bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of the Integrity of the Respiratory Chain in Rooster Sperm after Freezing
Authors: Nikitkina Elena , Krutikova Anna , Musidray Artem , Shapiev Ismail 
Published: April 2020

One of the causes of cell death is a violation of electron transfer through the respiratory chain of mitochondria. The aim of study was to assess the integrity of the respiratory chain sperm after freezing in roosters. The effect of 2.4 dinitrophenol (2.4 DNP) on NADH fluorescence in fresh and cryopreserved semen of roosters was investigated. If the transfer of electrons and protons along the respiratory chain functions normally 2.4 DNP enhances the nonphosphorylating oxidation of NADH and its fluorescence decreases. The sperm of 30 cocks was frozen. The NADH was measured on a LUMAM fluorescence microscope. The luminescence excitation wavelength of NADH is 365 nm. Registration of NADH was carried out at a wavelength 465 nm. The amount of NADH was expressed in conventional units and the change in fluorescence after addition of 2.4 DNP was calculated as a percentage of the initial value taken as 100 %. The fluorescence of NADH after adding 2.4 DNP to frozen semen was 9% less than in fresh semen. A relationship between the magnitude of the change in NADH fluorescence after the addition of DNP and egg fertilization was established (r =0.65; P<0.05). Respiratory stimulation by 2.4 DNP is an useful and informative indicator in assessing sperm quality.Support or Funding InformationThe study was supported by the Russian Science Foundation (project No. 18‐16‐00071)
No Sex Differences in Cerebral Blood Velocity Responses During Resistance Exercise
Authors: Sherman Sara R. , Rosenberg Alexander J. , Schroeder Elizabeth C. , Lefferts Wesley K. , Lima Natalia S. , Grigoriadis Georgios , Baynard Tracy 
Published: April 2020

IntroductionThe brain requires intricate regulation of blood flow to maintain adequate perfusion to the tissue. Resistance exercise (RE) causes large increases in mean arterial pressure that challenges the regulation of cerebral blood flow. It is unknown, however, if the sex differences in blood pressure regulation may alter how cerebral blood flow responds to the large increases in blood pressure induced during RE.PurposeTo determine if the cerebral blood velocity response during an acute bout of RE differs by sex.MethodsTwenty‐nine young, healthy adults (M/F: 17/12; 25 ± 5 yrs; 23.9 ± 3.3 kg/m2) performed 3 sets of 10 repetitions maximal isokinetic knee flexion/extension exercise. Systolic (SBP), diastolic (DBP), and mean (MAP) arterial pressure were measured using finger photoplethysmography and used to calculate pulse pressure (PP). Transcranial Doppler ultrasound (2 MHz) was used to measure right middle cerebral artery mean blood velocity (MCAmv) and calculate pulsatility index ([MCAsystolic ‐ MCAdiastolic velocity]/MCAmv). Conductance was calculated as MCAmv/MAP. The last 30‐sec of a 10‐min baseline and last 10‐sec of the 3rd RE set were averaged for analyses.ResultsOverall, females had higher MCAmv and conductance compared to males (p<0.05). SBP, DBP, MAP, PP, MCAmv increased and conductance decreased similarly for males and females during RE (p<0.05). Pulsatility index did not change during RE in either sex (p>0.05).ConclusionsThe blood pressure and cerebral hemodynamic response during RE was not different between sexes. These data suggest sex may not effect cerebrovascular responses to a large increase in blood pressure incurred during maximal RE in young healthy adults.
Blood pressure and cerebral blood velocity responses at baseline and during an acute bout of resistance exercise in young adults.

Male (n= 17)

Female (n= 12)

p for Interaction

Baseline

Resistance Exercise

Baseline

Resistance Exercise

SBP (mmHg)
*

124 ± 10

172 ± 18

123 ± 7

170 ± 27

0.92

DBP (mmHg)
*

73 ± 7

99 ± 10

74 ± 7

103 ± 17

0.58

MAP (mmHg)
*

93 ± 8

128 ± 13

96 ± 7

132 ± 10

0.80

PP (mmHg)
*

52 ± 7

72 ± 11

50 ± 6

67 ± 14

0.42

MCA Mean Velocity (cm/sec)
*,
‡

55 ± 9

65 ± 12

70 ± 15

81 ± 30

0.89

MCA Pulsatility Index

0.90 ± 0.16

0.82 ± 0.14

0.81 ± 0.15

0.82 ± 0.27

0.27

Conductance (cm/sec/mmHg)
*,
‡

0.60 ± 0.09

0.51 ±0.10

0.74 ± 0.17

0.62 ± 0.24

0.39

All data are presented mean ± standard deviation
*Main effect for time (p<0.05)
‡Main effect for sex (p<0.05)
Effects of “Junk‐Food” on Excitatory Transmission and NMDA Receptor Subunit Expression in the NAc of Females
Authors: France Amanda M. , Alonso-Caraballo Yanaira , Ferrario Carrie R. 
Published: April 2020

