bluuebunny/crossref_metadata_2025 huggingface dataset:
Suppression of γ‐secretase activity increases ganglioside levels at neuritic terminals of differentiated PC12
Authors: Oikawa Naoto , Goto Miho , Ikeda Kazutaka , Taguchi Ryo , Yanagisawa Katsuhiko
Published: June 2021

Mutations in presenilin genes cause the dominant type of familial Alzheimer disease. Presenilin mutations have recently been argued to work as loss‐of‐function; however, their pathological significance remains obscure. In this study, we investigated whether presenilins are involved in the regulation of the distribution of neuronal membrane lipids by treatment of neuronally differentiated PC12 with DAPT, an inhibitor of presenilin‐dependent γ‐secretase. Levels of membrane lipids in neuritic terminals and neuronal somata were analyzed using electrospray ionization tandem mass spectrometry. Through DAPT treatment, whereas cholesterol level stayed unchanged, the levels of phosphatidylcholine and sulfatide increased in both neuritic terminals and neuronal somata. Remarkably, the levels of gangliosides increased in neuritic terminals but decreased in neuronal somata by DAPT treatment concomitant with similar alteration for sphingomyelin. Considering the previous findings that accumulation of gangliosides and sphingomyelin at neuritic terminals induces Aβ amyloid formation, these results suggest that a decrease in γ‐secretase activity affects membrane lipids environment as leading to facilitation of Aβ amyloid formation in the brain. This study was supported by a Grant‐in‐Aid for Research Activity start‐up and for Scientific Research (B) from the Japan Society for the Promotion of Science.
Shorter forms of the proteoglycan, Perlecan; Adding complexity to an already complex molecule
Authors: Whitelock John Michael , Jung Moon Sun , Chuang Christine Yunang , Cheng Bill , Koo Tony , Lyons James (Guy) , Lord Megan Susan
Published: June 2021

Proteoglycans are a group of molecules that exist on cell surfaces and in the extracellular matrix and are characterized by their polydisperse glycosaminoglycan chains, which make this group of molecules one of the most complex in the proteome. The aim of our study was to investigate whether the proteoglycan, perlecan, the major heparan sulfate PG of basement membranes and basal laminae is alternatively spliced using both semi‐quantitative PCR and antibodies specific for the domains on the protein core. Our results indicated that is in some cell types, notably mast cells, transcripts from the C‐terminal region containing the endorepellin sequence, were over represented in the mRNA population suggesting either the use of an alternative promoter or transcripts that have arisen from mRNA splicing events. This data supports our hypothesis that the structure of proteoglycans is inherently more complex than first thought and this complexity changes dynamically in response to environmental and cell signals and being fundamentally important for many biological and pathological processes involving angiogenesis.
O‐GlcNAcylation and hyaluronan synthesis
Authors: passi alberto , vigetti davide , deleonibus sara , viola manuela , karousou evgenia , moretto paola , bartolini barbara , deluca giancarlo
Published: June 2021

The intracellular regulation of HA synthesis throughout O‐GlcNacylation is investigated. It was reported that the HA precursor UDP‐N‐acetylglucosamine (UDP‐GlcNac) and UDP glucuronic acid availability modulates of HA synthesis. In this study, we tested the role of O‐GlcNacylation, a type of cytoplasmic protein O‐glycosylation to ser/thr residues and its influence on hyaluronan synthase activity. We found that the inhibition of OGlcNacylation strongly reduced HA production whereas treatments that induced protein O‐GlcNacylation increased HA secretion. Gene expression studies revealed that HAS2 mRNA (i.e., the main HA synthesizing enzyme located in the plasma membrane) was the most sensible to O‐GlcNacylation and accumulated after its induction. We found evidences that the transcription regulator YY1 activates HAS2 expression after O‐GlcNacylation. At protein level, experiments with wheat germ agglutinin and recombinant 6myc‐HAS2 revealed that this enzyme was actually O‐GlcNacylated. Further, HAS2 O‐GlcNacylation increased HAS activity in purified microsomes leading to HA accumulation. Interestingly, the O‐GlcNacylation was able to delay the protein removal from cell membrane increasing the enzyme turn‐over, another way to induce increase of HA. This work was supported by FAR and Onlus Varesotto‐ITALY.
N‐(1‐Pyrenyl) maleimide induces apoptosis through mitochondrial pathway
Authors: Huang Pei-Rong , Wang Tzu-Chien
Published: June 2021

N‐(1‐pyrenyl) maleimide is a fluorescent compound that is frequently used as a derivatization agent for the detection of thio‐containing compounds. Reports on the biological effect of this compound in literature were rather scanty. We have previously shown that N‐(1‐pyrenyl) maleimide induces apoptosis in Jurkat T cells and displays great differential cytotoxicity against hematopoietic cancer cells. As yet, the molecular mechanism of N‐(1‐pyrenyl) maleimide‐induced apoptosis remains largely unknown. Here, we show that treatment with N‐(1‐pyrenyl) maleimide leads to loss of mitochondrial membrane potential (£G£rm) and release of cytochrome C from mitochondria to cytosol in Jurkat T. The expression of inhibitor of apoptosis protein (IAP) family, which includes c‐IAP1, c‐IAP2 and XIAP, was significantly decreased, while the caspase‐9 (a target of inhibition by IAP) was activated after treatment with N‐(1‐pyrenyl) maleimide. Surprisingly, the level of key regulators of mitochondrial apoptotic pathway, such as Bcl‐2, Bcl‐XL, Bax and Bad, was not noticeably changed in the treated cells, suggesting that N‐(1‐pyrenyl) maleimide can directly target mitochondria to trigger the intrinsic apoptotic pathway. These findings suggest that N‐(1‐pyrenyl) maleimide may be explored as a potential therapeutic agent for the treatment of leukemia. This work was supported by research grant NSC 99‐2311‐B‐182‐003‐MY3.
GSK3beta regulates Bcl2L12 and Bcl2L12A anti‐apoptosis signaling in glioblastoma and is inhibited by LiCl
Authors: Hong Yi-Ren , Lin Ching-Chih , Chou Chia-Hua , Howng Shen-Long
Published: June 2021

