, and mortality > 99% is harmful . Although uncorrected mortality was used in our analysis, the IOBC classification remains applicable due to the fact that control mortalities were low. This classification scheme has been widely adopted for the comparison of differential toxicities of pesticides to natural enemies in a number of laboratory studies .

In a second analysis, all acute mortality data were first corrected to account for variability among control groups , and then used to examine comparative effects of life stage and route of exposure. An initial full model included all four factors, pesticide, pesticide concentration, life stage and exposure route, and their interactions as categorical variables. Due to significant interactions, the corrected data set was then further separated by pesticide and pesticide concentration to examine the effects of life stage and exposure route on acute mortality of H. convergens. Additionally, the interaction for effects between pesticide and gender were tested for adults. Of the seven pesticides tested, only lambda-cyhalothrin and cyantraniliprole were either moderately harmful or harmful, causing greater than 80% mortality. Lambda cyhalothrin was either moderately harmful or harmful to both life stages of H. convergens when topically exposed at the 100% concentration or when residually exposed at either concentration. However, for oral exposure at both concentrations, it was moderately harmful or harmful to larvae, but not to adults. Cyantraniliprole was moderately harmful to larvae, but not to adults at the 100% concentration when orally exposed, but it was harmless or slightly harmful at the 10% concentration and for other exposure routes. Chlorantraniliprole, copper+mancozeb, novaluron, spinetoram, and sulfur were either harmless or slightly harmful to both life stages at both concentrations, regardless of exposure route. The acute toxicities of the seven pesticides to H. convergens in this study were classified according to guidelines initially established by the IOBC for laboratory bio-assays due to the widespread use of this classification.

However, a more recent revision of these guidelines for classifying pesticide selectivity may more accurately describe their potential toxicity to natural enemies in the field . The revision suggests reducing the number of categories from four to three; the harmless category remaining unchanged at less than 30% acute mortality, pesticides responsible for 30-79% acute mortality being changed from slightly to moderately harmful, and the top two categories of 80-99% and > 99% acute mortality being combined into a single harmful category. Although these revised categories have yet to be widely adopted, they offer a valuable improvement over the original classification as LC50 values would no longer be considered only slightly harmful and the artificial separation of > 99% mortality as a distinct category would be dropped. Under these revised guidelines, lambda-cyhalothrin would be considered harmful rather than moderately harmful to larvae for both concentrations and all three routes of exposure, and to adults at both concentrations for residual exposure and at the 100% concentration for topical exposure. Additionally, lambda-cyhalothrin would change from slightly harmful to moderately harmful for adults orally exposed to the 100% concentration and for adults topically exposed to the 10% concentration. Cyantraniliprole would also be reclassified from slightly harmful to moderately harmful at the 100% concentration for adults orally exposed and for larvae topically exposed, and from moderately harmful to harmful for larvae orally exposed to the 100% concentration. Spinetoram and novaluron would also change from slightly harmful to moderately harmful for topical exposure of larvae at the 100% concentration. Copper+mancozeb and sulfur would remain in the harmless category for all life stages, concentrations, and exposure routes. All further discussion of toxicity will be used in reference to the revised classification system. For corrected acute mortality, life stage had no significant effect for chlorantraniliprole, cyantraniliprole, spinetoram and sulfur at the 100% concentration or for copper+mancozeb at both concentrations. The most pronounced life stage effects were for cyantraniliprole, lambdacyhalothrin, novaluron, and spinetoram at the 100% concentration, and with larvae being more susceptible than adults in all cases. Similarly, novaluron has been shown to have a significant acute effect on larvae of C. carnea, but not on adults . In contrast, while sulfur had limited effects on both adults and larvae of H. convergens in this study,hydroponic nft gully for G. occidentalis, it had significant effects on larvae while being harmless to adults . However, immature life stages are not always more susceptible than adults to pesticide exposure. For instance, phosmet and dimethoate were harmless to eggs and pupae of C. carnea, but harmful to adults .

For instance, spinosad was harmless to both adults and nymphs of the predatory mirid bugs Macrolophus pygmaeusand Nesidiocoris tenuis, while indoxacarb was harmless to M. pygmaeus nymphs, but moderately harmful to adults . Though testing a single life stage can be a good starting point for assessing pesticide toxicity, it is important to identify the most susceptible life stage. For H. convergens, larvae were more susceptible to pesticide exposure than adults at the 100% concentrations, but not necessarily for the more dilute 10% concentrations. A similar pattern was found for exposure route; effects were more frequent at the 100% concentration, for larvae than for adults, and for cyantraniliprole, novaluron, lambdacyhalothrin and spinetoram rather than the other pesticides tested. For cyantraniliprole at the 100% concentration, oral exposure caused the greatest mortality for both larvae and adults, whereas for the other pesticides, when there were significant effects of exposure route, topical exposure caused the greatest mortality. Lambda-cyhalothrin proved to be an exception to this pattern with residual exposure having the greatest effect on adults at the 10% concentration, residual and topical exposure having a similar effect for adults at the 100% concentration, and all exposure routes having a similar effect for larvae at both concentrations. Other studies have also found that exposure route can affect mortality rates of natural enemies. For instance, the effects of spinetoram on adult females of the parasitoid Neochrysocharis formosawere classified differently for all exposure routes: harmful via oral exposure, harmless via topical exposure, and moderately harmful via residual exposure . Additionally, lambda-cyhalothrin was harmless to N. formosa females via oral exposure, but moderately harmful via topical or residual exposure. For spirotetramat, topical exposure was moderately harmful to adults of the parasitoid Tamarixia radiata, while residual exposure was harmless . At the maximum label rate, novaluron was harmful to nymphs of Podisus maculiventris Say via residual or oral exposure . In contrast, in our study, novaluron was moderately harmful to H. convergens larvae via topical exposure, but was harmless via residual or oral exposure. Given the variable effects of exposure route on mortality, bio-assays for the effects of reduced risk pesticides on natural enemies should include multiple routes of exposure rather than being restricted to residual exposure alone, the approach that is recommended in the first tier of the IOBC testing guidelines . As testing for all individual exposure routes can be considerably more time consuming, a more practical approach would be to use a combined exposure scenario, where insects are exposed via multiple routes simultaneously rather than each route individually. Another potential benefit to testing multiple exposure routes simultaneously rather than individually is that it would allow for any additive or synergistic effects. assessed the toxicity of reduced-risk insecticides to adults and larvae of two lacewing species, C. carnea and C. johnsoni, by simultaneously treating insects topically, residually, and orally. From measures of survivor ship 10 days after exposure, chlorantraniliprole and cyantraniliprole proved to be harmless to larvae, but harmful to adults. In addition, from the same study, spinetoram and lambda-cyhalothrin were moderately harmful to larvae and harmful to adults. In a similar study of the survivor ship of the predatory mite G. occidentalis from simultaneous routes of pesticide exposure, showed chlorantraniliprole and cyantraniliprole to be harmless and spinetoram and lambda-cyhalothrin to be harmful to both larvae and adults. While our study with H. convergens differed in being focused on acute mortality from individual exposure routes, it does help to explain the variation in toxicity among natural enemy species and life stages observed from these earlier studies on mortality effects from simultaneous routes of exposure. Chlorantraniliprole and cyantraniliprole were harmless to lacewing larvae and to larval and adult stages of G. occidentalis, but were harmful to lacewing adults and moderately harmful to harmful for larvae and adults of H. convergens. From our study, we found that for H. convergens the most important exposure route for these two insecticides was via oral ingestion, suggesting that natural enemy species or life stages with sucking mouth parts may escape direct oral exposure, whereas those with chewing mouth parts may experience more toxic effects. In contrast, for spinetoram and lambda-cyhalothrin the most important exposure routes for H. convergens were either topical or residual, which helps to explain the more consistent harmful effect of these two insecticides to a broader range of natural enemy species and active life stages. From our study with H. convergens, we have found that even reduced-risk insecticides can, in some cases, cause substantial acute mortality effects in laboratory bio-assays. We have also been able to demonstrate that pesticide, pesticide concentration, predator life stage and exposure route can all influence the acute mortality response of H. convergens in laboratory bio-assays. Of the seven pesticides tested, lambda-cyhalothrin caused the greatest acute mortality for H. convergens, and was classified as either moderately harmful or harmful at the full field rate concentration. Both cyantraniliprole and spinetoram were also either moderately harmful or harmful to H. convergens larvae at the full field rate concentration.

Accurate and time- or cost-efficient methods of diagnosing infected plants are important elements of a disease management program, both with respect to roguing to reduce pathogen spread , and the efficacy of pruning to clear plants of infection . Accurate diagnosis of PD in grapevines is complicated by quantitative and qualitative differences in symptoms among cultivars and other aspects of plant condition . Our results suggest that a well-trained observer can accurately diagnose PD based on visual symptoms, particularly for advanced cases of the disease. The small number of false positives in disease category 1 and 2 vines may have been due to misdiagnosis of other biotic or abiotic factors . Alternatively, false positives might indicate bacterial populations that are near the detection limit; conventional PCR has at least as low a detection threshold as other methods that rely on the presence of live bacterial cells . Regardless, although scouting based on visual symptoms clearly captured most cases of PD in the current study,hydroponic dutch buckets some caution should be used when trying to diagnose early disease stages to ensure that vines are not needlessly removed. There is no cure for grapevines once infected with X. fastidiosa, except for recovery that can occur in some overwintering vines .

The virulent nature of X. fastidiosa in grapevines, and the corresponding high mortality rate for early season infections, increases the potential value of any cultural practices that can cure vines of infection. Moreover, new vines replanted into established vineyards generally take longer to develop compared to vines planted in newly developed vineyards, potentially due to vine-to-vine competition for resources that limits growth of replacement vines. As a result, vines replanted in mature vineyards may never reach full productivity . Thus, management practices that speed the regeneration of healthy, fully developed, and productive vines may reduce the economic loss caused by PD . A multinomial logistic regression showed significant differences in the relative frequency of different grapevine growth outcomes between the two restoration methods . Chip-budded vines showed significantly lower frequency of strong growth and significantly higher frequencies of vines with developing growth and, especially, of no growth . Nearly 30% of chip-budded vines showed no growth in the following season, compared to 0% of vines on which established shoots were trained. These results indicate that training newly produced shoots from the remaining section of the scion was more likely to result in positive regrowth outcomes. As a result, of the two methods we evaluated, training of shoots that emerge from the scion of a severely pruned trunk is recommended for restoring growth.

