bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in the carboxylase profile are associated with early and late differentiation stages of osteoblasts and adipocytes from human mesenchymal stem cells
Authors: Cordonier Elizabeth , Kasputis Tadas , Mills Jessica D. , Han Zhongji , Pannier Angela K. , Zempleni Janos
Published: June 2021

Biotinylated carboxylases play essential roles in macronutrient metabolism. We tested the hypothesis that the abundance of biotinylated carboxylases increases in a temporal pattern during the differentiation of mesenchymal stem cells (MSC) into osteoblasts and adipocytes. Cells were collected during early (~10 d) and late stages (~20 d) of differentiation; non‐differentiating MSCs were used as control. The abundance of biotinylated pyruvate carboxylase (PC) and 3‐methylcrotonyl carboxylase (MCC) β‐chain was greater in both early and late stages of osteoblast differentiation compared with non‐differentiating controls; this was apparently caused by an (up to 180%) increase in the abundance of mRNA coding for PC and MCC during differentiation. In contrast, MSC differentiation into adipocytes was accompanied by a substantial (up to 700%) increase in the expression and biotinylation of acetyl‐CoA carboxylase 2, MCC α‐chain, and PC compared with controls, but the increased levels were limited to the early stages of differentiation. We propose that holocarboxylases play an essential role in cell differentiation and currently seek to demonstrate cause‐and‐effect by using biotin‐defined culture media and synthetic inhibitors of biotinylation events. (UNL ARD Hatch Act, NIH DK063945, DK077816, and DK082476)
Selenium‐enriched probiotics alleviate murine male fertility compromised by high fat diet
Authors: Ibrahim Hala A. M. , Liao Shengfa F. , Zhu Yongxing , Wu Cong , Lu Chenhui , Ezekwe Michael O. , Huang Kehe
Published: June 2021

A total of 75 male mice were randomly allotted to 5 groups of 15 each to study the effects of probiotics (PB), inorganic selenium (ISe) and selenium‐enriched probiotics (OSePB) on the male fertility of hyperlipidemic mice. Group 1 (negative control) were fed a normal basal diet and Group 2 (positive control) a high fat diet (HFD). Groups 3, 4 and 5 were fed the HFD supplemented with PB, ISe, and OSePB, respectively. The HFD was composed of 83.7% basal diet, 15% lard, 1% cholesterol and 0.3% cholic acid. After a 75‐day feeding trial, mice fed the HFD had significant (p<0.01) elevations of serum total cholesterol, triglycerides and low density lipoprotein, decreases of high density lipoprotein, decline in testosterone level, seminiferous tubule atrophy and degeneration, and reduced semen quality. PB, ISe or OSePB supplementation alleviated (p<0.05) the adverse effects of hyperlipidemia by increasing testosterone level, reducing testicular tissue injury, and improving sperm index. It was concluded that the adverse effects of hyperlipidemia on murine male fertility can be ameliorated by PB, ISe or OSePB supplementation. OSePB or ISe gave better results than PB and may be used to improve human male fertility compromised by hyperlipidemia or obesity. This study was funded by the Natural Science Foundation of China.
Effect of calcium intake on premenstrual symptoms
Authors: Ko Catherine , Burns-Whitmore B. , Spalding Thomas W. , Bidlack Wayne R.
Published: June 2021

Reports suggest an association of calcium supplementation and reduced severity of premenstrual symptoms. The aims of our study were to: 1) evaluate the association between dietary calcium intake and the level of premenstrual stress 2) determine whether an intake of calcium above the Dietary Reference Intake would reduce premenstrual symptoms.

Healthy, premenopausal women (19–45 years) were recruited. In the assessment phase (AP), premenstrual symptoms were documented over two menstrual cycles using a validated daily symptoms report, and five food records per person were collected.

Calcium was provided in increments of 250mg to all participants with the intent to reach 1200mg/d. During the calcium supplementation phase (SP), participants recorded symptoms for two cycles. A median split was used to divide participants into either a high‐ or low‐symptom group. Mean affective scores (MAS) and mean somatic scores (MSS) were used to conduct regression analyses.

Mean calcium intake increased from 614±195mg/d in AP to 1133±246mg/d in SP. No association was found between dietary calcium intake and level of affective or somatic premenstrual stress. After SP, a decrease of MAS was observed (p=0.07). The MSS was significantly lower after SP in the high symptom group (p=0.01). Calcium intakes slightly above the DRIs may be beneficial for women who experience mild to moderate premenstrual somatic symptoms.

Grant Funding Source: none
Effects of contaminated fish oil on aberrant crypt foci formation and colonic cell kinetics
Authors: Kang Brian , Kim Jinyoung , Hoh Eunha , Hong Mee Young
Published: June 2021

Fish oil is a rich source of n‐3 polyunsaturated fatty acids, which are known to prevent colon cancer. However this message is confounded by Food Drug Administration warnings which state that consumption of certain types of fish should be restricted due to contaminants such as polychlorinated biphenyls (PCBs). Therefore, this study examined the effects of fish oil contaminated with PCB (4000ng/ml fat) as compared to regular fish oil on the risk factors of colon cancer including aberrant crypt foci (ACF) formation and colonic cell kinetics (cell proliferation and apoptosis). Thirty Sprague‐Dawley rats were divided into three groups and fed corn oil, fish oil, or contaminated fish oil (PCB) for nine weeks and injected with saline or azoxymethane (2 times at wk 3 and 4). Fish oil‐fed rats had a lowered number of ACF (P=0.013) and high multiplicity ACF (P=0.024) compared to the corn oil and contaminated fish oil groups. Fish oil feeding resulted in lower cell proliferation index than corn oil and PCB groups (P=0.009). Cell apoptosis was not significantly different amongst any of the groups. These results suggest that dietary fish oil protects against colon cancer by lowering the number of early preneoplastic lesions through the inhibition of cell proliferation. PCB contaminated fish oil reduced the beneficial effects of fish oil on colon carcinogenesis. Supported by CHNR08‐810, SDSU UGP and NUTR 302L class.
Dairy‐derived bioactive compounds as modulators of stromal, adipocyte‐like and osteoblast‐like cell metabolism
Authors: Ilich Jasminka Z. , Kim Youjin , Kelly Owen
Published: June 2021

