bluuebunny/crossref_metadata_2025 huggingface dataset:
cMET inhibition potentiates the tumor‐selective damaging effects of NQO1‐bioactivatable agents by compromising DNA repair
Authors: Bhandare Snehal B. , Yeh I‐Ju , Poirier Jon‐Luc , Motea Edward A.
Published: May 2022

The overall prognosis for solid tumors overexpressing mesenchymal‐epithelial transition factor (cMET) receptor tyrosine kinase and NADPH:Quinone oxidoreductase 1 (NQO1) is poor, and innovative treatment strategies that selectively target these cancers are critically needed. Recent studies have implicated cMET in the activation of PARP1, a critical factor involved in DNA damage response and repair. Therefore, targeting cMET is an attractive strategy for cancer therapy. However, the overall efficacies of cMET inhibitors and NQO1 bioactivable agents are limited due to dose‐limiting toxicities and lack of tumor selectivity at high concentrations as a monotherapy. Here, we report that the combination treatment with sublethal doses of cMET inhibitors (some in clinical trials and FDA‐approved) and β‐lapachone (β‐lap, an NQO1‐bioactivatable drug in clinical trials) induced synergistic lethality in cancer cells in an NQO1‐dependent manner. Mechanistically, a sublethal dose of β‐lap creates reactive oxygen species (ROS) that damage DNA nucleobases (e.g., 8‐oxoguanine) but rapidly gets repaired by the ability of ROS‐activated cMET to enhance PARP1 activity for efficient DNA damage repair in NQO1+ cells. Thus, β‐lap in combination with cMET inhibition elevated DNA damage by compromising DNA repair, increased double‐strand break (DSB) formation and promoted tumor selective apoptosis in NQO1+cancer cells. We further determined that the combination treatment significantly inhibit tumor growth in 3D spheroids. Our results add a new strategy for personalized therapy: the targeting of cMET in NQO1+ cancers to potentiate the toxic effects of sub‐lethal doses of NQO1‐bioativatable agents and cMET inhibitors.
Efficacy of B‐Vitamin Supplementation in Stroke and Hypertension
Authors: Ahmed Amer , Field Martha S.
Published: May 2022

Stroke is the fifth leading cause of death in the United States, and hypertension is the leading risk factor for stroke. However, pharmacologic therapies have been shown to reduce stroke‐related complications, and nutrient supplementation is increasingly used in the stroke prevention treatment plan. This study aimed to collect and summarize evidence on the efficacy of B‐vitamin supplementation in the prevention and treatment of stroke and hypertension. Based on a preliminary literature search, it was predicted that B‐vitamin supplementation helps prevent stroke in hypertensive patients. A systematic review was conducted, searching through PubMed databases for randomized control trials (RCTs) of B‐vitamin interventions (vitamin B complex, vitamin B6, vitamin B12, riboflavin, thiamine, niacin, folic acid, or folate) and stroke and hypertension outcomes (hypertension, stroke, high blood pressure, vascular stiffness, or arterial stiffness). After search and screening by two independent individuals, we identified 69 RCTs for inclusion in the review. There are 30 RCTs with a direct B‐vitamin intervention and stroke and hypertension outcome relationship, 26 RCTs with an indirect relationship, 2 RCTs that came from the Vitamin Intervention for Stroke Prevention (VISP) Trials, 3 RCTs from the VITAmins TO Prevent Stroke (VITATOPS), 8 RCTs from the China Stroke Primary Prevention Trial. Preliminary findings reveal that folic acid supplementation was associated with a significant benefit in reducing the risk of stroke in hypertensive patients and a decrease in homocysteine concentrations. Concluding findings contribute to current knowledge regarding the use of B vitamin supplements in chronic disease prevention.
M ethod for Measuring Cervical Vagus Nerve Activity in Conscious and Freely Moving Rats
Authors: Miki Kenju , Ikegame Shizuka , Yoshimoto Misa
Published: May 2022

The vagus nerve innervates most of the organs in the thoracic and abdominal cavity and plays a central role in inter‐organ networks through afferent and efferent vagal nerve activity. However, few attempts have been made to measure vagal nerve activity in the conscious state; thus, the nature of vagal nerve activity remains unclear. In this study, we attempted to develop a method to measure cervical vagal nerve activity in conscious, freely moving rats. Using male Wistar rats, the right cervical vagus nerve was dissected approximately 1 mm under anesthesia, and a bipolar electrode made of twisted stainless‐steel wire was hooked onto the vagus nerve and fixed with silicone gel. A microcatheter for administering a long‐acting local anesthetic (levobupivacaine hydrochloride 0.75%) was placed at least 5 mm peripherally from the electrode. Measurements were recorded under conscious and free‐living conditions. The right cervical vagal nerve activity (rcVNA) showed continuous activity of approximately 20 μV at peak‐to‐peak potential. The rcVNA was reduced by approximately 80% after administering 30 μL of levobupivacaine through the microcatheter. This allowed us to block the afferent VNA and selectively measure the efferent VNA. Using this method, we observed that efferent rcVNA decreased during REM sleep compared with NREM sleep. Moreover, we succeeded in measuring rcVNA and renal SNA simultaneously and continuously in conscious rats.
Developing RNA Therapeutics for TDP‐43 Proteinopathy in ALS/FTD
Authors: Copley Katie E. , Smirnov Ashleigh , Shorter James
Published: May 2022