Bcl2L12 has been reported to be involved in post‐mitochondrial apoptotic events in glioblastoma, but the role of Bcl2L12A, a splicing variant of Bcl2L12, remains unknown. In this study, our data demonstrated that GSK3beta interacts with Bcl2L12, but not Bcl2L12A, which C‐terminal lacks a binding region. We found that a Bcl2L12153‐191 fragment located outside of the C‐terminus BH2 motif is responsible for GSK3beta binding. In vitro kinase and lamda‐phosphatase assays showed that GSK3beta phosphorylates Bcl2L12 at S156, while this site is absent on Bcl2L12A. Transiently expressed GFP‐fused proteins, including Bcl2L12, Bcl2L12(S156A) and Bcl2L12A, in U87MG cells. Staurosporine (STS) treatment demonstrated that both Bcl2L12 and Bcl2L12A may have anti‐apoptotic properties. In contrast, no anti‐apoptotic ability was seen in Bcl2L12(S156A). When STS was administered in combination with LiCl, the Bcl2L12‐expressing U87MG cells underwent apoptosis. This effect could be reversed by LiCl. In short, we established a model to demonstrate that GSK3beta interacts and phosphorylates Bcl2L12 and might also affect Bcl2L12A to modulate of the apoptosis signaling pathway in glioblastoma. These findings suggest that LiCl may be a prospective therapeutic agent against GBM
Golgi apparatus location and regulation of mTOR is regulated by novel Golgi recruiting protein GRP
Authors: Thomas Janice D. , Zheng X.F. Steven
Published: June 2021

Mammalian target of rapamycin (mTOR) is a central protein involved in the control of cell growth and metabolism. Dysregulation of mTOR signaling has been closely linked to many human diseases such as cancer, diabetes, obesity cardiovascular diseases, neurological disorders and age related diseases. Although many of the components in the mTOR pathway have been identified, little is known regarding the mechanism of the spatial arrangement of mTOR signaling. Previously, our lab reported that in several common cell lines, a significant amount of mTOR is localized to the membrane of particular sites called the endoplasmic reticulum (ER) and Golgi apparatus. mTOR localization to the ER and Golgi is mediated by novel motifs called the Golgi localization sequence (GLS) and an overlapping endoplasmic reticulum localization sequence (ELS), respectively. Mutation of the GLS or ELS, or over‐expression of these sequences alone, results in inhibition of both mTOR complexes. These results suggest that subcellular localization of mTOR to the Golgi and ER is essential for mTOR signaling. Because GLS lacks apparent membrane association property, we hypothesized that a Golgi membrane protein(s) is responsible for anchoring mTOR to Golgi through the interaction with GLS. We identified a protein that possesses a strong interaction with the GLS. This novel protein carries a Golgi membrane localization domain and directly interacts with and regulates mTOR activity. We therefore named it GLS Receptor Protein (GRP). My data exhibits strong evidence that GRP mediates mTOR localization to the Golgi and is involved in mTOR regulation. Support for this research was provided by the New Jersey Commission on Cancer Research and the National Institute of Health.
Novel selective estrogen receptor modulators induce LNCaP cell death via mechanisms involving apoptosis
Authors: Green Jennifer Evette , Cooperwood John S. , Stephenson Adrienne P. , Reams R. Renee
Published: June 2021

Selective Estrogen Modulators (SERMs) offer an alternative approach for treating prostate cancer that differs from the treatment options that are currently available. Using the LNCaP cell line we evaluated three novel SERMs, 3‐[2‐diethylamino]‐ethoxy]‐]‐D 1,3,5(10)‐estrien‐17‐one (DE), 3‐[2‐diisopropylamino]‐ethoxy‐ D1,3,5(10)‐estrien‐17‐one (DI) and 17‐b‐3‐[2‐piperidinyl]‐ethoxy]‐ ]‐D 1,3,5(10)‐estrien‐17‐ol (EPOH), which are aminoalkyloxy derivatives of 1,3,5(10)‐Estratrien. The cytotoxic effect (judged by cell viability) of DE, DI, EPOH and 4‐hydroxytamoxifen (4‐OHT) was determined using the Alamar Blue assay. The effect of the SERMs on apoptosis was determined by Acridine Orange and Ethidium Bromide (AO/EB) staining method. It was observed in comparison to the control, that the three experimental compounds caused a dose‐dependent decrease in cell viability (EC50 values ranging between 300 μM – 400 μM). AO/EB staining indicated the induction of apoptosis at the same concentrations that caused the dose‐dependent increase in cell death by DE, DI and EPOH. Taken together these results indicate that the novel SERMs cause cell death via underlying mechanisms that lead to apoptosis/programmed cell death and with further evaluation will reveal the potential of these novel SERMs for the treatment of prostate cancer. Supported by NIH/NCRR/RCMI Grant (G12 RR0 3020).
A Yeast‐Two Hybrid Screen Against HPV16 Capsid Proteins Indicates Dynactin 6 is a Potential Cellular Factor Involved in HPV16 Infection
Authors: Schlegel Angela M , Chapman Janice A , Campos Samuel K
Published: June 2021

