bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in serum citrulline during hematopoietic cell transplantation (HCT) in children
Authors: Gosselin Kerri , Bechard Lori J , Feldman Henry A , Kellogg Mark , Gura Kathleen , Sonis Andrew , Venick Robert , Gordon Catherine M , Guinan Eva C , Duggan Christopher
Published: June 2021

Children undergoing HCT may experience gastrointestinal side effects that limit oral intake and enteral absorption. Serum citrulline (CIT), a non‐essential amino acid produced by small bowel enterocytes, is a marker of intestinal mass. We sought to determine if serum CIT was associated with clinical markers of gastrointestinal health in children undergoing HCT. We conducted a multi‐center, prospective cohort study of 26 children to define time related changes in serum CIT over the course of HCT. Enteral intake, emesis, days on parenteral nutrition, stool volume, IL‐6 levels, and oral mucositis severity were measured. Weekly serum CIT concentrations were obtained from 10 days prior until 30 days after HCT. Mean baseline CIT concentration was 25.1 μmol/L (95% CI 7.5–83.5), decreasing to a nadir on day +8 that was lower with severe oral mucositis (6.2 μmol/L, 95% CI 3.1– 12.2) than without (11.5 μmol/L, 95% CI 5.5–23.9), before returning to baseline by day 30 (p=0.005). CIT was inversely associated with IL‐6 (1% lower CIT per 10% increase in IL‐6, p=0.016) but was not correlated with TNF‐alpha, leptin, or ghrelin. In children undergoing HCT, serum CIT is valid marker for oral mucositis severity and may indicate more extensive mucosal injury to the gastrointestinal tract. Supported by the Massachusetts Vitamin Litigation Grant and NIH grants M01‐RR02172, UL1 RR025758–01, M01‐RR00865, M01‐RR00188, and K24 HD 058795.
Three behaviorally‐based lifestyle interventions produce weight loss and decrease markers of cardiovascular disease risk in obese adults
Authors: Jones Jennifer Lynn , Welk Gregory , Stewart Jeanne , Walsh Sarah , Paulson Katie , Barclay Sally
Published: June 2021

We enrolled 78 obese adults (31 male and 47 female) and randomized them to one of the three following groups for an 8‐week intervention: guided weekly health coaching with a behavior change curriculum (HC), utilization of a physical activity monitor and associated web tool for self‐monitoring with minimal health coaching (AM), or combination of weekly health coaching and use of a physical activity monitor and associated web tool (COMBO). The objective was to determine the relative influence of the 3 interventions on weight loss and clinical risk factors for cardiovascular disease (CVD). Percent body fat was measured by handheld Bioelectrical Impedance Analysis (BIA), and waist circumference (WC) was measured at the umbilicus. Weight reduction was significant for all groups (−4.21 ± 3.08 kg; p < 0.0001). Significant intervention effects were observed for BMI (p < 0.0001), percent body fat (p = 0.0008), WC (p < 0.0001), diastolic blood pressure (p = 0.0004), total cholesterol (p = 0.0003), triglycerides (p = 0.0001), and fasting plasma glucose (p = 0.0001). Anthropometric and clinical outcomes did not differ across treatment groups (p >; 0.05); however, trends were observed for larger effects in the COMBO group. These results indicate efficacy of both a guided intervention and a self‐monitoring device for behavior change and subsequent weight loss, with the potential for a combination of the strategies to improve results.
Multi‐component access to a commercially available weight loss program: A randomized controlled trial
Authors: Johnston Craig Allen , Rost Stephanie , Miller‐Kovach Karen , Moreno Jennette P , Foreyt John P
Published: June 2021

This study examined weight loss between a community‐based, intensive behavioral counseling program (Weight Watchers PointsPlus®) that included 3 treatment access modes and a self‐help condition. A total of 292 participants were randomized to a Weight Watchers (WW; n=147) or a self‐help condition (SH; n=145). WW participants were provided with 3 ways to access the treatment: weekly meetings, WW mobile application, and WW online tools. Heights and weights were obtained at baseline, 3 and 6 months. Self‐report of access routes used was collected at 3 and 6 months. Data were available for 248 subjects at 24‐weeks (85.0% retention). A significant interaction of time and condition for BMI (F=36.7, p<.001) was found, indicating that WW participants significantly decreased their BMI at 6 months compared to SH participants. Mean weight loss for the WW and SH groups was 10.1 ± 11.7 lbs and 1.3 ± 8.1 lbs respectively. WW participants were 8.0 (95% CI, 3.9–6.2) times more likely to achieve a 5% baseline weight loss at 6 months than SH participants (p<.001). The use of the treatment access modes was also examined. Those using all 3 access routes to a high degree had the greatest weight loss (p<.001). This community‐based program produced clinically significant weight loss. Participants that used all treatment access modes were the most successful in the program. This study was supported by a grant from Weight Watchers International, Inc.
A Lower Calorie Egg Whites Breakfast is as Satiating as a Higher Calorie Whole Eggs Breakfast
Authors: Reimers Kristin , Meyer Michael , Ward Tabra , Andon Mark
Published: June 2021