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are both fatal neurodegenerative disorders characterized by neuronal loss. ALS is primarily characterized by motor impairments stemming from degeneration of motor neurons, whereas the main symptoms of FTD include changes in personality, behavior, and language stemming from degeneration of cortical neurons in the frontal and temporal lobes. ALS and FTD exist on a clinical continuum, where some patients present with features of both diseases. A molecular hallmark shared by almost all ALS patients and approximately half of FTD patients is the pathological aggregation of the RNA‐binding protein TDP‐43. It has recently been established that the solubility of TDP‐43 is increased by its binding to RNA. I hypothesize that aberrant aggregation of TDP‐43 in ALS/FTD models is due to alterations in TDP‐43:RNA interactions. I have utilized in vitroaggregation assays with purified TDP‐43 and short 34 nucleotide bait RNAs designed to bind TDP‐43. I find that bait RNAs can prevent aggregation of all tested disease‐linked TDP‐43 variants. I also find that disease‐causing missense mutations in TDP‐43 can alter the potency of bait RNAs against aggregation of TDP‐43. These results suggest that there are differences in the RNA interactions of TDP‐43 in disease. These findings also support the development of bait RNAs as a therapeutic strategy for ALS/FTD.
Consequent Endotoxemia Relieves Abnormalities in Renal Vasoconstriction and AT1/ACE/ACE2 Signaling in Preeclamptic Offspring: Roles of Sex and Antenatal Ang 1‐7 Therapy
Authors: Morgaan Hagar A. , Sallam Marwa Y. , El‐Gowelli Hanan M. , El‐Gowilly Sahar M. , El‐Mas Mahmoud M.
Published: May 2022

The renin angiotensin system plays a pathogenic role in the altered renal homeostasis in inflammatory states such as preeclampsia and endotoxemia. Here, we investigated whether preeclamptic fetal programming modifies hemodynamic and renal vasoconstrictor responsiveness to endotoxemia in adult offspring and whether these interactions are modulated by gestationally administered Ang 1‐7. Preeclampsia was induced by oral administration of L‐NAME (50 mg/kg/day) to pregnant dams. Adult offspring was challenged with lipopolysaccharides (LPS, 5 mg/kg) and changes in blood pressure (BP) and renal vasoconstrictor reactivity were assessed 4 hours later. Contrary to female counterparts, male offspring of PE rats exhibited significant rises in BP that were dramatically reduced after LPS administration. Vasoconstrictions induced by Ang II (0.25‐32 ng), but not phenylephrine (0.41‐900 ng), in isolated perfused kidneys of male and female offspring of PE dams were intensified compared with non‐PE offspring. The heightened Ang II responses were blunted by LPS in male, but not female, preparations. Further, gestational Ang 1‐7 supplementation had no effect on endotoxic hypotension in male offspring, but caused sex‐unrelated attenuation of Ang II and phenylephrine renal vasoconstrictions. Immunohistochemical studies showed that while the renal tubular expression of AT1 receptors and ACE was increased in PE offspring of both sexes, increased ACE2 expression was noted only in female offspring. The upregulation in these molecules was diminished after LPS exposure, an effect that was more apparent in the male gender. Additionally, gestational treatment with Ang1‐7 failed to modify expression patterns of AT1/ACE/ACE2 signaling in PE/LPS offspring. Together, sexual dimorphism exists in the ability of ensuing endotoxic challenge of adult offspring to offset imbalances in renal vasoconstriction and tubular AT1/ACE/ACE2 signaling caused by early PE insult. Further, antenatal Ang 1‐7 therapy rectifies the erupted vasoconstriction anomalies through yet unidentified mechanisms.
Impulsive behavior induced by Dopamine D3 receptor agonist treatment in Parkinson’s disease is mediated by β‐arrestin
Authors: Das Sanjay , Xu Wei , Kortagere Sandhya
Published: May 2022

Parkinson’s Disease (PD) is a movement disorder characterized by progressive degeneration and loss of mid‐brain dopaminergic neurons. Pramipexole (PRX), a Dopamine D3 receptor (D3R) agonist, has emerged as a potent therapeutic to treat motor deficits, but chronic therapy leads to impulse control disorders (ICDs) in PD patients. ICDs comprise a group of complex behavioral disorders characterized by failure to resist an impulse or temptation to perform an act that is harmful to the individual or to others. While the etiology of ICDs is not well understood, several clinical and animal studies have demonstrated that D3R agonist activity directly correlates with the development ICDs. D3Rs are highly expressed in the mesolimbic pathways and mediate motor and reward processes in the brain. D3R agonists can signal through multiple signaling pathways and predominantly through the activation of G‐proteins, β‐arrestin, or both. We hypothesize that activation of β‐arrestin signaling pathway by D3R agonists such as PRX can induce ICDs in PD. To test this hypothesis, PD rats were trained to criteria in a 5‐choice serial reaction time task (5CSRTT) and their baseline levels of impulsive responses (measured as premature responses), correct responses and omissions were recorded. Animals were then treated with vehicle or PRX or our G‐protein biased D3R agonist SK609 for 3 weeks (1x/day; i.p) and their responses in the 5CSRTT were measured at the end of week‐3. Results from the study demonstrated that PRX significantly increased omission, premature responses (PMR) compared to vehicle or SK609 treatment. SK609 reduced PMR and increased the correct responses and reduced omissions when compared to both PRX and vehicle treatment. The increase in PMR by PRX treatment correlated with an increase in anxiety as measured by time spent in the closed arms of the elevated plus maze. Current studies are aimed at understanding the molecular mechanisms of β‐arrestin mediated signaling that promote ICDs. PRX treatment was further found to enhance β‐arrestin expression whereas SK609 didn’t in PD rats. Our study will augment the understanding of biased D3R signaling and its role in modulating ICDs in PD. This study’s outcome will be directly translatable to optimizing therapy for PD without the risk of developing ICDs seen with existing therapies.
Anti‐hyperglycemic Activities of Korean Red Ginseng (Panax ginseng Meyer) and Arginyl‐fructose‐enriched Red Ginseng Extracts In‐vitro and In‐vivo Animal Models
Authors: Hong Ji‐Yoon , Lee Su‐young , Jang Ye‐Lim , Kim Gi‐Jung , Kim Tae‐Yang , Cho Cheon‐Weon , Sun Byung‐ryul , Kwon Young‐In
Published: May 2022