Human papillomaviruses (HPVs) infect genital and mucosal epithelium and include strains carrying different levels of risk for the host upon infection, including high‐risk strains that are potentially oncogenic such as HPV16. Little is known about the molecular events of HPV host cell invasion. HPV16 particles consist of two structural proteins: the major capsid protein L1 and the minor capsid protein L2. The goal of this project is to identify host cell proteins that interact with the HPV capsid proteins. A high stringency yeast two‐hybrid screen was performed to identify possible interactions and yielded >1000 colonies for the L2 bait and no colonies for the L1 bait. L2 clones were analyzed by PCR and sequencing to identify the interacting proteins, which included two clones encoding full‐length dynactin‐6. Mapping of the interaction site using the yeast two‐hybrid screen indicates a probable interaction site within residues 99–171 of dynactin 6. These results indicate that dynactin‐6 is a host cell factor involved in HPV16 infection through interaction with the L2 capsid protein. Current work seeks to confirm this interaction and determine the specific role this interaction plays in HPV16 infection. Support for this research comes from the Bio5 Institute (U. of Arizona) and the American Cancer Society, Research Scholar Grant, RSG‐117469.
Novel role of the antimicrobial peptide LL37 in the formation and stabilization of neutrophil extracellular traps
Authors: von Köckritz-Blickwede Maren , Neumann Ariane , Völlger Lena , Berends Evelien , Molhoek Margo , Nizet Victor , Naim Hassan
Published: June 2021

The antimicrobial peptide LL‐37, a member of the cathelicidin family, has been shown to be present in neutrophil extracellular traps (NETs). However, the function of LL‐37 within the NETs is still unknown, since LL‐37 loses its antimicrobial activity when bound to DNA in the NETs. The aim of this study was to explore the role of LL‐37 in NETs using confocal immunofluorescence microscopy. We could demonstrate that NETs treated with LL‐37 were distinctly more resistant to bacterial nuclease degradation as compared to non‐treated NETs. Furthermore LL‐37 was able to induce the formation of NETs by disruption of the nuclear membrane of stimulated human neutrophils. Biochemical assays utilizing a random LL‐37‐fragment library indicated that the ability of LL‐37 to block nuclease activity is based on its cationic character, whereas the NET‐induction is mediated by the hydrophobic character of the peptide. In conclusion, this study demonstrates a novel role of the antimicrobial peptide LL‐37 in host immune defence: the stabilization and induction of NETs.
Further Characterization of a Unique Mycobacterial Heme Uptake System
Authors: Goulding Celia Whitford , Owens Cedric , Chim Nick , Harmston Christine , Iniguez Angelina , Contreras Heidi , Morse Robert
Published: June 2021

Mycobacterium tuberculosis (Mtb), the primary agent of tuberculosis, is one of the most frequent causes of death by a single infectious agent. Mtb must import iron from its host for survival, and its siderophore‐dependent iron acquisition pathways are well established. Recently, we documented a newly characterized pathway, whereby Mtb can use free heme and heme from hemoglobin as an iron source.

Significantly, we identified the genomic region responsible for the passage of heme‐iron across the mycobacterial membrane. For the first time, key players of this heme uptake system were biochemically characterized including a secreted heme‐binding protein and two transmembrane proteins (MmpL3 ad MmpL11), all three specific to mycobacteria. Furthermore, within the Mtb cytosol, we have characterized a heme‐degrading protein MhuD, which releases iron from the heme tetrapyrrole ring for utilization by Mtb. We have extended our studies and will discuss further characterization of genes involved in this heme acquisition pathway, biochemical analysis of heme transfer from the secreted heme carrier protein to soluble domains of the putative heme transporters, as well as confirmation that the Mb essential gene which encodes for MmpL3, plays a role in mycobacterial heme transport.

The discovery of this unique mycobacterial heme acquisition pathway opens new avenues of exploration into mycobacterial therapeutics.

Sources of research funding are NIH NIAID/NIGMS and the American Lung Association.
Peculiarities of nutrition practice in population at risk of osteoporosis in Astana
Authors: Abduldayeva Aigul , Iskakova Saule
Published: June 2021

Urban population living in Astana aged 40 and above has been examined to study osteoporosis rate and its relationship with nutrition practice within the framework of the State Program for Public Health Development of Kazakhstan for 2011–2015 “Salamatty Kazakhstan” (“Healthy Kazakhstan”). Ultrasound densitometry method was used to detect changes associated with bone density. Population of Astana over 40 years old was examined using 24 hours recall to estimate the dietary intake indicating the weight of all consumed per day of food products, 100 questionnaires were studied. Data on family composition, ethnicity, occupational, age, and etc have been studied to ensure the representativeness of the sample by using the questionnaire. The analysis of dietary intake was found that the average caloric was about 1770 kcal, primarily at the expense of animal protein and carbohydrates. Carbohydrates are reimbursed in 73% of respondents mainly due to the “fast” carbohydrates. The content of dietary fiber in average was around 18 g, while at the recommended rate for the elderly should be not less than 30 – 35 g fiber per day. There was a vitamin and mineral deficiency in the elderly in 39.8% of cases, and the rest of their content was within normal limits.
Yogurt, dairy, calcium, and vitamin D intake are associated with lower body fat measures in US children: Results from NHANES 2005–2008
Authors: Keast Debra R , Albertson Ann M , Gugger Carolyn K , Holschuh Norton M
Published: June 2021

This study examined the associations between obesity measures, dairy and yogurt consumption, and intakes of calcium (CA), and vitamin D (VIT D) among American children. Using 1‐day dietary recall data from NHANES 2005–2008, children aged 8–18 yr (n=3,786) were classified by CA and VIT D intake tertiles, and dairy consumption: <1 (30%), 1‐<2 (29%), or 2+ servings (41%). Yogurt consumers (Y‐C) (8.5%) and nonconsumers (Y‐NC) (91.5%) were defined using 2‐day intake. After adjusting for gender, ethnicity, age, and energy (Model 1), dairy consumption was positively associated with intake of CA, and VIT D (p<0.01); and Y‐C had higher intakes than Y‐NC (p<0.01). In Model 1, Y‐C also had lower (p<0.05) BMI (21.3±0.3 vs. 22.0±0.2 kg/m2), overweight/obesity prevalence (27.2±4.0 vs. 36.1±1.9 %), waist circumference (74.5±1.0 vs. 76.9±0.5 cm), and subscapular skinfold (11.2±0.6 vs. 12.9±0.3 mm) than Y‐NC. With physical activity and other covariates added to Model 1, Y‐C had lower waist circumference (74.5±1.0 vs. 77.0±0.5 cm). Subscapular skinfold was lower for Y‐C than Y‐NC (11.3±0.6 vs. 12.9±0.3 mm), for consumers of 2+ vs. <1 dairy servings (12.3±0.2 vs. 13.4±0.3 mm), and for high vs. low tertiles of CA (12.2±0.3 vs. 13.4±0.3 mm) and VIT D (12.1±0.3 vs. 13.3±0.3 mm). The high VIT D tertile also had lower overweight/obesity prevalence (32.2±2.5 vs. 39.8±1.9 %). These results suggest that lower body fatness of yogurt and dairy consumers may be due to the high CA and VIT D content of dairy foods. Supported by the Bell Institute of Health and Nutrition, General Mills, Inc., Minneapolis, MN.
Freshman Health Initiative Survey: A Pilot Study
Authors: Schnoll Roseanne , Curran Robert , Burroughs Steven
Published: June 2021

