bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiresidual Pesticide Analysis of Four Sulfonylurea Pesticides by High‐Performance Liquid Chromatography with Diode‐Array Detection
Authors: Koo Yun‐chang , Lee Kwang‐Won , Yang Sung‐Yong , Kim Hyengkook , Shin Han‐seung
Published: June 2021

As the food trade is world‐widely increasing, it's necessary to set a regulation for some pesticides used in other counties but Korea. A method for the determinations of four sulfonylurea compounds, rimsulfuron, ethametsulfuron‐methyl, tribenuron‐methyl and chlorimuron‐ethyl was examined, and analyzed by HPLC with ODS column (250 mm ¡¿ 4.6 mm, 5 §‐ diameter particle size) which was maintained 35¡É. Mobile phase consisted of solvent A (20 mM KH2PO4, pH 2.5) and solvent B (acetonitrile). Isocratic elution of the column with 55 % solvent A and 45 % solvent B at a flow rate of 1 mL/min resulted in retention times of 5.92, 6.54, 9.28, and 14.35 min for rimsulfuron, ethametsulfuron‐methyl, tribenuron‐methyl, and chlorimuron‐ethyl, respectively. The limit of quantitation was calculated by the equation of ¡®LOQ area = (Area ¡¿ Noise ¡¿ 10)/Peak height¡, and the LOQ area was substituted into the standard curve. Four sulfonylurea pesticides were recovered at over 86% at the spike levels with 0.05, 0.1 and 0.5 mg/kg in rice, apple, and soybean. Relative standard deviations were less than 10% throughout all the recovery tests.
Children's vitamins and minerals intake from dietary supplements in Korea
Authors: Lee Ji Yeon , Jang Young Ai , Moon Jae Jin , Lee Haeng Shin , Lee Hae Jeong , Lee Yoonna , Kim Doo Hee , Koh Eunmi , Lee Hye Yeong , Yoon Hye Sung , Kim Cho‐il
Published: June 2021

With increasing intake of dietary supplements (DS) including health functional foods (FF) among children and a recent rescission of restrictions in shape/form of FF, children's excessive vitamins/minerals (V/M) intake became a matter of concern in Korea. A nationwide survey was conducted to estimate children's V/M intake from various dietary sources employing 3 representative samples of children for summer & fall of 2008 and spring of 2009 by stratified multistage sampling of 120 survey sites/season based on the 2005 census population. About 30 households from each survey site were screened for residing children of 0–19 years and @ 1,700 households remained as eligible sample per season. Trained dietitians visited households to perform face‐to‐face interview to children regarding DS and OTC V/M products consumption during 1 month prior to interview. Name & type of product, quantity & frequency of use, compliance with recommended dose, etc. were questioned. Out of 5,328 children responded, 19% reported 1 or more DS consumption. Consumption rate was higher in boys (20.1% vs. 17.8%) and youngsters (21.0% compared to 14.3% in adolescents). Although only a minor portion of children showed V/M intake higher than UL, most of them were 0–6 years. Because current maximum levels of V/M in DS were set for adults, using separate doses or directions for children is recommended. Supported by a grant 09082KFDA999 from KFDA in 2008–9.
A Multi‐Country Profile of Anemia in Infants less than 6 months of age
Authors: Sangha Jasbir Kaur , Kothari Monica Tandon
Published: June 2021

A term infant has an average hemoglobin level of 16.8 gm/dL and it is believed that infants have adequate iron nutritional status. However, various other factors such as family history of anemia, ethnicity, maternal and perinatal factors such as maternal anemia status, complications during labor and delivery, and jaundice in neonates and malaria can alter the hematological values in the critical period of infancy. In population based surveys such as the Demographic and Health Surveys (DHS) infants less than 6 months of age are usually not tested for anemia, however, the early phases DHS surveys, included anemia testing in children 0–59 months of age though not all countries reported anemia levels in children less than 6 months of age. In less developed countries newborn life is fraught with vicious cycle of infections and malnutrition, however, its impact on anemia levels in infants less than 6 months of age remains to be investigated from nationally representative samples from population based surveys. Therefore, this paper analyzes data from several countries in Central Asia, Southeast Asia, Africa and South America to determine the prevalence and distribution of anemia in infants less than 6 months of age. The paper will also discuss the socio‐demographic, maternal and nutritional factors that may be associated with anemia in infants less than 6 months of age in the developing world.
Green tea polyphenols improve serum lipid profiles and C‐reactive protein inflammation marker in DSS‐treated and atherogenic diet‐fed rats
Authors: Bornhoeft Julie , Averna Antoinette , Castaneda Debra , Nemoseck Tricia , Hong Mee Young
Published: June 2021

Acute and chronic inflammation play critical role on various disease development including atherosclerosis and heart disease. Green tea polyphenols possess potent antioxidative properties and have been shown the beneficial effects on heart health. The present study was carried out to determine if polyphenols found in green tea (Polyphenon E, generously provided by Polyphenon E International Inc., New York, NY) reduce dextran sodium sulfate (DSS)‐induced inflammation via their antioxidant capacity in atherogenic diet (high fat, cholesterol and sugar)‐fed rats. Green tea diet lowered total cholesterol and LDL cholesterol and increased HDL‐cholesterol (P<0.01). Green tea diet group also increased serum antioxidant capacity (P<0.01) and decreased C‐reactive protein (CRP) inflammation marker by 50% (P<0.01). Interestingly, green tea diet‐fed rats had lower amount of epididymal body fat (P<0.01). The data suggest that green tea polyphenols may contribute to a decreased risk of cardiovascular disease and atherosclerosis by its antioxidant activity, improved blood lipid profile and reduced inflammation. Supported by SDSU UGP and SDSU NUTR 302L class.
Kinetics of DNA unwinding by the RecD2 helicase from Deinococcus radiodurans
Authors: Shadrick William R. , Julin Douglas A.
Published: June 2021