Maillard reaction that occurs during the manufacturing processes of red ginseng produces a large amount of Amadori rearrangement compounds (ARCs), such as arginyl‐fructose (AF). The antioxidant, anti‐hyperglycemic immune enhancing effects of AF have been reported. Amino acid derivatives such as AF are formed through amadori rearrangement, the early step of maillard reaction from arginine and glucose in heat processed foods. Heat processed red ginseng have high content of AF and saponins. In this study inhibitory activities of red ginseng extract (RG) and AF‐enriched red ginseng extract (ARG) against rat intestinal α‐glucosidases (sucrase, maltase, and glucoamylase) and α‐amylase were evaluated using in‐vitro and Sprague Dawley (SD) rat models. The in‐vitro results showed that ARG have higher α‐glucosidase inhibitory activities than those of RG in a dose‐dependent manner. In SD rat model, postprandial blood glucose levels were measured for 3 hours after oral administration of 0.1 and 0.5 g/kg‐body weight of ARG or RG along with sucrose or starch. The postprandial blood glucose lowering effects of ARG and RG were compared to a known type 2 diabetes medicine (Acarbose®), a strong α‐glucosidase inhibitor in the SD rat model. In rats fed on sucrose, ARG significantly reduced the blood glucose spike after sucrose loading (p < 0.01). The maximum blood glucose concentration (Cmax) in ARG‐treated SD rats (0.1 g‐ARG/kg) was significantly lower than in RG‐treated SD rats (p < 0.01). These results suggest that AF‐enriched red ginseng could have a beneficial effect for blood glucose management relevant to diabetes prevention in normal and pre‐diabetic individuals via inhibition of the carbohydrate hydrolysis enzymes in the colon. AF, ARCs may have high potential as a material for the development of products, nutritious foods with antidiabetic activity. Our findings provide a pharmacological rationale and the basis for further clinical study for AF‐enriched natural products such as ginseng and red ginseng.
Heart Rate Fragmentation in Patients with Type 2 Diabetes Mellitus
Authors: Aguiar Mesquita Galdino Gabriela , Virgilio Silva Luiz Eduardo , Garcia Moura‐Tonello Silvia Cristina , Milan‐Mattos Juliana Cristina , Nogueira Linares Stephanie , Marques da Silva Thaís , Porta Alberto , Fazan Rubens , Beltrame Thomas , Catai Aparecida Maria
Published: May 2022

Heart rate fragmentation (HRF) is a recently proposed approach to evaluate ultra‐rapid variability of heart rate. HRF has been shown to be increased in the aging process and in conditions such as coronary artery disease. Moreover, it proved to be a strong biomarker of adverse cardiovascular events, mortality and was an independent predictor of atrial fibrillation. Type 2 diabetes mellitus (DM) is a disease known to affect heart rate variability (HRV) indices markedly; however, HRF in diabetic patients is to be studied. We aimed to evaluate HRF in patients with DM, without autonomic cardiovascular neuropathy, compared to their healthy counterparts. Electrocardiographic (ECG) recordings collected from patients with diabetes (n=102) and healthy subjects (n=96), aged 44 to 59 years, were retrospectively analyzed from the Cardiovascular Physical Therapy Laboratory database. ECG were recorded at rest (supine) for 10 to 15 minutes. RR interval (RRi) series were processed by the customized software PyBios. RRi values were symbolized as “‐1”, “0,” or “1” when the differences between successive RRi were negative, null, or positive, respectively. The percentage of inflection points (PI) was quantified and sequences of 4 consecutive symbols, so‐called “words”, were analyzed computing the occurrence of words with zero (W0), one (W1), two (W2), or three (W3) inflections points. The occurrence of words with only hard (‐1 to 1: WH), soft (‐1 to 0 or 0 to 1: WS), or mixed (WM) (i.e., both H and S) transitions were also quantified. Patients with diabetes presented higher PIP (58±1 vs 56±1, p=0.033), W3(17±1 vs14±1, p<0.001), WS(25±2 vs16±1, p<0.001) and WM(22±1 vs 17±1, p<0.001). On the other hand, patients with diabetes showed a lower percentage of W1 (31±1 vs 35±1, p=0.024) and WH (47±2 vs 61±2, p<0.001). The occurrence of W0 and W2 were similar between groups. In conclusion, diabetics patients without autonomic cardiovascular neuropathy have augmented fragmentation of heart rate as compared to healthy subjects. Clinical applicability: In the future, studies with this approach might be useful to assess heart rate dynamic abnormalities in order to detect cardiac risks in patients with diabetes mellitus.
Immunologically active cells and infertility ‐ Quantification of uterine Natural Killer cells in endometrial samples from patients with recurrent implantation failure
Authors: Varga Ivan , Lapides Lenka , Klein Martin , Babal Pavel
Published: May 2022

Background

Despite enormous advancements of reproductive medicine, recurrent implantation failure and habitual abortion remain an ongoing issue. One of the most important aspects of successful implantation is the intricate immune regulation necessary for the acceptance of the hemiallogenic embryo. The most numerous immune cells in the decidua are uterine Natural Killer (uNK) cells. Studies suggest that changes in the uNK cells count and physiology may be responsible for the aforementioned conditions. Thus, the uNK cells testing may provide valuable insights into their pathogenesis.