Nine hundred and seventy‐seven entering freshman were emailed surveys about their health, physical activity, and food intake. One hundred and thirty students completed the survey and 80 students completed a 24 hour dietary recall. Fifty percent of respondents felt that they did not meet their nutritional requirements. Sixty‐seven percent of the respondents consumed 2 servings or less of vegetables a day, and 80% consumed 2 servings or less of fruits a day. The 24 hour recall showed significant deficiencies in calcium, iron and fiber intake. Sixty‐three percent engaged in vigorous exercise less than 3X week and 52% did not engage in any strength training exercises at all. Seventy–nine percent of respondents felt that “lack of time” prevented them from meeting their activity goals and 47% felt that “lack of time” prevented them from meeting nutritional requirements, however, 47% of respondents report spending 16 hours or more per week on the Internet. Future goals include obtaining physical and biochemical data to identify students at risk and implement strategies to target our students’ health and wellness needs.
Dietary Intake of Omega‐3 Long‐chain Polyunsaturated Fatty Acids (n‐3 LCPUFA) by US Toddlers
Authors: Walsh Kelly Robert , Ye Qian , Piekarz Ana
Published: June 2021

Background

n‐3 LCPUFA, in particular docosahexaenoic acid (DHA), have critical roles in the nervous, visual and immune systems. While studies have addressed DHA intake and functional outcomes in infants, data on dietary intakes of US toddlers are scarce. The NHANES 1999–2000 study showed an average consumption of 20 mg of DHA per day which is less than recommended by international expert groups.

Objective

To evaluate the average daily consumption of DHA, eicosapentaenoic acid (EPA), and docosapentaenoic acid (DPA) from food and beverages among healthy US toddlers between 12 and 47 month of age based upon the NHANES 2007–2008.

Methods

Dietary intakes of 741 healthy toddlers from NHANES 2007–2008, a nationally representative survey, were analyzed. Intakes from dietary supplements were excluded from the analysis. Mean usual intakes of DHA, DPA and EPA from food and beverages were calculated using NHANES suggested methodology to ensure a representative sample.

Results

Mean (± SE) consumption of DHA, DPA and EPA among US toddlers is 19 (± 1.9) mg/day, 4.5 (± 0.3) mg/day, and 8 (± 0.9) mg/day, respectively.

Conclusion

Dietary intake of n‐3 LCPUFA by US toddlers remains below target recommendations established by international expert groups such as the FAO (2010). Dietary strategies for closing the gap between current intakes and recommended targets are needed.
Obesity and serum high sensitivity C‐reactive protein in Turkish immigrants with type 2 diabetes
Authors: Sukram Shiryn D. , Zarini Gustavo G. , Huffman Fatma G.
Published: June 2021

The study examined the associations of anthropometric indices of obesity with high sensitivity C‐reactive protein (hs‐CRP) in Turkish immigrants with T2D. A total of 110 subjects, physician‐diagnosed with T2D, aged 30 years and older were recruited from multiple sources from The Hague, Netherlands. Subjects with hs‐CRP levels >10 mg/L (n=17) were excluded from the analysis. Serum hs‐CRP were measured with immuno‐turbidimetric assay. Glycemic control (A1C) was determined by high‐pressure liquid chromatography. Analysis included descriptive statistics, Pearson's correlations and multiple linear regressions stratified by gender. Hs‐CRP was log transformed to achieve normality. Females had a higher BMI (P=0.007) and WHtR (P=0.011) as compared to males. Conversely, males had a higher WHpR (P<0.001) than females. Multiple linear regressions showed that after controlling for confounding variables log hs‐CRP was positively associated with WHtR (β=4.015, P<0.01), WC (β=0.025, P<0.05), and BMI (β=0.039, P<0.05) in females only. Our findings of gender specific obesity indices in relation with hs‐CRP need to be further investigated in the Turkish immigrant population. Hs‐CRP assessment may be added as a standard of care for T2D treatment within this population.
Xanthohumol lowers body weight and fasting plasma glucose in obese male Zucker fa/fa rats
Authors: Legette LeeCole L , Moreno Luna Arlyn Y. , Reed Ralph L , Miranda Cristobal L. , Bobe Gerd , Proteau Rosita R. , Stevens Jan F.
Published: June 2021

Metabolic syndrome is a leading public health concern with over 50 million Americans being affected and at increased risk for other chronic diseases including cardiovascular disease, obesity and type II diabetes. Xanthohumol, a prenylated flavonoid from hops (Humulus lupulus), was tested for efficacy on biomarkers of metabolic syndrome in 4 week old Zucker obese fa/fa rats. Rats received daily oral doses of xanthohumol at 0, 1.86, 5.69, and 16.9 mg/kg BW for 6 weeks. All rats were maintained on a high fat (60% kcal) AIN 93G diet for 3 weeks to induce severe obesity followed by a normal AIN‐93G (15% kcal fat) diet for the last 3 weeks of the study. Weekly food intake and body weight were recorded. Plasma cholesterol, glucose, insulin, and triglyceride levels were assessed using commercial assay kits. There was a dose‐dependent effect on body weight and plasma glucose levels. The highest dose group (n=6) had significantly lower plasma glucose levels compared to the control group (n=6) in male but not female rats. There was also a significant decrease in body weight for male rats in the highest dose group compared to rats that received no xanthohumol which was also not seen for female rats. Plasma cholesterol, insulin and triglycerides as well as food intake were not affected by treatment. Findings suggest that xanthohumol has beneficial effects on markers of metabolic syndrome but may induce gender‐specific responses.