RecD2 from Deinococcus radiodurans is a superfamily 1 DNA helicase that is homologous to the E. coli RecD protein but functions outside the context of RecBCD enzyme. We report here on the kinetics of DNA unwinding by the D. radiodurans RecD2 under single‐ and multiple‐turnover conditions. There is little unwinding of 20 bp substrates by preformed RecD2‐dsDNA complexes when excess ssDNA is present to trap enzyme molecules not bound to the substrate, using substrates with a 5′‐ssDNA overhang or a forked end. A 12 bp substrate is unwound rapidly under single‐turnover conditions. The unwinding time course can be simulated with a mechanism in which the DNA is unwound in two kinetic steps with rate constant = 5.5 sec1, kunw and a dissociation step from partially unwound DNA of koff = 1.9 sec1. These results indicate a kinetic step size of about 3 – 4 bp, an unwinding rate of about 15 – 20 bp/sec, and low processivity. The reaction time course data with the 20 bp substrates, determined under multiple‐turnover conditions, can be simulated with a four‐step mechanism and rate constant values very similar to those for the 12 bp substrate. Analysis of reactions done at several RecD2 protein concentrations indicates that the enzyme forms an inactive dimer or other oligomeric form at high enzyme concentrations.
Consumption of diets fortified with edible mushrooms alters IL‐6 secretion in vivo and in vitro and spleen cell proliferation in dextran sodium sulfate (DSS)‐treated mice
Authors: Christopher Lawrance , Traore Djibril , Kuvibidila Solo
Published: June 2021

Exotic mushrooms (e.i. shiitake – SM) have long been used in ancient Chinese medicine for treatment and prevention of chronic inflammatory diseases. However, little scientific data exist on the health benefits of white button (WM) and portabello (PM) mushrooms commonly consumed in the US. We investigated the capacity of WB, PM, and SM to modulate IL‐6 secretion in vivo and in vitro and spleen cell proliferation in two‐month old C57BL/6 mice fed either the AIN76 diet, or the same diet fortified with 5% dry mushrooms (16/group). After 6 weeks, 8 mice in each group received 3% DSS in drinking water (an experimental model for colitis) or plain water. Upon euthanasia, spleen cells were activated with Con A and LPS (2.5 μg/ml). In control, but not DSS‐treated mice, spleen cell proliferation was higher in mice fed WM and PM fortified diets than in those fed SM‐diet. In control mice, consumption of mushroom‐ diets was associated with lower plasma IL‐6 levels, with the greatest benefit observed with SM. In DSS‐treated mice, all mushrooms significantly increased plasma IL‐6 levels (p<0.05). Mushrooms reduced baseline in vitro IL‐6 secretion but had no effect in Con A‐treated cells. Reduced plasma IL‐6 levels in control mice may modulate CRP level, a risk factor for cardiovascular disease. Increased plasma IL‐6 levels in DSS‐treated mice may be beneficial because they would down‐regulate blood levels of proinflammatory cytokines.
Folate deficiency impairs brain oxygen delivery in rat
Authors: Troen Aron Milton , Hallacoglu Bertan , Sassaroli Angelo , Rosenberg Irwin H , Fantini Sergio
Published: June 2021

Dietary vascular risk factors, including poor folate status are potentially modifiable predictors of cognitive impairment among older adults. In mouse, we have shown consuming a diet deficient in vitamins folate, B12 and B6 to cause a 30% decrease in brain capillary density in association with significant cognitive impairment. Similarly, folate deficiency in rat impairs cognition and causes cerebral microvascular damage, without concomitant neurodegeneration. We hypothesized that folate deficiency might result in functional decrements in cerebral oxygen delivery and vascular reactivity. We fed rats folate deficient diets for up to 20 weeks and used non‐invasive near‐infrared spectroscopy to measure absolute brain concentrations of oxygen free and bound hemoglobin, tissue oxygen saturation, and related hemodynamic parameters at rest and during transient mild hypoxia and hypercapnia. Brain hemoglobin concentration and oxygen saturation were significantly lower in folate deficient than in control rats (73±10 vs. 95±14 μM and 55%±7% vs. 66% ±4% respectively). In contrast, arterial oxygen saturation did not differ by diet with baseline values of 96%±2% for both groups, indicating that the decrements in brain were independent of blood oxygen carrying capacity. Our results implicate microvascular rarefaction and diminished oxygen delivery as a mechanism of cognitive impairment in folate deficiency.
Dietary vitamin D3 at 10 fold the adequate intake may attenuate disease severity in the transgenic G93A mouse model of ALS
Authors: Gianforcaro Alexandro , Hamadeh Mazen J
Published: June 2021

Background

Amyotrophic lateral sclerosis (ALS) is a paralytic and fatal neurodegenerative disease due to motor neuron death. Vitamin D3 mitigates the severity of autoimmune and neurodegenerative diseases that share some common pathophysiologies with ALS.

Objective

To determine the effects of dietary vitamin D3 on functional and disease outcomes and lifespan in the transgenic G93A mouse model of ALS.

Methods

Starting at age 40 d, 24 G93A mice (7 F, 17 M) were assigned to a diet with either adequate (AI; 1 IU D3/g feed) or high (HiD; 10 IU D3/g feed) vitamin D3.

Preliminary Results

Around disease onset, HiD‐F had 60% higher ability to move and 40% higher body condition vs AI‐F (P < 0.001, for both). HiD‐F (19 ± 4 d) had 26% prolonged progression to endpoint from onset vs AI‐F (15 ± 0 d), albeit not significant (P = 0.179). HiD mice had 26% lower cumulative clinical scores vs AI mice (28 ± 2 vs 38 ± 1; P = 0.072). Within females, HiD had 43% lower cumulative clinical scores vs AI (21 ± 6 vs 36 ± 8; P = 0.097). Males reached disease onset 2.8 fold faster (HR, 95% CI: 2, 11; P = 0.004) and endpoint 2.4 fold faster (HR, 95% CI: 1, 12; P = 0.025) vs females.