Eggs are a highly satiating food. Egg whites (EW), due to greater protein and volume per kcal, may offer similar satiety with fewer calories than eggs (E). Fifty three men and women (ages 39 ± 10, BMI 22 ± 2) consumed each of two breakfasts: 2 slices toast, 10 g margarine, 284 mL non‐caloric beverage and either 141 mL EW (Egg Beaters® Original, ConAgra Foods), cooked (70 kcal, 15 g protein, 0.5 g fat) or 97 mL E, cooked (2 large eggs equivalent, 140 kcal, 13 g protein, 9.5 g fat). E breakfast provided 330 kcal, and EW breakfast provided 70 kcal less, 260 kcal. Ad libitum lunch was consumed 3 hours after the test meal. Equal satiety of the EW and E breakfast meals was observed as indicated by no difference (p >; 0.05) in 10 cm visual analog scale ratings before lunch for hunger (4.8 vs. 4.9), fullness (3.8 vs. 3.8), and lunch energy consumption (582 ± 305 vs. 575 ± 315 kcal), respectively. These data indicate that an EW serving providing half the calories, similar protein, and 45% greater volume is as satiating as E, resulting in reduced short‐term energy intake from breakfast and lunch combined.
Protease resistant soy proteins and their interactions to murine intestinal mucosa
Authors: Lim Jinkyu , Lee Ju‐Young , Lee Jinwoo , Yum Youjin , Kim Jong‐Sang
Published: June 2021

Some allergic foods we eat usually contain proteins resistant to proteases such as pepsin and pancreatic enzymes. We named it protease‐resistant food proteins (PRFPs). Soy bean as a staple food in many countries is consumed in various forms, such as, Tofu, soy milk, Miso, soy flour, soy sauce, Natto, and so on. We found that the major parts of soy proteins are resistant to proteases. With a hypothesis that the resistant soy proteins to the proteolytic enzymes from the stomach and small intestine can possibly interact with other nutrients in the intestine and the intestinal mucosa, we tested protein‐protein interactions and protein‐small molecule interactions with the protease‐resistant soy proteins using gel electrophoresis and MS/MS analysis. We found that the PRFPs sharing common compact beta‐sheeted structural characteristics and sequence similarities interact with mucosa proteins from small intestine. The lists of interacting proteins to PRFPs contain the functions of lowering cholesterol, controlling satiety, and many other interesting features. This study was supported by a research grant from KRF (2012–03520000).
Low‐fat dairy differentially improves metabolic syndrome parameters in male and female adults
Authors: Dugan Christine E , Barona Jacqueline , Fernandez Maria Luz
Published: June 2021

We evaluated the effect of low‐fat dairy (LFD) intake on markers of metabolic syndrome (MetS). Subjects (13 male, 22 female) with low dairy intake (<1.5 ser/dairy/day) were randomly assigned to receive LFD (296 mL low‐fat milk, 170 g yogurt, 510 g cheese stick) or isocaloric control foods (CON) (42 g granola bar, 177 mL juice) daily for 6‐wk. After a 4‐wk wash‐out, subjects received the alternate diet for 6‐wk. In women, LFD lowered diastolic blood pressure vs. baseline (p =0.02). LFD decreased body weight (p=0.02) waist circumference (WC) (p < 0.01) and body mass index (p<0.01) compared to CON. Change in WC was positively associated with change in plasma glucose (r=0.496, p=0.02) and negatively correlated with small LDL particles (r= −.482, p=0.03). In men, LFD lowered (p=0.01) systolic blood pressure vs. baseline and plasma glucose was lower compared to CON (p = 0.02). LFD tended to lower plasma triglycerides (TG) (p=0.08) vs. CON and changes in TG were positively associated (r = .791, p = 0.001) with the larger, more atherogenic VLDL particles. Fat intake was not different between groups, but protein was higher (p<0.001) and carbohydrate lower (p<0.001) during LFD. Energy intake was lower for females during LFD, thus LFD may promote beneficial anthropometric changes, possibly by triggering reduced energy intake. These data suggest that LFD may differentially lower Met S markers by gender. [Supported by the Dairy Council]
Differential effects of military training on tibia bone strength indices in male and female recruits
Authors: Gaffney‐Stomberg Erin , Lutz Laura J. , Murphy Nancy E. , Marchitelli Louis J. , Cable Sonya J. , Young Andrew J. , McClung James P.
Published: June 2021

Military training increases stress fracture incidence, especially in women. This study evaluated bone strength indices in men (n=15) and women (n=11) pre‐ and post‐basic combat training (BCT) using peripheral quantitative computed tomography and whether a calcium/vitamin D (Ca/vit D) supplement could improve bone health. Volunteers were randomized to receive either 2000 mg Ca and 800 IU vit D or placebo, daily for 9 weeks of BCT. Scans were performed on the non‐dominant leg at 4, 14 and 66% of the segment length proximal to the distal tibia end plate. Several bone indices changed after BCT, but there was no effect of Ca/vit D on those changes. Total density increased at the 4% site, resulting in an improved bone strength index (123 ± 33 to 125 ± 32 mg/mm*4; P<0.05). At the 14% site, cortical density increased in males (pre: 1117 ± 25, post: 1125 ± 22 mg/ccm) but not in females (pre: 1141 ± 22, post: 1140 ± 23 mg/ccm; sex‐by‐time, P<0.01). BCT affected stress‐strain index differently in males and females (sex‐by‐time, P=0.05) as SSI increased in males (pre: 1655 ± 375, post: 1679 ± 393 mm*3), and decreased in females (pre: 1322 ± 235, post: 1314 ± 220 mm*3) in our sample. These data suggest that sex differences in bone responses to BCT may, at least in part, explain the increased stress fracture incidence in females. Future research should focus on interventions, including nutritional countermeasures, to improve bone strength in females.
Mechanisms of protein quality control degradation in the nucleus
Authors: Gardner Richard , Fredrickson Eric , Rosenbaum Joel , Gallagher Pamela
Published: June 2021