Grant Funding Source : National Institutes of Health and USANA Health Sciences
Influence of Exercise and Weight Loss on Glucose Tolerance and Bone Mass in Young Growing Animals
Authors: Hembree Kelsey D , Rendina Elizabeth , Wang Yan , Clark Katie , Davis McKale R , Peterson Sandra K , Clarke Stephen L , Lucas Edralin A , Smith Brenda J
Published: June 2021

Childhood obesity has been associated with increased fracture incidence, but the effects of weight loss and exercise on the bone health of children remains unknown. Therefore, this study was designed to determine the effects of exercise and weight loss on bone mass and glucose homeostasis in a young growing animal model of diet‐induced obesity. Six wk old male C57BL/6 mice (n=42) were fed a control diet (CON=10% fat) or high fat diet (HF=60%) for 12 wks. The last 4 wks of the study, mice were further divided into exercise (Ex), no exercise (NoEx), or weight loss regimen (HF→CON). At end of the study, body weight was not significantly reduced in the HF+Ex group. In contrast, the 15% reduction in weight in the HF→CON group resulted in similar body weights to mice on the CON diet. Both the HF+Ex and the HF+NoEx groups experienced an increase (P<0.05) in fasting glucose, but the glucose levels of the HF→CON group were normalized to the CON groups. The HF+Ex and HF→CON groups demonstrated a similar response in their glucose tolerance test. The 4 wk exercise protocol increased whole body and femur bone mineral density (BMD), but the HF→CON mice significantly reduced their spine BMD compared to the non‐exercising groups. These data suggest that in obese animals weight loss without exercise results in a negative effect on spine bone density.

Grant Funding Source : Oklahoma Agricultural Experiment Station
The association between Serum Estrogen, Fat Mass and Type II Diabetes in the Boston Puerto Rican Health Study (BPRHS)
Authors: Landry Latrice , Tucker Katherine
Published: June 2021

Background

Research suggests that estrogen modulates adiposity and may be protective against diabetes (T2D). Data on the association between estrogen, obesity and diabetes is limited in Puerto Ricans. This study examines the cross‐sectional relationships between serum estradiol (E2) with gynoid and android fat mass and T2D in Puerto Rican adults in the Boston Puerto Rican Health Study (BPRHS).

Design

E2 and android and gynoid fat mass (from DXA) were measured in 538 BPRHS participants. Multivariate regression was used to assess cross sectional relationships between E2 and gynoid and android fat mass. Logistic regression was used to assess the associations with T2D. E2 and total energy intake were log transformed. Covariates included age, sex, pack years, physical activity score, menopausal status, hormone use, and intakes of total energy, fat, and carbohydrate.

Results

E2 was positively associated with gynoid (p<0.0001) and android (p<0.0001) fat and T2D (0.03). Android fat (p<0.0001) was positively associated with T2D. Gynoid fat (p<0.0001) was negatively associated with T2D.

Conclusion

Consistent with findings from other populations, our findings suggest a positive association between E2 and body fat and T2D. The inverse association between gynoid fat and T2D is novel. Longitudinal studies with adjustment for other metabolic measures are needed to clarify these potential associations.

Grant Funding Source : NIH‐P01 AG023394, Pfizer SNMA David E Satcher Fellowship
Reconsidering protein, energy, and diet‐induced thermogenesis
Authors: Moulton Christopher , Wilson Gabriel , Norton Layne , Layman Donald
Published: June 2021

Among the three macronutrients, dietary protein induces the highest energy expenditure per kcal consumed, which is often attributed to amino acid degradation and nitrogen disposal. However, this fails to account for protein's post‐prandial effects on metabolism. The branched‐chain amino acid leucine has a unique ability to activate muscle protein synthesis (MPS), which expends considerable amounts of cellular energy and is fundamental in active tissue maintenance. We have utilized a meal‐feeding model in adult rats to show that leucine‐induced activation of MPS depletes cellular ATP and activates the key energy sensor AMP kinase, which was associated with energy sparing via translation elongation inhibition and with energy production via acetyl‐CoA carboxylase (ACC) inhibition. Chronic feeding of leucine‐rich meals promoted favorable body composition outcomes and triggered changes in mitochondria consistent with support for the increased energy demand of MPS. These results indicate that dietary protein's impact on translation activity in skeletal muscle may account for a substantial portion of diet‐induce thermogenesis.
Grape seed extract and delta tocotrienol alone and in combination ameliorates histopathological features in C57BL/J6 mouse fed high‐fat diet
Authors: Sodhani Shradha , Juma Shanil , Vijayagopal Parakat , Prasad Chandan , Imrhan Victorine , Mills Nathaniel
Published: June 2021

The condition of non‐alcoholic steatohepatitis (NASH) presents a progressive pathology leading to steatotic fibrosis and potentially hepatic cancer. Polyphenols in grape seed extract (GSE) and fat soluble vitamin E (ä tocotrienol, ä3T) may play a beneficial role in hepatic fibrosis. In this study we examined the effects of dietary GSE and ä3T in C57BL/6J mice fed high‐fat diet (n=8/group) for 20 weeks. Post necropsy general histology and immunohistochemical analyses were performed with alpha‐smooth muscle actin (á‐SMA) as a biomarker in liver fibrosis. Mice in the high fat diet control group (HFD‐C) had increased fat accumulation and NASH‐like histopathological features. Histological data analysis of H&E stained liver sections showed that steatosis and lipid droplets were reduced in HFD+GSE group compared to HFD‐C group. Immunohistochemical staining and analysis of photomicrographs led to observable reduction of á‐SMA in HFD+GSE supplemented group compared to HFD‐C group. Our data shows that dietary GSE and ä‐tocotrienol alone and in combination may ameliorate the NASH‐like histopathological features including fibrosis in C57BL/6J mice.