Conclusion

Vitamin D3 at 10 fold the adequate intake may attenuate disease severity in the transgenic G93A mouse model of ALS, and improve functional outcomes in females. Supported by NSERC and Faculty of Health‐YORK U.
The role of hPNPase, an IFN‐inducible gene, in the hepatocellular innate immune response to HCV infection
Authors: Gonzalez Oscar , Yeh George K. , Koehler Carla M. , Teitiell Michael A. , French Samuel W.
Published: June 2021

Hepatitis C virus (HCV) infection has a prevalence of 3% worldwide, causing major liver disease to more than 170 million infected people. With 70% of acute infections progressing to persistence, it is the leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC). Viral infection is known to trigger host immune responses and initiate signaling pathways that induce IFN production. hPNPase (human polynucleotide phosphorylase) is a mitochondrial protein induced in response to type I interferon. To determine its involvement in antiviral response to HCV, we used an in vitro HCV cell culture infection system (HCVcc). We found hPNPase to be induced over time by infection with HCVcc. Further siRNA‐mediated knockdown of hPNPase increased cellular levels of the HCV Core protein during infection in Huh‐7 cells. This data implicates hPNPase as part of the hepatocyte's innate immune response to HCV infection.
Baseline and Serial Microparticle Concentration Analysis in Elderly Patients with Acute Hip Fracture Treated with Heparins
Authors: Azam Hamza Shahid , Litinas Evangelos , Hopkinson William , Hoppensteadt Debra , Lassen Michael , Fareed Jawed
Published: June 2021

Hip fractures in the elderly may lead to the development of venous thromboembolism and inflammation. Microparticles (MP) are small vesicles that are released upon inflammatory insults and are the main source of blood‐borne tissue factor and of pro‐coagulant phospholipids. This study was designed to measure the concentration of MPs in this patient population by ELISA methods. Thirty samples from a randomized multi‐center double blind control hip fracture study (PK‐532; n = 312) were used. The samples were collected prior to surgery (baseline) and at 1, 3, 5, and 7 days post op. Plasma from 36 healthy volunteers, normal human plasma (NHP) and pathologic pooled plasma were used as quality controls. The MP concentrations between the control groups was similar. The baselines samples demonstrated a 2.5‐fold increase in MP concentration (18.6 nM, ±10.2) in comparison to the control samples (6.7 nM, ±2.5). At post op days 1, 3, 5, and 7, three trends emerged. 15 out of 30 (50%) demonstrated a steady decrease in MP concentrations from baseline to 7 days. 5 out of 30 (16.6%) had a consecutive increase on these days. Finally 10 of 30 (33.3%) had a variable MP concentrations. These results demonstrate that patients with hip fracture requiring surgical intervention showed an up‐regulation of MPs which was down regulated by treatment with heparins. The marked fluctuations in MP levels in these patients and the lack of down regulatory responses may warrant additional pharmacologic intervention to achieve optimal anticoagulation.
DIABETES STIMULATES CEREBRAL NEOVASCULARIZATION: IMPLICATIONS FOR STROKE OUTCOME
Authors: Prakash Roshini , Ogbi Safia , Stern Javier E , Ergul Adviye
Published: June 2021

We have previously shown that ischemic brain injury results in greater hemorrhagic transformation (HT), a complication of acute ischemic stroke, in diabetic Goto‐Kakizaki (GK) rats. These rats also exhibited adaptive arteriogenesis characterized by increased tortuosity, collateral numbers and diameter. Based on these results we hypothesized that diabetes mediates cerebral neovascularization and these newly formed/remodeled vessels bleed easily. We evaluated the cerebral angioarchitecture by 3‐dimensional reconstruction of the vasculature using Z‐stacks images obtained by the confocal microscopy and measured stereological parameters viz: vascular volume, density and surface area. Angiogenesis was spatially characterized in different brain regions i.e. in cortex and striatum where infarcts and HT are observed. Cortical vascular volume and surface area were increased by 6 and 4 folds respectively (p<0.0001) in diabetic rats compared to control Wistars (n=5). Pronounced cortical neovascularization was also observed within the diabetic group. Significantly increased expression of vascular endothelial growth factor and matrix metalloprotease (MMP‐2) activity were also observed in macrovessels of diabetic rats. These results suggest that diabetes mediated cerebral neovascularization complicates disease pathology as seen in diabetic retinopathy.
Bowel Carcinoma Proliferative Activity in a model of experimental
Authors: Carvalho Sa Pedro , Cavalcanti Karen , Coelho Nuno , Souza Ivone Antonia , Cabrita Antonio Silverio
Published: June 2021

Insecticide 1‐naphtyl N‐methylcarbamate (1NC) has a wide spectrum action and is considered of low toxicity when applied according with the good practice rules. The present work was designed to evaluate morphology and proliferation rates of experimentally induced adenocarcinomas.

30 Wistar 8 weeks old male rats were randomly distributed into three groups to be submitted to dimethylhydrazine (DMH) 15mg/week during 8week. Group II, to 1NC 60mg/l in drinking water during 8weeks. Group III, and without manipulation â€“ Group I. All the animals were sacrificed by the end of the 28th week.

Group I rats had no colonic adenocarcinomas while groups II and III exhibited exophytic lesions with wider diameter between 2 and 4 cm. All were well differentiated adenocarcinomas except for one case 1NC induced that was moderately differentiated. Proliferation rate validated by PCNA immunohistochemical study showed to be higher in Group III: 6 negative cases in Group II against 3 negative cases in Group III; the cut off applied + (< 25% positive nuclei), ++ (25 % –75% positive nuclei) and 3+ (> 75% positive nuclei) demonstrated 5 cases of ++ and 3+ in Group III and 1 case 3+ in Group II as the other 3 cases had proliferation rate of +. Despite the small sample explored in this preliminary study, 1NC revealed to induce more aggressive adenocarcinomas validated by higher proliferation rate when compared with the DMH induced cases.
Colon Carcinogenesis p53 Dependent
Authors: Coelho Nuno , Sa Pedro , Alarcao Ana , Teixeira Lia , Cabrita Antonio Silverio
Published: June 2021

Insecticide 1‐naphtyl N‐methylcarbamate (1NC) has a wide spectrum action and is considered of low toxicity when applied according with the good practice rules. The present work was designed to evaluate morphology and proliferation rates of experimentally induced adenocarcinomas.

