bluuebunny/crossref_metadata_2025 huggingface dataset:
DNA Repair of Ovarian Cancer Cell Lines SKOV3 and OV90 Compared to Non‐Transformed PBMC Cells
Authors: Reese Megahn M. , Podltutsky Andrej
Published: May 2022

A defining characteristic of cancer is the increase in genomic instability along with the progression of carcinogenesis, in which DNA mutations are the biggest contributors. Further, most mutations related to the development of ovarian cancer are acquired instead of inherited, demonstrating that familial tumors are the minority of cases. Two ovarian cancer cell lines that are extensively used in research are OV‐90 and SK‐OV‐3. We used the comet assay (single‐cell gel electrophoresis) to quantify DNA repair abilities in these cell lines compared to PBMCs (peripheral blood mononuclear cells), in order to assess the sensitivity of these cell lines to radiation treatments. A difference in DNA repair ability is expected in the cell lines compared to PBMCs due to the genomic instability related to tumorigenesis. After being treated with X‐ray radiation and undergoing a designated repair time, individual cells were scored, using fluorescent microscopy, based on percent “head” and “tail” DNA. It appears that SK‐OV‐3 is repairing DNA faster than PBMC, while OV‐90 repairs DNA more similarly to PBMCs. This observation has important implications for the development of drugs to treat ovarian cancer, along with understanding the dynamics of DNA repair within cancer cells. Considering that most cases of epithelial ovarian cancer remain death sentences in spite of new treatments, insights regarding cell line sensitivity are of even more importance with every passing day.
Elastin Insufficiency Accelerates Age‐Related Impairment of Renal Hemodynamics
Authors: Dixon Alethia J. , Osei‐Owusu Patrick
Published: May 2022

Gradual loss of functional elastin fibers during aging leads to increased arterial stiffening and decreased compliance. Elastin insufficiency in mice and humans has been shown to be associated with arteriopathy and the development of hypertension. However, it is unclear whether elastin insufficiency in the resistance vasculature, as occurs in the renal vascular bed, contributes to age‐related decline in renal function. We hypothesized that elastin insufficiency exacerbates age‐related impairment of renal hemodynamics by accelerating functional and structural changes of the renal microvasculature. We assessed renal hemodynamics in anesthetized young (4‐6‐month‐old) and old (14–16‐month‐old) female wild‐type (WT) and elastin heterozygous (Eln+/‐) mice. Renal autoregulation was assessed by a stepwise increase in renal perfusion pressure (RPP) by simultaneously occluding the superior mesenteric and celiac arteries. Autonomic activity was clamped by continuous infusion of a cocktail containing vasoactive hormones. Baseline systolic blood pressure (SBP; 91.1±4.2 vs 90.1±3.1mmHg) and renal vascular resistance (RVR; 13.1±2.4 vs 10.0±1.5 mmHg/μL/min/g left kidney weight) were elevated in old versus young WT mice. Renal blood flow (RBF; 7.3±1.7 vs 9.8±1.4 μL/min/g left kidney weight), renal plasma flow (RPF; 5±1.2 vs 6±1mL/min/g left kidney weight) and glomerular filtration rate (GFR), all trended lower in old WT mice compared to young WT mice. However, in Eln+/‐mice, SBP was lower in old mice (78.2±9.4 vs 104.6±4.9mmHg), while RVR (13.5± 2.6 vs 14.4±1.1), RBF (6.7±1.2 vs 7.5±0.6), RPF (4.2±0.7 vs 4.5±0.3) and GFR were all similar between young and old mice. Increasing RPP caused a robust RVR and RPF response in young vs old WT mice. In contrast, the maximal changes in RBF (5.8±0.6 vs 5.5±1 μL/min/g left kidney weight), RVR (23.8±2.5 vs 21.8±7.0 mmHg/μL/min/g left kidney weight) were less robust and similar between young and old Eln+/‐mice. Autoregulatory index average values were closer to 0 in young and old Eln+/‐and old WT miceindicating greater impairment of renal autoregulation compared to young WT mice. Correlation analysis between pulse pressure and RBF showed that a high pulse pressure was more positively correlated to high RBF in young and old Eln+/‐mice as well as old WT mice. Together, our data demonstrate that elastin insufficiency accelerates age‐related impairment of renal hemodynamics and autoregulation likely mediated by functional and structural remodeling of the renal microvasculature.
Hepatoprotective effect of a Novel Functional Drink based on Ecuadorian Plant Species
Authors: Paladines‐Santacruz Geovanna L. , Orellana‐Manzano Andrea , Manzano Patricia , Zaruma‐Torres Fausto , Ortíz‐Ulloa Johana , Cárdenas Washington , Torres‐Arias Marbel , Seqqat Rachid , Vanden Berghe Wim
Published: May 2022