Supported by Research Enhancement Program, TWU
Consumption of dehydrated Opuntia ficus Indica (nopal) prevents the development of fatty liver by modifying hepatic lipid metabolism in obese Zucker fa/fa rats
Authors: Moran-Ramos Sofia , López-Romero Patricia , Tovar Armando R , Torres Nimbe
Published: June 2021

Opuntia ficus indica (nopal), a plant rich in bioactive compounds, has been widely used in the traditional Mexican medicine to treat diabetes. Therefore in the present study, we investigated the effect of nopal consumption in the prevention of fatty liver in obese Zucker rats.

Six weeks old male Zucker (fa/fa) rats (n=12 per group) were fed with a control diet (AIN‐93G; C) or C+ Nopal for 7 weeks. After 7 weeks of dietary treatment, rats fed C+nopal decreased by 50% hepatic triglycerides with respect to control and reduced hepatomegaly and macrovesicular steatosis. In addition, increased serum levels of adiponectin, mRNA expression of genes involved in lipid oxidation (PPARα, CPT1 and ACOX) and lipid export (MTTP and PLTP), hepatic CPT1 and MTTP protein levels. Furthermore, rats fed C+nopal decreased postprandial serum insulin levels; increased liver pAkt/Akt ratio in the postprandial state and significantly decreased serum ALT and AST levels.

This study shows that nopal consumption prevents the development of fatty liver in obese Zucker rats by increasing fatty acid oxidation and VLDL synthesis,, improving liver insulin signaling. Supported by Grupo Produce DF 2010 (to NT)

Grant Funding Source: Produce
Anti‐obesity and inflammatory effects of Lactobacillus sp. OPK‐3 isolated from kimchi in cell line and in animal model
Authors: Park Jeong-Eun , Moon Yeon-Jeong , Oh Suk-Heung , Cha Youn-Soo
Published: June 2021

This study investigated the inhibitory effect of Lactobacillus sp.OPK‐3 isolated from kimchi (L.sp.OPK‐3) on obesity and inflammatory cytokines using in vitro and in vivo animal model. 3T3‐L1 cells were treated with L.sp.OPK‐3 significantly reduced lipid accumulation. The mRNA levels of adipocyte‐specific marker genes were modulated by L.sp.OPK‐3. In vivo, five groups of C57BL/6J mice fed with a ND(Normal diet control), HFD(High fat diet control), HFD+ornithine, HFD+L.brevis, and HFD+L.sp. OPK‐3. All lactic acid bacteria supplemented groups had lower body weights and, more specifically, resulted in the Lactobacillus sp.OPK‐3 supplement group. Lipid profiles and inflammatory cytokines were improved by Lactobacillus sp. OPK‐3 when compared with HFD, ornithine and L.brevis groups. This study on regulation of gene expression which is related with inflammation, lipid and glucose metabolism, demonstrated the inter‐regulatory mechanism between transcription factor and sub‐gene, based on the gene expression change profiles by cDNA microarray. These results confirmed mRNA expression of enzymes related to lipid metabolism and inflammatory cytokines can be affected by the lactic acid bacteria. The results indicated that lactic acid bacteria from kimchi improve obesity and inflammatory cytokines through acting as a modulator of lipid metabolism (This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government(MEST) (NRF‐2008‐331‐ E00371).
NUTRITIONAL EVALUATION OF BLACK CUMIN SEED (NIGELLA SATIVA) SEED PROTEINS PRODUCTS
Authors: Osman Magdi A , Abdelrahman Ibrahim E
Published: June 2021

Black cumin, an oil seed plants, grain and extensively used in Middle East region. The seeds are used for oil extraction, while the meal by product is under utilized. Abette understanding of nutritional quality of seed protein products may lead to wide use of these proteins in food industry to meet the increasing demand for protein to support the growing world population. The purpose of this study is to determine nutritional quality of black cumin defatted flour (DF) protein concentrate (PC) and protein isolate (PI). Nutritional parameters investigated include chemical composition, amino acid antinutritional factors, in vitro protein digestibility (IVPD) and in vivo assay. Casein DF, PC or PI diets were fed to rats to determine true digestibility (TD), biological value (BV) and net protein utilization (NPU). The results revealed that the protein content PI was the highest (86.9%) followed by PC and DF. The protein products were good source of essential amino acid except methionine. PI showed the lowest level of enzyme inhibitors, phytate and tannin and high TVPD compared to DF and PC. Tats fed casein diet showed TD, BV and NPU followed by IP fed rats.
Effects of resistant starch of common and native potato on broiler performance
Authors: Ariza-Nieto Claudia , Rodriguez Daniel , Ariza-Nieto Magnolia , Afanador German
Published: June 2021

Resistant starch (RS) is that fraction of total ingested starch and its degradation products that escape from endogenous enzymatic action in the digestive tract and may contribute to the increase of beneficial bacteria in the cecum, thus contributing to improve the health status of the individual. The experiment involved 260 broilers randomly assigned to one of 4 treatments: control (C), mannan‐oligosaccharides (MOS), retrograded resistant starch from the S. Tuberosum potato (RST), and from the S. Phureja (RSP). Chickens were allocated in metabolism crates for digestibility study. Five birds from each dietary treatment were randomly selected at the end of the digestibility study for morphology measurements. Intestinal tissues were collected from the mid section of duodenum, jejunum and ileum to measure crypt depth and villus height. Data were analysis using complete randomized design. Average daily gain and feed intake were similar in all the groups (P>0.05). Birds fed RST and RSP had greater villus:crypt ratio in jejunum compared to C group (P<0.01) (10.48um and 10.08um vs 7.81um). In duodenum the greatest villus:crypt ratio was shown by the birds in the RST group (12.34um) compared to MOS (8.25um). The results suggest that potato resistant starch is a potential prebiotic.
Fetal overgrowth (FOG) is associated with early perturbations in amniotic fluid (AF) glucose, insulin and insulin like growth factors (IGF) in mothers with and without gestational diabetes (GDM)
Authors: Tisi Daniel K , Koski Kristine G
Published: June 2021