Thirty Wistar 8 weeks old male rats were randomly distributed into three groups (10 rats each), to be submitted to dimethylhydrazine (DMH) 15mg/week during 8weeks – Group II, to 1NC 60mg/liter in drinking water during 8weeks – Group III, and without manipulation – Group I. All the animals were sacrificed by the end of the twentyeighth week.

Group I rats had no colonic adenocarcinomas while groups II and III exhibited exophytic lesions with wider diameter between 2 and 4 cm. All were well differentiated adenocarcinomas except for one case 1NC induced that was moderately differentiated. Apoptosis was validated by p53/DO7 immunohistochemistry and showed to be higher in Group III cases (1NC induced): 6 positive cases against 1 positive case in Group II; the cut off applied + (< 25% positive nuclei), ++ (25 – 75% positive nuclei) and +++ (> 75% positive nuclei) demonstrated to be useful but only one case was ++ in Group III.

Despite the small sample explored in this preliminary study, 1NC carcinogenesis has to be someway related with apoptosis blockage.
Low cocaine responding rats have a greater basal number of hippocampal norepinephrine transporters than high cocaine responding rats
Authors: Richards Toni L , Zahniser Nancy R
Published: June 2021

Cocaine inhibits dopamine (DA) and norepinephrine (NE) uptake by blocking DA and NE transporters (DATs and NETs, respectively). We have classified outbred male Sprague‐Dawley rats given an acute low dose of cocaine (10 mg/kg, i.p.) as either low or high cocaine responders (LCRs or HCRs, respectively) based on the median split of the first 30 min of cocaine‐induced locomotor activity. This model of differential individual responsiveness to cocaine has revealed that LCRs, which exhibit greater cocaine‐induced locomotor sensitization, reward and reinforcement, have more striatal DATs at baseline than HCRs. To examine if basal NET numbers also differ, here we quantitated indirect saturation binding with a NET inhibitor ([3H]nisoxetine, 4 nM) in hippocampal membranes 35 min after acute cocaine. As expected, HCRs (n = 9) exhibited ~4 fold greater cocaine‐induced locomotor activity than LCRs (n = 7). Affinities of hippocampal NETs (Kd = 5 nM) did not differ significantly between LCRs and HCRs. In contrast, LCRs had 41% more hippocampal NET binding sites than HCRs (Bmax = 137 vs. 97 fmol/mg protein, respectively; p < 0.05, t‐test). Whether these differences in NET number result in corresponding functional differences still must be determined. Nonetheless, the greater basal number of NETs in LCRs is consistent with LCRs being less sensitive to the initial effects of low dose cocaine than HCRs. Supported by DA004216 & DA015050.
Change in down‐stream signaling of striatal dopaminergic transmission in methamphetamine‐sensitized mice
Authors: Tien Lu‐Tai , Lee Yih‐Jing
Published: June 2021

Repeated administration of methamphetamine has been known to produce a progressively enhanced and persistent behavioral response in rodents, a phenomenon called “behavioral sensitization”. The present study was designed to investigate the changes in down‐stream signaling of dopaminergic system after administration of methamphetamine. Mice were injected with saline or methamphetamine (1 mg/kg, i.p.) once a day for 7 consecutive days. The expression of behavioral sensitization was tested on day 11 (after 4‐day drug abstinence). The locomotor activity was increased in methamphetamine treated mice than that of the saline treated group. On day 11, methamphetamine‐induced locomotor activity was significantly higher than those of the same group of mice on day 1, suggesting the development of behavioral sensitization. Repeated injection of saline had no effect on locomotor activity in mice on any given day. The striatal neurons of saline and methamphetamine treated mice were collected after locomotor activity monitoring for DARPP‐32 (a dopamine and cAMP‐regulated phosphoprotein of Mr32) and phosphorylation of DARPP‐32 assay. Two sites of phosphorylation on DARPP‐32: p‐DARPP‐32 (Thr34) and p‐DARPP‐32 (Thr75) were detected. There were no significant differences in the protein levels of DARPP‐32, p‐DARPP‐32 (Thr34) and p‐DARPP‐32 (Thr75) between the saline and methamphetamine treated mice on day 1. The level of p‐DARPP‐32 (Thr75), but not DARPP‐32 and p‐DARPP‐32 (Thr34), is significantly higher than that of the saline treated mice on day 11. These data showed that phosphorylation of DARPP‐32 especially on Thr75 may be related to the expression of methamphetamine‐induced sensitization. (This study is supported by a grant NHRI‐EX99‐9815NC from National Health Research Institutes of Taiwan)
Effects of exendin in rats trained to discriminate between 22 and 2 hr food deprivation
Authors: Jewett David C. , Dunbar Andrew , Faile Shannon , Green Ryan , Bauer Dana , Bolek Barbara
Published: June 2021

Previous research had demonstrated that GLP‐1 agonists decrease food intake and energy expenditure. Previously we have found that drugs that alter energy expenditure also alter the discriminative stimulus effects of 22 hr food deprivation in rats. We tested the effects of exendin‐4, a GLP‐1 agonist, in 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 exendin‐4 (0.1–0.56 mcg/kg, i.p.). Thirty min later, the discriminative stimulus effects were assessed. Exendin‐4 did not alter the discriminative stimulus effects of 22 hr food deprivation. A larger dose (1.0 mcg/kg) eliminated responding and significantly decreased food intake. While exendin‐4 decreases intake and rate of responding, exendin‐4 did not alter the “hunger” discrimination.
Muscarinic Receptor Stimulation of D‐aspartate Uptake Into Human SH‐SY5Y Neuroblastoma Cells Is Attenuated By Hyposmolarity
Authors: Foster Daniel John , Heacock Anne M , Fisher Stephen K
Published: June 2021