Liver diseases are considered a severe public health problem involving disorders that alter liver function. Since the liver plays an essential role in the metabolism and detoxification of substances that enter the organism, it is susceptible to damage caused by xenobiotic agents such as viruses, alcohol, and drugs. Acetaminophen is an over‐the‐counter drug widely used as an antipyretic and analgesic. It is safe at therapeutic doses, but it is easily overused, causing severe liver diseases. In a previous study, our investigation group developed a functional drink from the aqueous extracts of Ilex guayusa, Vernonanthura patens, and cocoa husks. This combination presented various bioactive compounds with antioxidant properties that could help reduce oxidative stress in pathologic conditions. For the above mentioned, the study's objective was to evaluate the hepatoprotective effect of the beverage through an in vivo test. Male mice were treated with two formulations (pre‐formulated and microencapsulated) of the functional drink in three concentrations for seven days as a preventive oral treatment. After that, mice were induced to liver damage at a single intraperitoneal dose of acetaminophen (250 mg/kg of body weight). The evaluation was based on the liver damage biomarkers, ALT, and AST; anatomopathological studies were also evaluated. The results showed that the best formulation as an oral pre‐treatment was the pre‐formulated. The pre‐formulated beverage attenuated the injury caused by the acetaminophen by inhibiting serum ALT and AST activities and reducing liver macro and microscopic liver alterations. The successful doses were at 250 and 1000 mg/kg b.w. The microencapsulated did not present adequate liver protection at any doses evaluated because it has no statistical difference with the toxic model group.

In conclusion, the present study demonstrated for the first time that the functional drink effectively prevented acetaminophen‐induced liver injury in vivo. Our data indicated that the pre‐formulated was the most effective pre‐treatment to attenuate the hepatic injury. These results suggested that this beverage possessed antioxidative activity that likely contributed to its protective effect against acetaminophen liver damage.

*Corresponding Author: Orellana‐Manzano Andrea, email: akorella@espol.edu.ec
Nexinhib20 prevents myocardial ischemia‐reperfusion injury by inhibiting neutrophil adhesion and β2 integrin activation
Authors: Fan Zhichao , Liu Wei , Cronin Chunxia G. , Wang Chengliang , Ruan Jianbin , Johnson Jennifer L. , Catz Sergio , Sun Hao , Groisman Alex , Chen Yunfeng , Hu Liang , Vella Anthony T. , Liang Bruce
Published: May 2022

Primary percutaneous coronary intervention (PPCI)—a therapy intended to restore blood flow after acute myocardial infarction—can also induce myocardial ischemia‐reperfusion (I/R) injury, exacerbating cardiomyocyte death. Neutrophil‐mediated cardiac tissue damage contributes prominently to the process of myocardial I/R injury. Therefore, limiting neutrophil recruitment and exocytosis may prevent myocardial I/R injury after PPCI. Here, we found out that a neutrophil exocytosis inhibitor Nexinhib20 inhibits both neutrophil adhesion and exocytosis and prevents myocardial I/R injury in a mouse model. Using a microfluidic chamber, we found that Nexinhib20 inhibited interleukin 8 (IL‐8)‐induced β2 integrin‐dependent human neutrophil adhesion under flow. Using dynamic flow cytometry assay, we discovered that Nexinhib20 suppresses intracellular calcium flux and β2 integrin activation after IL‐8 stimulation. Western blots of Rac‐1‐GTP pull‐down assay confirmed that Nexinhib20 inhibited Rac‐1 activation in leukocytes. In vitro competing assay showed that Nexinhib20 antagonized the binding of Rac‐1 and GTP. Using a mouse model of myocardial I/R injury, Nexinhib20 administration after ischemia and before reperfusion significantly decreased neutrophil recruitment and infarct size. Our results highlight the translational potential of Nexinhib20 as a dual‐functional neutrophil inhibitory drug to prevent myocardial I/R injury.
The effect of novel second generation prebiotics (levan of β‐[2,6]‐glycosidic linkages) on the cardiovascular system in obesity
Authors: Sahyoun Amal , Kwon Young‐In , Munkhsaikhan Undral , Karboune Salwa , Kassan Modar
Published: May 2022

