bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oxygen Tolerance in the [FeFe] Hydrogenase Cb HydA1
Authors: Corrigan Patrick , Silakov Alexey 
Published: May 2022

Hydrogenases are enzymes that catalyze the formation and oxidation of hydrogen. [FeFe] hydrogenases have been shown to exhibit a high efficiency of H2production, which makes these systems attractive catalysts for renewable energy generating devices. Unfortunately, this class is known for its susceptibility to oxidative degradation, thereby limiting its ability to be used in next‐generation fuel source production devices. In this presentation, we describe the first investigations of the first example of an O2‐tolerant [FeFe] hydrogenase from Clostridium beijerinkii (CbHydA1).1–3. Our FTIR measurements and activity assays show that this enzyme is able to enter an inactivate state under oxidative conditions, is stable for long periods of time in this state, and can be reactivated under reducing conditions. Using FTIR spectroelectrochemistry, we showed that this enzyme is capable of inactivating at unusually low potentials and can inactivate in the absence of oxygen or sulfide, unlike other [FeFe] hydrogenases. Using a combination of FTIR measurements and density functional theory, we are able to propose two possible chemical identities of the inactive state. Finally, we utilized advanced EPR methods to investigate the electronic structure of the active center of CbHydA1, which shed light on the electronic‐structure prerequisites of O2‐tolerance in this unusual [FeFe] hydrogenase.(1) Morra, S.; Arizzi, M.; Valetti, F.; Gilardi, G. Oxygen Stability in the New [FeFe]‐Hydrogenase from Clostridium Beijerinckii SM10 (CbA5H). Biochemistry 2016, 55 (42), 5897–5900. https://doi.org/10.1021/acs.biochem.6b00780.(2) Corrigan, P. S.; Tirsch, J. L.; Silakov, A. Investigation of the Unusual Ability of the [FeFe] Hydrogenase from Clostridium Beijerinckii to Access an O 2 ‐Protected State. J. Am. Chem. Soc. 2020, 142 (28), 12409–12419. https://doi.org/10.1021/jacs.0c04964.(3) Winkler, M.; Duan, J.; Rutz, A.; Felbek, C.; Scholtysek, L.; Lampret, O.; Jaenecke, J.; Apfel, U.‐P.; Gilardi, G.; Valetti, F.; Fourmond, V.; Hofmann, E.; Léger, C.; Happe, T. A Safety Cap Protects Hydrogenase from Oxygen Attack. Nat. Commun. 2021, 12 (1), 756. https://doi.org/10.1038/s41467‐020‐20861‐2.
Orexin A enhances neuronal synchronization in adult rat hypothalamic culture: A model to study hypothalamic function
Authors: Mavanji Vijayakumar , Georgopoulos Apostolos P. , Kotz Catherine M. 
Published: May 2022