In the present study, the ability of muscarinic cholinergic receptors (mAChRs) to regulate the uptake of glutamate (monitored as D‐aspartate) into human SH‐SY5Y neuroblastoma cells under isotonic or hypotonic conditions has been examined. In isotonic media, agonist activation of mAChRs resulted in a significant increase (250–300% of control) in the uptake of D‐aspartate and concurrently, a cellular redistribution of the excitatory amino acid transporter 3 (EAAT3) to the plasma membrane. In hypotonic media, the ability of mAChR activation to facilitate D‐aspartate uptake was significantly attenuated (40–50%) and the cellular distribution of EAAT3 was disrupted. Reduction of mAChR‐stimulated D‐aspartate uptake under hyposmotic conditions could be fully reversed upon re‐exposure of the cells to isotonic media. Under both isotonic and hypotonic conditions, mAChR‐mediated increases in D‐aspartate uptake were dependent upon cytoskeletal integrity, protein kinase C (PKC) and phosphatidylinositol 3‐kinase (PI3K) activities and the availability of intracellular Ca2+. In contrast, a dependence on extracellular Ca2+ was only observed under isotonic conditions. The results suggest that although the uptake of D‐aspartate into SH‐SY5Y cells is enhanced following mAChR activation, this process is markedly attenuated by hyposmolarity. Supported by GM007767 (DJF).
Chracterization Of Biotin Operon In Pseudomonas Mutabilis
Authors: Pedapudi Sasanka , Clack Beatrice A.
Published: June 2021

Biotin (Vitamin H) is an essential nutrient for all organisms. The arrangement of the bio genes in the operon differ between gram positive and gram negative as well within a genus and between the species. This diversity is also extended to the number of bio genes present in the operon and their organization in an operon in bidirectional reading frame as in Escherichia coli. In Bacillus subtilis, all of the bio genes are organized within a single transcription unit, while in other cases, the genes are organized in multiple transcriptional units such as Baciillus sphaericus. The objective of this study was to clone, sequence and determine the order of the bio genes of Pseudomonas mutabilis. Polymerase Chain Reaction was performed with degenerate primers to clone the bio genes into pGEM®‐T Easy Vector for sequencing. Gene specific primers from the sequenced regions were used for modified DNA‐RACE (5′ & 3′) to identify the 5′ and 3′ regions of the bio genes. All five bio genes were successfully sequenced resulting in the order of the genes observed to be “B F H C D”; The bioA gene was sequenced and found to be separated from the primary operon. The analysis of sequenced data reveled that P. mutabilis is closely related to Pseudomonas putida. This work was funded through the Office of Research and Sponsored Projects at Stephen F. Austin State University.
PHARMACOLOGICAL CHARACTERIZATION OF A PURINERGIC RECEPTOR (P2X7) ANTAGONIST IN‐VITRO AND IN‐VIVO
Authors: Bhattacharya Anindya , Maher Michael P , Eckert William A , Olafson Traci , Wang Qi , Ao Hong , Wu Nyantsz , Neff Robert , Freedman Jamie M , Lebsack Alec D , Jones William M , Hawryluk Natalie A , Chaplan Sandra R , Wickenden Alan D
Published: June 2021

Purinergic receptor subtype 7 (P2X7) is expressed on immune/glial cells and is hypothesized to modulate both inflammatory and neuropathic pain as seen by genetic disruption and pharmacological intervention. In this study, we report the characterization of a novel P2X7 receptor antagonist discovered by GlaxoSmithKline (compound 16; Chambers et al., 2009; RSC‐SCI Medicinal Chemistry Symposium, Cambridge, UK). The compound blocked human, rat and mouse recombinant P2X7 receptors with mean pIC50 value of 6.8, 5.3 and 5.8 in FLIPR assay of intracellular calcium mobilization by Bz‐ATP. Compound 16 also attenuated Bz‐ATP dependent IL‐1β release in human and mouse whole blood with mean pIC50 value of 6.2 and 5.5, respectively. The block induced by compound 16 was surmountable in human whole blood and it displaced [3H]‐A804598 in membrane preparations from human P2X7‐1321N1 cells. In whole‐cell patch clamp assay, 1 μM of the compound blocked 300 μM Bz‐ATP‐induced inward current in a reversible manner. The compound was orally bioavailable when dosed in mice and produced a dose dependent inhibition of IL‐1β at 2 hours post oral dose in a blood ex‐vivo assay (ED50 ~ 1–3 mg/kg). At 15 mg/kg (oral) the compound also reversed mechanical allodynia in a mouse model of neuropathic pain (SNI). In addition, the compound also reversed CFA‐induced spontaneous inflammatory pain (weight bearing) in rats at 10 mg/kg tested at 1 and 2 hours post oral dose. Compound 16 is an efficacious P2X7 antagonist that maybe used as a tool to better understand the role of P2X7 in animal models of pain.
Effect of Nigella Sativa on Carbon Tetrachloride and Paracetamol Induced hepatotoxicity: Role of Antioxidant Enzymes and Cytokines
Authors: Barakat Mervat kamel , Oda Naglaa Raafat , Bayoumy Fatehia Ali , Bayoumy Fatehia Ali
Published: June 2021

Objective

To investigate the possible protective effect of Nigella Sativa oil (NSO) on carbon tetrachloride (CCL4) and paracetamol (Para) induced hepatotoxicity in view of correlation with antioxidant enzymes and cytokines.

Material and Method

40 male albino rats (150–200 gm) were divided equally into 5 groups:group A (control), B (Para treated), C (Para+ NSO treated), D (CCL4 treated), E (CCL4+ NSO treated). Blood samples for estimation of liver enzymes. Liver tissues were used for histopathology study and biochemical assays of antioxidant enzymes Superoxide dismutase (SOD), Catalase (CAT), and Glutathione S‐transferase (GST), and proinflammatory cytokine (IL‐1á).

Results

The plasma activity of Liver enzymes was significantly increased after treatment with CCL4, and Para. Their level was statistically decreased in rats pretreated with NSO. Tissue activity of GST was significantly higher in group of C as compared to B while SOD and Cat were significantly increased group E as compared to D. IL‐1 á was significantly increased in group E as compared to all groups.