Eating diets high in fats and sugars (i.e., “junk‐food”) produces alterations in the function of brain regions like the nucleus accumbens (NAc) that promote food‐seeking and over‐eating. For example, consumption of “junk‐food” increases the expression and function of high conductance calcium permeable AMPA receptors (CP‐AMPARS) in the NAc of obesity‐prone, but not obesity‐resistant male rats. These alterations persist for up to 30 days after initial “junk‐food” exposure. In addition, blockade of CP‐AMPARs within the NAc prevents enhanced food‐seeking in obesity‐prone rats. Thus, obesity‐prone rats are more sensitive to diet‐induced enhancements in NAc glutamatergic transmission that promote food‐seeking. However, whether similar experience‐induced alterations occur in females is unknown. Therefore, the goal of the studies presented here is to determine the effects of “junk‐food” on NAc glutamate receptor expression and function in females. Males were included for comparison and to replicate initial findings described above. Rats were given the “junk‐food” diet (10 days) followed by a return to ad lib standard chow for 14 days (i.e., “junk‐food” deprivation) in order to examine long‐lasting effects. Controls remained on standard chow throughout. Biotinylation or BS3 crosslinking followed by western blotting was then used to measure changes in the expression of AMPAR and NMDA receptor (NMDAR) subunits. In addition, whole‐cell patch clamp recordings from ex vivo brain slices were used to measure effects on AMPAR and NMDAR‐mediated transmission. Furthermore, the estrus cycle was monitored throughout and recordings were conducted in the metestrus/diestrus phase of the cycle. For biochemical studies, post hoc analyses are used to assess potential effects of the cycle on protein expression. In males, we found increases in GluA1, but not GluA2 surface expression and enhanced CP‐AMPAR‐mediated transmission following “junk‐food” consumption, replicating previous results. However, in females no effects on GluA1 or GluA2 surface expression or on AMPAR‐mediated transmission (CP‐AMPARs or non‐CP‐AMPARs) were found. However, there was a “junkfood” induced increase in the AMPA/NMDA ratio in females that was due to a reduction in NMDAR‐mediated transmission in the absence of any change in AMPAR‐mediated transmission. Thus, initial results show that there are sex differences in “junk‐food” induced alterations in CP‐AMPARs, and suggest a reduction in NMDA‐mediated transmission in females. Ongoing studies are examining effects of “junk‐food” on NMDAR subunit expression (GluN1, GluN2, GluN3). This will provide insights into which NMDAR populations may be affected. Together, these data shed light on potential sex specific mechanisms by which “junkfood” diet alters NAc function.Support or Funding InformationNIH R01 DK106188 (CRF)NIH F99 NS108549 (YAC)
HL‐60 Cells Alternative Response to an Ionomycin PMA Cocktail
Authors: Sternberg Lauren , O’Donnell Robert W. 
Published: April 2020

The combination of Ionomycin and phorbol 12‐myristate 13‐acetate (PMA) cocktail has been used to increase the proliferation of normal mouse and human T cells and PMA alone has been used to cause the differentiation of the myeloid leukaemia HL‐60 cells in culture to a macrophage‐like adherent population. Contrariwise, the cocktail was linked to apoptosis in acute HL‐60 cells. This distinction has also been noted in the literature by Wang, et al. (2014), who concluded that Jurkat T cells were inhibited at S‐phase and G2/M‐phase by the cocktail. When we cultured the HL‐60 cells for 48 hours in the presence of the cocktail, we also saw no growth and significant death. Originally, we suspected this distinction was due to the biological differences between normal cells and tumor cells; however, further investigation revealed a discrepancy within the literature. While HL‐60, Jurkat, and Glioma cells (Han 2013) are inhibited by the cocktail, breast cancer cell lines were MDA‐MB‐435 and MDA‐MB‐231 had enhanced invasion (Yiu 2006). Future experiments including a cell cycle analysis and apoptosis assay will be used to test other cancerous and non‐cancerous cell lines in hopes of elucidating a pattern for the effects of the ionomycin PMA cocktail.Support or Funding InformationCarl Savino and Olga Gerych‐Savino Endowment, The Geneseo Foundation, and David Wolf, D.O.
Pharmacology of Metabotropic Glutamate Receptor 2 Homo‐ and Heterodimers
Authors: McCullock Tyler William , Kammermeier Paul 
Published: April 2020