Whole fat dairy is rich in α‐linolenic acid (ALA), conjugated linoleic acid (CLA) and calcium (Ca), which are implied in promoting bone formation and decreasing adiposity. We examined individual and/or synergistic effects of ALA, CLA and Ca (at levels present in milk) in regulating bone/adipose cell metabolism. ST2 stromal, MC3T3‐L1 (L1) adipocyte like, and MC3T3‐E1 (E1) osteoblast like cells were treated with: a) LA:ALA=1–5:1; b) 80–90% c9, t11/5–10% t10, c12 CLA; c) 0.5–3 mM Ca; d) combination a), b), c) during cell growth. Local factors were also measured. LA:ALA=1–5:1 inhibited L1 proliferation/differentiation and promoted E1 differentiation. Best effect was found at 4:1 LA:ALA where insulin like growth factor 1 (IGF‐1) and IGF binding protein 3 (IGFBP‐3) production was lowest in L1 cells. All CLA blends decreased L1 and increased E1 differentiation. 1.5–2.5 mM Ca increased ST2 and E1, and decreased L1 proliferation. Combination of 4:1 LA:ALA+90:10% CLA+2.0 mM Ca lowered L1 and increased E1 differentiation. Overall, LA:ALA=4:1 may be the optimal ratio to enhance osteoblastogenesis and inhibit adipogenesis via IGF‐1/IGFBP3 modulation. This effect is strengthened by 90:10% CLA+2.0 mM Ca, indicating possible synergism of dairy components in promoting osteoblastic and inhibiting adipocytic differentiation, and implies preference for osteoblast vs. adipocyte growth. Funded by DMI#1673&FSUCRC# 119532
Predictors of anemia in women of reproductive age from Colombia: results from a national survey
Authors: Kordas Katarzyna , Centeno Zulma Yanira Fonseca , Tellez Paul Rene Ocampo , Pachon Helena
Published: June 2021

Anemia in women of reproductive age may have serious implications for reproductive health and caregiving. In 2005, 20% of Colombian women of reproductive age had anemia but only half of those cases could be explained by iron deficiency (ID). The objective of this study was to identify additional predictors of anemia in Colombian women. Data from the 2005 National Survey of the Nutrition Situation for 3571 nonpregnant women aged 13–49 y were used. Demographic factors and household food security were considered as potential predictors. In preliminary multivariate logistic regression, region of the country other than the Atlantic coast was related to 35–80% lower likelihood of anemia. Iron deficiency (800%), higher parity (40–60%), head of household older than 40 y (30%) and higher C‐reactive protein (90%) were related to higher likelihood of anemia (p<0.05). Women in households with moderate or severe food insecurity had higher (30%) likelihood of anemia but this did not reach statistical significance (p=0.08). The identification of predictors of anemia has important programmatic implications in the creation of targeted interventions for women of reproductive age in Colombia. Funding: ICBF, Pennsylvania State University, CIAT.
Determinants of dietary diversity and nutrient adequacy among post‐partum mothers in rural Kenya
Authors: Gewa Constance , Savaglio Lauren , Oguttu Monica
Published: June 2021

We examined dietary diversity and nutrient adequacy among post‐partum HIV‐ and HIV+ mothers in rural Kenya. One 24‐hour recall was administered to assess 301 mothers’ dietary intake in July‐August 2009. Information on socio‐economic, demographic and health factors were collected through interviews. A dietary diversity score was developed based on a minimum intake of 15 grams of each of the food groups: cereals/tubers; dark‐green leafy vegetables; vitamin A‐rich fruits/vegetables; other fruits/vegetables; dairy; meat/fish/poultry; eggs and legumes/nuts. Mean nutrient density ratio (MDR) was calculated as an average of the density ratios (truncated at 100%) of available iron, available zinc, calcium, vitamin A, vitamin B6, vitamin B12, riboflavin, folate, niacin, thiamin, vitamin C and vitamin E. Mother's diet was predominantly cereal‐based, lacked diversity and the MDR was only 50 percent. Mother's self‐perception of health status, mothers’ social interaction and household socio‐economic status and size were significantly associated with dietary diversity and MDR. High consumption of vitamin A‐rich and other fruits and vegetables and dairy and meat foods were significant predictors of MDR. Dark‐green leafy vegetables and legumes consumption did not predict MDR. This analysis contributes to the understanding of dietary practices and nutritional status of post‐partum mothers in Africa.
Stunting rates in young children aged 6 to 23 months in infants and toddlers born in metropolitan Quetzaltenango, Guatemala
Authors: Reurings Marieke , Doak Colleen M , Vossenaar Marieke , Solomons Noel W
Published: June 2021

Background

The Guatemalan national prevalence of stunting (chronic undernutrition) in children up to 60 months is 54%, which is the fifth highest in the world. As the stunting process continues out through the third year, stunting rates through 24 mo would expected to be lower.

Objective

To describe the rate of stunting in a sample of infants and toddlers recruited in Metropolitan Quetzaltenango.

Methods

A total of 300 young children (150 boys), ranging in age from 6 to 23 mo, equalized for months of life, were recruited at 2 public health clinics in metropolitan Quetzaltenango. Recumbent spine length and weight were measured according to standardized procedures and height‐for‐age (HAZ) were calculated. HAZ ≤2 SD of the WHO 2006 Growth Standards was considered stunting.

