bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of cadmium on aminotransferases serum activity in European carp
Authors: Krasnovskaia Marina , Karpenko Larisa , Polistovskaia Polina , Bakhta Alesya , Enukashvili Abram , Kozitcyna Anna , Ivanova Katerina 
Published: May 2022

In many densely populated areas of the Earth, there is an excess amount of heavy metals in soil, air and water, a significant part of which comes from industrial pollution. So it is important to evaluate possible effects of heavy metals in biological models.Aim of this study is to asset toxic influence of heavy metals in fish. This research was intended to analyze aminotransferases (ALT, AST) serum activity under the influence of various cadmium concentrations in European carp. Intracellular enzymes activity level elevation in serum is a sign of mass cell destruction and consequently is a sign of pathological process. In this experiment 3 groups of fish (European carp) were formed, 10 fish in each group: control group; 1‐experimental group – fish were contained in water with 0.05 mg/l cadmium acetate concentration (10 times higher than fishery waters permissible exposure limit according to the laws of the Russian Federation); 2‐experimental group – fish were contained in water with 0.5 mg/l cadmium acetate concentration (100 times higher than fishery waters permissible exposure limit according to the laws of the Russian Federation). Exposure time was 4 hours. The blood samplings were taken from heart by syringe and placed in dry test tubes. After samplings centrifugation in non‐hemolysed serum samples ALT and AST activities were assessed. Results presented as mean±standard error of the mean. Student's t‐test was used after proving normal distribution. Level of significance is labeled as p<0.05.The study revealed that there were meaningful elevation of ALT and AST serum activity (p≤0.05) by 44.22% (30,92±1,42 IU/l) and 15.36% (261,5±8,07 IU/l) accordingly in 1‐experimental group in compare with control group (21.44±0.72 IU/l и 226.68±4.35 IU/l). In case where the cadmium acetate concentration were 100 times higher than fishery waters permissible exposure limit were meaningful elevation of ALT and AST serum activity (p≤0.01) by 88.82% (38,34±0,19 IU/l) and 71.53% (388,82±14,57IU/l) accordingly in 2‐experimental group in compare with control group.After analyzing the received data there were established meaningful aminotransferases serum activity elevation under the influence of various cadmium concentrations in European carp. This may be due to the activation of gluconeogenesis, which is necessary to maintain an adequate glucose level in conditions of intoxication, which determines the direction of metabolic flows towards the predominance of catabolic reactions. This data is important to be taken into consideration not only during assessing physiological condition in fish, but also to increase understanding of the damage extent in biological models.
Arachidonyl‐2‐ Chlorethylamide alters the Gene Expression of Renin Angiotensin System Components in Primary Cultures of Astrocytes
Authors: O'Connor Ann Tenneil , Clark Michelle A. 
Published: May 2022

ObjectiveTo determine the of effect Arachidonyl‐2′‐chloroethylamide (ACEA) on the gene expression of renin angiotensin system (RAS) components in rat astrocytes.BackgroundWe have previously shown that activation of the Angiotensin type 1 receptor (AT1R) altered the expression of the cannabinoid receptor type 1 (CB1R) in rat astrocytes. Activation of the CB1R can potentially have synergistic or antagonistic effects on AT1R actions. Studies have shown that an overactive RAS contributes to pathological conditions. Thus, we determined the molecular effects of activation of the CB1R by ACEA on RAS components. To investigate this, we determined ACEA and Angiotensin (Ang) II effects alone, then combined, on the mRNA levels of RAS components in astrocytes isolated from the cerebellum of Wistar and Spontaneously hypertensive rats (SHRs). CB1R and RAS components are expressed in the cerebellum.MethodsAstrocytes were prepared from 2‐3 days rat pups. Quiescent cells were treated with 10nM ACEA and or 100nM Ang II for 12‐24 hours. mRNA levels of RAS components were measured by quantitative polymerase chain reaction.ResultsAng II predominantly downregulated RAS components in both Wistar and SHR astrocytes at the time points examined. ACEA downregulated AT1R mRNA in Wistar, increased angiotensinogen mRNA in both Wistar and SHR astrocytes, and increased angiotensin converting enzyme mRNA in SHR astrocytes. The effects of combined treatments were similar to Ang II single treatments.ConclusionCB1R activation altered the mRNA levels of RAS components but did not appear to have considerable impact on Ang II’s effects on these components. Since mRNA levels might not correlate with protein levels, determination of protein levels and/or activity would complement these findings.
Musculoskeletal Differences Between Amputated and Non‐Amputated Lower Limbs
Authors: Finnerty Caitlyn , Finco MG , Ngo Wayne , Holley Bethany , Menegaz Rachel A. 
Published: May 2022