Protein quality control (PQC) degradation systems protect the cell from the toxic accumulation of aggregation‐prone misfolded proteins. Because any protein can become misfolded, these systems must be able to distinguish abnormal proteins from normal ones, yet be capable of recognizing the wide variety of distinctly shaped misfolded proteins they are likely to encounter. How individual PQC degradation systems accomplish this remains an open question. In yeast, the nuclear PQC ubiquitin ligase San1 ubiquitinates misfolded nuclear proteins for proteasome degradation. Recently, we found that San1 directly recognizes its misfolded substrates via intrinsically disordered N‐ and C‐terminal domains. These disordered domains are punctuated with small segments of order and high sequence conservation that serve as substrate‐recognition sites San1 uses to target its different substrates. Within its misfolded substrates, San1 requires the exposure of a particular window of hydrophobicity and threshold of insolubility. While other cellular PQC degradation systems require chaperones for substrate degradation, chaperones are only required for San1‐mediated degradation if the misfolded substrate crosses an even higher threshold of insolubility required for San1 recognition. Altogether, our latest results shed new insights into how misfolded proteins are targeted for degradation in the nucleus
Se‐glutathione peroxidase mimic ebselen serves as a novel regulator of insulin secretion in mouse islets
Authors: Wang Xinhui , Lei Xin Gen
Published: June 2021

Our laboratory has previously shown impaired glucose‐stimulated insulin secretion (GSIS) in mice deficient in Se‐glutathione peroxidase 1 (GPX1−/−, Cu,Zn‐superoxide dismutase (SOD1−/−), or both enzymes (DKO). This study was to determine if and how the GPX1 (ebselen) and SOD1 (copper diisopropylsalicylate, CuDIPs) mimics rescued the impairment. Pancreatic islets were isolated from the three knockout mouse lines (3–6 mo, male) and their wild‐type (WT), treated with ebselen (50 μM), CuDIPs (10 μM), or Se (100 nM, sodium selenite) for 5 h, and incubated for 60 min with 2.8 or 16.7 mM glucose to collect medium supernatants and islets for assays. Ebselen, but not CuDIPS or Se, promoted (P < 0.01) GSIS in islets of all genotypes. Ebselen also rescued GSIS and improved glucose clearance in the GPX1−/− mice, and the rescue was associated with coordinated responses of four key functional proteins of GSIS. Bioinformatics and Q‐PCR analysis were followed to identify four potential signaling pathways for ebselen to regulate the gene expression of these four factors. A siRNA study verified PGC‐1α in the ARE signaling pathway as the key effector for the global regulation of ebselen on GSIS. In conclusion, we revealed a novel role and signal pathway of ebselen in rescuing GSIS of GPX1 deficient islets and mice, a distinct comparison of GPX1 and SOD1 in regulating GSIS, and a new strategy to treat insulin secretion disorders (NIH DK 53018).
Methylselenol, a selenium metabolite, inhibits colon cancer cell growth in vitro and in vivo
Authors: Zeng Huawei
Published: June 2021

Methylselenol is hypothesized to be a critical selenium (Se) metabolite for certain anticancer activities in vivo but its role in colon cancer prevention remains to be characterized. This study tested the hypothesis that methylselenol inhibits the growth of colon cancer cells and tumors. We found that submicromolar methylselenol exposure inhibited cell growth and led to an increase in the G1 and G2 fractions with a concomitant drop in the S‐phase, and an induction of apoptosis in cancerous colon HCT116 cells, but to a much lesser extent in noncancerous NCM460 colon cells. Furthermore, in agreement with the fact that blockade of extracellular‐regulated kinase1/2 (ERK1/2) and p38 mitogen activated protein kinase (p38 MAPK) pathways suppresses the growth of colon tumors, we demonstrated that methylselenol inhibited ERK1/2 and p38 MAPK in cancerous colon HCT116 cells but also to a lesser extent in NCM460 cells. Consistent with these in vitro observations, a daily single oral dose (1 to 3 mg Se/kg body weight) of methylseleninic acid, a methylselenol precursor, for 5 weeks inhibits the growth of colon cancer xenografts in a C57/BL mouse model. Taken together, methylselenol's strong potential of inhibiting colon cancer cell proliferation, may be an important mechanism by which Se exerts its anticancer activity in colon cancer prevention.
Genistein reduced colon cancer metastasis to liver through regulating the expression of metastasis‐related genes in xenographed mice model
Authors: Li Qian , Chen Hong , Lezmi Stephane , Chen Danni
Published: June 2021

This study aimed at investigating the effect of genistein (Gen) on liver metastases derived from colon cancer. Male athymic nude mice were fed with western diet (W; calories%, fat 39%, carb 50%), W diet supplemented with 100ppm Gen (GL) or 500ppm Gen (GH). After 16 weeks, all the animals were intrasplenically injected with metastatic human colon cancer cells SW620 and sacrificed 8 weeks later. GL and GH mice showed lower liver metastases (36% and 17%) compared to W group (55% of mice with metastases). Mean size of metastases in the W group (4.7 mm2) was larger than that in GL (2.1 mm2) and GH (2.0 mm2) groups. To understand the molecular mechanism of the anti‐metastatic function of Gen, the expression of multiple metastasis‐related genes were determined by real‐time PCR. mRNA of NDRG1 gene in livers was greatly decreased by 50% and 60% in GL (p=0.005) and GH group (p=0.001) compared to W group. As NDRG1 has been reported as a biomarker for metastasis and poor prognosis in hepatocellular carcinoma, the down‐regulation of NDRG1 by Gen supplementation was corresponding to decreased liver metastasis. In summary, our study indicated that dietary supplementation of Gen suppressed colon cancer metastasis, which is associated with altered expression of metastasis‐related genes by the treatment.