However, it is important to note that the current study did not estimate the amount of time required for severely pruned vines to return to full productivity. Moreover, the study did not include mature vines, in which growth responses may differ from young vines. Additional studies may be needed to quantify vine yield, and perhaps fruit quality, in severely pruned vines over multiple seasons.The usefulness of pruning for disease management depends on its ability to clear plants of pathogen infection . A comparison of symptom prevalence among severely pruned and control vines from different disease severity categories showed significant effects of the number of years after pruning , pruning treatment , and initial disease symptom category . The analysis also showed significant interactions between year and treatment and between treatment and symptom category , a non-significant interaction between year and symptom category , and a marginally significant three-way interaction . Overall, more vines had symptoms in the second year compared to the first , and there was a higher prevalence of returning symptom in vines from higher initial disease categories . Severe pruning showed an apparent benefit to reducing symptoms of PD after the first year, but this effect weakened substantially by the second year, with no differences for category 1 or 3 vines, and a slightly lower disease prevalence for severely pruned category 2 vines .

A survival analysis of severely pruned category 3 vines showed a significant difference in the rate of symptom return among plots . All vines in plots 1 to 3 had symptoms by autumn 2000, two years after pruning . In plots 4 and 5, more than 80% of vines showed symptoms after three years. Only plot 6 showed markedly lower disease prevalence; in plot 6, ~70% and 50% of severely pruned category 3 vines showed no symptoms after two and four years, respectively, versus ~36% of control vines overall, after two years. It is important to note that at the time of this study, disease pressure may not fully explain the return of symptoms in severely pruned vines. Surveys conducted during the first two years of the study throughout the entirety of the six research blocks showed that the prevalence of PD in control vines actually declined slightly from the first to the second year , but not due to an increase in replanting efforts or vine death , Rather, this decline in prevalence likely reflects overwinter recovery of mild cases of the disease . Thus, the observed return of symptoms in most severely pruned vines does not appear to be explained by reinfection with X. fastidiosa after clearing of infection during the severe-pruning process. Our results indicate that the apparent effectiveness of severe pruning depended on the initial disease severity, and the effectiveness weakened over time. This suggests at least two constraints exist regarding the general utility of pruning as a PD management tool. First, severe pruning does not appear to be useful for mild cases of PD, as many of those same vines would recover from the infection over the winter . Second, there appears to be little value in pruning severely diseased vines; the high frequency of symptom return within a few years indicates that even severe pruning does not clear most vines of X. fastidiosa infection. That leaves a statistically significant window with respect to intermediate severity cases, which may benefit from severe pruning. The apparent benefit for this category of diseased vines would stem from infections that are not so localized that they are highly susceptible to natural recovery over the winter, but also not fully systemic such that the infection has developed below the pruning point . Reliable identification of this narrow class of diseased vines may require substantial experience with PD scouting, detailed record keeping, and an appreciation for variability in symptoms or infection dynamics based on grapevine cultivar and environmental conditions . Research in other bacterial plant pathosystems has evaluated the potential benefit of pruning and whether pruning extent is related to its effectiveness at clearing hosts of infection . A study of the citrus disease huanglongbing, associated with infection by Candidatus Liberibacter spp.,bato bucket evaluated two levels of pruning severity, neither of which showed promise as a disease management tool . In this pathosystem, it is plausible that a very protracted incubation period may undermine the effectiveness of pruning, because by the time the first symptoms are visible, the infection may have already moved throughout much of the tree. Collectively, our results are more similar to a study of citrus variegated chlorosis . In this study, the presence of X. fastidiosa in plant tissues at different distances from symptomatic leaves was determined for varying levels of disease severity. X. fastidiosa was more widely distributed in trees with severe disease symptoms compared to those with early stage foliar symptoms. Although ColettaFilho et al. did not test whether pruning at various distances proximal to symptomatic leaves would eliminate X. fastidiosa infections, the current recommendation is to prune citrus material if early symptoms are present, and to not prune plants with severe disease symptoms . Citrus plant age is also an important consideration; Coletta-Filho and de Souza recommend that symptomatic citrus trees up to three-years-old be removed rather than pruned, whereas trees four-years-old or older should be pruned.

We did not examine vine age as a factor in this study, but the biology of citrus and grape differ in terms of the overwinter recovery that can occur in grape and the apparently slower movement of X. fastidiosa in citrus compared to grape. Anecdotally, the two most mature plots in our study showed the most rapid return of disease, and the youngest plot showed the slowest return. More studies of the effect of vine age are needed before concluding that interactive effects of plant age and pruning differ between the PD and citrus variegated chlorosis pathosystems.The plant microbiota, defined here as the community of bacteria, fungi, archaea, viruses, and other microscopic organisms that live on or in plant tissues , confer many services as well as disservices to their hosts, including disease development and defense , protection against herbivory , tolerance of abiotic stress , and aid in nutrient uptake . These microbial communities associate with all plant tissues , including seeds . Seeds play a major role in plant communities as agents of dispersal, genetic diversity, and regeneration , and they have significant economic and social value through agriculture . Seeds also are a major bottleneck in natural plant populations, as they face heightened mortality from abiotic stressors, pests, pathogens, and predators . As the initial source of inoculum in a plant’s life cycle, seed microbes are can be transmitted across plant generations and have lifelong impacts . Consequently, understanding how seeds acquire and interact with their microbiota, for example, via priority effects or according to the Primary Symbiont Hypothesis , has implications for improving seed health, seedling establishment, and plant community structure. Previous work on seed microbiota has primarily taken a pattern-based approach to studying assembly processes . Such an approach uses culturing and/or next-generation sequencing to compare, contrast, and correlate patterns in microbial community composition, diversity, and species co-occurrences. Typically, however, these community data provide limited insights into processes such as dispersal, microbe-plant interactions, and microbemicrobe interactions.

Given that seed microbial communities are highly variable across individual plants, plant species, and locations , such pattern-based data cannot always be used to predict assembly outcomes. Moreover, such studies often consider how these assembly processes occur at a single spatial scale . We hypothesize that a mechanistic, multi-scale approach would provide a more complete understanding of how microbial communities assemble in seeds, with the field of meta community ecology providing a theoretical framework for such an approach. Meta community theory accounts for the interaction between ecological processes and habitat heterogeneity across spatiotemporal scales to impact community patterns . This emphasis on multiple scales and heterogeneity can help explain the main drivers of community assembly and patterns of biodiversity and co-occurrence . Plant-associated microbial communities vary widely across environmental gradients and host genetics from the levels of tissues to populations . As such, treating individual plants as heterogeneous habitats for microorganisms that are embedded in a larger, heterogeneous landscape of multiple plants representing different species provides a new approach to observing, testing, and modeling drivers of microbial community variation . However, the study of microbiota through a meta community lens is still relatively new, both for animals and plants , and the plant seed represents a relatively understudied microbiome in this context. In this review, we address how mechanisms of seed microbial community assembly have been studied at different spatial micro-, meso-, and macro-scales , and advocate for a meta community-based approach to seed microbiology in future work. For this review, we use the definition of community assembly from Fukami : “the construction and maintenance of local communities through sequential, repeated immigration of species from the regional species pool.” Additionally, most studies that we cover in our review will be focused on fungi and bacteria . We acknowledge that archaea, viruses, and protists are frequent members of plant-associated microbial communities , many plant viruses are seed transmitted , and viruses can play a major role in the diversity and function of soil microbial communities . However, the ecological roles of these microbes in plant microbial communities, including those of seeds, are still largely unknown. As such, we cannot speak on their contributions to seed microbiota assembly here and recommend new research on these microbes in seeds. We will first summarize the modes of microbial acquisition into seeds, and how meta community ecology frames this assembly process. We then discuss studies of seed microbiome assembly which examine the processes of filtering, species interactions, dispersal, and ecological drift.

These data are consistent with the hypothesis that the adaptation of invertebrate-infecting nsRNA ancestor viruses to plants happened several times through independent events during the evolutionary history of nsRNA viruses infecting plants. It is worthy of note that rubodviruses, laulaviruses, and coguviruses, although phylogenetically related, have divergent genome structures and gene expression strategies. In fact, the members of the first two genera have a genome composed of three monocistronic nsRNAs encoding different proteins;RdRp, NP, and putative MP in the case of rubodviruses, and RdRp, NP and a protein of unknown function in the case of laulaviruses. Instead, coguviruses have a bipartite genome consisting of one nsRNA encoding the RdRp , and one ambisense RNA , in which the ORFs encoding the NP and the MP are separated by a long intergenic region . It has been shown that such an IR is AU-rich and self-complementary, thus assuming in both polarity strands a compact conformation containing a long hairpin predicted to serve as a transcription termination signal during the expression of viral genes.

Taking this into consideration, the question arises of how viruses with such different genomic organizations may have evolved from the same ancestor virus. In this respect,planting gutter it can be speculated that a recombination event between the viral and the vc strand of two genomic RNAs with long, AU rich, and almost identical 50 UTRs could generate ambisense RNAs containing an IR similar to those of coguviruses. Since nsRNA viruses with genomic RNAs showing structural features compatible with this possibility were not known previously, such a possibility appeared unlikely. However, the very long, AU-rich, and highly conserved 50 UTRs reported here for RNA 2 and RNA 3 of GMRV and GGDV, and also observed in the corresponding RNAs of ARWV1 and ARWV2, are the first clear evidence that nsRNA viruses with the structural features compatible with this evolutionary scenario may exist. Based on these considerations, the possibility that the bipartite genomes of coguviruses originated from a tripartite ancestor with genomic RNAs containing 50 UTRs similar to those observed in the rubodviruses appears feasible. No glycoprotein is encoded by rubodviruses , a feature previously reported also for coguviruses that, according to electron microscopy observations, are flexuous, non-enveloped viruses.