People with lower limb amputations frequently experience greater risks of musculoskeletal injury. Forces active during walking help to develop and maintain the shape, volume, and strength of musculoskeletal tissues. Conversely, altered walking patterns following limb loss may lead to atrophy of muscle and bone tissues. Reductions in joint spaces are indicative of excess stress placed on the limb, which may lead to osteoarthritis. Bone loss in high stress regions like the femoral neck can reduce the bone’s ability to resist compressive or rotational movements, making the bone more susceptible to fracture.The aim of this study was to measure musculoskeletal differences between an individual's residual (amputated) limb and intact (non‐amputated) limb to identify structures vulnerable to injury. We hypothesized that the residual limb, compared to the intact limb, would show: 1) less muscle mass and more fat as indicators of muscle atrophy, 2) wider hip and knee joint spaces as indicators of osteoarthritis in the intact limb, and 3) decreased femoral neck width as an indicator of fracture risk.CT scans of 10 males (42‐79 years) were obtained from the New Mexico Decedent Image Database. 3D Slicer software was used to measure gross skeletal properties, hip and knee joint dimensions, and cross‐sectional muscle and fat tissue areas at the midshaft. A Wilcoxon Signed‐Rank test was used to assess the differences between residual and intact limbs. The significance level was set at α ≤ 0.10 due to a small sample size. Compared to the intact limb, the residual limb had significantly less muscle tissue area (p=0.010) and a significantly narrower femoral neck width (p=0.077). No significant differences were found in hip or knee joint spaces between limbs.In agreement with hypotheses 1 and 3, these results suggest residual limbs are at increased risk of muscle atrophy and femoral neck fracture compared to intact limbs. Loading inequalities between the residual and intact limb likely contribute to these results. A better understanding of the structural properties associated with musculoskeletal atrophy could inform targeted therapies to reduce the likelihood of injury in this population. Future studies will assess biomechanical properties, such as moment of inertia, to better understand the residual limb’s ability to withstand torsional forces and fracture. Additional data on how musculoskeletal tissues respond to unloading at multiple structural levels can improve clinical interventions for lower limb strength and function in amputees.
The Gut‐Brain Connection: The Inflammatory Role of IL‐6 Caused by a Western Diet
Authors: Sotelo Alexis A. , Kania Barbie , Ty Darren , Wisco Jonathan 
Published: May 2022

IntroductionAn increase in chronic proinflammatory immune activity is recognized as an increasingly fundamental element in relation to neurodegenerative disorders. Alzheimer’s disease (AD) is one of the most common neurodegenerative disorders, with recent evidence suggesting the Western Diet as a source for systemic inflammation. The communication fusing the gut to brain axis has been investigated through hepcidin, derived from both hepatocytes and dendritic cells. Hepcidin could potentially play an anti‐inflammatory role in the process of gut dysbiosis through a means of either a localized approach of nutritional immunity, or a systemic approach. Similarly, IL‐6 is a part of the gut‐brain axis. The levels of IL‐6 expression are affected by the gut microbiota and therefore sought to interplay a key role in cognitive function and decline, which could ultimately lead to several neurodegenerative diseases such as Alzheimer’s. The production of IL‐6 is increased by inflammation and associated with sepsis, endotoxemia and treatment with pro inflammatory cytokines. The release of this cytokine peripherally influences the CNS by a ting on the microglia to stimulate a state of neuroinflammation. During inflammation, proinflammatory cytokine IL‐6 is released from the intestinal mucosa and activates the Vagus nerve afferents. Hence, it could conceivably be hypothesized that IL‐6 activates gastrointestinal submucosal neurons through the brain‐gut axis. The aim of this study is to conduct a systematic review of literature through the search of available online medical databases containing information pertaining to gut dysbiosis and the primary communication between the brain‐gut axis.MethodsA total of 80 articles were retrieved from the databases of Pubmed, Google Scholar and Frontiers. Our method of assessment analyzed prominent keywords in our study pertaining to the small intestines, brain, liver, hepcidin, TMAO, BDNF, iron, astrocytes, WD, microbiota, gut microbiome, gut dysbiosis, iron, inflammation, oxidative stress, IL‐6, and vagus nerve. After reading the full‐text then extracting the findings from each study, our team organized those findings of brain, liver and gut biomolecules in relation to maintaining homeostasis and prevention of inflammation. The databases were assessed fall of 2021 following exportation of articles to a bibliographical database for further screening processes.ResultsThe results of this study indicate the optimal state of a healthy brain is IL‐6, hepcidin, ferritin and iron are at baseline levels. Whereas, in a dysbiosis state likely caused by WD, the brain responds by upregulating IL‐6, hepcidin and ferritin levels.ConclusionsIt would be interesting for a future study to experimentally focus on whether hepcidin upregulation induces the downregulation of iron in the neuron, or if the upregulation of iron in the neuron induces the downregulation of hepcidin in the setting of neuroinflammation.
Innate Immune System Stimulation During Pregnancy Upregulates Thromboxane Synthesis in Rat Maternal Heart
Authors: Tucker Selina M. , Bradshaw Jessica L. , Cushen Spencer C. , Gardner Jennifer J. , Ricci Contessa A. , Goulopoulou Styliani 
Published: May 2022