Metabotropic Glutamate Receptors (mGluRs) are a family of class C g‐protein coupled receptors (GPCRs) that respond to glutamate as their native agonist. The eight members of the mGluR family are widely expressed across the nervous and are primarily charged with shaping and fine‐tuning neuronal excitability and synaptic efficacy. Like most class C GPCRs, mGluRs are obligate dimer proteins. All mGluRs maintain the capacity to homodimerize, but recent work has demonstrated that heterodimeric mGluRs can form and exist natively. Further, studies of the mGluR2‐4 heterodimer have demonstrated that this heterodimeric receptor exhibits a distinct and unpredicted pharmacological profile compared to either the mGluR2 homodimer or the mGluR4 homodimer. These findings underscore the importance of better understanding mGluR heterodimer pharmacology, particularly with preclinical studies showing promising results for leveraging mGluRs as therapeutic targets for a wide variety of conditions. To develop a better understanding of the behavior of mGluR2, we used a multifaceted approach to investigate mGluR2s dimerization preferences, whole cell signaling, and individual receptor signaling when expressed by itself, or with other mGluRs. To test dimerization preference, we utilize a split luciferase assay. To test whole cell signaling, we utilize a bioluminescent resonance energy transfer (BRET) based G‐protein Beta‐Gamma sensor. Finally, to detect and determine the pharmacological profile of specific mGluR2 heterodimers, we combine the split luciferase technique with BRET, allowing us to isolate signals from a specific dimer within a population of receptors. These studies will help define the pharmacological profile of various mGluR2 containing heterodimers, as well as develop new tools to help further interrogate the pharmacology of other dimer complexes. These new tools are also high‐throughput compatible, allowing for the development of more advanced drug screens to identify new molecules that specifically target mGluR heterodimers.Support or Funding InformationUniversity of Rochester Harold C Hodge Memorial Fund and the University of Rochester Drug Targets and Mechanism Pilot Award
Cell Death Induced by Airborne Particulate Matter in Murine Microglial Cells
Authors: Street Mikayla , Feiner Kailey , Mazzer Paula 
Published: April 2020

Airborne particulate matter has been linked to neurodegeneration in recent epidemiological studies. Previous work in our lab has shown that different types of particulate matter elicit different reactions. We set out to investigate two different particulate matter types in microglial cells to see whether they would have the same effect in different types of neurological cells. We combined caspase‐3 assay with lipid profiling to determine the mode of cell death induced by several different types of airborne particulate matter. Some types of particulate matter induced cell death via apoptosis and others induced cell death by neuroinflammatory methods.Support or Funding InformationNIH P20GM103443
RIP‐3 and HMGB‐1 are upregulated in adult hypertensive male spontaneously hypertensive rats (SHR).
Authors: Abdelbary Mahmoud A. , Gillis Ellen E. , Musall Jacqueline , Sullivan Jennifer C. 
Published: April 2020