In contrast, glycoproteins are expressed by most nsRNA plant viruses transmitted by arthropods . The lack of glycoprotein in the genome of rubodviruses and coguviruses opens the question on the existence of vectors, if any, involved in their transmission. In this respect, it is worthy of note that ophioviruses and varicosaviruses, which are plant-restricted or transmitted by fungi, also do not code for any glycoprotein. Vegetative propagation has been proposed as the prevalent transmission mechanism for ARWV1 and ARWV2. Whether, this is also the case for GMRV and GGDV needs further investigation. Most plant viruses code for viral suppressor proteins counteracting the plant antiviral defense mechanisms based on RNA silencing. Further studies are needed to ascertain whether one or more of the three proteins encoded by GGDV and GMRV and other rubodviruses may interfere with RNA silencing, thus showing multifunctional role, as already reported for VSRs of other viruses. GMRV and GGDV were identified in two different grapevines cultivars, Muscat rose and Garan dmak, that were tested by HTS; moreover, both viruses were found in association with other viruses and viroids. Interestingly, no obvious symptoms were observed in the two grapevines. Although infectivity of both viruses was ascertained by graft-transmission, only a Cabernet franc grapevine infected by GGDV developed symptoms post-grafting; further HTS analysis on this indicator plant reveals the presence of GRSPaV, GYSVd-1, and HSVd. Therefore, it was not possible to ascertain whether GGDV is associated with symptoms and additional studies on its pathogenicity are needed, likewise GMRV. An initial survey using several accessions of grapevine located in three different collections in California resulted in the identification of two and one plants infected by GGDV and GMRV, respectively; like the original sources of the viruses , these grapevines were symptomless.

A more extensive survey, including other grapevine-growing regions in California and the USA, is necessary to determine the real distribution of these novel viruses. In that sense, the detection method developed in this study could be useful for virus testing and certification programs. Finally, during the review process of this manuscript, two novel mycoviruses related to coguviruses and rubodviruses were reported, which extends the host range of phenui-like viruses.Fetal tumors are rare. Autopsy evaluation of stillborn fetuses has revealed a 0.5% rate of fetal tumors. Of liveborn children, prenatally diagnosed masses are most commonly lymphatic malformations and sacrococcygeal teratomas. Cervical teratomas, although accounting for only 3% of teratomas diagnosed in the neonatal period, can cause airway compromise and hydrops, and may be managed with ex-utero intrapartum treatment . Fetal renal tumors are less common, occurring in 7 of 100,000 live births, and there have been reports of fetal neuroblastoma as well. Fetal tumors with associated hydrops portend a particularly poor prognosis, with survival rates below 20%. Early prenatal diagnosis is essential for identifying patients who may benefit from fetal, rather than postnatal intervention, as is the case for certain teratomas. However, in cases where fetal intervention is unlikely to improve survival, knowledge of the wide differential diagnosis of a fetal tumor can significantly improve education and counseling for families.We report a case of a fetus diagnosed with a large renal mass with diffuse metastatic lesions and hydrops. A 34-year-old G5P2002 woman who presented to labor and delivery at 30 weeks and 4 days gestation with several days of abdominal pain and tightness. She had been referred for an ultrasound by her midwife after measuring larger than dates. The ultrasound was concerning for a fetal mass and polyhydramnios, and she was referred to Labor & Delivery for evaluation. Repeat ultrasound at our institution showed fetal hydrops with scalp edema, pericardial and pleural effusions, and a solid 10.4 × 8.2 cm mass with vascular flow extending from the right cheek into the right chest . There was evidence of polyhydramnios with an amniotic fluid index of 27.4 cm. The fetal stomach and airway were not visualized, raising concern for obstruction by the mass. Fetal magnetic resonance imaging confirmed a large 9.8× 8.9 × 8.4 cm soft tissue mass involving the right lateral neck, right axilla, right upper extremity, and right chest wall , with notable compression of the airway. Multiple additional small subcutaneous nodules were identified throughout the fetal torso and extremities, including a 1.8 × 2.8 cm lesion on the left thigh, a 2.7 × 2.5 cm lesion on the right cheek, and a 2.9 × 2.1 cm lesion on the right buttock. Hydrops was confirmed. A fetal echocardiogram was done which showed normal cardiac anatomy, normal biventricular systolic function, diffuse skin edema, bilateral pleural effusions, and a trivial pericardial effusion. The combined cardiac index was normal at 430 ml/min/kg. The differential diagnosis for this large soft tissue tumor presenting in a fetus was broad . Initially, there was concern primarily for a cervical teratoma or lymphatic malformation causing airway impingement and hydrops, and consideration was given to in utero resection to alleviate the hydrops. Ex-utero intrapartum treatment was also a potential management option. The findings of widely disseminated lesions on MRI, however, indicated that the fetus likely did not have a cervical teratoma or lymphangioma amenable to surgical resection. The multiplicity of the lesions raised concern for widely metastatic neoplasm or multiple vascular malformations. Although the fetus was not in imminent distress, the findings of fetal hydrops portended an extremely poor prognosis.

In addition, the rapidly accumulating polyhydramnios was physically disabling for the mother. Given the severity of these findings,gutter berries extensive multidisciplinary discussions were held between the patient, maternal fetal medicine, and the fetal surgery teams. There was no identifiable fetal or postnatal intervention for the fetus, and given the high likelihood of morbidity and fetal or neonatal death, it was decided to proceed with elective termination via intra-fetal cardiac potassium chloride injection followed by induction of labor to avoid the maternal morbidity of a Cesarean section. On hospital day 3, the fetus was delivered vaginally. Immediately notable were innumerable subcutaneous masses . On autopsy, the primary tumor was found to arise from the right kidney with extensive subcutaneous and internal metastatic disease. The largest metastatic focus was the originally identified tumor on the right chest, extending to the right back, measuring 10.5 cm in largest dimension. Additional metastatic subcutaneous disease was identified on the face, extremities, and abdominal wall . Microscopic examination revealed sheets of tumor cells invading the soft tissue, muscle, lymph nodes, right kidney, adrenal glands, thymus, heart, and liver. Tumor cells demonstrated epithelioid to spindle cell morphology in a loose fibrous stroma arranged in sheets and scattered nests. There were variable amounts of eosinophilic cytoplasm, round to oval nuclei, and prominent nucleoli. Additionally, rhabdoid cells were present with abundant eosinophilic cytoplasm, eccentric nuclei, and scattered intracytoplasmic inclusions. Intravascular and perineural tumor was noted in multiple sections. Immunohistochemistry revealed near diffuse positivity for cytokeratin , diffuse positivity for vimentin, focal CD34 staining, retained SMARCA4 and SMARCB1 . However, within the tumor cells there was focal loss of SMARCB1/ INI-1. Of note, WT1 had patchy positivity in the cytoplasm but was not found in the nucleus. Staining for synaptophysin, CD99, HMB45, myogenin, MYOD1, SMA, desmin, SALL4, Glypican-3, and PHOX2B were negative. The results of immunohistochemical analysis were consistent with the diagnosis of malignant primitive epithelioid sarcoma with features of malignant rhabdoid tumor. This is the first report of a fetal diagnosis of malignant primitive epithelioid sarcoma with features of malignant rhabdoid tumor in the literature. Additionally, retention of SMARCB1/ INI-1 expression is exceedingly rare in epithelioid sarcoma. Congenital malignancies are extremely uncommon, affecting fewer than 13 per 100,000 live births, and the most prevalent etiologies include teratomas, neuroblastoma, soft tissue tumors, or leukemia. Epithelioid sarcoma is a rare malignant neoplasm of mesenchymal origin most commonly seen in young adults and occasionally young children. It is most often slow-growing, painless, and difficult to diagnose given its ambiguous nature. Histologically, it demonstrates epithelioid morphology with deletion of SMARCB1/INI-1. SMARCB1/INI-1 is a tumor suppressor gene. Its native function is poorly understood, although it is known to be a member of a family of genes encoding chromatin-remodeling complex. Loss of SMARCB1/INI-1 expression is seen in the majority of epithelioid sarcomas and the vast majority of rhabdoid tumors. In one series of rhabdoid tumors, only 16% had retained expression of SMARCB1/INI-1. Another series of two siblings with classic rhabdoid tumor also retained expression of SMARCB1/INI-1, suggesting that rhabdoid tumor may arise by a separate mutational locus. One additional case of epithelioid sarcoma with retained SMARCB1/INI-1 expression has been reported in a 15-month old infant presenting with an isolated heel mass. Malignant rhabdoid tumor is a highly aggressive subtype of sarcoma. It is most commonly seen in young children and originates in the kidneys, central nervous system , or soft tissue, but has been previously reported in fetal and neonatal patients. When diagnosed in utero, the mean gestational age is around 33 weeks. Rhabdoid tumor is identified histologically by the presence of rhabdoid tumor cells with round vesicular nuclei, prominent nucleoli, and eosinophilic inclusions within the cytoplasm, and usually with deletion of the SMARCB1/INI-1 gene on chromosome 22q11. They exhibit positivity for cytokeratins AE1/AE3, vimentin, and epithelial membrane antigen , although there is some variability. In a 40-year review of the literature, 12 fetal cases and 60 neonatal cases were identifie. Most were of renal origin without CNS involvement, one-third were extrarenal tumors not involving the CNS, and 16.7% were of CNS origin. Overall survival was 9.7%. More than half had distant metastasis at diagnosis, and all patients with metastatic disease at diagnosis died. Multiple subcutaneous nodules resembling the “blueberry muffin” presentation of neuroblastoma were present in this case as well as several others. Placental metastases may also be seen, but all reports describe placental metastases of the tumor without evidence of maternal disease. This tumor is aggressive and rapidly fatal with survival ranging from a few minutes to 3 months in one case series of 8 fetuses and neonates with malignant rhabdoid tumor. Treatment options are extremely limited.

Fallen Fruit’s website points out that the video probes “at the correspondence between the public walking on the tour and the anonymous public of the internet.” This year, the group was also invited to do a project with TED Active. They devised “The Banana Hotline,“ wherein the public is invited to follow a set of instructions, record a memory, and email the audio or video to the group, who will then put together “a living monument of sound.” The exhibit at Art+Sci will also include The Loneliest Fruit in the World . This video portrays the human activities, interactions, and explorations that spring up around a stand of arctic berries growing near Tromsö, Norway: “Against a beautiful, spare landscape peppered with tiny blueberries,” according to the website description, “the video follows a group of Norwegians who while picking negotiate the relation between solitude, gleaning and company.” Integration of chemical signals at the peripheral sensory system remains one of the least understood mechanisms of insect olfaction, particularly in mosquitoes. Despite the great progress made in the last 2 decades in understanding how receptors form the basis of chemosensory perception in insects,bato bucket how olfactory signals integrate at the periphery remains an enigma . ‘‘It is as if a new continent has been discovered but only the coastline has been mapped’’ .