BackgroundInfections during pregnancy are associated with adverse maternal and fetal outcomes. We previously showed that exposure to immunostimulatory CpG oligonucleotides (ODN2395, synthetic Toll‐like receptor 9 agonist) during pregnancy induces maternal vascular inflammation and enhances vascular tone in pregnant rats. These outcomes were mediated in part by activation of the cyclooxygenase/thromboxane A2 (COX/TxA2) pathway. The objective of this study was to investigate the impact of ODN2395‐induced immune system stimulation on maternal hearts during pregnancy.HypothesesExposure to TLR9‐mediated immune system activation during pregnancy upregulates the COX/TxA2signaling pathway in maternal cardiac tissues.MethodsRats were treated with a synthetic CpG DNA (ODN2395, 1 mg/kg, intraperitoneal injection) or vehicle (saline) in late pregnancy (gestational day (GD) 14, 16, and 18) and euthanized on GD 20 (term 22‐23). Fetoplacental biometrics were recorded after euthanasia and maternal hearts were collected to assess COX‐1 and COX‐2 expression via western blotting and 6‐keto PGF1α (PGI2 metabolic byproduct) and TxB2 (TxA2 metabolic byproduct) production use ELISA.ResultsMaternal heart weights did not differ between groups (ODN2395: 0.26 g/100 g BW vs. Vehicle: 0.23 g/100 g BW, n≥7, p=0.11). Left ventricular tissues from dams treated with ODN2395 released higher concentrations of TxB2compared to cardiac tissues from vehicle‐treated dams (ODN2395: 0.56 ± 0.06 ng/mg total protein vs. Vehicle: 0.31 ± 0.04 ng/mg total protein, n≥5, p=0.0041) but there were no differences in cardiac 6‐keto PGF1α release between groups (p=0.16). COX‐2 expression was lower in left ventricles from ODN2395‐treated rats compared to vehicle‐treated rats (p=0.009). There were no differences in cardiac COX‐1 expression between groups (p=0.27). Exposure to ODN2395 during pregnancy did not affect fetal weights (ODN2395: 2.28 ± 0.03 g vs. Vehicle: 2.25 ± 0.06 g, n≥7, p=0.9) or placenta weights (ODN2395: 0.44 ± 0.02 g vs. Vehicle 0.49 ± 0.01 g, n≥7, p=0.06), but increased fetal‐placental weight ratio (ODN2395: 5.3 ± 0.22 vs. Vehicle: 4.7 ± 0.15, p = 0.04). COX‐2 expression was greater in placental tissues from ODN2395‐treated rats (p=0.004) but there were no differences in placental 6‐keto PGF1α(p=0.51) and TxB2 production (p=0.32).ConclusionExposure to immunostimulatory CpG ODN during pregnancy induces upregulation of TxB2 synthesis in maternal cardiac tissues coupled with a reduction in COX‐2 expression. We suggest that cardiac inflammation during pregnancy may increase maternal risk for future cardiovascular events.
Development of Field‐Based Prediction Equations For Determining Leg Limb Occlusion Pressure
Authors: Neph Sylvia , Plouffe Audrey E. , Vondrasek Joseph E. , Cox Benjamin , Vranish Jennifer E. , Montoye Alexander E. 
Published: May 2022