BackgroundThe regulation of sleep/wake behavior and energy homeostasis is maintained in part by the lateral hypothalamic (LH) neuropeptide orexin A (OXA, hypocretin). A reduction in orexin signaling is associated with sleep disorders and obesity, whereas higher LH orexin signaling and sensitivity promotes obesity resistance. Similarly, dysregulation of hypothalamic neural networks is associated with onset of age‐related diseases, including obesity and several neurological diseases. Despite the increase in obesity with aging, and that adult populations are the subjects in the majority of pharmaceutical and obesity studies, conventional models for neuronal networks utilize embryonic neural cultures rather than adult neurons. Synchronous activity, a feature of normal brain function, and a measure of functional connectivity, describes correlated changes in neuronal activity between neurons, Synchronization determines the nature of the final output from a given neural structure. Neural synchrony is altered by behavioral perturbations, in embryonic neurons obtained from obesity‐resistant rats, and following application of OXA onto embryonic hypothalamic cultures. Synchronous network dynamics in adult hypothalamic neurons remain largely undescribed.MethodsWe established an adult rat hypothalamic culture in multi‐electrode‐array (MEA) dishes and recorded the field potentials. Then we studied the effect of exogenous OXA on network synchronization of these adult hypothalamic cultures. In addition, we studied the wake promoting effects of OXA in vivo when directly injected into the rostral lateral hypothalamus (rLH). To study the wake promoting effects of LH OXA, rats were implanted with a radiotelemetric transmitter and EEG/EMG electrodes to record vigilance states (F40‐EET, Data Sciences International [DSI], St. Paul, MN), and a cannula targeting the rLH. Following surgical recovery, either OXA (250 pmol/0.5 μl) or saline was infused into the rLH (single injection) and sleep/wake states were measured for 2h post‐injection.ResultsThe results show that adult hypothalamic cultures are viable for nearly 3 months in vitro, good quality MEA recordings can be obtained from these cultures in vitro, and finally, that cultured adult hypothalamus is responsive to OXA. In addition, LH administration of OXA enhanced wakefulness in rats, indicating that OXA enhances wakefulness for up to 2h post‐injection.ConclusionsThese results support that adult rat hypothalamic cultures could be used as a model to study the neural mechanisms underlying obesity, and that obesity resistance is associated with increased hypothalamus synchronization. Increased wake following rLH OXA suggest the possibility that OXA promotes wake partly by promoting neural synchrony in the hypothalamus.
Mitochondrial Uncoupling Decreases Sarcopenic Obesity By Activation of Skeletal Muscle Mitochondrial Quality Control and Attenuated ER Stress
Authors: Dantas Wagner S. , Zunica Elizabeth R. M. , Heintz Elizabeth C. , Hoppel Charles L. , Axelrod Christopher L. , Kirwan John P. 
Published: May 2022

BackgroundSarcopenic obesity is a highly prevalent disease with poor survival and ineffective medical interventions. Mitochondrial dysfunction is purported to be central in the pathogenesis of sarcopenic obesity by impairing both organelle biogenesis and quality control. We have previously identified an orally available mitochondrial‐targeted furazano[3,4‐b]pyrazine named BAM15 that selectively lowers respiratory coupling efficiency and protects against diet‐induced obesity in mice. Here, we tested the hypothesis that mitochondrial uncoupling simultaneously attenuates loss of muscle function and weight gain in a mouse model of sarcopenic obesity.Methods80‐week‐old male C57BL/6J mice with obesity were randomized to 10 weeks of high fat diet (CTRL) or BAM15 (BAM15; 0.1% w/w in high fat diet) treatment. Body composition, muscle function, energy expenditure, and locomotor activity were determined after treatment. Skeletal muscle was harvested and evaluated for histology, gene expression, protein signaling, and mitochondrial structure and function.ResultsBAM15 decreased body weight (~25% reduction, P<0.001) which was attributable to increased energy expenditure (~20% increment, P<0.001). BAM15 increased muscle mass (~13% increment, P<0.001), strength (~37% increment, P<0.0001), and locomotor activity (~25% increment, P<0.001). Improvements in physical function were mediated in part by reductions in skeletal muscle inflammation (IL‐6 and gp130, both P<0.05), enhanced mitochondrial function, and improved endoplasmic reticulum homeostasis and reduced inflammation. Specifically, BAM15 activated mitochondrial quality control through AMPK (PINK1‐ubiquitin binding and LC3II, P<0.01), increased electron transport chain activity (citrate synthase and complex II activity, all P<0.05), restricted endoplasmic reticulum (ER) misfolding (decreased oligomer A11 insoluble/soluble ratio, P<0.0001) while limiting ER stress (decreased PERK signaling, P<0.0001), apoptotic signaling (decreased cytochrome C release and Caspase‐3/9 activation, all P<0.001), and muscle protein degradation (decreased 14‐kDa actin fragment insoluble/soluble ratio, P<0.001).ConclusionsMitochondrial uncoupling agents such as BAM15 may mitigate age‐related decline in muscle mass and function by molecular and cellular bioenergetic adaptations that confer protection against sarcopenic obesity through activation of mitochondrial quality control and attenuation of ER stress.
Structural and functional analysis of cancer related proteins: A critical target for drug development
Authors: Sharma Ruby , Saxena Ajay K. 
Published: May 2022