Results

The overall mean HAZ‐score was −1.89±1.11 and 135 (45%) children were stunted. Children of women classified of Mayan ascent were significantly (p<0.001) shorter (HAZ‐score −2.20±1.03, and 54% stunted) than others (HAZ‐score −1.71±1.11, and 40% stunted).The age‐specific HAZ‐scores were −1.71±1.13, − 2.07±1.16 and −1.94±0.99 for the 6–11 mo, 12–17 mo and 18–23 mo old age groups, respectively and stunting rates were 36%, 54% and 48%, respectively.

Conclusion

Stunting rates are high, progressing with aging through the first 2 years of life.

Funded by Sight and Life, Switzerland
Obesity and Sedentary Life Style are not associated with increasing risk of Colorectal Cancer in Jordan
Authors: Waly Mostafa , Arafa Mostafa , Jriesat Sahar , Sallam Sunny , Al-Kafajei Ahmed
Published: June 2021

Methods

A case‐control study included 440 Jordanian subjects (220 recently diagnosed CRC patients and 220 age and gender matched healthy subjects as a control group). The body mass index (BMI) was calculated for all study participants. The physical activity levels (low, moderate or high) were determined according to World Health Organization (Global Physical Activity Questionnaire) (WHO, Physical activity/GPAQ v2.0).

Results

There were 220 (118 males and 102 females) CRC cases and 220 matched controls for age and gender. The CRC cases and controls were similar in age (56.3 ± 12.3 years for males and 53.7 ± 6.8 years for females) and gender (53.6 % males & 46.4% females). The mean BMI for all study participants was similar (ranged from 28 to 29.5). The CRC female and male patients and their matched controls had no significant difference with regards to BMI (P>0.05). CRC patients and controls had comparable low to moderate physical activity levels. All study subjects were not maintaining a routine exercise practice.

Conclusion

Inconsistent with the reported studies at developed nations, our study indicated that obesity (BMI<30) and a trend of decreased physical activity were prevalent among the study subjects (patients and controls) with no significance differences.
Antioxidant‐rich berries exert modest bone protective effects in postmenopausal smokers
Authors: Kaume Lydia , Gbur Edward E , DiBrezzo Ro , Devareddy Latha
Published: June 2021

Smoking aggravates bone loss in postmenopausal women due to increased oxidative stress and inflammation. Berries rich in phenolics have shown bone protective affects in animal studies partially due to their anti‐oxidant properties. We assessed effects of: (i) smoking and (ii) consumption 45g/day of blueberries and blackberries for 9 months on bone health. Postmenopausal smokers (n=31) and nonsmokers (NS) (n=18) were recruited. Smokers were randomly assigned to one of three treatment groups: smoker control (S‐C) (n=12); smoker + 45 g freeze‐dried blackberries (S‐BB) (n=6); or smoker + 45 g freeze‐dried blueberries (S‐BL) (n=13). We measured: bone mineral density (BMD) of total body, and other regions at 0 and 9m, and urinary deoxypyridinoline, bone alkaline phosphate, osteocalcin, thiobarbituric acid reactive substances and highly sensitive C‐reactive protein at 0, 3, 6, and 9 month. Total body BMD decreased by 0.0359 g/cm2 in S‐C group compared to 0.0041 and 0.0226 g/cm2 in S‐BB and S‐BL groups respectively, and increased by 0.0031 g/cm2 in NS group (p=0.0284). No significant changes were noted on other parameters. These findings indicate that antioxidant rich berries modestly protect against smoking‐induced bone loss in postmenopausal women. The mechanisms by which berries prevent bone loss need further exploration. This study was funded by Arkansas Biosciences Institute.
Differential effects of white button mushrooms (WBM), portabella (PM), and shiitake (SM) on IL‐18 secretion by lipopolysaccharide (LPS)‐treated mice: implication on risk of infection and chronic inflammation
Authors: Kuvibidila Solo
Published: June 2021

IL‐18, a cytokine previously called IFN‐gamma inducing factor and primarily secreted by macrophages and dendritic cells, plays a crucial role in immunity against viruses and intracellular pathogens. IL‐18 blockade increases infection severity. Mushrooms (i.e. SM) have been used in ancient medicine for treatment and prevention of infections. However, their effect on IL‐18 secretion is unknown. We hypothesize that one of the mechanisms by which mushrooms protect from infections is by IL‐ 18 induction. Eight week‐old C56BL/6 mice were fed either the AIN76 diet or the same diet fortified with 5% lyophilized WBM, PM, or SM (n=11–12) for 6 weeks. Mice were euthanized 2 h after receiving an ip. injection of 50 μg LPS. Plasma IL‐18 was assayed by ELISA. In LPS‐untreated mice, while PM and SM reduced IL‐ 18 median plasma levels by 20% and 10% respectively, WBM had no effect. In LPS‐treated mice, PM reduced median IL‐18 levels by 25%. LPS injection increased plasma IL‐18 levels by ~ 3 fold (p<0.05). Mushrooms ± LPS did not alter the weights of body, spleen, thymus, heart and kidneys. Multiple regression analysis suggested that spleen and kidney weights were positively (p <0.05) associated with IL‐18 levels probably due to macrophages. Data suggest that mushrooms may not modulate the risk of infection via IL‐18. However, PM may modulate the risk of inflammatory conditions induced by chronic infection through IL‐18 reduction. Supported by Mushroom Council/USDA Grant 580790706 and Oklahoma State University
Glucagon synthesizing alpha‐cells secret Islet Homeostasis protein (IHoP) for regulation of hormone synthesis and proliferation of insulin producing beta‐cells
Authors: Oh Seh-Hoon , Petersen Bryon E
Published: June 2021

Objective

We have identified a novel protein in bone marrow‐derived insulin‐producing cells. We now characterize a role for this protein, termed Islet Homeostasis Protein (IHoP), in the pancreatic islet.