Grant Funding Source: university of illinois
Dietary ellagic acid suppresses atherosclerotic lesion formation and vascular inflammation in apoE‐deficient mice
Authors: Park Sin‐Hye , Kang Young‐Hee
Published: June 2021

High levels of plasma LDL cholesterol are an important determinant of atherosclerotic lesion formation. The disruption of the process of cholesterol efflux or reverse cholesterol transport (RCT) of peripheral cells may promote atherogenesis. The aim of the current study is to investigate athero‐protective effects of ellagic acid administration on inflammatory atherosclerosis in apoE knockout (KO) mice fed athero‐control diet for 12 weeks. ApoE KO mice were divided into two groups and one group received 10 mg/kg ellagic acid orally during the same experimental periods. Ellagic acid supplementation to apoE KO mice reduced the number of eosinophils and basophils and improved serum lipid profile by decreasing atherosclerosis index and increasing HDL level. Treating with ellagic acid enhanced peritoneal macrophage expression of ABCA1 and ABCG1 responsible for cholesterol efflux associated with RCT. Atherosclerotic lesion formation in the aortas were suppressed by supplying ellagic acid, as evidenced by Oil red O staining. Moreover, plasma levels of soluble VCAM‐1 and C‐reactive protein, biomarkers of atherosclerotic inflammation, were diminished in ellagic acid‐supplementation. In addition, ellagic acid reduced aortic levels of VCAM‐1, MCP‐1, PECAM‐1, and F4/80. Therefore, dietary ellagic acid may be an athero‐protective agent retarding atherosclerotic lesion formation and vascular inflammation.
Yuzu (Citrus junos Tanaka) improves cognitive function and glucose homeostasis in β‐amyloid infused rats
Authors: Park Sunmin , Kim Da Sol , Yang Hye Jeong , Hwang Jin Taek
Published: June 2021

Our preliminary study revealed that dementia induced by β‐amyloid accumulation itself exacerbates peripheral glucose homeostasis in rats. We were interested to study that long‐term oral consumption of Yuzu extract improves cognitive dysfunction and glucose homeostasis in the β‐‐amyloid‐induced rats. Male rats received intracerebralventricular infusions of β‐amyloid (25–35) (AMY) or β‐amyloid (35–25) (RAMY), at a rate of 3.6 nmol/day for 14 days. AMY‐infused rats were divided into two groups: one group received 3% lyophilized 70% ethanol extracts of Yuzu in a high fat diet and the other group received 3% dextrin. After 6 weeks of treatment, AMY increased β‐amyloid deposition in the hippocampus and the accumulation was not detected in RAMY. AMY attenuated hippocampal insulin signaling to induce hyperphosphorylation of tau. Yuzu treatment prevented the attenuation of the hippocampal insulin signaling in AMY‐infused rats and reduced tau phosphorylation. As a result, AMY induced cognitive dysfunction in comparison to RAMY while yuzu reversed cognitive impairment. AMY decreased body weight without lowering visceral fat accumulation by decreasing food intake and yuzu treatment blocked the decrease. However, yuzu treated rats had less visceral fat. AMY did not disrupt glucose tolerance due to increased insulin secretion although AMY exacerbated insulin resistance as indicated by HOMA‐IR and insulin tolerance tests. Yuzu treatment tended to improve the first phase of glucose tolerance and it was related to improved insulin sensitivity since it decreased insulin secretion. In conclusion, yuzu treatment alleviated cognitive dysfunction and impaired energy and glucose homeostasis by improving insulin sensitivity in rats with AMY‐induced cognitive dysfunction.
Characterization of the neuroprotective potential of Passion Fruit
Authors: Tal Yael , Samach Alon , Tirosh Oren , Troen Aron M
Published: June 2021

Neurodegenerative diseases including Alzheimer's and Parkinson's disease share common mechanisms of neurotoxic damage from reactive oxygen species. Evidence from animal and in vitro models suggests that natural compounds found in a variety of edible berries and fruits can mitigate such damage, and can therefore play an important role in a comprehensive approach to neuroprotection. Passiflora comprises a genus of perennial climbing vines that produce large attractive flowers resembling a crucifix and yield aromatic juicy fruit. Although anxiolytic properties of some Passiflora species have been described, their potential neuroprotective properties are not well characterized. We bred hybrid lines related to the commercially grown ‘Passion Dream’ variety and screened them for in‐vitro cytoprotective properties. Cultured HT4 neurons were exposed to chemicals that normally cause cell death: low levels of hydrogen peroxide flux or elevated levels of extracellular glutamate. This screen identified a line with greater protective activity than either the commercial “Passion Dream” line or N‐acetylcysteine, a known neuroprotective antioxidant. Results extending these findings in vivo will be presented.
Omega‐3 fatty acids are potential chemo‐sensitizing agents for the treatment of B‐cell Chronic Lymphocytic Leukemia
Authors: Fahrmann Johannes Francois , Hardman Elaine
Published: June 2021

B‐Cell chronic lymphocytic leukemia (CLL) is the most common form of leukemia in the United States. Clinical treatment of CLL is limited due to drug resistance and therapy‐induced toxicities. We hypothesized that omega 3 fatty acids (n‐3), eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA), would increase the sensitivity of B‐leukemic cell lines EHEB, JVM‐2 and MEC‐2 to doxorubicin, vincristine and/or fludarabine in vitro and that enhanced sensitivity is due, in part, to the formation of lipid peroxides and generation of reactive oxygen species (ROS).

Results indicated that 1) EPA and DHA differentially sensitized EHEB, JVM‐2 and MEC‐2 to doxorubicin, vincristine and fludarabine in vitro, 2) lipid peroxidation increased in the presence of n‐3, 3) MEC‐2 illustrated increased lipid peroxidation in presence of DHA and doxorubicin versus either alone, 4) n‐3 increased generation of ROS in MEC‐2, 5) the addition of á‐tocopherol abrogated the increase in chemo‐sensitivity of MEC‐2 to doxorubicin by DHA.

Omega 3 fatty acids present a promising non‐toxic approach to sensitize malignant cells to anti‐cancer drugs.