In the largest majority of reported cases , antennal neurons of Cx. quinquefasciatus displayed excitatory responses , but evidence for inhibitory responses , already known for Ae. aegypti , is now emerging for Cx. quinquefasciatus . It has been observed in moths , beetles , the fruit fly , and mosquitoes that activation of one neuron interferes with signaling of other olfactory receptor neurons . It has also been reported that a single compound caused reduction of nerve impulse followed by a transient post-stimulus excitation . Although Carlson and collaborators elegantly demonstrated that in the fruit fly lateral inhibition is most likely mediated by ephaptic coupling , the complete ensemble of the molecular mechanism of inhibition at the peripheral olfactory system of mosquitoes remains terra incognita. A simple explanation of the ephaptic coupling is that, upon stimulation of an ORN, the potential declines. Consequently, per channel current generated by a cocompart mentalized neuron is reduced . This scenario argues that the firing of a neuron causes reduced spike frequency by a colocated neuron because of the close apposition of their neuronal processes. Although ephaptic coupling could explain lateral inhibition, other mechanisms of intraneuron inhibition may exist. While de-orphanizing odorant receptors expressed predominantly in Cx. quinquefasciatus female antennae, we serendipitously recorded currents from an OR that generate inhibition in response to certain odorants. Further studies unraveled a hitherto unknown mechanism of peripheral, intrareceptor inhibition in mosquito olfaction.

In our attempts to de-orphanize ORs from the southern house mosquito, Cx. quinquefasciatus, we challenged Xenopus oocytes coexpressing CquiOR32 along with the obligatory coreceptor Orco with a panelof more than 200 compounds, including mostly physiologically and behaviorally relevant compounds . Because CquiOR32 is predominantly expressed in female antennae , we reasoned that this receptor might be involved in the reception of attractants or repellents. CquiOR32- CquiOrco-expressing oocytes generated dose-dependent inward currents when challenged with various odorants, including cyclohexanone, methyl salicylate, and 2-methyl-2-thiazoline . Interestingly, however, eucalyptol, fenchone, DEET, picaridin, IR3535, PMD, and other compounds generated currents in reverse direction thus resembling inverse agonists. These unusual currents of reverse direction were reproducible, and no indication was found of adaptation . No currents were recorded when oocytes alone or oocytes expressing only CquiOR32 or only CquiOrco were challenged either with methyl salicylate or eucalyptol . However, CquiOR32-CquiOrco-expressing oocytes responded to both compounds. Methyl salicylate elicited inward currents, and eucalyptol generated currents in the reverse direction in a dose-dependent manner .

To gain better insights into possible mechanism of these currents in reverse direction, we obtained current-voltage curves from oocytes expressing wild-type CquiOR32-CquiOrco with voltage clamped at 80,60, 40, 20, 0, +20, and +40 mV and using methyl salicylate and eucalyptol as stimuli . Similar recordings were performed using N-methyl-D-glucamine chloride as a source of bulky, impermeable mono cation or with sodium gluconate buffer instead of NaCl . These data corroborate that OR32-Orco forms nonselective cation channels , as indicated by the reversal potential shift to more negative voltage upon replacement of Na+ by less permeable NMG . An examination of the first and third quadrants for each graphic in Figure 2 shows that I-V curves elicited by methyl salicylate are above baseline and below control I-V curves for positive and negative voltages, respectively , consistent with a potentiation of the control current by methyl salicylate. By contrast, I-V curves generated with eucalyptol, albeit not robust, are below and above control I-V curves for positive and negative holding potentials, respectively , suggesting an inhibition effect. In the presence of eucalyptol, replacing Na+ with NMG had a minor effect on the I-V curves , whereas replacing Cl with gluconate seems to suggest that Cl is implicated in this inhibition .We then surmised that these ‘‘inhibitors’’ might modulate responses to odorants when these two types of stimuli are simultaneously delivered to receptors. First, we challenged a CquiOR32-CquiOrco-expressing oocyte with methyl salicylate and then eucalyptol and recorded regular and reverse currents, respectively . Then we challenged the same oocyte preparation with mixtures of methyl salicylate and eucalyptol. Binary mixtures with higher doses of eucalyptol elicited reverse currents. Inhibition was also observed with lower doses of eucalyptol, which generated dose-dependent reduced inward currents. A continuous trace displayed in Figure S4 shows no difference in the responses to methyl salicylate before and after these tests thus ruling out adaptation. Taking together, these findings suggest that ‘‘intrareceptor’’ inhibition occurred consistently with outward currents previously recorded from antennal neurons of the vinegar fly, Drosophila melanogaster .w1118 flies gave very weak responses to eucalyptol at high doses, but interestingly Orco-Gal4/UAS-CqOR32 flies generated dose-dependent, inverse EAG responses . To further scrutinize this unusual reverse EAG responses, we used gas chromatography with electroantennographic detection . In GC-EAD analyses, injected mixtures are separated by GC and subjected to antennal preparations under the same condition thus ruling out any possibility of mechanical interference and minor sample contamination. Here, methyl salicylate responded with regular EAG responses, i.e., with the first phase , which is referred to as rise of the receptor potential, and the second phase starting at the end of the stimulus, commonly referred to as the decline of the receptor potential . This is analogous to the depolarization, repolarization, and hyperpolarization of a nervous impulse. As opposed to methyl salicylate, eucalyptol consistently gave inverse EAD responses thus corroborating what we observed in EAG analyses . Next, we recorded EAG responses when flies were challenged with odorants and an inhibitor. First, we compared the response of w1118 and Orco-Gal4/UAS-CqOR32 flies to -2-hexenal when it was delivered alone or in combination with eucalyptol. EAG responses from w1118 flies to 0.1% -2-hexenal alone or in combination with 10% eucalyptol did not differ significantly .

By contrast,dutch bucket hydroponic EAG responses from Orco-Gal4/UAS-CqOR32 flies to 0.1% -2-hexenal plus 10% eucalyptol were significantly lower than those elicited by 0.1% -2-hexenal alone . We then examined the dose-dependent effect of this inhibition by using Orco-Gal4/UAS-CqOR32 flies. Robust responses to 0.1% methyl salicylate were reduced in a dose-dependent manner with the addition of eucalyptol but remained unchanged at the end of the tests. Likewise, EAG responses to 0.01% -2-hexenal were reduced when coapplied with eucalyptol . Of note, -2-hexenal does not activate CquiOR32 . Such inhibition presumably results from CquiOR32 indirectly inhibiting responses of the fly endogenous receptors to -2-hexenal. In these continuous experiments, a small difference between EAG responses before and after costimulus tests may be due to loss of this volatile semiochemical from the cartridge rather than adaptation. Similar inhibition was observed when 2-heptanone was applied alone or coapplied with eucalyptol . Taken together, these results further suggest that intrareceptor inhibition occurs in vivo as indicated by the inhibitory effect of eucalyptol on methyl salicylate responses. Additionally, the effect of eucalyptol on the response to -2-hexenal suggests that intraneuronal inhibition occurred. A few lines of evidence support this hypothesis. First and foremost, eucalyptol does not cause inhibition in control flies and -2-hexenal does not activate CquiOR32 . The simplest explanation is that, in Orco-Gal4/UAS-CqOR32 flies, all endogenous receptors are coexpressed with CquiOR32. Thus, CquiOR32 response to eucalyptol interferes with the response of DmelOR7a to -2-hexenal. In short, inhibitor and agonist are likely to be acting on different receptors in the same neurons, thus an intraneuron inhibition. To further test the notion of intraneuronal inhibition, we turned to single sensillum recordings .The best ligand for ab4A, the neuron in ab4 sensilla with a large spike amplitude, is -2-hexenal , although ab4A is also very sensitive to other ligands, including hexanal . Contrary to ab4B, ab4A houses only one OR, namely, DmelOr7a . Because expression of CquiOR32 was driven by DmelOrco, ab4A neurons in our transgenic flies house both DmelOr7a and CquiOR32. Coexpression was confirmed by a significantly stronger response to methyl salicylate recorded from Orco-Gal4/UAS-CquiOR32 than from WT flies , while retaining response to hexanal . It is known that methyl salicylate is the best ligand for DmelOr10a in ab1D but elicits only very low response in ab4A . The low response of WT flies to methyl salicylate did not differ significantly when the odorant was delivered alone or codelivered with eucalyptol . By contrast, responses recorded from Orco-Gal4/UAS-CquiOR32 flies were significantly lower when the two stimuli were delivered simultaneously from two different cartridges . Next, we tested whether CquiOR32 response to eucalyptol would affect DmelOR7a response to a cognate ligand, hexanal. Responses of WT flies to hexanal did not differ significantly when comparing hexanal alone with hexanal plus eucalyptol . Recordings from ab4 sensilla in the Orco-Gal4/UAS-CquiOR32 flies showed a slight, albeit not significant, increase in response to hexanal. This is unlikely to be due to hexanal activation of CquiOR32 . When hexanal and eucalyptol were delivered simultaneously firing of DmelOR7a was completely abolished . We also recorded from ab7 sensilla, which expresses DmelOR98a, in ab7A and for which butyl acetate is one of the best ligands . In the transgenic flies both methyl salicylate and butyl acetate generated excitatory responses , which were inhibited by eucalyptol . Because methyl salicylate and eucalyptol elicit inward and reverse currents in CquiOR32, this in vivo inhibition is not surprising. However, the consistent observation that eucalyptol inhibits the response of an endogenous receptor to a cognate ligand supports the notion that intraneuronal inhibition occurs when receptors are colocated in a neuron. Specifically, the inhibitory responses of CquiOR32 interferes with the activation of a collocated receptor by a cognate ligand. For example, activation of DmelOR7a in ab4A neuron by hexanal and activation of DmelOR98a in ab7A neuron by butyl acetate were both inhibited by eucalyptol upon interaction with CquiOR32. Contrary to the fruit fly, which expresses only one receptor per neuron , mosquitoes can coexpress multiple ORs in the same neuron .In the traditional view, the skin serves as a protective barrier between the body and the external environment. Yet, more and more evidence suggests that cutaneous function extends far beyond mere protection. In fact, cutaneous function regulates a wide spectrum of cutaneous and systemic functions. Compromised epidermal function has been linked to the development of a variety of cutaneous and extracutaneous disorders. For example, disruption of epidermal permeability barrier not only provokes the release and production of proinflammatory cytokines, but also induces infiltration and activation of inflammatory cells in the skin, suggesting that defective epidermal permeability barrier predisposes to the development of inflammatory dermatoses. But barrier disruption also stimulates barrier homeostasis responses, including epidermal proliferation and lipid synthesis. Moreover, defects in epidermal permeability barrier allow the penetration of microbial pathogens into the skin. Importantly, recent studies showed that the epidermal dysfunction-induced elevations in cutaneous cytokines lead to increased levels of proinflammatory cytokines not only in the skin, but also in circulation, supporting not only a pathogenic role for epidermal function in cutaneous and extracutaneous inflammation, but also suggesting a link between cutaneous function and inflammation-associated systemic disorders. In addition to epidermal permeability barrier homeostasis, other epidermal functions, such as pH and stratum corneum hydration, also regulate cutaneous functions. Accordingly, improvements in epidermal function could benefit multiple cutaneous and extracutaneous functions. Because of the importance of cutaneous function, much recent attention has focused on the identification of active ingredients that could lead to the development of products that improve cutaneous function.