Blood flow restriction and ischemic preconditioning are occlusion techniques used in clinical and athletic populations as rehabilitation techniques and ergogenic aids. Both require determination of limb occlusion pressure (LOP) to know how much pressure to put in a limb‐worn cuff. Measurement of LOP takes sophisticated equipment and designing a low‐cost way of predicting LOP may allow for these occlusion techniques to be more readily used in field‐based settings.PURPOSETo determine demographic and other variables that can be used to predict LOP.METHODSParticipants (n=23, 57% female) aged 18 to 53 years reported to the laboratory for a single session. Participant age, height, weight, body mass index (BMI), body fat percentage, thigh circumference of both legs, systolic, diastolic blood pressure of both arms and mean arterial pressure (MAP), resting heart rate, and single‐leg wall sit times for both legs were assessed. Additionally, participants lay in a supine position and were fitted with an automated system (Delfi) on the proximal thigh, one leg at a time, for determining LOP. LOP was confirmed for each leg using a Doppler ultrasound (GE logique) on the popliteal artery. Then, correlation and stepwise regression analyses were used to determine which variables were significant predictors of LOP.RESULTSRight arm MAP (r=0.648) and BMI (r=0.539) had the highest correlations with LOP. Stepwise regression yielded two potential predictive equations to predict LOP. Equation 1: LOP = 2.117*MAP ‐ 6.678 (r2=0.42; standard error of estimate (SEE)=17.6 mmHg); Equation 2: LOP = 1.892*MAP + 2.441*BMI ‐ 52.059 (r2=0.62; SEE=14.6 mmHg).CONCLUSIONArm blood pressure was most closely associated with leg LOP, and MAP along with BMI might serve as useful measures for predicting LOP in field settings. Cross‐validation is needed to confirm these preliminary findings, and further analysis should explore other variables that may allow for more accurate prediction of LOP in field settings.
Dynamic Muscle Fatigue and Potentiation in Human Plantar Flexors
Authors: Zero Alexander M. , Paris Michael T. , Rice Charles L. 
Published: May 2022

Activation history can enhance or impair subsequent contractile responses. Following a brief high‐intensity voluntary contraction, the contractile response for the same submaximal input stimulus will be enhanced (post‐activation potentiation; PAP). However, prolonged voluntary activation can impair contractile responses due to neuromuscular fatigue. Because both effects are dependent on immediate prior activation, a co‐existence between these two competing processes is observed. Specifically, submaximal frequencies are enhanced (PAP) while maximal frequencies are simultaneously depressed (fatigue). Due to preferential enhancement of low frequency activation, PAP can mitigate the effects of prolonged low‐frequency force depression (PLFFD), ‐ a long‐lasting reduction of force evoked at low‐frequencies following fatiguing exercise. Usually, this relationship has been evaluated using isometric contractions. The purpose was to assess the relationship between PAP, muscle fatigue and recovery during and following an isotonic (dynamic) fatiguing protocol through which power (force and velocity) are assessed. It is hypothesized that PAP will attenuate PLFFD during the fatiguing protocol and throughout 30 minutes of recovery.The plantar flexors of 5 healthy young adult males (n=4) and females (n=1) to date have been tested while lying prone in a Cybex dynamometer. The ankle was at 90° and knee joint at 180° (straight leg). Inelastic large Velcro straps firmly secured the hips, shoulders and thighs to the dynamometer. Participants performed isotonic plantar flexion contractions over a 20° range of motion (90° to 110°) of the ankle joint with a load of 20% of maximal voluntary isometric contraction (MVC) until peak power was reduced by 20%, 50% and 75% of baseline values. At these intervals of power loss, electrical stimulation of the tibial nerve in the popliteal fossa was done to record unloaded power responses at 10 Hz and 50 Hz. Following task failure (75% power loss), recovery was assessed immediately and at 3, 5, 10, 20 and 30 minutes. For these, voluntary contractions (10 concentric at 20% MVC) were done to induce PAP. Evoked unloaded dynamic measurements were done prior to and following the 10 concentric contractions to assess any effect of PAP.PLFFD is a preferential loss of low frequency force assessed by comparing a stimulated force ratio between responses at 10 Hz to 50 Hz. During dynamic contractions we used a peak power 10:50 Hz ratio to incorporate the factor of velocity. During fatigue the peak power 10:50 Hz ratio was enhanced by ~35% at 20% power loss, by ~15% at 50% power loss, and minimally enhanced (~9%) at task failure (75% power loss). During recovery this ratio was enhanced follow the potentiating contractions compared to non‐potentiated; task failure (~3%), 3 min (~26%), 5 min (~45%), 10 min (~20%), 20 min (~25%) and 30 min (~38%).These results indicate that during dynamic muscle fatigue and recovery, PAP may mitigate PLFFD to varying degrees.
ClC‐K2 Cl‐ Channel exhibits discrete actions on acid‐base transport of A‐ and B‐Type Intercalated Cells of the Collecting Duct
Authors: Pyrshev Kyrylo , Stavniichuk Anna , Tomilin Viktor N. , Zaika Oleg , Pochynyuk Oleh 
Published: May 2022