Methods

We compared Pipelle endometrial sampling with conventional curettage to find out whether the less invasive Pipelle method is a viable alternative of tissue collection.

Results

We examined tissue samples from 9 patients obtained by both methods. The average size of tissue samples obtained with pipelle was 10,27 mm2, samples obtained with curettage had on average 18,74 mm2. Using immunohistochemical visualization of CD56 (NK cells) and granzyme B antigens (serine protease‐expressing activation state of NK cells), we found out that the average total count of CD56 / mm2 was for pipelle 90,13 and 84,04 for curettage. We also proved a correlation between granzyme B positivity and identification of NK cells clusters. The results showed that Pipelle endometrial sampling is a suitable method of tissue harvesting for the purpose of uNK cells examination. Discussion: Pipelle endometrial sampling is safe, cost‐effective and can be performed on an outpatient basis without the need of anaesthesia or analgesia. Several issues remain to be solved: how to standardize the subsequent uNK cells testing, how to interpret the results and finally yet importantly, how to use this knowledge in personalized treatment protocols.
Vascular Function and Sugar Sweetened Beverage Consumption in Hispanic and Non‐Hispanic Males
Authors: Papastratakos Eleni , Piero Rachel , Cuy Castellanos Diana , Crecelius Anne R.
Published: May 2022

Sugar sweetened beverages (SSBs) account for a significant portion of daily caloric intake for many individuals, especially Hispanic males. Hispanic males also have one of the highest rates of obesity in the United States. Both factors, individually or combined, put Hispanic males at an increased risk of cardiovascular disease. The purpose of this study was to assess the differences between vascular function in the Hispanic male population and the non‐Hispanic male population, as well as the cause for any differences. The diets of all the participants (11 Hispanic males and 11 non‐Hispanic males from the Midwest) were evaluated specifically to identify SSB intake. Vascular function was determined by post‐occlusion reactive hyperemia (RH) using venous occlusion plethysmography (VOP). Additionally, dietary, physical activity, and medical questionnaires were administered for each participant to identify other factors that could influence vascular function. No significant differences were found in peak forearm blood flow (FBF) between the age‐matched pairs of Hispanic and non‐Hispanic males (36.9 ± 8.5 vs 36.4 ± 12.2 ml/dl/100 ml, respectively; p=0.924). Peak forearm vascular conductance, which takes into account any potential differences in perfusion pressure, were also not different between Hispanic and non‐Hispanics (34.7 ± 7.6 vs 34.9 ± 11.4 units; p=0.957). The dietary and physical activity habits of these two groups did not differ significantly, and overall anthropometrics and medical histories were also similar between groups. Thus, we conclude that in this population, ethnicity alone has limited impact on vascular function, dietary, and physical activity behaviors. Rather, additional environmental and lifestyle factors play a more significant role in the determination of vascular function and ultimate cardiovascular disease risk than one’s ethnicity.
Orchidectomy exacerbates respiratory instability induced by intermittent hypoxia in C57BL6/J mice
Authors: Ganouna‐Cohen Gauthier , Marcouiller François , Joseph Vincent
Published: May 2022

Introduction

Sleep apnea (SA) is characterized by obstructions of the airways or cessation of breathing during sleep leading to intermittent hypoxia (IH). In rodents, exposure to IH induces strong oxidative stress in the peripheral chemoreceptors (the main oxygen sensors inducing cardiorespiratory responses to IH), which increase their activity leading to instability of the respiratory control system, ultimately increasing the frequency of apnea during sleep. In human, patients with SA have low circulating testosterone levels and the severity of SA in overweight patients is negatively correlated with testosterone levels. However, testosterone can reduce oxidative stress in certain animal and clinical models. We therefore tested the hypothesis that testosterone modulates the respiratory instability induced by HI.

Method

We used intact (Sham) or orchidectomized (ORX) C57BL/6J male mice exposed for 14 days to IH (12h / day, 10 cycles / h, 6% nadir oxygen) or in normoxia (Nx). We used whole body plethysmography on freely behaving and non‐anesthetized mice to measure the frequency of apneas and sighs as well as the length of the apneas during sleep. The antioxidant activity of superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were then measured in plasma.

Results

IH increases the frequency of post‐sigh apneas as well as the length of post‐sigh and spontaneous apneas in Sham and ORX mice. In ORX‐IH mice, the length of post‐sigh apneas was further increased, and there was a higher proportion of multiple apneas after a single sigh. In Nx mice, GPx activity was increased by ORX, this effect was not present in mice exposed to IH.