At baseline and at the end of 2 weeks of intervention period, body weight, body mass index and composition were determined. Body composition was measured using the Tanita-BC418 bio-electrical impedance analyzer . Height was measured without shoes using a stadiometer and recorded to the nearest 0.1 cm. In the meantime, subjects reported on their overall well-being at baseline and at the end of two weeks by completing the SF-36 general wellness questionnaire. Aliquot of stool samples was diluted, acidified, and filtered, and SCFAs were quantified by gas chromatography flame ionization detection, as previously described. SCFA standard mix was purchased from Sigma-Aldrich . Urinary free rosmarinic acid as a biochemical marker of adherence to spice supplementation was measured by high-performance liquid chromatography according to published methods. Measures of beta-diversity were computed using the weighted and unweighted UniFrac distance metric using phyloseq in R. Differences in whole communities across groups were determined by Permutational Multivariate Analysis of Variance using the Adonis command provided by Vegan in R.

The results were visualized via Principal Coordinate Analysis ordination . OTUs differentially abundant between follow up and baseline visits in subjects receiving spice and placebo interventions were identified using DESeq2. This algorithm performs normalization using size factors estimated by the median-of-ratios method,blueberry packing boxes employs an empirical Bayesian approach to shrink dispersion, and fits the data to negative binomial models. Differential abundance was determined using the Wald test with automatic filtering of low abundance OTUs and automatic calculation of adjusted p-values , and the enriched OTUs were visualized using the ggplot2 package in R. As Bonferroni correction is often considered overly conservative, an extended set of OTUs significant at p < 0.2 was listed. Differentially abundant OTUs between spice and placebo groups were also detected by DESeq2. Null and test models were constructed for each OTU using DESeq. Each model includes variables “time” and “intervention” . Differentiating the two models was an interaction between “time” and “intervention”, which was only present in the test model. To identify OTUs with abundance changes that occurred specifically with spice but not placebo intervention, we used a likelihood ratio test to test for a significant additional contribution of the interaction between time and spice vs. placebo exposure on OTU abundance in the test model when compared to the null model. Here, we report that daily intake of 5 g of mixed spices for 2 weeks in healthy subjects resulted in a significant reduction in the relative abundance of the phylum Firmicutes , and a trend of increasing in phylum Bacteroidetes as compared with a matched control group.

Most healthy adult micro-biota are dominated by these two bacterial phyla, which together make up about 95% of gut micro-biota. A significantly higher abundance of Firmicutes and a higher Firmicutes/Bacteroidetes ratio has been frequently demonstrated in obese individuals, whose gut micro-biome is characterized by an increased capacity for energy harvest, inflammation, and gut barrier disruption. In addition to the shifts observed in Firmicutes and Bacteroidetes abundance, a number of OTUs in the family of Bifidobateriaceae and Bacteroidaceae, as well as the family of Streptococcaceae, Lachnospiraceae, Ruminococcaceae, Veillonellaceae, and Erispelotrichaceae, were significantly altered between groups . Spice intervention significantly enhanced two Bifidobacterium OTUs of the Bifidobacteriaceae family—B. animalis and B. pseudolongum—as well as one Lactobacillus OTU compared with control group. Bifidobacterium has been shown to associate with the production of a number of potentially health promoting metabolites, including short chain fatty acids, conjugated linoleic acid, and bacteriocins. The abundance of B. animalis was reported to negatively associate with the body mass index. Results from the present study are consistent with published reports in that consuming polyphenol rich foods increases the relative abundance of Bifidobacterium and Lactobacillus and reduces pathogenic Clostridium species. Bifidobacterium and Labtobacillus are considered “healthy bacteria”, and members of the Bifidobacterium and Lactobacillus, as well as Streptococcus, are frequently used as probiotic strains with evident health benefits such as immune-modulation, cancer prevention, inflammation management, and the control of diverse bacterial consortia infections.

We noted that the abundance of all three Ruminococcus spp. were increased. In humans, Ruminococcus spp. were found as abundant members of a “core gut micro-biome” in a majority of humans. Some Ruminococcus spp. in our gut micro-biomes play an important role in helping us degrade and convert complex polysaccharides into a variety of nutrients for their hosts. The slow digestion of these special carbohydrates has been associated with numerous health benefits such as reversing infectious diarrhea and reducing the risk of diabetes and colon cancer. Interestingly, the Bacteroides spp., such as B. fragilis, also have the ability to recognize and metabolize plant- and host-derived polysaccharides, and to produce polysaccharides as well. Polysaccharide A, e.g., synthesized by B. fragilis, can promote immunological tolerance to pathogenic species such as Helicobacter hepaticus and protect the host from inflammation and associated colorectal cancer.SCFAs constitute approximately 10% of the energy source in healthy people. These microbial-derived products are utilized by the host and exert a range of health-promoting functions. Butyrate is used preferentially as an energy source by the gut mucosa, is anti-inflammatory, and protects against colorectal cancer, whereas propionate is largely taken up by the liver and is a good precursor for gluconeogenesis, and promotes satiety and reduction in cholesterol liponeogenesis and protein synthesis. Acetate is the most predominant gut-produced SCFA in peripheral blood and plays a role in prevention of weight gain through an anorectic effect, inflammation, and metabolic dysregulation. Valerate is present in substantially low amount, and research on this SCFA is very limited. A recent study, however, showed that valerate significantly inhibited the growth of C. difficile in vitro and in vivo, suggesting valerate can potentially be used as a safe, microorganism-free method to treat C. difficile infection. In the present work, the level of all four SCFAs was trending higher after spice intervention but the difference did not attain statistical significance due perhaps to the large interpersonal variations and small size of the study population. Nonetheless, our results indicated that spice supplementation can change SCFA production. Human studies investigating the effects of spice supplementation on gut micro-biota composition are very limited. Kang et al., reported that intake of capsaicin powder in healthy subjects increased the Firmicutes/Bacteroidetes ratio, accompanied with increased plasma levels of glucagon-like peptide 1 and gastric inhibitory polypeptide and decreased plasma ghrelin level. In animal models, Van Hul et al., reported that cinnamon bark extract lowered fat mass gain and adipose tissue inflammation in mice fed a high-fat diet leading to reduced liver steatosis and lower plasma nonesterified fatty acid levels that were associated with change on the microbial composition. Spices have been shown to alter serum biochemical parameters related to inflammation or low-grade inflammation induced by high-fat diet and have protective effects against chronic diseases. This evidence indicates that spices may play important role in modulating the growth of gut micro-biota and promoting human health. The observed lack of significant difference in microbial richness is largely attributed to the brevity of the intervention,package of blueberries which is also believed to account to for the lack of change in species evenness as well as of overall microbial composition. Another limitation of the study is that to avoid the sustained effects of spices on micro-biota, our study design was a randomized placebo-controlled and not a crossover clinical trial, therefore, there are large interpersonal variations in gut micro-biota composition and metabolite formation.

Lastly, since this is not a controlled feeding study, dietary recall data analyzed by a dietitian was the only method used to assess participants’ compliance with the beige diet. Analysis of dietary data may have an impact on interpretation of the results. The human gut micro-biome mediates many key biological functions and its imbalance, termed dysbiosis, is associated with a number of inflammatory and metabolic diseases from inflammatory bowel disease to asthma to obesity and insulin resistance . How to effectively shift the micro-biome and restore balance is a key question for disease prevention and treatment. The gut micro-biome is influenced by a number of factors including the nature of the initial colonization at birth , host genotype, age, and diet. As diet is a readily modifiable factor, it is an obvious target for interventions. Several studies have confirmed high inter-individual variability in the bacterial composition of the gut micro-biome in healthy individuals . Despite this high variability at the species level, enterotypes, or distinct clusters at the genus level, were described as core micro-biomes that are independent of age, gender, nationality, or BMI . Although the concept of enterotypes is itself controversial, diet has been shown to play a key role in determining enterotype . Although the core micro-biota within each person are stable over longer time scales , community composition is highly dynamic on shorter time scales . In fact, major shifts occur within 1 day of a significant dietary change . “Blooms” in specific bacterial groups were observed in response to controlled feeding of different fermentable fibers . Dietary changes affect both the structure and function of the gut micro-biome in animals , and humans under controlled feeding conditions . Rapid shifts in micro-biome composition are observed in response to change from a vegetarian to an animal based diet . An ecological perspective helps to delineate the complexity and multi-layered nature of the relationships between the micro-biota, the human host, and both the nutritive and non-nutritive compounds we ingest . The concept of the human gut micro-biome as a distinct ecosystem or collection of micro-ecosystems allows us to identify and characterize the components of the system, including its inputs and outputs. In this case, the inputs of the system include all of the various ingested compounds that can either serve as food substrates or that can be metabolized by or that affect the metabolism of the micro-biota . Some of these inputs, such as probiotics have been studied extensively. It has been well documented that certain sugars such as galactooligosaccharides, fructooligosaccharides, and oligosaccharides found in milk act as prebiotics that support the establishment and growth of certain commensal microbial species . Research has also documented the effects of antibiotics, and pathogens on the micro-biota composition, its recovery or lack of recovery to baseline following resolution, and the various immunological and physiological effects of these perturbations . Yet, little is known about the effects of ingested microorganisms on gut micro-biota composition or function, and even the basic questions of which microbes, how many of them, and how much they vary from diet to diet and meal to meal, have not been answered. We do know about the microbial ecology of various specialty foods where fermentation, colonization, ripening, and/or aging are part of the preparation of these foods, for example pancetta and of course cheese . The microbial ecology of the surfaces of raw plant-derived foods such as fruits and vegetables has also been characterized . Furthermore, it is known that the microbial ecology of endemic microbes found on food surfaces can affect mechanisms by which pathogens colonize these foods . A recent article showed that certain ingested microbes found in foods such as cheese and deli meats were detected in the stool of individuals who consumed them, and that furthermore they were culturable and thus survived transit through the upper intestinal tract . However, the microbial ecology or microbial assemblages of different meals and diets, as well as the total number of live microorganisms ingested in these meals and diets are largely unknown. In fact, studies of the effects of diets and foods on the gut micro-biota rely on dietary recalls and other dietary reporting instruments that were not designed to capture the potential variability in aspects of foods other than their basic macronutrient and micronutrient content. Specifically, current instruments for collecting individual dietary data do not capture the provenance of foods or their preparation, both of which would likely influence certain compositional aspects of the foods, especially the microbes on those foods. We performed a preliminary study designed to generate hypotheses about the microbes we eat, and how they vary in terms of total abundance and relative composition in different meals and dietary patterns typical of American dietary intakes.