Objectives

Bacteria dysbiosis is very well documented in obesity and is known to regulate the cardiovascular system. Recovering the bacteria composition is a key element in improving the cardiovascular function during obesity.

Hypothesis

In this study we investigate the effect of levan, a novel second generation of prebiotic (β‐[2,6]‐linkages, on the cardiovascular system in obesity.

Methods

Eight‐week‐old C57/b6 mice were fed with high fat diet (HFD) for 12 weeks in the presence and absence of levan (β‐[2,6]‐linkages). Treatment was administered by gavage (250 mg/kg) for the last 30 days of HFD feeding. Body weight, blood glucose, body composition and lipid profile were determined. Vascular function and endothelial function markers were studied in aorta and mesenteric resistance arteries (MRA).

Results

Levan (β‐[2,6]‐linkages) treatment reduced the body weight in obese mice. Blood glucose, % of fat mass, total cholesterol, and LDL levels were significantly reduced and the % of lean mass was significantly increased after Levan (β‐[2,6]‐linkages) treatment. Aortic and mesenteric arteries vascular response to sodium nitroprusside (SNP) was similar among groups. However, the vascular response to acetylcholine (Ach) was improved in the treated group.

Conclusions

Treatment with Levan (β‐[2,6]‐linkages) attenuated the increase in body weight in obese mice and restored the lipid profile and the vascular function.
Developmental Regulation of Sarcomere Branching in Cardiomyocytes
Authors: Ajayi Peter T. , Kim Yuho , Bleck Christopher K. E. , Glancy Brian
Published: May 2022

Cardiac muscles undergo constant contractions to continuously supply oxygenated blood to both central and peripheral organs. To sustain such large demand, cardiac muscles must convert biological fuel to functional force via the contraction of their sarcomeres. Using nanoscale 3D imaging analysis, our group demonstrated that sarcomeres within the cardiac muscle of mature mice (≥ 2 months) are frequently branched to form a unified myofibrillar matrix across the entire cell, thereby providing the architecture needed for both longitudinal and lateral transmission of active force. How these contractile networks are formed during postnatal development remains unclear. Here we used 3D focused‐ion beam scanning electron microscopy (FIB‐SEM) to evaluate myofibrillar ultrastructure within cardiac muscle cells in mice at postnatal (P) days 1, 7, 14, and 42, respectively. 3D reconstruction of myofibrillar structures showed highly connected networks at all time points. Quantitative assessment of sarcomere branch‐points revealed that branching frequency is varied by developmental process. The percentages of sarcomeres with at least one branch‐point are as follows: 57 ± 2.4% (mean ± SE, 14 cells, 129 myofibrils, 802 sarcomeres) in P1, 64 ± 2.1% (17 cells, 161 myofibrils, 795 sarcomeres) in P7, 49 ± 3.9% (4 cells, 50 myofibrils, 250 sarcomeres) in P14, and 27 ± 1.1 % (7 cells, 236 myofibrils, 2320 sarcomeres) in P42. Data was further separated based on whether a single sarcomere is branched into two (single‐branch, Fig 1E, F) or more (multi‐branch, Fig 1G, H) structures to delineate the nature and overall magnitude of branching within cardiomyocytes (Fig 2B‐ D). Across the postnatal development (P1 to P42), there was a 0.4‐fold change in the magnitude of branching within ten sarcomeres (Fig 2D; 7.9±0.41, 11±0.45, 6.5±0.58, and 3.3±0.16 for P1, P7, P14, and P42, respectively). All changes were statistically significant at P<0.05. In conclusion, the frequency of sarcomere branching was highest at P7 and progressively declined until maturation. Findings indicate that while the architecture needed for active force transmission across the length and width of the cell is already formed at birth, the connectivity of cardiac contractile networks is dynamic throughout postnatal development. These results suggest that sarcomere branching dynamics may provide a target for modulating the contractility of cardiac muscle cells.
Proteomic analysis of urinary exosomes from patients with lithium‐induced nephrogenic diabetes insipidus
Authors: Grønkjær Camilla , Apolito Luciano D. , Rej Soham , Fenton Robert , Trepiccione Francesco , Christensen Birgitte M.
Published: May 2022