Conclusion

The lack of testosterone in ORX mice amplifies the effects of IH on respiratory stability during sleep and alters the regulation of GPx activity in plasma. Further studies are needed to better understand the underlying mechanism
Investigating the Link Between the Gut Microbiome and Epilepsy in Zebrafish
Authors: Woodis Kristina M. , Uno Jennifer K.
Published: May 2022

Epilepsy is the fourth most common neurological disorder affecting over 50 million people worldwide. Recent research indicates that modulation of the gut microbiome can alter the course of the disease in animal models as well as in humans. The gut microbiome is an ecosystem of fungi, viruses, and bacteria that is influenced by an individual’s genetics, diet, and lifestyle. The impact that the gut‐brain axis may have on epilepsy is not well understood. Dietary modifications are often used as treatment options for epileptic patients that are unresponsive to other anticonvulsant therapies. In fact, current research indicates that ketogenic diets are associated with a decrease in microbial diversity and diminished seizure activity. It is not known whether this dietary modification works through the gut‐brain axis. Zebrafish are a commonly used animal model to study human disease, including epilepsy and gut microbiome mechanisms. The aim of this study is to validate the use of zebrafish to study the interactions between the gut‐brain axis and epilepsy, as well as further explore the mechanism by which the microbiome alters the course of the disease. We hypothesize that antibiotic‐treated fish will experience diminished seizure responses to Pentylenetetrazol (PTZ) due to a decrease in intestinal bacterial abundance. Zebrafish were treated with broad‐spectrum antibiotics to alter the diversity of the microbiome. Fish were then given PTZ to induce a seizure‐like state. Seizure activity was recorded and evaluated using EthoVision tracking system and software. Intestinal and brain tissue were harvested for bacterial DNA and RNA respectively. Bacterial abundance was significantly decreased in antibiotic treated fish. Seizure activity was significantly increased in PTZ‐treated fish and was surprisingly augmented in antibiotic‐treated fish (n= 5, p < 0.05). In addition, we observed an increase in glutamate along with a decrease in GABA receptor expression in PTZ‐treated fish; these results were reversed in fish treated with both antibiotics and PTZ. Collectively these results indicate that zebrafish are an effective model for studying gut‐brain communication involving epilepsy and modulating the gut microbiome may influence seizure activity in PTZ‐treated fish.
Fear Processing in Mice with PMAT Deficiency
Authors: Beaver Jasmin N. , Kassis Sarah K. , Weber Brady L. , Ford Matthew T. , Anello Anna E. , Gilman T. L.
Published: May 2022

The plasma membrane monoamine transporter (PMAT, Slc29a4) is a polyspecific cation transporter that, in the brain, predominantly takes up monoamine neurotransmitters like dopamine and serotonin. PMAT function is hypothesized to emerge when other monoamine transporter function is impaired, such as under conditions of heightened monoamine neurotransmitter release in response to stressors. In humans, common genetic polymorphisms can reduce PMAT function, but these have exclusively been studied in the context of metformin treatment response under diabetic conditions. Nothing is currently known about how reduced PMAT function affects fear processing, nor any other aspects of emotional learning or memory. Our lab sought to begin examining the effects of PMAT deficiency on fear processing by using mice with constitutive reductions in PMAT function (i.e., heterozygotes) and comparing these to wildtype mice. Specifically, we evaluated both cued and contextual fear processing. Based on evidence that chronic pharmacological blockade of serotonin transporter‐mediated monoamine uptake decreased both cued and contextual fear expression, we hypothesized heterozygote mice would display lower levels of both cued and contextual fear relative to wildtypes. Surprisingly, we only observed a trend (p=0.08) specifically in male heterozygotes for attenuated cued fear expression. Otherwise, heterozygotes of both sexes exhibited cued fear acquisition, extinction retention, and renewal that was indistinguishable from wildtypes, as was their contextual fear processing. Four weeks after fear processing behavior tests concluded, mice were subjected to a brief swim stress, and blood was collected for stress hormone quantification. These data are currently being analyzed, and we hypothesize that heterotypic stressor exposure will unmask the behavioral sequelae of reduced PMAT function. Our current findings are consistent with previous reports of sex‐selective effects of PMAT deficiency on behavioral responses to stressor exposure. Continued investigation into how reduced PMAT function affects emotion processing and stress responsivity could have translational implications for people with functional PMAT polymorphisms.
Heat stress‐induced changes in the myocardium
Authors: Roths Melissa , Rudolph Tori , Freestone Alyssa , Baumgard Lance , Selsby Joshua
Published: May 2022

Prolonged exposure to heat can lead to environment‐induced heat stress (EIHS), which may be a threat to human health. How EIHS affects cardiac morphology and the myocardium are unknown. We hypothesized that EIHS would alter cardiac morphology and cause cellular dysfunction. To test this hypothesis, crossbred female pigs were exposed to thermoneutral (TN; 20.6 ± 0.2 ºC; n=8) or EIHS (37.4 ± 0.2 ºC; n=8) conditions for 24 h. Rectal temperature (RT), skin temperature (ST), and respiratory rate (RR) were recorded every 4 h during the environmental challenge. Pigs were euthanized following the environmental challenge and hearts were collected. Hearts were weighed and heart length (apex to base), width (left/right dimension), and left ventricle (LV) and right ventricle (RV) wall thickness were measured with calipers. Portions of LV and RV were lyophilized to measure tissue water content or homogenized for protein extraction and western blotting. Environment‐induced heat stress increased RT by 1.3 ºC (p<0.01), ST by 11 °C (p<0.01) and RR by 72 breaths per minute (p<0.01) compared to TN. Heart weight tended to be decreased (7.6%; p=0.07) and heart length was decreased (8.5%; p=0.01) by EIHS, but heart width was similar between groups. Left ventricle wall thickness was increased (22%; p=0.02) and RV thickness was decreased (26%; p=0.04) in EIHS compared to TN. Water content in the RV was similar between groups, however, in LV it was increased (8.6%; p<0.01) in EIHS compared to TN, suggesting edema contributed to increased LV thickness. Lastly, using a western blot approach we discovered that pathways regulating energy homeostasis were impacted by EIHS and sometimes differed between RV and LV. In RV and LV, phosphorylation (p) of AMP‐activated protein kinase (AMPK) was decreased by EIHS (RV – 73%, p<0.02; LV – 54%, p=0.04), and in the RV was accompanied by increased protein phosphate type 2A (PP2A; 15.5%, p<0.01), which regulates pAMPK, and decreased p‐Acetyl‐CoA carboxylase (ACC; 40%, p<0.01), a client protein of pAMPK, whereas these were similar in LV. In RV, mTOR signaling appeared to be blunted, mitochondrial content increased, and markers of mitophagy increased by EIHS compared to TN, however, LV was resistant to these changes. In total, these data demonstrate that a single bout of EIHS caused cardiac morphological changes and biochemical changes in the myocardium and that EIHS affects the LV and RV differently.
The NuA4 Acetyltransferase, Acetylation of Histone H4 and the H2A.Z Variant Histone are Required for Appropriate RNA Splicing in Saccharomyces cerevisiae
Authors: Bolle Camila , Snyder Christopher , Lin Mengjia M. , Kress Tracy L.
Published: May 2022