FOG occurs in mothers with and without GDM. Few biochemical markers have been identified. We investigated the possibility that perturbations in 2nd trimester AF glucose, insulin, IGF 2 or its binding proteins (BP) 1 or 3 might underscore FOG and that these early AF disturbances might differ in mothers with or without GDM. Our study population (n=701) was stratified as follows: mothers were classified as GDM or NON GDM and their offspring as appropriate (AGA) or large for gestational age (LGA). ANCOVA, multiple linear and logistic regressions were performed and, using pairs of AF constituents, 2D Bayesian probability plots were created to predict LGA. In the GDM population AF insulin was positively and IGF 2 was negatively associated with growth centile, while elevated AF glucose increased the odds of neonates being born LGA. In the NON GDM population AF insulin was positively and BP 3 negatively associated with growth centile, but the likelihood of LGA was not increased by AF glucose. Bayesian probability plots showed that specific AF constituents separated LGA from AGA infants in GDM mothers but were less discriminating in NON GDM mothers. In conclusion, fetal exposure to increased AF glucose in the first trimester increases the risk of delivery of an LGA infant in GDM mothers. Lower BP 3 in NON GDM mothers and IGF 2 in GDM mothers provide further evidence that perturbations in growth factors may underscore FOG. [Funded by CIHR]

Grant Funding Source: CIHR
Differential binding of low molecular weight heparins with antithrombin and platelet factor 4 as measured by surface enhanced laser desorption ionization (SELDI)
Authors: Kahn Daniel , Hoppensteadt Debra , Hejna Matthew , Jeske Walter , Walenga Jeanine M , Fareed Jawed
Published: June 2021

Introduction

Heparin (H) and low molecular weight heparins (LMWHs) are composed of multiple chain components varying in their physicochemical and biologic profile. LMWHs are derived by depolymerization of heparin and differ in their composition and biologic profiles. Antithrombin (AT) and platelet factor 4 (PF4) represent two of the major modulators of heparin oligosaccharides. Protein chip array technology is useful in the binding of proteins to various ligands. In this study SELDI is utilized to investigate the differential binding of H and various LMWHs.

Materials and methods

H and various branded LMWHs such as Enoxaparin, Dalteparin, Tinzaparin and generic versions of Enoxaparin were studied for their complexation with purified human AT and PF4. All agents were mixed in Tris HCl buffer matrix at equigravimetric amounts. Gold chips were employed to determine the molecular profile and the binding ratios were determined by measuring peak intensities. The relative suppression of peak intensities were proportional to the binding profile of each of these agents.

Results

For both the AT and PF4 binding on a gravimetric basis, H was found to bind much higher than the LMWHs. Significant differences in the binding profiles for the AT and PF4 were noted among different LMWHs. These binding differences may be due to the compositional differences in these drugs.
Overexpression of claudin‐6, ‐7 or ‐9 modifies the activation of MMP‐2 and MMP‐9
Authors: Martinez Ana Cristina Torres , Estrada Luis Felipe Montaño , Rendon-Huerta Erika P.
Published: June 2021

Tight junctions (TJ) have a critical role in the control of paracellular transport and in the maintenance of cell polarity. Claudin family is essential to confer different properties to epithelial cell permeability; however their expression is altered in several tumors and is associated with increased invasive capacity as a result of increased activity of metaloproteinases (MMPs). Claudins ‐1, ‐2, ‐3 and ‐4 can recruit and promote the activation of MMP‐2. Decrease in claudin 10 expression diminishes cell invasiveness through down‐regulation of MMP‐2 and MT1‐MMP in hepatocellular carcinoma. Our group has reported that CLDN 6, 7 and 9 are overexpressed in human gastric adenocarcinomas and these claudins promote cell proliferation and invasiveness, however the mechanisms involved are unknown. To determine whether overexpression of claudins‐6, ‐7 or ‐9 leads to activation of MMPs, we evaluate changes in metalloproteinases expression and activation by immunoprecipitation and zymography assays in claudin‐transfected human gastric adenocarcinoma (AGS) and breast cancer (MCF‐7) cells. The results show a probable interaction of claudin‐9 with MMP‐2 and MMP‐9 in AGS cells and a correlation between overexpression of claudin‐7 and ‐9 with a decrease in the activity of MMP‐2. Similarly colocalization of claudin‐6 and ‐7 with MMP‐2 and claudin ‐6, ‐7 and ‐9 with MMP‐9 was observed in MCF‐7 cells. These results suggest an important role for claudin‐7 and ‐ 9 in the activation of these metalloproteinases.