Protein kinases are involved in cancer metabolism and have been targeted for structure‐based drug development. The crystals structures of several clinically important protein kinase‐inhibitor complexes have been determined, which provide the mechanism of inhibition of protein kinases. In our laboratory, we are working on structure and function analysis on two protein kinases e.g. (i) a non‐receptor tyrosine kinase expressed in hepatocellular carcinoma (TEC) and Class I Phosphoinositide 3‐Kinase (PI3K). We have cloned and expressed both kinases in E. coli and looking forward for biochemical and structural characterization of both proteins.In addition, we are also working on structural and biochemical characterization of Ergp55 protein belongs to Ets family of transcription factor. The Ergp55 is involved in various development processes and regulation of cancer metabolism. Using biochemical assay, we have identified the part of Ergp55 protein that mediates the auto‐inhibition using E74 and cfos promoter DNA sequences. We have also crystallized and solved the crystal structure of ETS domain of Ergp55 in native and its complexes with E74 and cFOS promoter DNA sequences. The structure analysis provides the insights into DNA sequence discrimination by ETS domain of Ergp55 protein. These studies will aid in designing the compounds, which stabilize the inhibited form of Ergp55 and inhibits its binding to DNA. Based on crystal structures, we have designed the DNA binding compounds, which interfere in ETS‐DNA binding and proved to be potential drugs against prostate cancer.
A 24‐Hour Fast Increases Liver Cytotoxic T Cell Populations in Genetically Obese Mice: A Preliminary Study
Authors: Graham Elliot L. , Trikha S. Raj J. , Risk Briana , Weir Tiffany L. , Gentile Christopher L. 
Published: May 2022

BACKGROUNDRecent data suggests that regulated fasting protocols can improve metabolic parameters associated with obesity. However, questions remain regarding the appropriate fasting duration needed to attenuate obesity‐associated complications such as glucose intolerance, arterial stiffness, and immune cell dysfunction. Therefore, the primary purpose of this study was to examine if an acute 24‐hour fast improves detrimental consequences of obesity and alters tissue‐specific immune cell counts in obese mice.METHODSTen 12‐week‐old male and female genetically obese mice (Ob; n=20) and wild‐type littermates (WT; n=20) were fed a standard diet and randomized to either ad libitum feeding or a 24‐hour fast (WT‐CON, WT‐Fast, Ob‐CON, Ob‐Fast). Mouse weight was measured before and after feeding protocols. Following feeding protocols, arterial stiffness was assessed via arterial pulse wave velocity (aPWV); liver and mesenteric adipose tissue (MAT) immune cell populations were assessed via spectral flow cytometry and normalized by tissue weight. For statistical analyses, sex‐adjusted regression models were fit to account for the confounding effects of sex.RESULTSThere was not a significant interaction between genotype and intervention on changes in body weight, plasma glucose concentrations, or aPWV (p=0.16; p=0.33; p=0.37, respectively). After adjusting for genotype, acute fasting did not significantly alter changes in plasma glucose concentrations or aPWV (p=0.95; p=0.22). However, after adjusting for genotype, acute fasting promoted significant weight loss (CON 0.4g ± 0.56; Fast ‐2.6g ± 0.72; p<0.01). Compared to Ob‐CON, Ob‐Fast mice had significant increases in liver total immune and cytotoxic T cells (p<0.01) yet no significant changes in either liver regulatory T or helper T cells nor in MAT total immune, cytotoxic T, helper T, and regulatory T cell populations. WT‐Fast mice had significant reductions in liver total immune, cytotoxic T, helper T, and regulatory T cell populations while these same cell populations were increased in MAT when compared to WT‐CON mice.CONCLUSIONSAn acute 24 hr. fast in mice was insufficient to significantly modulate glucose intolerance and aPWV in Ob mice compared to WT. However, fasting differentially affected liver and MAT T cell populations in mice. These results suggest acute fasting could be a robust mediator of T cell populations with a particular emphasis on expanding liver cytotoxic T cell communities in obese mice.
Astrocyte‐to‐neuron reprogramming for spinal cord repair
Authors: Niceforo Alessia , Fortino Tara , Shah Yashvi , Zholudeva Lyandysha V. , Qiang Liang , Lane Michael A. 
Published: May 2022