Conclusions

our results suggest the potential protective role of NSO in CCL4 and Paracetamol induced hepatotoxicity through selective increase of antioxidant enzymes.
Arginase II Deletion Improves Diabetes‐Induced Neurogenic and Endothelial Dysfunction in Mice Corpora Cavernosa
Authors: Toque Haroldo Alfredo Flores , Xu Zhimin , Tostes Rita C. , Webb R. Clinton , Caldwell Ruth B. , Caldwell R. William
Published: June 2021

Diabetes‐induced erectile dysfunction involves elevated arginase (AG) activity. Because NO synthase and AG share and compete for their substrate L‐arginine, NO production is likely linked to regulation of AG. We hypothesized that AG II isoform deletion enhances corpora cavernosal (CC) smooth muscle relaxation in a streptozotocin diabetic (D) model using wild type (WT) and AG II knockout (KO) mice. AG activity and expression and functional response of CC were assessed. CC strips from AG II KO mice exhibited enhanced relaxation to both nitrergic stimulation (1–32 Hz) and the endothelium‐dependent (ED) acetylcholine (84±4%) compared to WT mice (70±3%). WT+D mice showed a significant reduction of nitrergic and ED maximum relaxation (44±8%), but this impairment was prevented in AG II KO+D mice (69±4%). Alpha‐adrenergic agent‐induced contractile responses were increased in CC from WT+D compared to non D controls. Contractile responses were significantly decreased in AG II KO control and D mice vs the WT groups. Diabetes in WT mice increased AG activity and expression (II) and decreased nNOS and phospho‐eNOS levels. AG II KO mice did not exhibit these changes. Additionally, the AG inhibitor BEC (50 μM) enhanced nitrergic and ED relaxation in CC of WT+D mice. Thus, we show for the first time that AG II deletion prevents D‐impairment in CC relaxation. Our findings indicate that AG II plays a role in impaired erectile function.
A novel proadrenergic effect of cardiac natriuretic peptides: inhibition by histamine H3‐receptor activation
Authors: Chan Noel Yan‐Ki , Seyedi Nahid , Levi Roberto
Published: June 2021

Natriuretic peptides (NP) are generally viewed as cardioprotective. Yet, NP were recently found to increase myocardial infarct size in mice, while their deletion decreased it. Further, a recombinant NP (nesiritide) was shown to increase death risk in heart failure patients. Excessive norepinephrine release may have been pivotal in these unexpected events. Thus, we questioned by which mechanisms NP could exert a pro‐adrenergic effect and whether this could be inhibited by histamine H3‐receptor (H3R) activation. NP treatment of sympathetic nerve endings isolated from guinea‐pig hearts (cardiac synaptosomes) and NGF‐differentiated PC12 cells markedly increased cAMP levels and PKA activity, thus promoting catecholamine exocytosis. These pro‐adrenergic effects derived from a cGMP/PKG‐mediated prevention of cAMP hydrolysis by PDE3. Activation of H3R, constitutively expressed in synaptosomes and permanently transfected in PC12 cells, inhibited NP‐induced catecholamine release and prevented the increases in cAMP levels and PKA activity. These effects most likely resulted from a H3R‐mediated enhancement of PDE3 activity and/or PKG inhibition. These findings elucidate new means to control catecholamine exocytosis in the heart, and may foster the design of new agents (e.g., selective H3R agonists) to enable a safe and effective treatment of congestive heart failure with NP.

This work was supported by NIH Grant HL 034215 and by a PhRMA Pre‐doctoral fellowship.
Hypothalamic‐Pituitary‐Adrenal Feedback Regulation
Authors: GOVINDAN MANJAPRA VARIATH
Published: June 2021

The hypothalamic‐pituitary‐adrenal axis (HPA) is regulated by corticotrophin‐releasing hormone (CRH), arginine vasopressin (AVP) and by glucocorticoids. In response to stress, the neuroendocrine system releases CRH and AVP to regulate adrenocorticotropin (ACTH) hormone secretion from the anterior pituitary and glucocorticoids from the adrenal cortex. Increasing levels of circulating glucocorticoids downregulate the synthesis of the glucocorticoid receptor (GR) in various tissues in an endocrine feedback loop and produce inhibitory effects on the secretion of CRH and ACTH (4, to bring back the system to its resting state. Despite this wealth of information, the precise mechanism of feedback regulation remains elusive.

We have identified FBR sequences in the human GR gene promoter and examined the roles of GR, CREB and HDAC‐6 in the negative feedback regulation associated with promoter elements of the human GR gene. We have demonstrated that the enhancer and suppression activities contained within the promoter elements of the GR gene occur through the interaction of GR and the active recruitment of HDAC‐6 and phosphorylated CREB (CREB‐P) by the ligand bound GR. We demonstrated a direct involvement between GR, CREB and HDAC‐6 in the feedback regulation.

In the cortisol depleted state, phosphorylated CREB binds to the CRE in the composite feedback regulatory element and CBP/p/CAF complexes. This interaction results in the acetylation of core histones and chromatin decondensation which turns the GR gene on. In the presence of active glucocorticoids, nGRE recruits GR‐ligand complexes which tether HDAC‐6 and phosphatase complexes. The GR‐DEX bound HDAC‐6 deacetylates core histones and the GR‐DEX tethered phosphatase dephosphorylates CREB‐P. Deacetylation leads to chromatin condensation and dephosphorylation of CREB‐P which results in its removal from the CRE which turns GR gene off.
Triarylmethane‐34 (TRAM‐34), inhibitor of Ca2+‐activated K+ channels of intermediate conductance, is a potent blocker of gap junctions
Authors: Figueroa Xavier F , Gaete Pablo S , Sáez Juan C , Riquelme Manuel
Published: June 2021

Ca2+‐activated K+ channels of intermediate conductance (IKCa) and gap junctions (GJs) between endothelial and smooth muscle cells have been proposed to play a central role in the vasodilator response mediated by the endothelium‐derived hyperpolarizing factor (EDHF). Although TRAM‐34, the IKCa channels blocker, has emerged as an important pharmacological tool for studying EDHF‐mediated responses, its effect on GJs has not been described. We analyzed intercellular communication and cellular distribution of the GJ protein subunit, connexins (Cxs), in HeLa cells transfected with Cx43 or Cx43 conjugated to EGFP. Treatment with 10μM TRAM‐34 induced a time dependent reduction in dye coupling to Lucifer yellow; the average number of coupled cells observed in positive tests was reduced by ~50% after 20 min and abolished after 70 min. Dye coupling was recovered to control 30 min after TRAM‐34 wash out. The reduction in GJ communication was not associated with changes in levels or distribution of Cx43‐EGFP. Therefore, TRAM‐34, in addition to inhibit IKCa channels, also potently blocks the cell‐to‐cell communication mediated by the GJ protein Cx43 that is an important connexin involved in the coordination of resistance vessel function.