Lithium (Li)‐induced Nephrogenic Diabetes Insipidus (NDI) is a large healthcare problem, which affect around 40% of patients with bipolar disorder undergoing Li treatment. NDI is characterized by the inability in the kidney to concentrate urine in response to vasopressin in the kidney collecting duct (CD). In animals, Li causes a downregulation of aquaporin‐2 and a cellular remodeling of the CD in rats resulting in a reduced fraction of principal cells relative to intercalated cells. Whether Li has similar effects in the human kidney is, however, not known. Urinary exosomes are extracellular nanovesicles derived from intracellular multivesicular bodies fused with the plasma membrane of epithelial cells lining the kidney tubular system and the urinary tract. The purpose of the first part of this study is to analyze whether the urinary rat proteome reflects the Li‐induced changes in the kidney. This is followed by proteomic analyses of urinary exosomes isolated from Li‐NDI patients.

Rats were divided in three groups; controls (n=8), 2‐week Li treatment (40 mmol/kg/food, n=8) and 4‐week Li treatment (40 mmol/kg/food, n=8). Treatment with Li showed a significant increase in urine output after 2‐weeks (P<0.01) and 4‐weeks (P<0.05). Additionally, the rats developed severe polydipsia, with water intake significantly increased after 2‐weeks Li treatment (P<0.001) and 4‐weeks Li treatment (P<0.0001).

Early morning urine samples from the placebo group in a previous published 12‐week placebo‐controlled pilot trial (Fotso Soh et al, 2021. A double‐blind, randomized, placebo‐controlled pilot trial of atorvastatin for nephrogenic diabetes insipidus in lithium users. Bipolar Disorders 23, 66–75) involving Li‐NDI patients (n=9) were collected. The Li‐NDI participants have been on a stable dose of Li (˃2 months) and determined to have a urinary osmolality <600 mOsm/kg at baseline. In addition, urine from random healthy controls (n=6) were collected. Urine samples from the rodent study and human study were subjected to ultracentrifugation for exosome isolation and will be further processed for mass spectrometry analysis (MS/MS).
Erythropoietin M2 macrophage Signaling Promotes Schwann Cells Clearance and Functional Recovery Following Peripheral Nerve Injury
Authors: Govindappa Prem Kumar , Elfar John C.
Published: May 2022

Background

Traumatic peripheral nerve injury (TPNI) obliterates axons and Schwann cells (SCs) that must be cleared through phagocytosis to allow surviving SCs to de‐differentiate and divide and guide the regeneration process. In TPNI, macrophage (MØ) infiltration and early phenotypic transitions (M1 to M2) are critical for the phagocytosis of SCs debris. Delayed phagocytosis critically aggravates inflammation and impairs MØ function, which leads to failure to advance SC orchestration and neuronal regeneration. Erythropoietin (EPO), a pluripotent hormone, helps to guide MØs and SC transition post‐TPNI. The significance of the effects of EPO on debris clearance, apoptosis, myelination, and functional improvement is unknown, despite much effort motivating clinical translation of EPO for TPNI. We hypothesized that a role in supporting M2 phenotype macrophages after TPNI might explain EPO’s neuroprotective function through SCs debris clearance.