BackgroundNecrotic cell death is a hallmark of hypertension. We previously published that male SHR have greater renal necrosis compared to age matched female SHR. Consistent with this, inhibition of necrosis attenuated maturation induced increases in blood pressure (BP) in male, but not female SHR. However, the mechanisms underlying sex differences in renal necrotic cell death in SHR are unknown. The goal of the current study is to test the hypothesis that maturation in male SHR drives increases in necrosis and upregulation of key proteins mediating necrotic cell death, receptor‐interacting protein kinase 3 (RIP‐3) and high mobility group box 1 (HMGB‐1).MethodsMale and female SHR were euthanized at 6 (pre‐hypertensive, sexually immature; n=6) or 15 weeks of age (hypertensive, sexually mature; n=6). The right kidney was isolated and processed for flow cytometric analysis of cell death using 7AAD. The left kidney was isolated for Western blot analysis of total RIP‐3, phosphorylated RIP‐3 and HMGB‐1. Protein expression was normalized to β‐actin. All data are expressed as mean ± standard error.ResultsRenal necrosis was comparable between 6 week old male and female SHR (expressed as % total kidney cells: 2.4±0.2 vs. 1.8±0.1, respectively) and increased with age only in males (5.4±0.2 vs. 1.7±0.3, respectively; Psex<0.0001; Page<0.0001; Pinteraction<0.0001). To gain insight into what mediates sex differences in necrosis in 15 week old SHR, protein expression of RIP‐3 and HMGB‐1 were determined in 6 vs. 15 week old male SHR and in 15 week old male and female SHR. Protein expression of RIP‐3 (1±0.1 vs. 0.7±0.1; P=0.008; n=5–6), p‐RIP‐3 (1±0.1 vs. 0.7±0.03; P=0.008; n=5–6) and HMGB‐1 (1±0.06 vs. 0.7±0.1; P=0.005; n=5–6) were all greater in 15 week old vs 6 week old male SHR. 15 week old male SHR also had greater protein expression of RIP‐3 (1±0.1 vs. 0.4±0.1; P=0.003; n=6), p‐RIP‐3 (1±0.2 vs. 0.4±0.1; P=0.02; n=6) and HMGB‐1 (1±0.2 vs. 0.6±0.1; P=0.02; n=6) compared to age matched females.ConclusionIn contrast to adult, hypertensive SHR, sex differences in renal necrosis were absent in young, sexually immature pre‐hypertensive SHR. Consistent with greater renal necrosis, key proteins of necrotic cell death machinery were also upregulated in hypertensive male SHR compared to either age matched hypertensive females or young, pre‐hypertensive male SHR. Future studies are needed to test the hypothesis that RIP‐3 activation and the subsequent HMGB1 release potentially underlie greater renal necrosis in hypertensive male SHR compared to age matched females.
Spatiotemporal Patterning of ABC Multidrug‐Resistance Transporters During Germ Layer Specification
Authors: Schrankel Catherine S. , Vyas Himanshu , Espinoza Jose A. , Hamdoun Amro 
Published: April 2020

Xenobiotic transporters are important for the protection of differentiated somatic cells, yet less is known about the role of these proteins in rapidly changing embryonic cells. A surprisingly large number of ABC transporters (80%) are expressed from fertilized egg to gastrulation, but their spatial and temporal patterns of expression during early embryogenesis remain incompletely understood. Here I report a pattern of ABC transporter expression that emerges during the early differentiation of mesoderm, endoderm, and ectoderm of the sea urchin. These included orthologs of six major xenobiotic transporters, including ABCB1, ABCB4, ABCC1, ABCC4, ABCC5, and ABCG2. In‐situ hybridization (ISH) revealed that transporters followed one of three patterns of early developmental expression: The first, seen with protective and homeostatic genes such as ABCB1 and ABCC1, was the early and ubiquitous expression across all three germ layers, and later enrichment in ectoderm‐derived sensory and feeding structures of the larva. The second, seen with the transporters ABCC4 and ‐C5 that are known to transport signal molecules, is a spatially restricted expression domain within a ring of mesodermal cells of the early blastula; ABCC4 then becomes restricted to primordial germ cells (PGCs) and their embryonic niche during gastrulation. Third, ABCB4 and ABCG2 show expression only in endoderm fated cells, particularly in the hindgut (presumptive intestines) at the onset of gut development; both genes eventually expand to the midgut region (presumptive stomach). To further refine the ISH map, I used fluorescent substrates to assay the functional outputs of transporter mRNA localization. ABCC1 and ‐C4 specific substrates are strongly effluxed out of the ring of mesodermal cells. This overlaps with robust ABCC5 transport activity in blastulae, suggesting a potential ABCC‐mediated signaling center in early mesoderm, where the PGCs are born. In the endoderm, ABCB4 and ABCG2‐specific efflux activity begins several days before the larva feeds. Collectively, this work is the first systems‐level description of the emergence of xenobiotic transporter expression patterns across the germ layers during embryogenesis. The resulting map of transport reveals unanticipated developmental functions for these genes and provides a framework for understanding the genetic regulatory controls of transporter expression. This will have additional impact on predicting and mitigating the effects of xenobiotic exposures during vulnerable windows of early development.Support or Funding InformationSupported by NIH R01ES027921 and R01ES030318 to AH, and NIH F32ES029843 to CSS.
Press Pulse Therapy in MMTV‐PyMT Murine Breast Cancer
Authors: Poff Angela , Moss Sara , Soliven Maricel , Noboa Karina , D’Agostino Dominic 
Published: April 2020