VRAID 15/2009, FONDEF D07I1086.
Do Ovulatory Hormones Protect Flow‐Mediated Dilation Against The Insult Of A High‐Fat Meal?
Authors: Harris Ryan A , Tedjasaputra Vince , Richardson Russell S
Published: June 2021

The fluctuation in female sex‐steroid hormones across the menstrual cycle has been associated with changes in flow‐mediated dilation (FMD), such that an increase in estrogen contributes to improvements in FMD. In contrast, ingestion of a high‐fat meal has been documented to reduce FMD. Therefore, it was hypothesized that elevated estrogen, during the ovulatory and luteal phases, would prevent the reduction in FMD following ingestion of a high‐fat meal. FMD was determined at baseline and 4 hours following the ingestion of a high‐fat meal in women during the menses (M), follicular (F), and luteal (L) phases. Men participated as control subjects and were studied once. Estradiol was elevated (p<0.05) during the F (5.3±0.7 pg/ml) and L (5.2±0.6 pg/ml) phases compared to the M (3.9±0.5 pg/ml) phase, whereas progesterone was significantly elevated during the L (141±23 pg/ml) phase compared to M (69±11 pg/ml) and F (69±9 pg/ml) phases. Post‐prandial FMD was similar (p>0.05) in the women during any menstrual cycle phase; however, men exhibited ~50% reduction (p<0.05) in FMD following the ingestion of the high‐fat meal (6.4±1% to 3.3±1%). Interestingly, delta FMD and concentrations of estradiol or progesterone were not correlated. Although this study reveals a sex difference in the vascular response to a high fat meal, female sex‐steroid hormones could be questioned in affording women vascular protection from such an insult.
AMP‐activated protein kinase (AMPK) mediated relaxation is enhanced in aorta from old versus young Sprague‐Dawley Rats
Authors: Ford Rebecca J , Durham Kristina , Rush James WE
Published: June 2021

Activation of AMPK induces vasorelaxation in young, healthy arteries, but it is unknown if this response is intact in arteries from old animals. Here we investigate AMPK‐mediated relaxation in aorta from aged rats. Relaxation dose‐response curves to the AMPK activator 5‐aminoimidazole‐4‐carboxamide‐1‐β‐D‐ribofuranoside (AICAR) were generated in phenylephrine pre‐contracted aortic rings from 12 young (10 wks) and 12 old (85 wks) male Sprague Dawley rats using vascular myography. Area under the dose‐response curves (AUC, arbitrary units) was determined as an index of relaxation. Mechanisms of relaxation were dissected in rings with intact (E+) or mechanically disrupted (E‐) endothelium and with or without pharmacological inhibition of nitric oxide synthase (using N‐nitro‐L‐arginine methyl ester; L‐NAME). Endothelium‐dependent and ‐independent relaxation to AICAR occurred in aortic rings from old (AUC E+: 196±27, E‐: 130±9) and young (AUC E+: 94±5, E‐: 53±4) rats, and responses were enhanced in old versus young rings (P < 0.01 E+, P < 0.01 E‐). In both young and old aorta, the endothelium‐dependent component of relaxation was removed with L‐NAME (AUC OLD: 98±10 P =NS, Young: 52±3 P = NS vs E‐). We demonstrate for the first time that AMPK‐mediated relaxation occurs and is enhanced in aorta from old animals, and that AMPK may be a target to enhance vasomotor function in aged arteries. Funded by NSERC
Nitric oxide and renal haemodynamics during postnatal maturation: Role of angiotensin receptors
Authors: Vinturache Angela Elena , Smith Francine Gabriel
Published: June 2021

To investigate whether renal haemodynamic effects of nitric oxide (NO) in the newborn are mediated by activation of angiotensin type 1 (AT1) or/and type 2 (AT2) receptors, three experiments were carried out in conscious, chronically instrumented lambs aged ~one (N=5) and ~six weeks (N=6). Measurements were made before and after I.V. administration of antagonists selective to the AT1 receptor (ZD 7155, experiment 1), the AT2 receptor (PD 123319, experiment 2), and both receptors (ZD 7155+PD 123319, experiment 3) before and after administration of the L‐arginine analogue, L‐NAME. After ZD 7155 and ZD 7155+PD 123319, but not PD 123319 alone, renal vascular resistance (RVR) decreased in both age groups by ~25%. At one week, the RVR increase after L‐NAME was ~20% with ZD 7155, and ZD 7155 plus PD 123319, and ~16% with PD 123319 alone. At six weeks, the RVR increase after L‐NAME was ~40% with ZD 7155, and ZD 7155+PD 123319, and ~30% with PD 123319 alone. Therefore, ANG II modulates baseline renal haemodynamics as well as the renal haemodynamic effects of NO in an age‐dependent manner, through activation of both AT1 and AT2 receptors.
P‐Rex1 regulates lung microvascular permeability
Authors: Naikawadi Ram Prasad , Cheng Ni , Wu Dianqing , Ye Richard
Published: June 2021