Gene expression is a tightly regulated process that involves properly synthesizing an RNA transcript from DNA, as well as RNA processing to correctly prepare it for proper translation into a protein. One key step in RNA processing is RNA splicing, the process by which non protein coding regions, known as introns, are removed from the RNA. It has been previously shown that splicing can occur co‐transcriptionally, and therefore there is the potential for coupling between transcription and RNA splicing. Mechanisms of coupling are currently being explored and one group of candidate proteins for coupling are histone modification enzymes and the histones that they modify. Chemical modifications to these histone proteins can alter histone association with the DNA, thereby regulating access of the transcription machinery to the DNA and impacting gene expression according to the needs of the cell. Therefore, these histone modification enzymes/modified histones are present at the right time and place to also regulate co‐transcriptional splicing. Here we tested the hypothesis that the NuA4 histone acetyltransferase and targeted acetylation of histone H4, which is known to alter chromatin structure and promote gene expression, is also important for regulating RNA splicing in Saccharomyces cerevisiae. NuA4, along with the chromatin remodeling enzyme Swr1, are also important for the insertion of and acetylation of the histone variant H2A.Z (a.k.a Htz1p), which is required for transcriptional activation. We investigated whether the NuA4 complex function and HTZ1 impacted splicing. Using genetic interaction studies, we revealed interactions between mutations that impact H4 acetylation/H2A.Z insertion and several splicing factor genes, implicating H4 acetylation and H2A.Z in RNA splicing. Furthermore, we show that point mutation of H4 or mutations that impair the function of the NuA4 complex both impact RNA splicing. We are now testing whether NuA4 or H4 acetylation are required for recruitment of splicing proteins to an RNA during transcription. Interestingly, our genetic interaction studies revealed a novel interaction between HTZ1 and genes involved in 3’ cleavage and polyadenylation machinery, which we are continuing to explore. Taken together our data support a role for the NuA4 enzyme, histone H4 acetylation and H2A.Z in RNA splicing, with potential connections to additional steps in RNA processing.
Chronic binge alcohol‐mediated alterations in myotube EV profile affect recipient myotube miRNA expression
Authors: Bourgeois Brianna , Levitt Danielle E. , Molina Patricia E. , Simon Liz
Published: May 2022

The average age of people living with HIV (PLWH) is now over 50 years, and this increased life expectancy is associated with increased prevalence of comorbid conditions including myopathy. At‐risk alcohol use is twice as prevalent in PLWH as the general population, and loss of muscle mass is a concern in PLWH and those with at‐risk alcohol use. Our group has previously shown that chronic binge alcohol (CBA) in SIV‐infected macaques decreases myoblast differentiation and is partly mediated by altered microRNA (miR) profile. miRs transported between cells in extracellular vesicles (EVs) mediate numerous physiologic and pathologic cellular responses and can alter the function of target cells through miR delivery. We tested the hypothesis that EVs derived from myotubes isolated from CBA‐administered female macaques would alter miR expression in recipient myotubes. Seven SIV‐infected female rhesus macaques received either CBA (2.5 g/kg/day, N=4) or water (VEH, N=3) for 14.5 months. Three months following the initiation of CBA or VEH, the animals were vaginally inoculated with SIVmac251. Antiretroviral therapy was initiated 2.5 months later. Animals were sacrificed 9 months later, and primary cells were derived from skeletal muscle tissue 24 hours following the last dose of alcohol (blood alcohol=0 mM). EVs were isolated by ultracentrifugation from the conditioned cell culture supernatant of three 10 cm plates of differentiated primary myoblasts (i.e. myotubes). EVs were transferred to naïve (prior to SIV or CBA) recipient macaque myotubes in serum‐free media and incubated for 48 hours. Following incubation, cells were washed, and RNA extracted to determine relative expression of miRs 133a, 133b, 206, 16, and 23a. EVs derived from CBA myotubes increased expression of miR‐206 in target myotubes (p<0.05, Cohen d=2.68). There was no difference in the expression of other measured miRs; however, there were large effect sizes for CBA‐derived EVs to increase miR‐133a and miR‐23a expression (Cohen d=1.13 and 1.51, respectively) in recipient myotubes. These results indicate that CBA‐mediated alterations in myotube EV profile affect recipient myotube miRNA expression. Ongoing studies will determine if this altered recipient miR expression reflects altered EV miR expression and/or host cell miR expression and determine the functional relevance of increased miR‐206 on myotube differentiation and hypertrophy.
Amelioration of acute lung injury by peptides targeting scavenger receptor CD36
Authors: Ke Yunbo , Li Yue , Bocharov Alexander , Birukova Anna , Birukov Konstantin
Published: May 2022