Traumatic spinal cord injury (SCI) disrupts neural pathways and results in the devastating tissue damage and loss of function, with subsequent reactive gliosis and tissue scarring that limits further loss, but also limits endogenous repair. One therapeutic strategy gaining some attention in recent years is cellular reprogramming, which harnesses advances in cellular engineering to directly convert cells from one phenotype to another. The present work explores strategies for converting either mature or developing astrocytes into neurons. Our initial experiments have begun exploring viral targeting of glial cells to promote doxycycline‐inducible overexpression of neurogenic transcription factors (TFs) Ascl1 or a combination of microRNAs (miRs) miR124, miR9/9*, for neuronal conversion. Our primary goal was to establish optimal conditions for consistent and efficient reprogramming of developing spinal glia and mature reactive spinal astrocytes.Primary astrocyte cultures were generated from postnatal days 2‐3 (P2‐3) rat cortex and spinal cord, and then were activated with TGFβ (Transforming growth factor beta) one day prior to viral transduction. Fifteen days following doxycycline administration, astrocytes took on a typical neuronal morphology. Immunocytochemistry confirmed that these cells were positive for neuronal markers MAP2, NeuN and TAUR1, and negative for GFAP. Multielectrode array recordings of converted cells revealed spontaneous firing consistent with neuronal activity.These preliminary results confirm that cortical and spinal astrocytes can be efficiently reprogrammed into neurons by either Ascl1 or miRs. Advances in cellular reprogramming are providing novel ways to manipulate the cytoarchitecture of the injured cervical spinal cord. Our ongoing work is now using this same approach to modify developing glial precursors that can be harness for their pro‐reparative properties for spinal cord repair.
The 2021 Marine Pharmacology and Pharmaceuticals Pipeline
Authors: Wu Alex C. , Jelielek Karolina K. , Le Henry Q. , Butt Maryam , Newman David J. , Glaser Keith B. , Pierce Marsha L. , Mayer Alejandro M. 
Published: May 2022

ObjectiveMarine organisms continue providing significant chemical biodiversity that contributes to both the global preclinical and clinical pharmaceutical pipelines. A recently (2021) redesigned website www.marinepharmacology.org allows for easier and more frequent updates, thus highlighting the continuous progress in this area of marine pharmacology/pharmaceutical research and drug development.HypothesisThere has been progress in the approval and development of marine‐derived drugs in 2021.MethodsThe following four websites were researched to determine new marine‐derived drugs approved in 2021: (a) USA: https://www.accessdata.fda.gov/scripts/cder/daf/ United States Food and Drug Administration (FDA); (b) Europe: https://www.clinicaltrialsregister.eu/ctr‐search/search European Clinical Trials register; (c) China: https://www.chictr.org.cn/enindex.aspx Chinese Clinical Trials register; and (d) www.clinicaltrials.gov to determine the current status of marine‐derived compounds in Phase I, Phase II, and Phase III of drug development.ResultsOur research supports our hypothesis because in 2021 there has been continued progress in the approval and development of marine‐derived drugs. In 2021 the clinical marine pharmaceutical pipeline consisted of 15 marine‐derived drugs approved by the FDA, 2 approved in Australia, and 1 approved in China: for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969); for Herpes Simplex Virus, vidarabine (Vira‐A®, FDA‐approved 1976); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia, omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004, Vascepa®, FDA‐approved 2012, Epanova®, FDA‐approved 2014); for cancer, eribulin mesylate (Halaven®, FDA‐approved 2010), brentuximab vedotin (Adcetris®, FDA‐approved 2011), trabectedin (Yondelis®, FDA‐approved 2015), panobinostat (Farydak®, FDA‐approved 2015), plitidepsin (Aplidine®, approved in Australia 2018), polatuzumab vedotin (Polivy™, FDA‐approved 2019), enfortumab vedotin‐ejfv (PACDEV™, FDA‐approved 2019), lurbinectedin (Zepzelca™, FDA‐approved 2020, Australia 2021), belantamab mafadotin‐blmf (Blenrep™, FDA‐approved 2020), disitamab vedotin (Aidixi™, approved in China 2021), and tisotumab vidotin (Tivdak™, FDA‐Approved 2021). Furthermore, we determined that there were 33 marine‐derived compounds in active clinical trials in 2021: 5 marine‐derived compounds were in Phase III, 12 compounds in Phase II, and at least 16 compounds in Phase I. Many of compounds classified as auristatin‐containing antibody drug conjugates.ConclusionsIn 2021, new marine‐derived drugs were approved by FDA, as well as in Australia and China. The clinical development of marine‐derived drugs remained very robust and active with several advancing to Ph III, Ph II and entering at Ph I.
Human transcription factor protein interaction networks
Authors: Varjosalo Markku , Göös Helka , Kinnunen Matias , Salokas Kari , Liu Xiaonan 
Published: May 2022