The collecting duct is a highly adaptive terminal part of the nephron, which is essential for maintaining systemic homeostasis. Principal and intercalated cells perform different physiological tasks and exhibit distinctive morphology. However, acid‐secreting A‐ and base secreting B‐type of intercalated cells cannot be easily separated in functional studies despite virtually mirror opposite localization of their transport acid‐base systems.Here, we used BCECF‐sensitive intracellular pH (pHi) measurements in split‐opened collecting ducts followed by immunofluorescent microscopy in WT and intercalated cell‐specific ClC‐K2‐/‐ mice to demonstrate that ClC‐K2 inhibition enables to distinguish signals from A‐ and B‐ types of intercalated cells.We show that ClC‐K2 Cl‐ channel is expressed on the basolateral side of intercalated cells, where it governs Cl‐‐dependent H+/HCO3‐ transport. ClC‐K2 blocker, NPPB, caused acidification or alkalization in different subpopulations of intercalated cells in WT but not ClC‐K2 ‐/‐ mice. Immunofluorescent assessment of the same collecting ducts revealed that NPPB increased pHi in AE1‐positive A‐type and decreased pHi in pendrin‐positive B‐type of intercalated cells. Induction of metabolic acidosis led to a significantly augmented abundance and H+ secretion in A‐type and decreased proton transport in B‐type of intercalated cells, whereas metabolic alkalosis caused the opposite changes in intercalated cell function, but did not substantially change their relative abundance.Concluding, we show that inhibition of ClC‐K2 can be employed to discriminate between A‐ and B‐type of intercalated cells in split‐opened collecting duct preparations. We further demonstrate that this method can be used to independently monitor changes in the functional status and abundance of A‐ and B‐types in response to systemic acid/base stimuli.
Developing RNA‐Seq and ChIP‐Seq Pipelines on an Oxford Nanopore Technologies Platform to Study Notch1 Transcriptional Dynamics
Authors: Klier Nikola , White Brandon 
Published: May 2022

Notch signaling plays a crucial role in cell development and proliferation. The Notch receptor is an inducible transcription factor found on the cell surface which is cleaved resulting in the release of the intracellular domain (NICD). The NICD enters the nucleus and complexes with CBF1 and Mastermind (MAM) to activate target genes. CBF1 recognizes the binding motif RTGRGAR (R=G or A), and this is referred to an NRE (Notch Response Element). Recent studies have shown that a paired CBF1 binding site is also important for target gene activation. While individual genes regulated by SPS are known, the extent to which an SPS vs individual NREs are responsible for Notch‐mediated transcription on a genomic level remains an active area of research. It is hypothesized that the majority of Notch‐mediated transcription occurs as a result of a dimeric Notch complex in the promoters or enhancers of transcribed genes. Recently, high‐throughput techniques have allowed the study of Notch complex assembly and their effects on transcription. Here, we use an Oxford Nanopore Platform was used.To identify genes regulated by Notch transcriptional complexes, RNA‐Seq was performed. Total RNA was isolated from SUPT1 cells treated with 5 µM of the Notch inhibitor DAPT for 24 hours, or DMSO as a control. Poly‐A RNA was purified using an oligo‐dT column and double‐stranded cDNA generated. PCR adapters were nonspecifically ligated to cDNA and amplified to increase library yield. Sequencing was performed using an Oxford Nanopore Minion, and results were mapped to the human reference genome to identify genes significantly downregulated when Notch was inhibited. To identify NREs and SPS in the regulatory elements of these genes, ChIP‐Seq was performed for Notch. ChIP was performed in DAPT and DMSO treated cells, and ChIP DNA used to prepare a sequencing library as above. Sequenced fragments were aligned to a known regulatory elements dataset, and genes with significant Notch binding identified.RNA‐Seq data shows significant downregulation of known Notch targets genes, such as Hes1, Hey1, and Hes4. Sequenced mRNA fragments, however, averaged 1kb suggesting size selectivity during the purification or amplification steps of library preparation. Additionally, underrepresentation of other Notch known targets such as Hes5 may suggest that computational stringency may have to be adjusted. We are currently pursuing ChIP‐Seq experiments that will be presented at the conference to identify the locations of SPS in the genome. Identified SPS in regulatory elements will then be correlated with the genes they activate, and the downstream effects on transcription analyzed. It is expected that a large number of genes that are inactivated under Notch inhibition conditions will exhibit a SPS in a corresponding regulatory element.