CD36 belongs to scavenger receptor class B and is a fatty acid translocase. Our previous studies have suggested that the receptor may be involved in regulation of responses to endotoxin and inflammation of the lungs in mice. In current studies, we have designed a group of helical peptides that interact with and modulate the activities of CD36 and tested their activities against endothelial barrier dysfunction and acute lung injuries in mice. Peptides that inhibit CD36 activities including L37pA, ELR‐B and ELR‐BP reversed or mitigated the barrier disruptive activities induced by LPS, HKSA, truncated phospholipids and histone H3 as measured by TER (trans‐endothelial electric resistance). In consistence with this, the peptides also enhanced the barrier protective activities of OxPAPC. The peptides were delivered intravenously to mice treated with LPS or HKSA to induce acute lung injuries. It has been observed that among peptides that antagonize the activities of CD36, L37pA, ELR‐B and ELR‐BP have demonstrated inhibitory effects on acute lung injuries induced by LPS characterized by reduced extravagance of inflammatory cells from circulation to bronchoalveolar lavage (BAL) and a decrease in protein concentration in BAL fluid from the peptide‐treated mice. The observation indicates that modulation of CD36 activities by peptide inhibitors is a novel strategy to reduce acute lung injuries induced by pro‐inflammatory agents.
Cinnamon effects on PPARγ and Adiponectin in 3T3‐L1 adipocytes
Authors: Stockert Amy , Mild Sean , Swartz Cole , Gadkar Oshra , Eskender Neb , Aulthouse Amy
Published: May 2022

Peroxisome‐proliferator activated receptor (PPARγ) plays a role in tumor growth inhibition and metabolic activity. Cinnamon extract components have been proposed as a possible PPARγ ligand and we sought to investigate if cinnamon treatment of cells affected the PPARγ and adiponectin expression. 3T3‐L1 cells were cinnamon treated for 72 and 120 hours. Media was collected for ELISA quantification of Adiponectin levels. Copy DNA was produced from the total RNA and expression levels of PPARγ and Adiponectin were examined by TaqMan real‐time PCR assays. Expression of PPARγ increases significantly in both post‐differentiated cells at 72 hours and in non‐differentiated cells at 120 hours. Expression of AdipoQ is increased in post‐differentiated cells significantly when compared to non‐differentiated cells. AdipoQ expression increases at 72 hours post differentiation. Post differentiation cells demonstrated high levels of adiponectin in the media. No significant difference was observed between the cinnamon treated cells and the vehicle control cells. Higher levels of PPARγ in the post‐differentiated cells correlate with the increase in adiponectin expression and adiponectin secreted in the post‐differentiation cells. A lag between the adipoQ expression increases at 72 hours post‐differentiation and adiponectin in the media was detected at high levels at 120 hours. Although there is no significant difference in the adiponectin in the media between vehicle samples and cinnamon treated samples, the expression data compares the control to the cinnamon treated cells, therefore the expression changes observed result from cinnamon treatment. This data suggests that perhaps a mechanism exists that stops adiponectin at the translation level.
A reduced complexity cross between BALB/c substrains identifies Zhx2 as a candidate gene underlying oxycodone metabolite brain concentration and state‐dependent learning of opioid reward
Authors: Beierle Jacob A. , Yao Emily J. , Goldstein Stan I. , Scotellaro Julia L. , Sena Katherine D. , Averin Olga , Moody David E. , Reilly Christopher A. , Emili Andrew , Peltz Gary , Ferris Martin T. , Bryant Camron D.
Published: May 2022

Understanding the pharmacokinetic profile of an opioid drug is vital to therapeutic success, and mutations in human PK genes can drastically alter therapeutic efficacy of opioids. We observed that at 30 min post‐oxycodone administration (1.25 mg/kg, i.p.) BALB/cJ mice showed a higher whole brain concentration of oxycodone, and female specific increase in noroxycodone, and oxymorphone compared to BALB/cByJ. This observation could explain the sex‐specific increase in oxycodone state‐dependent conditioned place preference in BALB/cJ female mice. To potentially link behavioral differences with PK differences, we conducted quantitative trait locus (QTL) mapping of whole brain oxycodone and metabolite concentrations in a reduced complexity cross (RCC). Because BALB/cJ and BALB/cByJ substrains differ by ~8,500 SNPs/indels, large genetic loci identified in an F2 cross are offset by a dramatic reduction in potentially causal variants. QTL mapping in 133 BALB/cJ x BALB/cByJ F2 mice (68F, 65M) revealed a single QTL on chromosome 15 associated with brain oxymorphone concentration that explained 29% of the phenotypic variance in females. Oxymorphone is a full agonist at the mu opioid receptor, with 8x the potency of oxycodone, and likely contributes to oxycodone addictive properties. Hippocampal and striatal cis‐eQTL analysis revealed genetically regulated expression of Zhx2, a transcriptional inhibitor known to harbor a private BALB/cJ retroviral insertion that dramatically reduces protein expression and leads to sex specific dysregulation of CYP450 genes within the liver. Whole brain mass spectroscopy proteomics in the parental strains corroborated these eQTL findings. We hypothesize that decreased Zhx2 expression leads to increased CYP450 expression, increased brain oxymorphone, and increased oxycodone‐induced behaviors. Interestingly, human GWAS of nicotine consumption identified a nominal association (10^‐7) with ZHX2, indicating that this transcriptional repressor could influence metabolism of multiple drugs of abuse.
Fundamentals of Biochemistry, a free and new LibreText book for Undergraduate Courses
Authors: Jakubowski Henry , Flatt Patricia , Agnew Henry , Larsen Delmar
Published: May 2022