Grant Funding Source: NASA WVSGC Fellowship Grant
Effects of Mung‐Bean and Black‐Bean ethanol extracts on inflammation related to obesity in the high fat induced obesity model
Authors: Lee Myoungsook , Kim Minjin , Kim Soyoung , Wi Haeri , Park Hyejin , Choi Wonhyung
Published: June 2021

Obesity is known to induce an inflammation status that is implicated in clinical complications such as insulin resistance, diabetes and atherosclerosis. The leguminous plants are known as diet food because mung bean(MB) has lower glycemic Index and anthocyanins of the black bean(BB) has physiological effects on antioxidant, anticancer and anti‐inflammatory activity. Our purpose of this study was to determine the effects of MB and BB on inhibition of inflammation‐related to adipogenesis. C57BL/6 male mice(4wk) were fed with 45% high fat diet for 10 weeks to induce obesity and the control(CON), 45% high fat diets(HF), HF with Orlistat 0.03%(O), HF with MB ethanol extracts(1g/kg), HF with BB ethanol extract(1g/kg) were orally administrated for 12weeks. The weight of epididymal fat was significantly low in MB group. PPARγ, C/EBP and ACC expression, related to lipid synthesis, on MB and BB were lower than other groups in liver. IL‐10, inhibitor of inflammation related to adipocyte differentiation, significantly increased in BB group. HSL and AMPK genes, fat degradation enzyme, were significantly increased in BB group. ACC gene, key enzyme of fat synthesis, was significantly decreased in BB group. In conclusion, MB and BB may develop an anti‐obesity food because they have effects on inhibition of inflammation related to fat synthesis and activation of lipolysis.

Grant Funding Source: Agriculture Science&Technology Development(PJ907089), Technology Commercialization Support Program, Ministry for Food, Agriculture, Forestry and Fisheries(811003031SU000), Republic of Korea.
Sex‐Dependent Depot Differences in MMPs and Inflammation of Adipose Tissue Remodeling in Mice
Authors: Wu Yuanyuan , Lee MiJeong , Fried Susan K.
Published: June 2021

We reported (Grove et al, IJO, 2010) gonadal (GON) and inguinal (ING) sc adipose tissues of high fat (HF) fed obese female compared to male mice (C57/BL6) are less inflamed. Microarray and RT‐qPCR studies revealed significant sex and depot differences in the expression of metalloproteinases (MMPs), enzymes that remodel adipose tissue in obesity. We followed up by comparing the mRNA expression of MMPs, 3 and 12, and their inhibitors (TIMPs) in age‐matched mice fed low compared high fat (45% calories) diets for 14 weeks, and extended studies to the mesenteric, a visceral depot and dorsal Sc (DSC), which in contrast to the ING, does not include mammary gland. As expected, HF diets induce a robust inflammatory response that includes increases in MMP12 in males, but not females and the response was greater in the visceral than sc depots. In females, in contrast, MMP3 mRNA was higher in sc than visceral (both ING and DSC) with low fat (LF) feeding and was not affected by the HF diet. However, HF diet increased MMP3 in all depots in males. MMP3 and MMP12 were mainly expressed in the stromal cell fraction. Flow cytometry and qRT‐PCR analysis suggested MMP3 transcripts were made from Lin− CD29+Sca1+CD34+/− population enriched for preadipocyte markers, while MMP12 was expressed in macrophages. We conclude that there are major sex dependent depot differences in gene expression of MMPs and TIMPs and those differences may contribute to sex‐ and depot‐differences in adipocyte size and adipose tissue expansion.

Funding sources: Evans Center for Interdisciplinary Biomedical Research and NIH(R24DK087669).
Inflammatory and antioxidant response in obese septic shock patients
Authors: Victorino Vanessa Jacob , Barbeiro Hermes Vieira , Barbeiro Denise Frediani , Silva Fabiano Pinheiro , Souza Heraldo Possolo
Published: June 2021

There is no consensus about the influence of obesity on sepsis. Hence, we evaluated the inflammatory and antioxidant response in obese patients (body mass index >; 30) with septic shock compared to non‐infected obese and non‐obese septic patients. Blood samples were obtained from 27 critically ill patients admitted to ICUs in Clinics Hospital, Universidade de Sao Paulo. Cytokines were measured by ELISA Milliplex and antioxidant activity by colorimetric methods. There are small differences in the cytokine profiles in obese septic patients (n=6), compared to obese non‐infected ones (n=10). Only FGF2, TGF‐α, IFN‐α2, IFN‐γ, IL‐10, MCP‐3, IL‐13 and IL‐15 presented significantly higher levels in septic patients. Interestingly, there was a marked increase in superoxide dismutase (SOD) and catalase (CAT) activity in erythrocytes from the septic group. Compared to their non‐obese septic counterparts, septic obese patients presented significantly lower levels of FGF2, IL‐4, TNF‐β and VEGF. SOD activity was higher in this group, compared to non‐obese patients. We concluded that obese patients with septic shock maintain cytokine levels similar to the ones observed in their non‐obese counterparts, while increasing their antioxidant activity.
EP4 antagonist suppresses MMP‐9 expression in aortas derived from apoE‐null mice
Authors: Khan K.M. Faisal , Falcone Domenick J.
Published: June 2021