Methods and Results

We evaluated EPO’s effect on sciatic nerve crush injury (SNCI) and found EPO treatment (5000 IU/kg, intraperitoneally, post‐surgery days 1, 2, and 3,) increased myelination and sciatic functional index (SFI) and augmented anti‐apoptosis and phagocytosis of myelin debris via CD206+ macrophages when compared to saline treatment in the mouse. EPO increased efferocytosis of apoptotic sciatic nerve derived Schwann cells (SNSCs) both in bone marrow derived macrophages (BMMØs) and peritoneal derived macrophages (PMØs) in‐vitro, as well as in PMØs in‐vivo by immunofluorescence (IF) and flow cytometry. EPO treatment significantly attenuated BMMØ pro‐inflammatory genes (IL1b, iNOS, and CD68) and augmented anti‐inflammatory genes (IL10, and CD163) in addition to the cell surface marker CD206 expression after lipopolysaccharide (LPS) induction. EPO also showed anti‐apoptotic (Annexin V/ 7AAD) effects in BMMØs. Our data demonstrate that EPO promotes the M2 phenotype macrophages to ameliorate SN apoptosis and efferocytosis of dying SCs and myelin debris and improves SN functional recovery following SNCI.

Conclusions

To the best of our knowledge, this is the first study to demonstrate EPO’s ameliorative effect on the progression of post‐TPNI via M2 phenotype macrophage phagocytosis of SCs debris with early anti‐apoptotic and anti‐inflammatory effects. These data may support the findings of other researchers and might shed light on the role of EPO in improving functional recovery in the traumatized limb.
Identification of Deubiquitinases (DUBs) that modulate PAR1‐mediated p38 MAPK inflammatory signaling in endothelial cells
Authors: Cheng Norton , Patwardhan Anand , Trejo JoAnn
Published: May 2022

Protease‐activated receptor‐1 (PAR1), a GPCR for thrombin, induces endothelial dysfunction via multiple signal transduction pathways including p38 mitogen‐activated protein kinase (MAPK). We showed that thrombin‐stimulates K63‐linked ubiquitination of PAR1 that drives recruitment of transforming growth factor‐β‐activated kinase‐1 binding protein‐2 (TAB2), an adaptor protein that binds TAB1, which triggers p38 activation on endosomes and promotes endothelial barrier disruption. However, the regulatory processes that control thrombin‐stimulated PAR1 ubiquitin‐dependent p38 inflammatory signaling are not known. To address this gap in knowledge, we sought to identify specific deubiquitinases (DUBs) that counter thrombin‐induced ubiquitin‐mediated p38 signaling by conducting an unbiased genome‐wide siRNA library screen targeting all 96 human DUB genes in human cultured endothelial cells. We hypothesize that a PAR1‐specific DUB is essential for controlling proper dynamics of thrombin‐mediated p38 signaling and might be altered in endothelial dysfunction. To systematically identify DUBs, endothelial cells were transfected with pools of 4 siRNAs targeting each of the 96 DUBs and thrombin‐stimulated p38 phosphorylation was assessed by immunoblotting. In parallel, a genome‐wide DUB siRNA library screen was conducted in HeLa cells. We identified 8 different DUBs that preferentially cleave K63‐linked ubiquitin and regulate thrombin‐p38 signaling in both endothelial and HeLa cells. A time‐course analysis of thrombin‐stimulated p38 signaling confirmed that depletion of each of the 8 DUBs alone caused an increase in the magnitude and/or duration of p38 signaling. Among the candidate DUBs, USP34, USP47 and CYLD exhibited the highest expression in both endothelial and HeLa cells detected by RT‐qPCR and were further examined. Using individual siRNAs targeting each of the 3 candidate DUBs, we validated that knockdown of either USP34, USP47 or CYLD alone was sufficient to markedly enhance thrombin‐induced p38 signaling in endothelial cells. Currently, I am examining the impact of USP34‐mediated modulation of thrombin‐p38 signaling on endothelial inflammatory responses. I will present new data examining the role of USP34 on thrombin‐induced endothelial barrier disruption and induction of cytokine expression. While ubiquitin‐dependent signaling is widespread, our knowledge of the processes that control ubiquitin‐mediated GPCR signaling is limited, especially related to the function of GPCR‐specific DUBs. Here, we report the first genome‐wide DUB siRNA library screen on GPCR signaling and identified a subset of DUBs that specifically regulate PAR1‐p38 inflammatory signaling that may serve as important therapeutic targets for vascular inflammation.
Functional Extraction and Spectroscopic Investigation of Structural Dynamics in E. coli ATP Synthase
Authors: Dowdall Owen , Defnet Amy , Steed Ryan
Published: May 2022