Methods

Detection of IHoP was performed using RT‐PCR, immunocytochemistry and in‐situ hybridization. IHoP functions were utilizing proliferation, insulin secretion by in vitro assays, and following siRNA protocols for suppression of IHoP.

Results

IHoP did not homologue with known pancreatic hormones. IHoP expression was seen in both BM‐derived IPCs and isolated pancreatic islets. Immunohistochemistry on pancreatic islet revealed that IHoP localized to the glucagon synthesizing alpha‐cells. Inhibition of IHoP by siRNA resulted in the loss of glucagon expression, which induced low blood glucose levels (63–85mg/dL). IHoP is secreted from the glucagon synthesizing α‐cells, and that it has a role in regulating cellular proliferation of beta‐cell under high glucose conditions. Subsequently, cellular apoptosis was observed throughout the islet, including the insulin‐producing beta‐cells. Islets of pre‐onset diabetic patients showed normal expression of IHoP and glucagon; however IHoP was lost upon onset of the disease.

Conclusion

These data suggest that IHoP could be a new functional protein required in the homeostasis of the islet and it may also play a role in our understanding of T1D progression.
Sun1 and Sun2 are required for nuclear positioning and nuclear migration of neuronal cells in postnatal brain development
Authors: Chi Ya-Hui , Wang Jing-Ya , Huang Chien-Chi , Wang Wang-Ping
Published: June 2021

Establishment of polarity, which involves orientation and relocation of the cell nucleus, is essential for proliferation and differentiation of a variety of cells including neurons, epithelial cells and myocytes. Current view pictures both actin‐ and microtubule‐mediated nuclear positioning are required for this process. Although cytoskeleton drives different cellular functions, questions remain concerning the orchestration of nuclear positioning or migration relative to other organelles within the cell. The SUN‐domain proteins are components of the LINC complex that reside with the inner nuclear membrane. Depletion of the SUN‐domain proteins is associated with tissue‐specific developmental defects. Here we created Sun1 and Sun2 knockout mice and found both genes are important for the development of cerebellum and hippocampus. More than 70% of Purkinje cells in the Sun1−/− Sun2+/− cerebellum fail to present at the surface of the internal granule layer (IGL). In addition, granule neurons in the Sun1−/− hippocampus are less efficient for retrograde nuclear migration. These results input the current understanding of the SUN‐domain proteins for postnatal brain development.
Impaired mTOR/S6 signaling pathway contributes to neurodegeneration in early postnatal Nbs1‐deficient mouse cerebral cortex
Authors: Liu Bo , Tong Wei-Min
Published: June 2021

Nijmegen breakage syndrome (NBS) is a genomic instability disease caused by hypomorphic mutations of NBS1 gene encoding Nbs1 protein. The Nbs1 protein is a component of the Mre11/Rad50/Nbs1 (MRN) complex that acts as a DNA double‐strand break sensor and functions in response to DNA damage. We have generated a mouse model (termed as NbnCNS‐del mice) by inactivating Nbn gene in the mouse central nervous system (CNS) with nestin‐Cre system. Our previous study has indicated that the neurons in the cerebral cortex, hippocampus and thalamus of NbnCNS‐del mice displayed neurodegeneration changes that consist of size reduction, numerical decreasing and disorganization. However, the molecular mechanisms of the phenotype remain unknown. In the present study, we report that in cerebral cortical neurons, Nbs1 deficiency affects glial cell function, for the reduced expression level of nerve growth factor (NGF) mRNA and protein. In turn, altered glial cell function results in the down‐regulation of neuronal mTOR and S6 ribosomal protein expression in postnatal NbnCNS‐del mice. This result demonstrates that the interplay between neuron and glia contributes to the neurodegeneration, and implies an important role of DNA damage response in neuron.

This work is supported in part by National Novel Drug Development Fund (2009ZX09303‐008), and by 111 project (B08007).
Adaptive Single Neuron Hypertensive Gene Expression Programs in the Nucleus Tractus Solitarius
Authors: Brureau Anthony Yannick Bernard , Vadigepalli Rajanikanth , Paton Julian , Schwaber James
Published: June 2021

We describe the initial adaptive response process of single neurons following elevation of the blood pressure that identify processes that may evolve over time to alter central blood pressure set point control in neurogenic hypertension. Our initial approach is to study single A2 neurons in the nucleus tractus solitarius NTS. We then study the network of NTS A2 neurons with central amygdale (CeA) and ventrolateral medulla (VLM) by injecting Retrobeads in CeA and VLM in order to identify NTS A2 neurons with projections in this network. We lift NTS A2 neurons by Laser capture Microdissection while mapping their origin in the 3‐ dimensional NTS structure to assay several hundred neurons per brain. Our 96 qPCR gene expression assays are refined from previous global gene expression studies to select a cohort that appear co‐regulated and coordinated in response to blood pressure and relevant to the A2 phenotype and to the angiotensin II AT1R receptor activated responses. Analysis of the resulting high‐dimensional dataset reveals a remarkable complexity in the response to an increase of blood pressure. These results demonstrate differential expression programs for each anatomically distinct neuronal group and neuronal type.