Acute Lung Injury (ALI) is a major cause of death in sepsis resulting from Gram‐negative bacterial infection. Polymorphonuclear leukocytes (PMNs) respond to septicemic infection by releasing superoxide anions, which is a major mechanism for bacterial clearance. However, uncontrolled activation of the NADPH oxidase leads to injury to the host tissue, which underscores the importance for the proper regulation of this enzyme. PIP3 dependent Rac exchanger 1 (P‐Rex1), a Rac guanine nucleotide exchange factor, is known for its role in regulating chemoattractant‐induced superoxide production in PMNs through G‐protein coupled receptor (GPCR) signaling. Using P‐Rex1 knockout mice, we found that the LPS‐induced lung injury was significantly reduced as compared to the wild type mice. In addition, we also observed P‐Rex1 expression in lung microvascular endothelial cells, suggesting a function of P‐Rex1 in endothelial cell signaling. siRNA‐mediated knock‐down of P‐Rex1 expression in endothelial cells prevented the increase in endothelial barrier permeability following stimulation with TNF‐alpha, which is a major cytokine released in LPS‐induced sepsis. The altered endothelial permeability is accompanied by a severe ablation of TNF‐alpha‐induced Rac activation and reactive oxygen species (ROS) generation in endothelial cells. From these results, we conclude that TNF‐alpha contributes to lung microvascular permeability through ROS‐mediated disruption of the endothelial junction and P‐Rex1 plays a central role in this process. (Source of research support – NIH)
Isoproterenol‐induced heart failure model in the rat: cardiac autonomic tone, left ventricular function and baroreflex sensitivity
Authors: Dias Daniel Penteado Martins , Salgado Helio Cesar , Beltz Terry G , Xue Baojian , Johnson Alan Kim
Published: June 2021

We investigated the cardiac autonomic tone, left ventricle function and baroreflex sensitivity in rats subjected to isoproterenol‐induced heart failure (HF). Male Sprague Dawley rats received 2 injections (separated by a 24‐hour interval) of isoproterenol (ISO; 170 mg/kg/day, sc; n=8) or vehicle (V; saline; sc; n=8). After 4 weeks, the rats were implanted with femoral artery and vein catheters and tested 3 days later. Cardiac autonomic tone, left ventricle function and baroreflex sensitivity were examined with intravenous injections of atropine (1 mg/kg) and propranolol (2 mg/kg), using Millar catheters and the sequence method. ISO treatment decreased basal HR (ISO 332 vs. V 351 bpm), intrinsic HR (ISO 327 vs. V 347 bpm) and reduced the tachycardia elicited by atropine (ISO 389 vs. V 427 bpm). However, propranolol elicited similar bradycardia in both groups. Rats treated with ISO showed decreased dP/dt+ (ISO 8047 vs. V 10167 mmHg/s) and dP/dt‐ (ISO 8149 vs. V 11428 mmHg/s), but no differences were observed in the baroreflex sensitivity (ISO 1.50 vs. V 1.46 ms/mmHg). Our results extend the characterization of the ISO‐induced HF model and demonstrate a decrease in the basal and intrinsic HR without any change in baroreflex sensitivity.
Induction of Nogo‐A in Cardiomyopathy
Authors: Martin Jody L , Kartje Wendy , Goldspink Paul , Akar Joseph , Paul Caitlin , Sarkey Jason
Published: June 2021

The Nogo isoforms A, B, and C are part of a superfamily of proteins termed the reticulons, from the same gene (Nogo/RTN4), share a c‐terminal domain, and are thought to have preferential ER localization. Nogo‐A has been traditionally described as a myelin associated inhibitor of axonal sprouting, with a molecular weight of ~160 kilodaltons (kDa) limited tissue distribution, and multiple transmembrane domains. There is a vast amount of data examining the function of the Nogo reticulon protein family, particularly with respect to neuronal plasticity however, very little is known about the Nogo proteins in the heart. In failing human heart samples we saw a 3 fold increase in Nogo‐A, vs nonfailing by western immunoanalysis (n=5). RT‐PCR of isolated human cardiac fibroblasts showed a 3 fold increase in nogo‐A mRNA with 2 hours of cyclic stretch. Western analysis of PKC epsilon overexpressing heart extracts showed increased levels of Nogo‐A, corresponding to the development of the dilated phenotype. Isolated cardiomyocyte experiment demonstrate an increase in Nogo‐A with mechanical stretch but not with hypertrophic agonist treatment. We propose an increase Nogo‐A in the heart is an adaptive mechanoresponse.
AcSDKP Reduces Collagen Crosslinking, MMP and NFKB Expression in Angiotensin II Induced Hypertension
Authors: Gonzalez German Esteban , Nakagawa Pablo , Liu Yunhe , Liao Tang‐Dong , Yang Xiao‐Ping , Rhaleb Nour‐Eddine , Carretero Oscar A.
Published: June 2021

We hypothesized that N‐acetyl‐Ser‐Asp‐Lys‐Pro AcSDKP not only reduces fibrosis but also crosslinking. Moreover we tested whether it increases MMP, which has antifibrotic effects and whether it reduces NFκB in the heart. Lewis rats were divided in 4 groups (n=6, each): 1 (control); 2 (AngII 750…g/kg/day); 3 (AcSDKP 800…g/kg/day); and 4 (AngII+AcSDKP). Systolic blood pressure (mmHg) was recorded for 3 weeks. After 3 weeks crosslinking was quantified by measuring total (TC), insoluble (IC) and soluble (SC) collagen (μg/mg dry weight tissue) and MMP2 activity (% respect to 1) in cardiac tissue. NFκB activity was quantified (EMSA assay).

Results

(X±SEM)

Control
AngII
AcSDKP
AcSDKP+AngII

BP
125±3
202±15*
122±3
205±8*

TC
11.7±1.3
19.0±1.8*
12.5±1.5
13.0±0.9#

IC
9.5±1.3
16.3±1.6*
10.7±1.3
10.0±0.9#

SC
2.0±0.2
2.7±0.7
1.7±0.4
2.9±0.7

MMP2
100.0±19.2
101.9±13.0
74.9±13.1
139.6±19.5†

NFκB
11.5±1.4
24.4±1.8*
15.0±3.5
15.7±2.0#

p<.05 vs G1 and 2 respectively;

p=.094vsG2

Conclusion

AcSDKP decreased crosslinked collagen which is important for reducing myocardial stiffness in hypertension. MMP2 activity tends to increase, and may have a small role in the antifibrotic effect of AcSDKP. Also, AcSDKP decreased NFκB suggesting that its antiinflammatory effect may be mediated by reducing this factor. Grant#: NIH HL028982 to OAC