We are developing a new fee online biochemistry text, Fundamentals of Biochemistry, within the LibreTexts project (Libretexts.org) as part of a Department of Education funded grant to produce textbooks for all chemistry courses required for a four‐year ACS certified degree. This text covers a full year sequence of biochemistry and will be available in Fall 2022. The book could serve traditional, ASBMB‐certified and ACS‐certified majors, as well as students taking a one‐semester course. Adopting instructors could create a custom text from the content. Characteristic of all LibreText books, Fundamentals of Biochemistry will be open to everyone via the internet and completely free, as all should have access to knowledge. It is comprehensive and topical, as new materials are easily added. It has and will be constructed with a community of active collaborators. The LibreText collections of books have served 223 million students and saved them $31 million, an undue burden for our students and society.

Some of the features of Fundamentals of Biochemistry include

Interactive 2D mathematical graphs with sliders that allow change of constants with immediate update of graphs;

Interactive molecular models using iCn3D (https://www.ncbi.nlm.nih.gov/Structure/icn3d/full.html) that allows students to view and interact with rendered structures;

Imbedded VCell (https://vcell.org/) models that allow users to display time course graphs for simple to complex reactions;

Animations to accompany the VCell progress curve graphs;

Chapter sections on metabolic control analyses that will include Vcell models;

Addition of new materials such as RNA glycans, SARS‐CoV‐2 structures, etc.

The text will be available for adoption by Fall 2022. We continue to need help in writing key features and in a continual process of updating, especially in these areas:

end‐of‐chapter and chapter‐imbedded problems (hopefully based on research literature)

mechanisms of your favorite enzymes or topics you feel not presented well in other texts

inclusive and diverse content

climate change, health disparities, addiction, etc.
The Effect of Cold and Warm Water Ingestion Prior to and During Passive Heating on Local Sweat Rate
Authors: Leach Caileigh , Newhouse Douglas , Ravanelli Nicholas
Published: May 2022

BACKGROUND

During heat stress, sweating is initiated to mitigate the rise in core temperature. Relative to warm water, previous evidence has demonstrated that cold water ingestion prior to exercise‐induced heat stress in compensable conditions delays the onset of sweating, and produces a reduction in local sweat rate immediately following ingestion. However, it remains unknown whether ingestion of cold water will elicit a similar delay in the onset of sweating and reduction in local sweating during more extreme thermal challenges. It is hypothesized that cold water ingestion prior to and during passive heat stress will result in a delayed onset of sweating and a reduction in local sweat rate in comparison to warm water.

METHODS

To date, three male participants (25±4 years, 82±8 kg, 1.9±0.1 m) have completed two passive heating sessions using a water perfused blanket system to increase rectal temperature by 1˚C above baseline. Participants ingested 3.2 ml‧kg‐1 (263±27 mL) of either cold (1.5˚C, CW) or warm (37˚C, WW) water five minutes before passive heating, and following a 0.5˚C and 1.0˚C increase in rectal temperature. Local sweat rate of the forearm and mean skin temperature were measured throughout baseline and passive heating.

RESULTS

Baseline rectal temperature was not different between CW (37.0±0.2˚C) and WW (36.9±0.1˚C, p=0.17) trials. Mean skin temperature was not different at baseline (CW: 33.5±0.2˚C, WW: 33.1±0.4˚C, p=0.60), and following a 0.5˚C (CW: 38.9±0.5˚C, WW: 38.9±0.6˚C, p = 0.99) and 1.0˚C (CW: 39.3±0.5˚C, WW: 39.2±0.4˚C, p=0.58) increase in rectal temperature. Mean body temperature at the onset of sweating on the forearm was not affected by the water temperature (CW: 37.1±0.2˚C, WW: 37.1±0.3˚C, p=0.79). When compared to the 5‐minutes before 37˚C water ingestion (PRE), mean local sweat rate of the forearm was not different during the 10‐minutes after (POST) at 0.5˚C (PRE: 0.98±0.23 mg‧cm‐2‧min‐1, POST: 1.05±0.36 mg‧cm‐2‧min‐1, p=0.48) and 1.0˚C (PRE: 1.11±0.37 mg‧cm‐2‧min‐1, POST: 1.15±0.49 mg‧cm‐2‧min‐1, p=0.74) increase in rectal temperature. Similarly, mean local sweat rate of the forearm was not different POST compared to PRE ingestion of 1.5˚C water following a 0.5˚C (PRE: 0.89±0.28 mg‧cm‐2‧min‐1, POST: 0.88±0.27 mg‧cm‐2‧min‐1, p=0.74) and 1.0˚C (PRE: 0.93±0.42 mg‧cm‐2‧min‐1, POST: 0.92±0.41 mg‧cm‐2‧min‐1, p=0.71) increase in rectal temperature. However, CW ingestion following a 0.5˚C increase in rectal temperature resulted in a lower local sweat rate of the forearm in comparison to WW for the remainder of heating (p<0.03).

CONCLUSION

Preliminary data suggest cold water ingestion may alter the local sweating response in young healthy males during passive heating.