Research Support: NIH GM 083108
Altered neddylation in the heart of a mouse model for cardiomyopathy
Authors: Suzuki Osamu , Koura Minako , Noguchi Yoko , Uchio-Yamada Kozue , Matsuda Junichiro
Published: June 2021

We reported that a strain (4C30 strain) of mice with sialyltransferase (ST3GalII) transgenes would be a new mouse model for adult‐onset dilated cardiomyopathy (Proc Jpn Acad Ser B Phys Biol Sci. 2011; 87(8):550–562). Overexpression of ST3GalII perturbed protein glycosylation in 4C30 hearts, suggesting alteration of folding, translocation, and degradation of various proteins in 4C30 hearts. To study a possible involvement of protein degradation mechanisms in the cardiomyopathy, we compared neddylation statuses of heart proteins in our cardiomyopathic 4C30 mice with those in normal controls (C57BL/6NCr). Quantitative Western blots (QWB) for NEDD8 with GAPDH as a loading control revealed that two bands (100 and 105 kDa), but not four additional bands (90, 70, 60, or 40 KDa), were significantly denser in 4C30 hearts than in control hearts (p<0.05). QWB for culins with GAPDH indicated that culin‐2, but not culin‐1 or ‐3, was significantly more abundant (p<0.05). These results suggest that the protein degradation is altered in cardiomyopathic hearts.
A novel mellitin‐derived peptide nanoparticle delivery system for STAT3 siRNA mediated killing of B16 melanoma cells
Authors: Hou Kirk , Wickline Sam
Published: June 2021

Exogenously delivered siRNA has the potential to form a new class of cancer therapeutics based on sequence specific silencing of oncogene expression. Previous work has shown that the transcription factor, Signal Transducer and Activator of Transcription 3 (STAT3), is constitutively activated in the B16 melanoma cell line and mediates resistance to apoptosis by driving expression of anti‐apoptotic genes. In this study, we sought to establish the efficacy of novel peptide‐based siRNA delivery agents derived from the bee‐venom component, melittin, for STAT3 knockdown in B16 cells. Modifications to melittin (GIGAVLKVLTTGLPALISWIKRKRQQ) include truncation of its N‐terminus to reduce cytotoxicity and addition of basic residues to its C‐terminus to improve electrostatic binding to siRNA. The resulting peptide, p5RHH (VLTTGLPALISWIRRRHRRHC), complexes siRNA to form a nanoparticle delivery system, and mediates sequence specific knockdown of GFP in B16‐GFP cells. Confocal microscopy reveals that Cy3 labeled oligonucleotides are taken up by cells with diffuse cytoplasmic staining after 2 hours. Notably, p5RHH‐mediated transfection of 50nM STAT3 siRNA reduces B16 viability by 25% without vector associated cytotoxicity. These results indicate that p5RHH is a high efficiency, low toxicity siRNA transfection agent with implications for cancer therapy. Supported by Sigma‐Aldrich Predoctoral Fellowship.
Computational Characterization of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5b]pyridine at Androgen Receptor: Mechanistic support for its role in Prostate Cancer
Authors: Glass-Holmes Mashunte , Aguilar Byron , Oriaku Ebenezer , Goodman Carl B.
Published: June 2021

The cooked meat‐derived carcinogenic heterocyclic amine 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5b]pyridine (PhIP) has been shown to produce tissue specific tumors in animal models. The mechanism for this specificity has not been fully elucidated and has been attributed to organ differences. In order to evaluate the modifying effects of PhIP on carcinogenesis we analyzed its ability to activate or inhibit an androgenic response. The present study seeks to evaluate the interactions of PhIP and identify molecular events associated with PhIP‐mediated disruption of the androgen receptor (AR) function. We demonstrate that PhIP binds with the ligand‐binding domain by computational analysis. Using molecular docking simulations, of ArgusLab and Molegro, we have identified a possible binding mode of PhIP to the AR based on the predicted binding free energy, compared to that of DHT at the AR. This molecular docking simulation gives insight into the molecular mechanism by which PhIP is able to initiate transduction of the activation signal. Additional studies using receptor‐binding experiments will be used to confirm modeling data. Our Docking results indicate tht PhIP may play an important role in the modification of AR gene expression and initiation of prostate cancer. Research supported by NCRR/RCMI G12 RR03020, NIGMS/MBRS/SCORE GM08111, and HRSA SD34HP0 4018.
Effects of amphetamine in rats trained to discriminate between 22 and 2 hr food deprivation
Authors: Jewett David C. , Johnson Amy R. , Dobbe Jennifer E. , Kron Benjamin A. , Vargo Joseph P. , Hoida Emily J. , Van Asten Ryan M. , Nelson Courtney L. M.
Published: June 2021

Amphetamine decreases food intake in several species. Previously Corwin, Woolverton, and Schuster (1990) trained food‐restricted rats to discriminative between food ingested 3 or 22 hr before training sessions. In this procedure, amphetamine did not reliably produce discriminative stimulus effects similar to those of recent food ingestion. We tested the effects of amphetamine in non‐restricted rats trained to discriminate 22 hr food deprivation from 2 hr food deprivation in a two‐lever, operant choice task. After rats acquired the discrimination, subjects were food restricted for 22 hr and administered saline or amphetamine (0.032 to 1.0 mg/kg, i.p.) before generalization tests. Food intake was measured for 1 hr after generalization tests. Amphetamine (0.32 mg/kg) significantly decreased the discriminative stimulus effects of 22 hr deprivation. Larger doses of amphetamine eliminated responding. Food intake was significantly affected by amphetamine (0.32 and 1.0 mg/kg). The findings are consistent with the notion that amphetamine affects feeding stimulated by energy needs, and suggest differences between the two discrimination techniques.
Enhancement and perceptual masking of a nicotine discriminative stimulus in rhesus monkeys
Authors: Cunningham Colin Seamus , McMahon Lance Richard
Published: June 2021