The magnitude of the potential acidity of the soil depends on the type and amount of clay and organic matter. Although there is some inverse parallelism between acidity saturation and base saturation, these terms are not complementary because they have different denominators . Contrary to Al toxicity and acidity saturation, there is no direct association between base saturation and crop yields. Instead, a minimum base saturation threshold is defined such that, above it, no soil acidity problems are detected . Therefore, recommended target base saturation levels must be defined locally for each crop type . Most lime requirement methods used in temperate regions target soil pH , estimating with locally-calibrated models the lime rate required to raise the soil pH to a specific level . In acid tropical soils, maximum yields can be obtained with a pH as low as 5, depending on other soil chemical properties , and raising the pH to higher values can result in a loss of soil structure and other problems . Therefore, a target pH level is seldomly used, and when used, it must be defined locally . There is a negative exponential relationship between exchangeable acidity and soil pH . Very high exchangeable acidity values are only found in soils with a low pH,garden grow bags but not all soils with a low pH have high exchangeable acidity.

Exchangeable acidity approaches 0 at a pH above 5.5, and there is virtually no exchangeable acidity at a pH above 6 . Therefore, a target pH between 5.5 and 6 should be high enough to address Al toxicity problems.Acid tropical soils have very low plant-available phosphorus because of the high P fixation capacity of Fe and Al oxides often present in weathered tropical soils. Liming has the associated benefit of increasing P availability, which might result in significant yield responses, particularly when P fertilization is low . However, liming can only provide short-term relief to P deficiencies in soils with low P reserves . Therefore, phosphorus availability is not considered a direct target of liming, and lime requirement models do not consider it. However, the increase in P availability can be an important reason for observing a yield response to lime.The lime recommendation models were evaluated using data from four soil incubation studies that measured the effect of liming on exchangeable acidity and ECEC or acidity saturation . Studies only measuring the effect of liming on pH were not considered. Soil incubation studies are experiments in which soil samples are mixed with different lime treatments and incubated under controlled conditions for about a month to ensure that all lime reacts with the soil. The liming effect is assessed by measuring chemical soil properties before and after each treatment. The data from Kamprath , Cochrane et al. , and Ananthacumaraswamy and Baker were readily available, but Teixeira et al. soil data were not. The Teixeira et al. data was reconstructed in two steps: the initial soil properties were back solved from lime requirement formulas and lime rates, and the final soil properties were estimated using the regression formulas provided in the supplementary information .

Table 1 describes the main features of these four data sets. We have shown important differences in lime rates and prediction accuracy depending on the target soil property and model . When the target is to ameliorate the Al toxicity of the soil by neutralizing its acidity saturation to a certain level, both Kamprath and Cochrane et al. models provided reasonable accuracy . Nevertheless, the new model formulated in Eq. 12 offers improved accuracy and the advantage of being sustained by a formal mathematical derivation that can be expanded . Similarly, the base saturation model also has great prediction accuracy, particularly for target base saturation levels of around 50% . In contrast, no model based on a target pH can deliver accurate results without additional soil tests, and they need to be developed locally . The model presented here is the only model based on a target acidity saturation with greater accuracy than the original Cochrane et al. model . The authors of the ACID4, NuMaSS, and MG5 models claimed that they modified the Cochrane et al. model to improve the accuracy for their target region. However, there are no available data supporting those statements. The decreased accuracy that we found may partly be because we did not have access to these data. However, we believe that these more complex models suffer from overfitting to the datasets used to build them. In other words, they may perform better in particular regions, but this has come at the expense of general validity. Conversely, the new model is more robust than previous models because it is based on mathematical foundations and strong empirical relations. These relations are consistent through a wide range of soils from different regions.

However, we observed a small incubation study effect in the relations shown in Figure 3, which might be a consequence of the soil region or, more likely, because of the incubation study per se . Experimental results have an error component, including systematic errors that are consistent within one experiment but differ between experiments, introducing statistical bias. This bias can be reduced with standardized procedures. However, lime incubation studies are not fully standardized, and they differ in the incubation time and temperature, liming materials, and water additions, among other variables. For instance, we excluded data from an incubation study that used tap water rather than distilled water to keep the soil samples moist during the incubation because control treatments had significantly more exchangeable Ca2+ and less exchangeable acidity than the initial conditions. A more thorough standardization of experimental procedures for measuring liming effects would help the development of general models for lime requirement estimation. A novel feature of the model formulated in Eq. 12 is that the lime factor is a continuous function of TAS. The Cochrane et al. model modifies the lf depending on TAS and the initial acidity saturation, using a discontinuous rule with two fixed lf . However, the proposed rule does not always improve accuracy, not even in their data, as shown by the points with incorrect lf . In the MG5 and NuMaSS methods, the lf depends on clay content or activity. Our review does not show evidence for a need to adjust the lf as a function of clay, despite the wide range of clay content and soils included in the four soil incubation studies used here. Adjusting the lf and lime rates by clay content might be a work-around to account for differences in soil bulk density when the method returns lime rates in tons per hectare without directly including the soil bulk density in the formulas. Nevertheless, clay type and content could be considered in future corrections of the TAS method, particularly if there are high deviations in the association between lime rate and Δexch. acid and Δexch. bases. It seems counter intuitive that, while both the acidity saturation and base saturation models are highly accurate on their target,tomato grow bags the lime requirement they predict can be so sharply different . These differences highlight the importance of identifying the soil chemical property most associated with the crop yield response to liming. It might be that reaching a given level for some property, such as a base saturation of 70% or a pH of 6, guarantees that all soil acidity problems are solved without leading to over liming problems. However, this approach can also result in lime requirement estimates that are much too high , which might be particularly problematic when lime is expensive, and its manipulation cumbersome. The alternative is to target the most limiting factor for crop yield, which is frequently Al toxicity in acid tropical soils . However, this approach can under predict lime requirements when Al toxicity is the only target but not the acidity problem most limiting crop yields.

A comprehensive approach would predict the lime rate needed to tackle every potential acidity problem while considering other management alternatives. However, crop responses to other acidity problems, such as calcium and magnesium deficiencies, are unclear, and their liming requirements have not been defined. Thus, more research on crop responses to lime in soils with these specific acidity problems is needed to develop a lime requirement method that tackles them all. Lime requirement models can be useful for strategic research on potential lime use in tropical regions where liming is still a rare practice and the experimental evidence is scarce . These models estimate the lime rate needed to reach a target soil condition based on readily available standard soil data . Such information could be used together with the crop response to that soil condition to estimate the effect of liming on crop yield. For instance, there is ample evidence of the association between acidity saturation and crop yields . Therefore, the expected yield response to lime can be predicted by estimating what fraction of the maximum yield is observed at the current acidity saturation level while assuming that the final yield after liming is the inverse of that fraction. If data on lime and grain prices is available, such functions can be used to get a first approximation of the potential profitability of liming. Such analysis can help identify regions where liming investments might be more successful, pinpointing national governments and private sector efforts. However, this does not mean that the readily available soil data used by the models reviewed here has sufficient quality for farm-level recommendations . Therefore, farm-level lime requirements would be more accurate when based on soil properties measurements or additional local soil-quality indicators, such as soil color, soil texture, or presence of specific plant species . The soil properties used by the lime requirement models reviewed here are wet-lab measurements, which are costly and may be inaccessible for farmers in the tropics. Therefore, farmers in the tropics could benefit from cost effective, quick tests for lime requirement prediction, but these need to be developed locally.Worldwide, biodiversity is declining at unprecedented rates, threatening species persistence as well as the benefits humans gain from ecosystems. These benefits, known as ecosystem services, have become an increasingly important argument for biodiversity conservation. The economic and other benefits from ecosystems can motivate conservation action, and are more and more being used in payment for ecosystem service schemes. Once an economic value of the service has been determined, it can be captured in commercial markets or quantified in terms comparable with economic services and manufactured capital. These economic values can then potentially be used to support biodiversity conservation within policies. The use of ecosystem services arguments for justifying biodiversity conservation is, however, not without risk or controversy. Many experimental studies show that biodiversity increases the magnitude and/or stability of ecosystem functioning , and that most species contribute to ecosystem functioning in some way. However, such studies do not consider the costs of maintaining or promoting biodiversity, even though costs are generally a limiting factor for implementing real-world conservation policies. When the economic pay-off from ecosystem services is the main factor motivating conservation, the cost-effective action is to conserve the subset of species that provide the greatest return at relatively short timescales. Because real-world communities are almost invariably dominated by a small number of species that often respond readily to conservation management, we hypothesize that in real-world landscapes the majority of the services is provided by a relatively small number of species; that these species are generally common, and that threatened species rarely contribute to present ecosystem service delivery; and that the most important ecosystem-service-providing species can be easily enhanced by simple management actions that are insufficient to support threatened species. Support for these hypotheses would suggest that delivery of ecosystem services is insufficient as a general argument for biodiversity conservation. Here we test these hypotheses using data from 90 studies and 1,394 crop fields on crop-visiting bee communities from five continents. Pollination is an important ecosystem service. The economic contribution of pollinators to crop production is significant, and bees are generally considered the most important pollinators of crops. We find that wild bee communities contribute on average over $3,000 ha 1 to the production of insect-pollinated crops. However, a limited subset of all known bee species provides the majority of pollination services because, across different crops, years and large bio geographical regions, crop-visiting bee communities are dominated by a small number of common species and rarely contain regionally threatened species.