ATP synthase is a membrane protein complex that synthesizes adenosine triphosphate (ATP), an integral molecule that provides energy to all forms of life. The complex is comprised of two rotary motors, the soluble F1 sector and the membrane embedded Fo sector. ATP synthesis is catalyzed by conformational changes within F1, which are driven by the rotation of the c‐ring (rotor) of Fo. The Fo motor is powered by the electrochemical gradient of H+ across the cell membrane, and movement of H+ through Fo is facilitated by two aqueous half‐channels within subunit a (stator) and at the rotor‐stator interface. The mechanism of how H+‐driven rotation in Fo is generated is not fully understood. Cryo‐electron microscopy has revealed structures of E. coli F1Fo ATP synthase, but inconsistencies between this structural data and previous cross‐linking data within subunit a suggest the existence of multiple conformations. In order to gain insight into suspected conformational dynamics of the stator, we have utilized site‐directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy to observe backbone mobility at several points in subunit a. We demonstrate the feasibility of membrane extraction using mixed micelles of dodecylmaltoside/phosphatidylcholine or styrene maleic acid lipid particles (SMALPs). Furthermore, we found high backbone mobility at sites on the cytoplasmic surface of the subunit a stator.
Low output syndrome after cardiac surgery is mediated through extra corporal circulation induced RAGE activation
Authors: Velten Markus , Hilbert Tobias , Dewald Oliver , Meyer Rainer , Duerr Daniel
Published: May 2022

Objective

Coronary artery bypass grafting (CABG) is a surgical procedure routinely performed using extracorporeal circulation (ECC). This on‐pump CABG is known to be associated with severe postoperative organ failure including cardiac dysfunction and low cardiac output syndrome. Mechanisms inducing cardiac dysfunction are complex but may include on‐pump CABG induced inflammation due to exposure of the blood to the bypass circuit, potentially increasing serum concentrations of inflammatory mediators. High‐mobility group box 1 (HMGB1) functions as an inflammatory mediator and has been shown to induce cardiovascular dysfunction in sepsis and impair cardiomyocyte contractility in experimental models. However, the impact of HMGB1 on on‐pump CABG induced myocardial dysfunction is not known. Therefore, we tested the hypothesis that on‐pump CABG associated increases in serum concentrations of inflammatory mediators induce cardiomyocyte dysfunction in vitro and investigated potential mechanism including the role of the HMGB1‐RAGE (receptor for advanced glycation end products) axis.

Approach

After institutional revenue board approval, informed consent, and IACUC approval, serum samples (n = 18) were collected from CABG patients pre, 1, 6, and 24 h after aortic de‐clamping. TNFa, IL‐1b, IL‐6, HMGB1, and sRAGE serum concentrations were analyzed by ELISA. Primary murine cardiomyocytes were incubated with patient serum and single cell contractility was assessed at frequencies between 0.5 and 10Hz using an optical contractility system (IonOptics LLC, Westwood, MA).

Results

IL‐6, HMGB1, and sRAGE serum levels were increased after CABG (Figure 1). In preliminary studies we incubated murine cardiomyocytes with human serum collected from healthy volunteers and determined the highest serum concentrations that did not affect cardiomyocyte contractility. Cardiomyocytes were incubate with human serum and serum collected pre‐surgery had no effects on murine cardiomyocyte contractility. However, primary murine cardiomyocytes incubated with serum collected 1h and 6h after aortic de‐clamping showed a significant decrease in % peak shortening. Pre‐incubation with the RAGE antibody FPS‐ZM1 (Cayman, Hamburg, Germany) prevented the serum induced cardiomyocyte dysfunction (Figure 2).