Bupropion and varenicline are two pharmacotherapies (Zyban® and Chantix®, respectively) for smoking cessation. Bupropion is a monoamine reuptake inhibitor that is also a nicotinic antagonist. Varenicline has lower efficacy than nicotine at α4β2 nicotinic receptors in vitro, and is expected to antagonize the effects of nicotine. The extent to which these drugs interact with nicotine has not been fully established. To examine the ability of nicotinic as well as non‐nicotinic compounds (cocaine and midazolam) to alter the discriminative stimulus effects of nicotine, 5 adult rhesus monkeys were trained to discriminate nicotine (1.78 mg/kg base) under a fixed ratio 5 schedule. Bupropion produced 23% nicotine lever responding and antagonized the nicotine discriminative stimulus, as evidenced by a rightward shift in the nicotine dose‐response curve. Varenicline fully substituted for nicotine and the combined effects of nicotine and varenicline were synergistic. The monoamine reuptake blocker cocaine and the benzodiazepine midazolam produced nicotine lever responding when given alone (44% and 4%, respectively) and antagonized the nicotine discriminative stimulus. Varenicline produced synergistic effects with nicotine, inconsistent with low efficacy at a common site in vivo. Antagonism of the nicotine discriminative stimulus by bupropion, cocaine, and midazolam is consistent with perceptual masking.
N‐palmitoylethanolamide Treatment Exhibits Antidepressant Effects In A Mouse Model Of Anxiety/Depressive Like Behavior
Authors: Crupi Rosalia , Mazzon Emanuela , Impellizzeri Daniela , Esposito Emanuela , Cuzzocrea Salvatore
Published: June 2021

Palmitoylethanolamide (PEA) is a member of fatty‐acid ethanolamide family. This endocannabinoids is an endogenous neuromodulator with a broad spectrum of pharmacological properties, including analgesic, anti‐inflammatory, anticonvulsant and antiproliferative effects. The aim of this study was to investigate the effects of PEA (5,10 mg/kg, i.p.) in the neural development in the adult hippocampus and the behavioral changes in a depressive‐like state. The antidepressant‐like effect of PEA was evaluated by using a mouse model of anxiety/depressive‐like behavior induced by chronic corticosterone treatment. 129 Sv/Ev mice were administered corticosterone for 7 wk, and behavioral tests as novelty‐suppressed feeding, open field, forced swim were performed. Cell proliferation in hippocampal dentate gyrus (DG) was investigated by 5‐bromo‐2‐ deoxyuridine and doublecortin immunohistochemistry techniques. Golgi‐ impregnated method was used to evaluate the changes of dendritic spines in DG neurons. The depressive‐like state caused by corticosterone treatment was reversed by exogenous administration of PEA for 3 wk; the proliferation of progenitor cells was reduced under chronic corticosterone treatment, whereas treatment with PEA prevented the corticosterone‐induced reduction in hippocampal cell proliferation. Corticosterone‐treated mice exhibited a reduced spine density, which was ameliorated by PEA administration. These findings suggest that PEA may play an important role in the near future in the treatment of depression.
The Role of Brain Derived Neurotrophic Factor in Depression In a Mouse Model
Authors: Overacre Abigail , Sakata Kazuko
Published: June 2021

Background

Brain derived neurotrophic factor(BDNF) levels have been thought to be a link to depression. It is not clear in which brain region BDNF mRNA levels are reduced in BDNF‐KIV mice. While BDNF gene expression is regulated by at least 9 promoters, promoter IV is of interest because it is suggested to be decreased in depressed patients.

Objective

In our research, we generated mutant mice that lack promoter IV‐driven BDNF expression and show depression‐like behavior (BDNF‐KIV). We aimed to determine 1) which brain regions show substantially decreased total BDNF mRNA levels, and 2) whether there is a correlation between brain and blood BDNF levels using the mouse model of depression. We also screened for what gene expression was affected by lack of promoter IV‐driven BDNF expression.

Methods

Mouse RNA was extracted from various neuroanatomical structures of the brain as well as PBMC cells in the blood using QIA‐Shredder and RNeasy column systems. A cDNA was prepared using the Transcriptor First Strand cDNA Synthesis Kit and a sample was preamplified using PreAmp Master Mix. A primer‐probe plate was designed to express genes, focusing on the BDNF exons. Gene expression was calculated with a reference gene, Cyclophilin D. The Biomark HD system by Fluidigm expressed the genes using qRT‐PCR, comparing BDNF levels with other genes in WT versus BDNF‐KIV mice.

Results

Gene expression showed BDNF levels were lower in brain tissues of KIV mice than in WT mice, especially in the prefrontal cortex and hippocampus. BDNF levels were higher in PBMC cells of KIV mice in comparison with WT mice. The results were calculated directly from the gene expression in Exon IX.

Conclusion

BDNF levels are linked to depression and are found to be lower in KIV mice, especially in the PFC and the HIP. However, BDNF levels are reversed in PBMC cells, suggesting that BDNF levels in PBMC do not correlate to those in the brain and cannot be used as a biomarker of depression.
Acetaminophen induces oligodendrocyte precursor cell death and reactive astroglia in a primary mixed glial cell culture
Authors: Perez María Julia , Tallis Silvina , Novak Analia , Delfante Amalia , Rubio Modesto Carlos , Ghanem Carolina Inés
Published: June 2021

Brain damage in acetaminophen (AP) overdose is associated with liver failure but its direct effects has not been evaluated.

Aim

To study the effect of toxic and non‐toxic doses of AP on astrocyte and oligodendrocyte precursor cell (OPC) viability in a primary mixed glial cell culture.

MYM

Primary cultures of mixed glial cells were carried out according to McCarthy and de Vellis (1980). Cells were treated with vehicle (DMSO) or AP (1 and 20 mM). After 48 hs cell viability was assessed; astrocyte marker (GFAP) and OPC marker (PDGFR‐alpha) expression was evaluated by western‐blot (WB) and immunocytochemistry.

Results

The 1 mM dose of AP did not produce significant differences in cell viability, while 20 mM exposure decreased in 40% cell viability (P<0.001). The 20 mM dose produced the death of the 90% of the OPC in the culture and a decrease of 89% of PDGFR‐alpha expression (P<0.001) with an increase in GFAP optical density/cell and WB expression (47% and 97% respectively) (P<0.05). The 1mM dose did not produce significant changes in GFAP but produce a 25% significant decreased in PDGFR‐alpha expression (p<0.05).