Cox‐2‐dependent synthesis of PGE2, and PGE2 engagement of the EP4 receptor induces macrophage MMP‐9 expression, which is thought to destabilize occlusive and aneurysmal lesions. In these studies, we compared the effects of a short regimen of orally administered EP4 antagonist or celecoxib on MMP‐9 expression in aortas derived from apoE‐null mice fed a Western diet. After 12 weeks, mice received by gavage DMSO, celecoxib (10 mg/kg) or EP4 antagonist AH23848 (10 mg/kg) for 5 days. On day 6, mice were euthanized and blood samples collected for plasma cholesterol. Aortas were recovered, flash frozen in liquid nitrogen, pulverized and solubilized in lysis buffer. MMP‐9 levels in the aortic extracts were evaluated by Western blot. Serum cholesterol levels, which were markedly elevated (1056±317 mg/dl; mean ± SD), did not differ between the treatment groups. MMP‐9 antigen was not detected in aortic extracts from apoE‐null mice fed a normal diet. In contrast, MMP‐9 was detected in aortic extracts recovered from all mice fed the Western diet for 12 weeks, and in extracts recovered from mice treated with DMSO for 5 days. In marked contrast, MMP‐9 antigen was reduced or barely detectable in extracts derived from mice treated with celecoxib or the EP4 antagonist. Taken together, these data confirm that MMP‐9 expression in atherosclerotic aortas is attenuated by blocking PGE2 binding to EP4. These studies were supported by HL093331.
Reconstruction and evolution of metabolic networks across a phylogenetic tree
Authors: Vitkup Dennis
Published: June 2021

Due to inherent uncertainty of computational annotations, predictions of biochemical function need to be treated probabilistically. We present a first global probabilistic approach to annotate genome‐scale metabolic networks that integrates sequence homology and context‐based correlations under a single principled framework. The developed method for Global Biochemical reconstruction Using Sampling (GLOBUS) not only provides annotation probabilities for each functional assignment, but also suggests likely alternative functions. GLOBUS is based on statistical Gibbs sampling of probable metabolic annotations and is able to make accurate functional assignments even in cases of remote sequence identity to known enzymes. Taking advantage of the developed methods for metabolic reconstruction, we performed a comparative study of the genotype to phenotype relationship in many hundreds of different species throughout the bacterial phylogenetic tree. Our results, which are in excellent agreement with available experimentally measured phenotypes, reveal significant differences in metabolic properties of closely related strains, and a marked contribution of bacterial lifestyle to phenotypic diversity. The analysis allows an understanding of phenotype‐to‐genotype relationships, and their evolution, on a scale not currently accessible to experimental approaches.
Regulation of liver growth by Glypican 3, CD81, Hedgehog, and Hhex
Authors: Bhave Vishakha , Mars Wendy , Donthamsetty Shashikiran , Luo Jianhua , Bowen William , Michalopoulos George
Published: June 2021

Previous studies from our lab have shown Glypican 3 (GPC3) as a negative regulator of growth. CD81 was found to be a binding partner for GPC3 and its expression and co‐localization with GPC3 increased at the end of hepatocyte proliferation. However, the mechanisms through which these two molecules might regulate liver regeneration are not known. We tested the hypothesis that GPC3 downregulates Hedgehog (HH) signaling pathway by competing with Patched‐1 for HH binding. We found decreased GPC3‐Indian HH binding at peak proliferation in mice followed by decrease in Glioblastoma 1 (GLI1) protein. We performed a yeast two‐hybrid assay and identified Hemopoietically expressed homeobox (Hhex, a known transcriptional repressor) as a binding partner for CD81. We tested the hypothesis that Hhex binding to CD81 keeps it outside the nucleus. However when GPC3 binds to CD81, CD81‐Hhex binding decreases, resulting in nuclear translocation of Hhex and transcriptional repression. In support of this, we saw decreased GPC3‐CD81 binding at hepatocyte proliferation peak, increased CD81‐Hhex binding, and decreased nuclear Hhex. GPC3 transgenic mice were used as an additional tool to test our hypothesis. Overall, our data suggest that GPC3 downregulates cell proliferation by: 1) binding to HH and downregulating HH pathway; and 2) binding with CD81 making it unavailable to bind to Hhex and causing nuclear translocation of Hhex.
Mitochondrial unfolded protein response (mtUPR) participates in hepatic dysfunction in sepsis
Authors: Huang Li‐Ju , Ho Jia‐Jing , Yang Rei‐Cheng
Published: June 2021

Mitochondrial dysfunction‐induced bioenergetic failure seems to be a primary event in sepsis‐ensued multiple organ failure. Accumulation of ROS leads to a stressed situation and increase the pool of unfolded and misfolded proteins in mitochondria. The mitochondrial unfolded protein response (mtUPR) is one of the mitochondria‐nuclear communication, that activates transcription of nuclear‐encoded mitochondrial chaperone (hsp60/10) genes to promote protein homeostasis within the organelle. Therefore, the mtUPR is the most important protein quality control system in oxidative stress‐ related pathology. From our previous studies in a cecal ligation and puncture (CLP) sepsis model, we have found that content of ATP decreased, membrane potential decreased, apoptosis increased, etc. Nevertheless, we also found most of the adverse effects could be reversed by previous overproduction of chaperone proteins. These results encourage us to further investigate whether disarrangement of the mtUPR may contribute to the result of a cell energetic failure that correlates with hepatic dysfunction in sepsis. The results are positive and showed that the response could be induced 3 hours after sepsis induction but then these chaperone proteins decreased along with the progress of sepsis. In conclusion, we suggested that mtUPR failure occurred during sepsis and the index of mtUPR may be a valuable additional measurement in assessing the risk of organ dysfunction in the clinical setting.
A one, two finish of β‐catenin: targeting its activity and synthesis in hepatocellular carcinoma by novel small molecules and antisense peptide‐nucleic acids
Authors: Delgado Evan R , Bahal Raman , Ly Danith , Mustata Gabriela , Monga Satdarshan P. S.
Published: June 2021