Besides reactive oxygen species , ATs also inhibit the production of reactive nitrogen species , particularly nitric oxide , as well as their associated oxidative processes. Furthermore, release of pro-inflammatory mediators and adhesion molecules is suppressed by ATs via targeting of the respective signaling pathways, e.g., the arachidonic acid and the tumor necrosis factor -α, nuclear factor -κB pathways. Aside from cardioprotection, the anti-inflammatory property of ATs also contributes to their anti-obesity effect and action in adipose tissue. Studies using AT-rich fruit extracts showed that ATs prevented the upregulation of inflammatory response in adipose tissue, when triggered with consumption of a high-fat diet, by regulating the NF-κB stress signaling pathway. ATs also reduce the level of pro-inflammatory cytokines and modulate the expression of adipocytokines. Although studies using cell lines and animal models support the anti-obesity effect of ATs, a causal relationship between AT consumption and reduction of body mass index has yet to be conclusively established by clinical studies. This could be due to one or more parameters that are associated with different clinical studies,plastic plant pot such as the amount and types of ATs ingested, the food matrix of ATs, and the inherent difference of the study subjects.

In addition to their antioxidant and anti-inflammatory activities, ATs act in cancer chemoprevention by inducing terminal differentiation of tumor cells and thus impeding tumorigenesis. Prevention of malignant cell transformation and inhibition of cancer cell proliferation by ATs have also been reported. ATs can further interfere with cancer development by activating caspases and inducing apoptosis of cancer cells. However, the cancer chemotherapeutic effect of ATs still requires strong supporting evidence from clinical studies. In addition, the bio-conversion of ATs by human intestinal micro-biome should be rigorously investigated. Similar to other AT-rich fruits, the pomegranate also has a cardioprotective effect by targeting two major causal factors of atherosclerotic lesion and cardiovascular disease, namely the accumulation of cholesterol and oxidized lipids, and arterial macrophage foam cell formatio. Pomegranate juice and fruit extracts are also associated with antidiabetic activities, largely due to the antioxidant properties of HTs and ATs in reducing oxidative stress and lipid peroxidation. Supplements of pomegranate juice or fruit extracts at 1 g/kg/day for five weeks, effectively enhanced endothelial NO synthase expression, as well as plasma nitrate and nitrite levels in plasma in obese Zucker rats fed with an atherogenic diet. In addition, supplements of juice at 100 mg/kg/day or 300 mg/kg/day for four weeks prevented the development of high blood pressure in diabetic rats. Consumption of pomegranate juice rich in ATs and HTs also lowered systolic and diastolic blood pressure in hypertensive patients.

The bioactivities of pomegranate ATs have mostly been studied in the context of pomegranate juice and fruit extracts. However, these AT-rich sources are abundant in vitamin C, carotenoids, and HTs, which can also contribute to the ascribed bioactivities. To this end, “white” pomegranate cultivars are available, which lack the accumulation of ATs in leaves, fruits and flowers. Comparative analysis of the white and AT-rich pomegranate cultivars will be informative as to the specific role of ATs in the health-promoting activities of pomegranate juice and fruit extracts. ETs are abundant in pomegranate fruit peel, as well as juice and extracts that are produced commercially from the whole fruit. Besides pomegranate, ETs are also present in a wide range of medicinal and food plants, such as many berries, nuts, and herbs. As such, the knowledge of bio-transformation and bio-conversion of pomegranate ETs by human and microbial enzymes will inform nutritional and pharmacological studies on ETs that are isolated from other plants. EA, a hydrolysis product of ETs , can be detected in the blood circulation of healthy volunteers within half an hour of ingesting pomegranate juice or extracts. The human gut micro-biota can further convert EA into urolithins prior to their absorption by the intestinal cells. It was shown that urolithin concentrations reached up to 18.6 µM in the plasma of healthy volunteers after consuming pomegranate juice for five consecutive days. Aromatase that converts androgens to estrogens has been considered to be a therapeutic target for treating the hormone-sensitive type of breast cancer. Although several urolithins exhibited anti-aromatase activities in a placental microsome-based enzyme assay, only UB competitively inhibited and most effectively suppressed the aromatase activity in an aromatase-over expressing breast cancer cell line.

Consistent with “in cell” aromatase-inhibitory activity, UB also significantly arrested testosterone-induced proliferation of MCF-7aro cells, suggesting an underlying mechanism of aromatase inhibition. Interestingly, estrogen-induced proliferation of MCF-7aro cells was also inhibited by urolithins. However, inhibition was was likely effected through mechanisms independent of the aromatase activity. Besides breast cancer, urolithins can also reduce the risks of another sex steroid hormone-related cancer, endometrial cancer. UA arrested cell cycle at the G2/M phase and regulated the expression of cell cycle-related proteins at this phase. On the other hand, UA was shown to act as an estrogen agonist and modulate the estrogen-receptor α -dependent gene expression in the ER-positive endometrial cancer cells. It remains to be determined whether and how estrogen signaling could be involved in the suppression of endometrial cancer cell growth by urolithins. The canonical Wnt/β-catenin signaling pathway has been shown to activate T-cell factor transcription and function in colon carcinogenesis. To determine the potential involvement of urolithins in colon carcinogenesis through Wnt signaling, phenolic extracts of several fruits rich in ETs, including pomegranate, strawberry , and Jamun berry , as well as pure EA and UA compounds, were tested in the human embryonic kidney 293T cells that express a reconstructed canonical Wnt signaling pathway. Although the fruit extracts and chemicals all inhibited Wnt signaling, only UA exhibited an IC50 value that was physiologically relevant in the lumen of colons when taking enterohepatic circulation of urolithins into consideration. During the initiation stage of the HT-29 colon cancer cells, urolithins A–D inhibited TCDD-induced, CYP1-mediated EROD activities. At the cancer progression stage, UA and UB impaired the proliferation of HT-29 cells and led to cell cycle arrest at the G2/M phase, as well as activation of CDKN1A expression. In addition, treatment with urolithins resulted in activation of caspases 3, 8, and 9, suggesting that urolithins induced both the extrinsic and intrinsic apoptotic pathways in HT-29 cells. UA, UB, and mUA inhibited the proliferation of the T24 human bladder cancer cells in vitro, with IC50 values of 43.9, 35.2, and 46.3 µM, respectively, comparable to the UA inhibition of Wnt signaling at an IC50 of 39 µM. The transcript and protein levels of Phospho-p38 mitogen-activated protein kinase were increased by the urolithin treatment, while those of MAP kinase kinase kinase1 and Phospho-c-Jun were decreased in the T24 cells. Furthermore, these urolithins reduced the level of H2O2-induced oxidative stress and induced apoptosis through activation of caspase 3 and PPAR-γ protein expression. Inflammatory responses involving the activation of neutrophils and monocytes play a central role in the development of cardiovascular diseases. Intriguingly,nursery pots the number of free hydroxyl functional groups on urolithins appeared to impact how they modulate the inflammatory functions of neutrophils.

Among the urolithins tested , UA exhibited the most potent antioxidant activities against the release of ROS from the pro-inflammatory triggered neutrophils. UB significantly affected several inflammatory biomarkers that are associated with cardiovascular events, by inhibiting the production of interleukin 8 and metalloproteinase-9 , and preventing the shedding of selectin CD62L triggered by the pro-inflammatory factor cytochalasin A/formyl-met-leu-phenylalanine . UC , on the other hand, inhibited the release of elastase, a pro-inflammatory mediator responsible for extracellular matrix degradation, from the f-MLP-stimulated neutrophils. In addition to recruitment of neutrophils, monocyte adhesion to endothelial cells represents another key event in inflammatory responses. A mixture of UA and UB restricted the adhesion of Tamm-Horsfall protein-1 monocytes to the human umbilical vein endothelial cells. UA glucuronide , but not UA, UB or UB glucuronide, inhibited monocyte adhesion to TNFα-stimulated human aortic endothelial cells ; a dosage-dependent inhibition of TNFα-induced migration of endothelial cells was also shown for the above-mentioned urolithins. NO plays multifaceted roles in combatting cardiovascular diseases. Although UA, UB, and UB glucuronide did not show any effect individually at 15 µM on NO bio-availability, a mixture of the three urolithins at equal concentrations activated the expression of eNOS after a 5-min incubation and increased NO production in primary HAECs after a 24-h incubation. Overall, these in vitro studies with neutrophils, monocytes, and NO suggested the potential anti-inflammatory and cardiovascular-protective functions of urolithins. Based on the urinary excretion of urolithins by healthy volunteers after ingesting ET-rich foods or fruit extracts, three urolithin metabolic types have been defined, including A , B , and 0. Interestingly, the population of the human gut bacteria Gordonibacter urolithinfaciens correlated positively with the in vivo production of UA, but inversely with that of UB and isoUA. A recent study observed an interlinked relationship among gut dysbiosis , ET metabolism, and obesity. A relatively high percentage of metabotype B was found in the overweight-obese group, while metabotype A had a higher presentation in the normo weight than the overweight-obese group. In addition, G. urolithinfaciens levels were higher in the metabotype A than the metabotype B individuals. Further investigations should provide a mechanistic understanding of how consuming the polyphenol precursors of urolithins, in the presence of UA-producing bacteria, may reduce the risks of diseases associated with obesity. Large inter-individual variations in the cardiovascular risk biomarkers were observed in healthy overweight-obese individuals after consuming pomegranate supplement. However, after clustering the different urolithin metabotypes in these individuals, improved blood lipid profiles were evident in the metabotype B group, in a dose-dependent fashion, while there was no significant effect in the metabotype A group. Interestingly, several metabotype 0 individuals shifted to metabotype A or B after consuming pomegranate extracts. Together with the study by Selma et al. , these results suggest that consumption of pomegranate extracts may have personalized effects that are associated with the gut micro-biota and the urolithin metabotypes of the individuals. Of the various pomegranate phenolic metabolites exhibiting anti-Alzheimer activities in in vitro assays, only urolithins, including UA, UB, mUA, and methyl-urolithin B , were predicted to be capable of crossing the blood-brain barrier. Methylation of urolithins by the mammalian enzymes may further improve their lipophilicity and facilitate the penetration of the blood-brain barrier.Corroborating with the computational predictions, urolithins reduced the production of the neurotoxic, fibrillogenic β-amyloid peptide in vitro. Significant improvement of the survival/mobility of Caenorhabditis eleganswas also achieved by mUB treatment. However, empirical evidence is still needed to verify the roles of urolithins in preventing Alzheimer’s disease in vivo. Advanced protein glycation products have been implicated in the development of chronic diseases, such as diabetes and Alzheimer’s disease. The inhibitory effects of pomegranate fruit extract, punicalagin, EA, gallic acid , UA, and UB toward the early, middle, and late stage of protein glycation were compared using in vitro assays. Although all phenolic metabolites showed anti-glycation activities, pomegranate extract, punicalagin, and EA were more effective than GA, UA, and UB in preventing the glycation of proteins by fructose. The mechanistic basis underlying UA’s anti-aging activities was explored recently using C. elegans, mammalian cells, and rodent models. Feeding C. elegans with UA led to a dosage-dependent extension of the worm’s lifespan. It was shown that UA could activate mitophagy and eliminate damaged mitochondria in C. elegans, mammalian cells derived from muscle or intestinal tissues, and muscles of young and old rodents. UA was also found to be bio-available in the skeletal muscle and could improve muscle function in mice and rats. The role of urolithins in enhancing mitochondrial function and muscle quality presents promising dietary leads for improved mobility in the elderly population. Pomegranate fruits, leaves, flowers, and seeds have all been used in traditional herbal medicine for treating various illness. However, bio-active metabolites present in pomegranate fruit peel and juice have received considerable more attention than those found in other tissues. Bio-activities of the phytochemicals accumulating in tissues other than fruit should be investigated more rigorously in the future. In addition, detailed metabolite profiling and characterization of different pomegranate cultivars, grown under non-standard conditions , can be carried out to explore further the phytochemical diversity in pomegranate.