Conclusion

AP produces direct toxic effects on glial cells characterized by oligodendrocyte death and the induction of reactive astroglia.
Establish a cell viability assay to identify novel cytotoxic Organic Anion Transporting Polypeptide 1B3 substrates
Authors: Obaidat-Hays Amanda , Kindscher Kelly , Timmermann Barbara , Hagenbuch Bruno
Published: June 2021

Organic anion transporting polypeptides (OATPs) mediate the uptake of numerous structurally diverse compounds. The normally liver‐specific OATP1B3 is also expressed in different cancers, suggesting that it can be a therapeutic target for anticancer drug delivery. Because OATP1B3 also transports chemotherapy drugs, we established a cell based assay to screen for novel cytotoxic OATP1B3 substrates. We screened a library of Kansas plant extracts to isolate and chemically identify novel cytotoxic compounds that could be developed as lead compounds to treat OATP1B3‐expressing cancers. In this study, we used Chinese Hamster Ovary (CHO) cells that stably express OATP1B3 and measured cell viability of wild‐type and OATP1B3‐expressing CHO cells in the presence of Kansas plant extracts of different polarities using a luminescent cell viability assay. We identified the butanol fractions of Rhus aromatica (V345) and Rhus glabra (V585) that kill OATP1B3‐expressing CHO cells at much lower concentrations than wild‐type CHO cells, suggesting that these extracts contain cytotoxic OATP1B3‐substrates. We will further sub‐fractionate these extracts to isolate and chemically identify the cytotoxic natural products. In conclusion, this cell‐based viability assay might be a good tool to identify potential cytotoxic OATP1B3 substrates that can be delivered into OATP1B3‐expressing cancers.
Depolarization potentials in mouse lateral septal nucleus neurons mediated by TRPC4‐like channels
Authors: Tian JinBin , Zhu Yingmin , Lu Yungang , Zhu Michael Xi
Published: June 2021

The lateral septal nucleus (LSN) neurons express high levels of Canonical Transient Receptor Potential type 4 (TRPC4). In this study, we examined electrophysiological properties evoked by manipulations known to activate TRPC4 channels in LSN neurons from mouse brain slices using whole cell current clamp recordings. We show that: 1) pressure injection of (S)‐3,5‐DHPG to LSN neurons induced an instant burst firing on top of a prolonged depolarization plateau, 2) superfusion of the slice with DHPG caused membrane potential oscillations and repetitive burst firing of LSN neurons, 3) depolarization current injection increased the occurrence of depolarization plateau induced by DHPG and prolonged the plateau duration, 4) pretreatment of brain slices with pertussis toxin suppressed DHPG‐induced depolarization plateau in majority of the LSN neurons, 5) the DHPG‐induced depolarization plateau was blocked by a novel TRPC4 channel blocker, shown to be selective for TRPC4 and TRPC5 in heterologous expression systems, suggesting that the major component of the depolarization plateau is mediated by TRPC4 channels. These results demonstrate the involvement of TRPC4 channels in the integration of excitatory inputs of LSN neurons through metabotropic receptor and G protein signaling. Supported by NIH grant DK081654
A pharmacodynamic comparison of piperidine and pyridine alkaloid actions at fetal muscle‐type nicotinic acetylcholine receptors (nAChR)
Authors: Green Benedict T , Lee Stephen T , Panter Kip E
Published: June 2021

Piperidine and pyridine alkaloids are found in many species of plants including Lobelia spp., Conium spp., Nicotiana spp., and Lupinus spp. Some of these alkaloids cause multiple congenital contracture deformities (MCC) and cleft palates in cattle, pigs, sheep, and goats. The mechanism behind MCC defect formation in livestock is thought to be the inhibition of fetal movement due to stimulation followed by desensitization of fetal muscle‐type nAChR. A pharmacodynamic comparison of piperidine and pyridine alkaloids was made in TE‐671 cells that express fetal muscle‐type nAChR. TE‐671 cells were cultured in DMEM supplemented with fetal bovine serum and penicillin/streptomycin. The membrane depolarization responses were measured by changes in fluorescence of a membrane potential‐sensitive dye. Alkaloids with a carbon side‐chain of at least three carbons or larger attached to the carbon alpha to the piperidine nitrogen were more potent and effective. The presence of a double bond adjacent to the nitrogen of the piperidine or pyridine ring increased potency. The rank order of potency was: anabasine > nicotine > coniine > lobeline > myosmine > ammodendrine. This research supported by ARS‐USDA.
G Protein Estrogen Receptor (GPER) Regulates Kv11.1 Ion Channel Activity in ERneg Breast Cancer Cells
Authors: Gentile Saverio
Published: June 2021

Breast cancer is a major cause of death among women worldwide, and novel strategies need to be developed to lessen its morbidity. Although, the mechanism of breast cancer proliferation is poorly understood, it has been established that in cancer cells displaying an E2 receptor positive (ERpos) or E2 receptor negative (ERneg) phenotype, proliferation can be mediated by an E2‐dependent mechanism. Interestingly, recent studies have demonstrated that E2 effects on proliferation of ERneg breast cancer cells can be mediated by the G protein‐coupled receptor, GPER. However, very little is known about the biochemical signaling cascade activated by E2 via GPER.

Clinical heterogeneity of breast cancers indicates new subsets of gene expression that can create novel molecular pathways mediating estrogen's (E2) effects on proliferation. The human ether‐a‐go‐go related gene (hERG) encodes the voltage‐gated potassium channel Kv11.1, which is primarily expressed in electrically excitable cells.