Hepatocellular Carcinoma (HCC) is the 3rd most common cause of cancer related death throughout the world due to a lack of an effective therapy. There has been increasing evidence that about 30% of all clinically diagnosed HCC cases are a result of mutations in the β‐catenin gene. Several factors are regulated by β‐catenin, which play a role in cell proliferation, migration, angiogenesis, and survival, which can lead to tumor formation. We have developed a small molecule (SM) inhibitor of β‐catenin activity (PMED‐1), as well as a novel, cell permeable, anti‐sense oligonucleotide peptide nucleic acid (PNA) that specifically targets the β‐catenin transcription start site (T1) and inhibits its protein and mRNA expression. Either SM or PNA treatment is able to reduce β‐catenin activity at 24–72 hours as shown by decrease in β‐catenin‐TCF luciferase reporter in multiple HCC cell lines. Concomitantly, decreased β‐catenin activity led to a reduction in cell viability (MTT) and proliferation (Thymidine Incorporation), while increasing the number of apoptotic cells (TUNEL). Following treatment of either SM or PNA also results in decreases in β‐catenin driven pro‐survival, proliferation, and pro‐angiogenesis genes such as Cyclin D1, GS, and VEGF‐A. Thus we report two independent strategies, one reducing β‐catenin overactivity in HCC by targeting β‐catenin‐β interactions in nuclei, and second decreasing β‐catenin gene expression. Both strategies lead to down regulation of β‐catenin target genes in HCC cells, thus impacting tumor biology and hence have significant therapeutic implications.
From Animal Models to Clinical Practicality: Lessons Learned from Current Translational Progress in Diabetic Peripheral Neuropathy Research
Authors: Li Chengyuan , Barnard Neal
Published: June 2021

Diabetic peripheral neuropathy (DPN) is a leading risk factor for diabetes‐related mortality and morbidity. Despite promising effects in animal models, none of the pharmacological agents has been successfully translated into effective treatments in humans. Challenges faced by investigators seeking to translate results from animal models into human trials include marked species differences in genotype and behavior, nerve structure and metabolism, duration of diabetes, and tissue vulnerability. While animal experiments have provided much of our understanding of the disease biology, these species differences necessitate a careful reevaluation of the validity of data obtained from experimental approaches. Therefore, we examined these pathogenetic treatments and summarized their clinical outcomes in treating DPN. Moreover, we examined the most commonly used rodent models during preclinical drug screening, including STZ‐diabetic rats and db/db mice. By putting previous observations generated by animal studies into perspective, we investigated the salient features and predictability of these models in relation to the pathophysiological and pharmacological outcomes in diabetic patients. In particular, we identified the molecular, cellular and physiological differences that significantly impair the translation from experimental approached to human clinical setting. We conclude that these findings should be taken into critical consideration by researchers as limitations for efficient pharmacological translation.
NECROPSY: USEFUL TOOL TO STUDY THE LIFE HABITS AND THE PARASITISM BY Dioctophyme renale IN MANED WOLF (Chrysocyonbrachyurus), BRAZIL
Authors: Duarte Janaina , de Mello Ribeiro Claudia , Hyodo Vivian Cristina Costa Castilho , Costa Adriana Miranda Bezerra , Martins Jessica Amancio , Oliveira Maria Elena
Published: June 2021

The maned wolf is a wild canid found in cerrado areas or in open fields in Brazil. A large number of these animals host parasites such as Dioctophyme renale, Dipylidium sp., Toxocara sp., Ancylostoma caninum and coccidia which may lead to death due to stressing situations. Although scarce, epidemiological studies of wild canids have cited that parasitic diseases can be the first causes of maned wolf mortality. Therewith, necropsy technique can be a useful tool to understand important characteristics these wild animals with risk of extinction. In this work, a case of a maned wolf parasitized by D. renale is reported. A young male maned Wolf was found dead and in rigor mortis in a road at Pindamonhangaba Municipality, São Paulo, Brazil. The animal underwent necropsy in the Veterinary Hospital of Universidade Paulista – UNIP, São Jose dos Campos Campus, São Paulo, Brazil. During necropsy, intense epistasis, oral cavity bleeding, hemothorax and hemoperitoneum were observed, suggesting that the cause of death of this maned wolf was traumatism, probably due to road killing. In the stomach, several rats and Diptera larvae were found partially digested. The Diptera larvae detection suggests a meat‐based diet in a state of putrefaction what can be explained by diet restriction in urban areas. The right kidney showed renal parenchyma atrophy, bloody content and three D. renale specimens: one female, one male and one larva. The bloody content present in the kidney was analyzed under an optical microscope. The identified eggs were bioperculate, elliptical, brown and had a thick layer, characteristic of D. renale, confirming the literature findings. The necropsy showed be an important understanding tool of life habits of wild animals that have undergone many changes with increasing urbanization.
Role of Ets‐2 in CD144 expression in human peripheral blood mononuclear cells and early‐outgrowth cells
Authors: Tejerina Teresa , Medina Ursula , Redondo Santiago , Ostos Fernando , Navarro‐Dorado Jorge , Ramajo Marta
Published: June 2021

In atherosclerotic patients undergoing coronary artery‐bypass grafting (CABG) it has been shown decreased numbers of endothelial progenitor CD144+ cells, compared to coronary artery disease‐free valvular patients (1). At the same time, Ets‐2 is a transcription factor involved in endothelial differentiation and basic and clinical models of vascular remodelling. We aimed to determine the expression of CD144 in peripheral blood mononuclear cells from CABG and valvular patients, and the role of Ets‐2 inhibition on CD144 expression. CD144 expression was determined from peripheral blood mononuclear cells obtained from 15 CABG and 16 valvular patients. Ets‐2 expression was silenced by siRNA and checked by Western blot (WB). In both models, CD144 expression was assessed by WB. In patients, CD144 expression was lower in CABG patients, compared to valvular (1.145±0.2165 vs. 1.834±0.1832, p=0.0209). However, inhibition of Ets‐2 expression siRNA produced a trend to a higher expression of CD144 in cultured early‐outgrowth cells from healthy donors (1.112 ± 0.07387 vs. 1.269 ± 0.05976). In conclusion, the expression of Ets‐2 remains lower in CABG patients, whereas Ets‐2 inhibition increases CD144 in early outgrowth cells from healthy donors, FISS (PI080920) and Spanish Network CV RECAVA (RD06/0014/1007), Spanish Ministry of Health
Activation of Rac1 at adherens junctions promotes VE‐cadherin trans interaction
Authors: Daneshjou Nazila , Komarova Yulia A. , Malik Asrar B.
Published: June 2021