This work was supported by PAPIIT‐IN208910‐2.
Cleaved high molecular weight kininogen stimulates JNK/FOXO4/MnSOD pathway for induction of endothelial progenitor cell senescence
Authors: Zhu Xuemei , Yang Aizhen , Xie Zhanli , Wu Yi
Published: June 2021

Recently we have reported that cleaved high molecular weight kininogen (HKa) accelerates the onset of endothelial progenitor cell (EPC) senescence (Arterioscler Thromb Vasc Biol. 2011;31:883), here we explored the underlying mechanisms. It has been known that phosphorylation of transcription factor FOXO4, mediated by c‐Jun N‐terminal kinases (JNK), increases the expression of Mn‐superoxide dismutase (MnSOD), therefore regulating cell senescence. Treatment of human EPCs with 30 nM HKa for 72 hours stimulated JNK phosphorylation at Thr183/Tyr185, and FOXO4 phosphorylation at Thr451, and its effect became the maximum at 100 nM, about 15% of plasma HK level. Concomitantly, as measured by Western blotting and real time PCR, HKa treatment for 72 hours upregulated the expression of MnSOD at protein and mRNA levels in a concentration‐dependent manner. To narrow down the functional domain of HKa, we generated recombinant proteins of human HK heavy chain (HC, 19‐380aa) and light chain (LC, 390‐644aa) and found that HC, but not LC, at 100nM displayed a similar activity with HKa in increasing the percentage of senescent EPCs (63%±6.6 v.s. 77.5%±6.02). Moreover, HC at 100 nM increased FOXO4 phosphorylation at Thr451 and the protein level of MnSOD in EPCs. These results demonstrate that HKa accelerates the onset of EPC senescence, at least in part, by stimulating JNK/FOXO4/MnSOD path way, its effect is mediated by the HC.
Changes In The Expression Of Phosphate Transporters, Inflammatory Markers, and Bone Cytokines With Increasing Age
Authors: Loayza Marco S. , Shum Laura , Ferguson Virgina , Levi Moshe , King Karen
Published: June 2021

Aging is a complex biological process with many factors1. Regulation of a single ion such as phosphorus (Pi) can change with age. Pi takes part in cell signaling, energy metabolism and nucleic acid synthesis2. It is systemically regulated by the intestine and kidney3. Pi regulation declines with age3 and in diseases like diabetes mellitus and chronic renal disease. Disruption of Pi regulation can lead to disease in skeletal and cardiac systems, as well as further renal issues4. Since 85% of Pi is stored in the skeleton5, we focused on aging bone where the effects of systemic Pi dysfunction are apparent3. Mice aged 12, 80, and more than 100 weeks were euthanized. Femurs were harvested for gene expression analysis of Pi transporters, cytokines, and inflammatory markers. Fold‐changes in expression were determined using 2−ΔΔCτ method. The difference between age groups was analyzed using one way ANOVA. The results show that Pi homeostasis is not efficient with older age. Expression of Pi transporters and bone health markers both change with age. Understanding the link between chronic diseases and bone health is vital to the aging population for future management and prevention.
[Dmt1]N/OFQ(1‐13)‐NH2, a potent NOP/MOP receptor mixed agonist
Authors: Calo’ Girolamo , Molinari Stefano , Camarda Valeria , Rizzi Anna , Marzola Giuliano , Salvadori Severo , Marzola Erika , Molinari Paola , McDonald John , Ko Mei-Chuan , Lambert David , Guerrini Remo
Published: June 2021

i.t. nociceptin/orphanin FQ (N/OFQ) evokes antinociceptive effects in rodents. Studies in monkeys demonstrated that i.t. coapplication of N/OFQ and morphine elicits synergistic antinociceptive actions. Novel N/OFQ related peptides were synthesised in order to identify and characterize mixed NOP/MOP agonists. The following in vitro assays were used: Ca2+ mobilization in cells expressing hNOP or hMOP receptors and the chimeric protein Gαqi5, receptor and [35S]GTPS binding, guinea pig ileum bioassay. In vivo experiments were performed in mice and monkeys using the tail withdrawal assay. From Ca2+ mobilization studies [Dmt1]N/OFQ(1‐13)NH2 was selected as the most potent and least selective compound. The mixed NOP/MOP full agonist activity and high affinity of [Dmt1]N/OFQ(1‐13)NH2 was confirmed at recombinant receptors in receptor and [35S]GTPS binding studies, at rat spinal cord receptors in [35S]GTPS binding experiments, and in the guinea pig ileum. In vivo in mice and monkeys i.t. morphine and N/OFQ evoked similar antinociceptive effects. [Dmt1]N/OFQ(1‐13)NH2 was also able to elicit a robust antinociceptive action being more potent than N/OFQ (by 30 fold) and morphine (by 3 fold). These results demonstrate that [Dmt1]N/OFQ(1‐13)NH2 behaves as mixed NOP/MOP agonist and substantiate the proposal that such mixed ligands are worthy of development as spinal analgesics.
Defective migration in Activator of G protein Signaling 3‐null leukocytes in response to CXCL12 and CCL19 stimulation
Authors: Branham-O'Connor Melissa , Maher Ellen M. , Zhang Xian , Lanier Stephen M. , Blumer Joe B.
Published: June 2021

Activator of G protein Signaling 3 (AGS3/Gpsm1), an accessory protein for G protein signaling, has functional roles in the kidney and CNS. Here we show that AGS3 is also expressed in spleen, thymus and bone marrow‐derived dendritic cells (BMDCs), and is upregulated in B‐ and T lymphocytes and BMDCs upon activation. We also observed that AGS3 – Gαi1 complexes are regulated by chemokine receptor activation and AGS3 appears to form a chemokine‐regulated, Gαi‐dependent complex with CXCR4 and CCR7 suggesting that AGS3 may integrate signals emanating from chemokine receptors. As part of a broad effort to define the extent of functional diversity of AGS‐regulated events in vivo, we generated AGS3 null mice. Defects in Gαi signaling effect lymphocyte differentiation and motility; therefore, we measured lymphocyte populations as well as chemokine‐directed chemotaxis of lymphocytes and BMDCs from wild‐type and AGS3‐null mice. We discovered defects in chemotaxis of AGS3‐null BMDCs as well as purified B‐ and T lymphocytes. This defect is not due to alterations in receptor expression, as WT and AGS3‐null cells appear to express similar levels of CXCR4. These studies indicate a role for AGS3 and other GPR proteins in the regulation of G‐protein signaling in the immune system, providing unexpected venues for the potential development of therapeutic agents that modulate immune function by targeting these regulatory mechanisms.