Transcription factors (TFs) interact with several other proteins in the process of transcriptional regulation. Here, we identify 6703 and 1536 protein–protein interactions for 109 different human TFs through proximity‐dependent biotinylation (BioID) and affinity purification mass spectrometry (AP‐MS), respectively. The BioID analysis identifiesmore high‐confidence interactions, highlighting the transient and dynamic nature of many of the TF interactions.By performing clustering and correlation analyses, we identify subgroups of TFs associated with specific biological functions, such as RNA splicing or chromatin remodeling. We also observe 202 TF‐TF interactions, of which 118 are interactions with nuclear factor 1 (NFI) family members, indicating uncharacterized cross‐talk between NFI signaling and other TF signaling pathways. Moreover, TF interactions with basal transcription machinery are mainly observed through TFIID and SAGA complexes.This study provides a rich resource of human TF interactions and also act as a starting point for future studies aimed at understanding TF‐mediated transcription.
Effects of High Frequency Resistance Training on RNA Expression Related to Inflammation: A Case Report
Authors: Saffold Keith , Kelley Christian , Buford Thomas 
Published: May 2022

BACKGROUNDResistance training (RT) is the main modality of exercise used to induce hypertrophic and neuromuscular adaptations. RT done at high intensities (HI‐RT) is also capable of inducing epigenetic modifications leading to altered protein expression. HI‐RT increases protein expression of many enzymes that play key roles in disease prevention and treatment. Inflammation is one pathology that is associated with many disease states that can be altered by the effects of HI‐RT. Both pro‐ and anti‐inflammatory cytokines are upregulated after a bout of exercise. Long term training leads to increases in anti‐inflammatory cytokines such as muscle‐derived interleukin‐6 (IL‐6). Current recommendations state that each muscle group should be exercised 2‐3 nonconsecutive days per week to allow time for recovery. HI‐RT protocols including training much higher training volumes and frequencies have been used by advanced athletes for decades, the effects of these protocols on health outcomes are not reflected in the literature.STUDY OBJECTIVEThe objective of this case report is to determine the effects of high‐frequency and high‐volume HI‐RT on RNA expression linked to inflammatory cytokines.HYPOTHESISIt is hypothesized that both the pro‐inflammatory and anti‐inflammatory cytokines will be upregulated following a high‐frequency high‐volume HI‐RT program due to increased muscle damage and macrophage infiltration.METHODSThe participant (N = 1; 24 years; 192 cm; 107.7 kg; 25.6% body fat) had leg circumference measured at 50% of the length of the femur. Then, a muscle biopsy was performed on the right leg vastus lateralis (VL). Muscle samples were stored at ‐80°C for later analysis. The participant completed a high‐frequency, high‐volume HI‐RT program for one month, which included barbell squats 5 days per week. After the training program was completed, leg circumference was measured again and a second biopsy of the right leg VL was taken. RNA was isolated from the samples for real‐time polymerase chain reaction (RT‐PCR) analysis of IL‐6 and tumor necrosis factor alpha (TNF‐α).RESULTSThe participant experienced a 4 cm increase in thigh circumference (6.4%). Due to time constraints, RT‐PCR analysis was delayed. Analysis will be concluded prior to the conference meeting.CONCLUSIONSTo be updated after analysis is complete.