The monomeric RhoGTPases Rac1, Cdc42, and RhoA play a critical role in regulating endothelial junctional permeability; however, the causal relationship between Rac1 activity at adherens junctions (AJs) and stability of Vascular Endothelial (VE)‐cadherin adhesion, the main adhesive complex of AJs, is not well understood. Utilizing a photo‐activatable Rac1 probe (PA‐Rac1) as a tool to control the spatial and temporal activity of Rac1 we demonstrated that photo‐activation of PA‐Rac1 at the level of AJs induced VE‐cadherin accumulation at a rate constant of 0.36 ± 0.22 min−1 in human dermal microvascular endothelial cells. Pretreatment of cells with inhibitory peptide (SP) that binds to a VE‐cadherin extracellular 1 module and prevents trans interaction abolished PA‐Rac1 effect, suggesting that Rac1 stabilizes VE‐cadherin adhesion. Consistently, PA‐Rac1 activation decreased the dissociation rate of wt‐VE‐cadherin, but not adhesion‐defective Trp2Ala and Trp4Ala mutant, from AJs as demonstrated by dynamics of photo‐switchable protein Dendra2‐tagged VE‐cadherin. In addition, PA‐Rac1 activation at AJs decreased mechanical tension across VE‐cadherin adhesion, which was directly measured using VE‐cadherin FRET‐based tension biosensor. These data cumulatively demonstrate that Rac1 signaling stabilizes VE‐cadherin trans interaction and AJ integrity by reducing tension across cell‐cell junction.
Adenosine A1 receptor signaling inhibits BK channels
Authors: Kunduri Swati Sarada , Dick Gregory M , Nayeem Mohammed A , Mustafa S. Jamal
Published: June 2021

Adenosine receptor (AR) contracts and relaxes mouse aorta through all four subtypes (A1, A2A, A2B, and A3) linked to different signaling mechanisms. Deciphering complex tissue responses remains difficult because relationships of individual receptor subtypes and various end‐effectors (e.g., ion channels) are yet to be identified. A1AR is linked to cytochrome P450 4a (Cyp4a), an enzyme producing 20‐HETE from arachidonic acid. 20‐HETE can inhibit large conductance Ca2+/voltage‐sensitive K+ (BK) channels that are important in adenosine‐induced smooth muscle relaxation. We tested the hypothesis that activation of A1AR blunts adenosine‐induced vascular relaxation by coupling 20‐HETE production to inhibition of BK channels. Patch clamp recordings and Western blots were conducted by using wild type (WT) and A1AR knockout (A1KO) mice aortic myocytes. We observed no difference in total BK current or BK channel proteins (BK‐α and BK‐β1) in aortic myocytes from WT and A1KO mice. Non‐selective adenosine agonist NECA (10 μM) increased BK current in myocytes from both WT and A1KO mice, but had greater response in aortic myocytes from A1KO mice (p < 0.05). 20‐HETE (100 nM) inhibited BK current in myocytes from WT and A1KO mice and no difference was observed in the inhibition. Our data suggest that A1AR limits adenosine‐induced vascular relaxation by inhibiting BK channels. (HL027339, HL094447).
Impaired caveolin‐1 contributes to hepatic ischemia and reperfusion injury via suppression of sphingosine kinase/sphingosine‐1‐phosphate receptor pathway
Authors: Kang Jung‐Woo , Pyo Suhkneung , Lee Sun‐Mee
Published: June 2021

Caveolae are membrane structure enriched in glycosphingolipid and cholesterol and caveolin‐1 (Cav‐1) has been recognized to be pivotal in ischemic tolerance. Sphingosine‐1‐phosphate (S1P), one of the sphingolipid metabolites, is well known for its anti‐apoptotic properties to counteract against ischemia/reperfusion (I/R) injury and it acts through G protein‐coupled S1P receptors (S1P1‐S1P5). Recently, sphingosine kinases (SphK1 and SphK2) which phosphorylate sphingosine to S1P have been a target to regulate S1P pathway. Here, we investigated the cytoprotective role of Cav‐1 against I/R injury focusing on S1P signaling pathway. Mice underwent partial hepatic ischemia for 60 min followed by reperfusion. Recombinant peptides mimicking Cav‐1 scaffolding domain (CSD, 2.5 mg/kg, i.p.) was administered 1 h prior to ischemia. Ischemia itself reduced hepatic Cav‐1 protein level and this decrease persisted by 1 h of reperfusion. However, this decreased Cav‐1 protein level recovered to basal level 6 h of reperfusion and maintained by 24 h of reperfusion. The loss of caveolae structure and decreased Cav‐1 protein level was restored by CSD 1 h of reperfusion. Cav‐1 induction with CSD attenuated I/R‐induced increases in serum ALT and TNF‐α activity. The levels of SphK1 and Sphk2 mRNA decreased 1 h of reperfusion. CSD attenuated the decrease of SphK1 mRNA level while it did not affect SphK2. At 1 h of reperfusion, the levels of S1P1 and S1P2 mRNA decreased and CSD attenuated the decrease of S1P2 mRNA level while it did not affect S1P1. CSD also decreased phosphorylation of p38 MAPK and JNK. Moreover, I/R resulted in increases of cytochrome c release and apoptotic hepatocytes and CSD attenuated these increases. Our findings suggest that decreased Cav‐1 expression might contribute to hepatic I/R injury through down‐regulation of S1P pathway.