BackgroundTumors can be viewed as populations of cells which may succumb to eradication principles observed in nature. For example, population reduction or eradication occurs when populations are exposed to simultaneous press and pulse pressures. Press disturbances are pressures which produce chronic stress in populations, while pulse disturbances produce acute stress. We recently proposed a novel Press‐Pulse therapeutic strategy for the metabolic management of cancer that is based on these principles (
https://nutritionandmetabolism.biomedcentral.com/articles/10.1186/s12986‐017‐0178‐2). We hypothesize that specific press‐pulse protocols will be effective in managing even aggressive cancers, while allowing for the use of low dose and low‐ or non‐toxic metabolic‐targeted therapies.MethodsIn this project, we are testing the press disturbances of the ketogenic diet and ketone supplementation, and the pulse disturbances of 2‐deoxyglucose (glycolytic inhibitor), sodium phenylbutyrate (glutamine targeting agent), hyperbaric oxygen therapy, and pharmacological ascorbic acid, in the MMTV‐PyMT mouse model of breast cancer.Preliminary ResultsTo date, we have tested the proposed metabolic treatments as monotherapies and combination therapies. Tumor growth rate and survival time were largely unaltered by the treatments tested. Ketogenic diet and sodium phenylbutyrate were the only treatments to affect survival time as a monotherapy. Ketogenic diet treated mice lived 8.7% longer than standard diet fed mice (p<0.05). Sodium phenylbutyrate treated mice had a decrease in survival of 8.1% from the standard diet fed mice (p<.05). Data analyses pending for the effects of combination therapies.ConclusionsWe are not yet able to determine if Press‐Pulse targeting is effective in this model system as we have only tested and analyzed monotherapies. Ketogenic diet therapy appears to elicit a minor therapeutic effect as a stand‐alone, but the other metabolic‐targeted treatments utilized were not effective as monotherapies. Studies to test combinations of these therapies have been completed, and the analyses of the data is ongoing.Support or Funding InformationDonner Foundation, Florida High Tech Corridor
SPARC: Time Series Transcriptomic Analysis of the Nucleus of the Solitary Tract During the Development of Hypertension
Authors: Moss Alison , Kuttippurathu Lakshmi , Achanta Sirisha , Schwaber James S. , Vadigepalli Rajanikanth 
Published: April 2020

Essential hypertension affects millions of people and is known have a neurogenic component where regions in the brain and brainstem involved in the autonomic nervous system govern the regulation of blood pressure set point. While some work has been done to profile gene expression levels in the nucleus of the solitary tract (NTS) in males, to date no work has specifically looked at how gene expression changes in the NTS drive hypertension development in females. We have conducted a time series mRNA profiling analysis in female spontaneously hypertensive rats (SHR) in comparison to wild type Wistar Kyoto (WKY) allowing us to compare and contrast the results with previously obtained data in males. Rats were sacrificed over the development of and during chronic hypertension at 8, 12, 16, and 24 weeks of age and brainstems were removed and stored in OCT. Brainstems were sectioned on a cryostat and the NTS was extracted using a micropunch. Total RNA was extracted and samples were submitted for RNAseq. We used a data analysis approach focusing on comparative pattern counts (COMPACT) to exhaustively identify the dominant and subtle differential expression patterns (Kuttippurathu et al., BMC Genomics, 2016). Over 2000 genes that show both strain and age dependent differential expression were examined further to interrogate differences in expression patterns between SHR and WKY. Broad analysis reveals that genes involved in the adaptive immune response are differentially regulated in the SHR compared to WKY. Using COMPACT to examine specific patterns, we found that expression levels of many genes involved in transport and secretion of catecholamines do not change over time in WKY but are specifically downregulated in SHR before the onset of hypertension at 8 weeks of age. Interestingly, pathways such as arachidonic acid metabolism that have been implicated in hypertension development in males did not show significant differences between SHR and WKY in females. These results provide valuable insight into the regulatory networks governing hypertension development in females and allows us to compare and contrast this to what is known in males.Support or Funding InformationSupported by the NIH Common Fund SPARC Program award OT2 OD023848 and NIH NHLBI Multiscale Modeling award U01 HL133360.