Interestingly, recent translational research in cancer biology has revealed that Kv11.1 is robustly expressed in cancers of varying histogeneses, including breast cancer. Although, it is well established that Kv11.1 plays a major role in regulating membrane potential nothing is known about its role in breast cancer and its contribution in proliferation.

Our experiments show that changes of Kv11.1 activity determine variation of proliferation rate in ERneg breast cancer cells. Electrophysiological, biochemical analyses show that Kv11.1 ion channel activity is stimulated by an E2‐activated signaling cascade via GPER in ERneg breast cancer cells. In addition, stimulation of Kv11.1 activity led to increase of intracellular calcium. Understanding the role of hormonal regulation of Kv11.1 ion channel will lead to discovery of a myriad of new potential drug target for cancer treatment.
Effects of metformin on bethanechol‐induced bladder contractions
Authors: Peuler Jacob Dale , Curtis Jeremy K , Phelps Laura E
Published: June 2021

By contracting detrusor smooth muscle bethanechol can treat urinary retention in patients with diabetic bladder dysfunction. Previously, we reported that millimolar levels of the antidiabetic drug metformin can markedly inhibit such contractions in the rat bladder. In the present work, we find that levels ≤ 200 micromolar do not do so suggesting that therapeutically‐relevant levels of metformin (as seen in plasma of patients) will not interfere with the ability of bethanechol to contract and thus empty the bladder in patients who need it. The mechanism whereby metformin (at ≥1 millimolar) inhibits bethanechol‐induced bladder smooth muscle contractions is unknown. We also found that, at its EC50 of 5 millimolar, metformin's inhibition of bethanechol contractions was significantly antagonized (p<0.05) by 4‐aminopyridine (4AP) and tetraethylammonium (TEA). Therefore, we suspect that such metformin inhibits bethanechol contractions in bladder by activating potassium (K) efflux through both 4AP‐sensitive and TEA‐sensitive K channels. Support: MWU Biomedical Science Masters Program.
Contraction of rat vena cava by endothelin‐1 is dependent on phospholipase‐Cβ, but independent of IP3 receptor activation
Authors: Tykocki Nathan R. , Wu BinXi , Jackson William F. , Watts Stephanie W.
Published: June 2021

In vascular smooth muscle, inositol 1,4,5‐trisphosphate (IP3) regulates excitation‐contraction coupling by activating the release of sarcoplasmic Ca2+ stores. IP3 is produced through the activity of phospholipase Cβ (PLCβ), which cleaves phosphatidylinositol into diacylglycerol (DAG) and IP3. While the vasoconstrictor ET‐1 activates PLCβ and increases IP3 formation in arteries, it is unclear if this is so in veins. We hypothesized that contraction to ET‐1 in rat aorta (RA) and vena cava (RVC) depends upon IP3‐mediated Ca2+ release. To test if PLCβ was activated by ET‐1, isometric contraction was measured in RA and RVC rings exposed to vehicle, the PLCβ inhibitor U‐73122 or its inactive analog U‐ 73343 (1 μM), or the IP3 receptor antagonist 2‐APB (100 μM). While U‐73343 did not significantly inhibit contraction to ET‐1, U‐73122 significantly reduced maximum contraction to ET‐1 in RA (86±6% of control) and RVC (49±11% of control). Contrary to our hypothesis, 2‐APB significantly reduced maximum contraction to ET‐1 only in RA (55±4%) and not RVC (82±8%). These data were supported by Western blot analysis, showing 63.1 times more IP3R‐1 and 12.6 times more IP3R‐2 protein in RA than RVC, as measured by densitometry. These findings suggest that ET‐1 does activate PLCβ in RA and RVC, but only contraction to ET‐1 in RA is regulated by IP3. Rather, DAG and not IP3 may regulate contraction to ET‐1 in RVC. Supported by NIH P01HL70687.
ALIPHATIC ALCOHOLS PROPAGATE OXIDATIVE STRESS‐RESPONSIVE CELL DEATH SIGNALS IN THE MOUSE LIVER AND KIDNEYS BY PERTURBING EXPRESSION OF PRO‐ AND ANTIAPOPTOTIC GENES
Authors: Lawana Vivek , Patel Ami , Ray Sid D
Published: June 2021

Toxic effects of aliphatic alcohols [methanol (ME), ethanol (ET), isopropyl alcohol (ISO) and tertiary‐butanol (tBut)] remains an important focus of mechanistic investigations; however, their mechanisms of actions at the genetic level and relative potential to induce apoptotic and necrotic cell deaths remains unclear. The main study objectives were to compare: i) hepato‐ and nephrotoxic potentials, ii) abilities to produce oxidative stress (OS), and iii) impact on the expression of proapoptotic (bax, APAF‐1, Cyt c & caspase 3) and antiapoptotic (bcl‐2, bcl‐xL) genes in the liver and kidneys by various alcohols. A single LD40 dose of ME, ET, ISO & tBut were orally gavaged to 4 groups of female ICR mice (12–15 wks old). Control group received only water, and all animals were sacrificed at 24hrs. Blood was collected for serum chemistry and organs were collected (frozen at −70 °C) for westerns and tissue biochemistry. Compared to control (100%), several organ injury biomarkers considerably increased [serum ALT (388%); BUN (462%); liver MDA (215%); kidney MDA (206%)], whereas, reduced glutathione content significantly decreased in the liver (24%) and kidneys (39%) by t‐Butanol. Pro‐apoptogenic genes showed increased expression, whereas, antiapoptotic group showed a dramatic decrease. ME, ET and ISO also showed similar trends. These results suggest a clear‐cut mediation of various alcohol‐induced oxidative stress in inducing cell injury and cell death via cell death signaling pathways, and it was observed that higher the number of carbon atoms, the greater was the toxicity (tBut>ISO>ET>ME). Suppo. By AMS Coll of Pharm & HScs.