Our central claim is that the causal status effect derives from the supporting role that causal information plays in explanation. Although this idea is a long-standing theme in the study of categorization , relatively little research has directly examined how explanatory goodness relates to categorization. In this paper, we study the relationship between explanatory goodness and categorization. We demonstrate that the magnitude of the causal status effect depends on the quality of the explanation in which the causal information is embedded. We first briefly review the causal status hypothesis and present arguments that the causal status effect depends on explanatory goodness. Then we present a study to support this claim. Finally, we discuss the implications of this study for categorization more broadly. Intuitively,plastic flower buckets wholesale some components of a concept do indeed appear more important for categorization judgments. As Medin and Shoben point out, it is much easier to imagine a robin that is not red than it is to imagine a robin that lacks the appropriate genetic structure of robins: imagining a robin with, say, zebra DNA, would make many other typical properties of robins implausible .

There is a long history of research on categorization demonstrating that some features associated with category members are particularly important for categorization judgments . One prominent perspective on centrality is based on the theory- or knowledge-based view of conceptual structure . According to this framework, knowledge of a category is, in important ways, like a scientific theory, comprising a “host of mental explanations” . In this view, category knowledge is seen, not in terms of a prototypical member or collection of exemplars, but rather in terms of “an explanatory principle common to category members . Accordingly, categorization is typically viewed as an inference to the best explanation. Murphy and Medin illustrate this idea with a well-known example: a man at a party who jumps into a pool fully clothed in a business suit would probably be classified as intoxicated, not because he is similar to the prototype or to instances of a “drunken behavior” category, but because being intoxicated is the best explanation for his behavior . Despite the importance of the theory-based view for orienting research on categorization and centrality, the framework leaves unspecified the specific constraints related to background knowledge that influence classification decisions.

One important response to this challenge is the idea that information that participates in causal relations is of greatest centrality . In Ahn’s causal status hypothesis, causes are weighted more heavily in categorization decisions than are the corresponding effects. For example, in the simple case of a single causal relationship between two features, such as having wings and flying , having wings would be given greater weight than flying, and would more greatly influence categorization decisions. Why should causes play a more important role than the corresponding effects in the underlying principles of a category? According to Ahn, et al.,causal properties may be seen as generating other features: e.g., DNA produces external features such as hair color. Features such as DNA may thus be regarded psychologically as most defining or diagnostic of category membership, because they form part of the essence or core of a concept. It follows, Ahn, et al. claim, that possessing the most central features would provide better evidence for category membership than more superficial features. While not disputing the empirical validity of the causal status effect, and the critical role of causal information for many if not most knowledge-dependent categorization decisions, we worry that the exclusive focus on causal knowledge risks obscuring the supporting role that causal information plays in explanatory processes.As Lombrozo notes, explanations can’t be reduced to just the supporting causal information, as explanations entail a set of factors that go beyond the causal information per se .

For one thing, there are many kinds of causal structures —e.g., causal chains versus feedback systems—that draw on different kinds of domain knowledge. An additional factor is the preference for simple or parsimonious causal explanations over those that would invoke more assumptions . In a recent study that illuminates the relationship between the causal status effect and explanatory structure, Lombrozo has shown that the strength of the causal status effect depends partly on the type of explanation— mechanistic or teleological —in which the causal knowledge is embedded. In one experiment, participants were presented with a novel category characterized by two features and were told that one feature causes the other . Participants were then asked to explain why the animal has the second feature , and to estimate the probability that an object missing one of the features was a member of the category. Participants who provided a mechanistic explanation showed a larger causal status effect than those who provided a teleological explanation . In addition to the type of explanation, the causal status effect may also depend on the quality of an explanation. We suggest that some of the puzzling results obtained in studies of the causal status hypothesis might be best explained in terms of explanatory goodness. In one such study , Ahn, Kim, Lassaline, and Dennis gave participants a standard and two alternatives. Participants were instructed to choose which of the two alternatives should be categorized with the standard. One standard read: “This object has a high-intensity light bulb and a pouch that can contain liquid because it was designed to kill bugs.” The first alternative shared the cause, but not the effects ; the other alternative shared the effects, but not the cause . Ahn, et al. predicted that people would prefer to match an alternative to the standard on the basis of the shared cause, rather than the shared effects. Although the overall results confirmed the prediction, inspection of the results reveals that only half the items conformed to the prediction. Table 1 shows the materials used in the study and, for each item, the percentage of choices for the matching cause. While there is a noticeable causal status effect for three of the items,black flower buckets two items exhibit chance responding, and one even suggests a preference for the shared effect alternative. We suggest that the item variability can best be understood in terms of the explanatory role of the causal property being asserted for each item. One hallmark of a good functional explanation is that the intended function is causally connected to the effects to be explained. Ideally, the facts can then be understood as subsumed under a general causal law . In addition, good explanations exhibit breadth—the extent to which an explanation accounts for most if not all of the available facts ; and depth—the extent to which the local explanation fits within a larger explanatory framework, and can itself be explained . These considerations can shed light on why some items in Ahn et al.’s study conformed to their prediction, while others did not. Consider the item with the strongest causal status effect, the first item . In this case, a clear, general causal relationship links the intention with the effects: it’s generally known that bugs are attracted to light, and that bug-killing agents typically take the form of liquid pesticides. Second, the explanation is broad and deep .

Now, consider the worst-performing item: “This painting has four pillars and is red because the painter intended to draw a dog.” The causal relation between four pillars and the intention to draw a dog is extremely weak, and there is no clear explanatory framework that would link the intention to paint a dog with any of the facts. Of course, these are after-the-fact suggestions. To make this account plausible, what is needed is a manipulation of explanatory goodness. In the current study, we test the hypothesis that the magnitude of the causal status effect depends on explanatory quality: we predict that the better the explanation in which causal information is embedded, the stronger the causal status effect. The design was a 2 × 2 mixed factorial with explanatory quality , manipulated within-subjects and task order,manipulated as a between-subjects counterbalancing variable.The materials consisted of 24 stimulus sets, each based on a standard “fact-explanation” pair, which comprised a set of facts joined with a potential explanation for those facts. In the explanation-rating task the facts were presented as short sentences describing a particular situation, which participants were instructed to assume as true; and the explanation was presented as a separate sentence describing a potential explanatory account of those facts. For example, one set of facts read: “This object has a high-intensity light bulb and a pouch that can contain liquid”; and the associated explanation read: “This object was designed to kill bugs.” For the categorization task, the facts and explanation were combined into a single sentence, following Ahn, et al. . Of the 24 standards, 16 were test items, and 8 were fillers. The fillers were constructed to minimize the likelihood of demand characteristics in the categorization task . Of the 16 test items, 6 standards were taken without modification from the materials used in the Ahn, et al. study described above. In addition to the six standards from Ahn et al.’s study, we constructed an additional ten items, five of which were designed to express good explanations; and five to express poor explanations. The good-explanation items were designed so that the explanation specified a general and plausible causal relation that accounted for all of the stated facts . . These additional ten standards were adapted from materials used in studies of category-based inference by Sloman and by Patalano, Chin-Parker, and Ross . The experiment consisted of two paper-and-pencil tasks, counterbalanced for order across participants: evaluating explanatory quality and carrying out a two-alternative forced-choice categorization task. For the explanation rating task, participants were given all 24 standards, each arranged on a separate page of the test booklet. The facts were presented at the top of each page in a separate sentence, with the candidate explanation presented immediately below the facts. Participants were instructed to read the facts and the explanation, and then to provide a rating of how good or satisfying they found the explanation to be on a scale from 1 to 7 . They were then instructed to briefly explain their response. The categorization task was identical to that of Experiment 4 in Ahn, et al. as described earlier.Specifically, it was a forced-choice task between two alternatives, in which participants were instructed to choose the one alternative that should be categorized with the target. Each triad—a standard together with both alternatives—was presented on a separate page of the test booklet, with the standard at the top of the page, and the alternatives below it. For the test items, one alternative shared the cause with the standard, but not the effects; the other alternative shared the effects, but not the cause. Sample items are presented in Table 2.The eight filler items were constructed to vary the pattern so as to minimize the possibility of task demands. To this end, for four fillers, the alternatives both shared the cause with the standard, but differed on the effects; and for the remaining four fillers, both alternatives shared the effects, but differed on the cause. For each participant, item order was randomized in both tasks; for the forced choice task, left-right presentation of alternatives was also randomized.Participants completed the tasks at their own pace: the categorization task required roughly 15 minutes to complete; the explanation-rating task, 25 minutes. The results of this study provide support for the main claim of this paper: that the causal status effect derives from the role that causal information plays in an explanation; and specifically, that the magnitude of the causal status effect depends on the goodness of the supporting explanation. This finding adds support to the position that explanatory structure is important in categorization . According to this explanation-based perspective, categorization is treated as an inference to the best explanation. This perspective suggests that a primary research focus should be on investigating properties that characterize the quality of explanations, and the role that those properties play in cognitive tasks, such as categorization. A possible objection to our study would be that we simply constructed explanations that were inconsistent with participants’ prior knowledge. Ahn et al., found that the causal status effect can be eliminated if the stated causal relationship contradicts background knowledge.