Conclusion

on‐pump CABG increased protein serum concentrations of inflammatory cytokines including HMGB1. HMGB1 has been shown to cleave and activate RAGE. sRAGE serum levels have been increased after on‐pump CABG. Deactivating RAGE with the high‐affinity inhibitor FPS‐ZM1 prevented serum induced cardiomyocyte dysfunction, suggesting that the HMGB1‐RAGE axis is mediating on‐pump CABG associated low‐cardiac‐output syndrome.
4‐Aminopyridine promotes cutaneous reinnervation and accelerates wound healing
Authors: Jagadeeshaprasad M G , Govindappa Prem Kumar , Nelson Amanda , Noble Mark D. , Elfar John
Published: May 2022

Skin wound healing is a tightly orchestrated process between epithelial, dermal, neuronal and vascular cells, all working in concert to restore tissue integrity and skin barrier function. Here, we report the therapeutic regenerative effects of FDA‐approved 4‐aminopyridine (4‐AP) to promote skin wound healing by accelerating re‐epithelialization, wound induced hair neogenesis (WIHN), dermal collagen deposition, angiogenesis, and reinnervation. We demonstrate that 4‐AP enhanced wound closure by promoting keratinocyte proliferation and migration as well as collagen deposition through myofibroblast differentiation. 4‐AP treatment increased both epidermal thickness, as well as the number of hair follicles and blood vessels in healed wounds, suggesting better restoration of skin function and architecture compared to simple wound scar formation. Moreover, 4‐AP enhanced reinnervation of healed wounds by promoting Schwann cell (SC) de‐differentiation and secretion of neuromediators (NGF, SP) associated with regeneration. In‐vitro studies using human cells demonstrated that 4‐AP enhanced proliferation and migration of keratinocytes, and SCs and that 4‐AP enhances cellular interactions between neuronal and non‐neuronal cells to further accelerate wound healing. 4‐AP induced secretion of NGF from both SCs and keratinocytes. 4‐AP enhances many of the key attributes of successful wound healing and is a promising therapeutic adjuvant for skin wound healing and tissue regeneration.
Effect of adrenalectomy and insulin resistance on non‐shivering thermogenesis in the corpulent rat
Authors: Tulp Orien L. , Einstein George P. , Awan Aftab R.
Published: May 2022

Glucocorticoid hormones have been reported to impede optimal insulin sensitivity and glucose uptake in peripheral tissues via interactions with GLUT transporters, leading to insulin resistance and impaired glucose uptake via GLUT transporters. The LA/Ntul//‐cp rat expresses obesity in 25% of the offspring by 5 to 6 weeks of age but remains non diabetic throughout much if not all of its lifespan when normally reared. Groups of congenic lean and obese LA/Ntul//‐cp rats were fed low glycemic Purina Chow diets from 6 to 9 weeks of age or overfed with a high energy cafeteria diet (Café) from 9 until 12 weeks of age. Subgroups of obese animals were adrenalectomized (ADX) at 6 weeks of age (WoA) and subjected to the same dietary regimen to determine if the Café overfeeding could enhance the normal thermic response to norepinephrine (NE) in the obese phenotype and to determine if ADX would result in a restoration of glycemic status and NE‐induced thermogenesis in the obese phenotype. Measures of Oral Glucose Tolerance (OGT), Area under the glucose curve (AUC), and NE‐stimulated VO2 were determined (NE, 200 µg, s.c.) at 6, 9 and 12 WoA in all treatment groups and Insulin:Glucose ratios (I:G) at 12 WoA. Obese animal demonstrated hyperphagia at all ages studied, and Obese ADX hyperphagia from 9 to 12 WoA. OGT and AUC indicated modest glucose intolerance in the obese phenotype, and which was restored following ADX at 6 and 9 WoA. NE resulted in a progressive increase in VO2 in lean rats at each age studied, but NE‐stimulated VO2 was lower in the obese phenotype and decreased further after the Café feeding regimen. ADX was associated with a restoration of OGT, I:G and NE stimulated VO2 equivalent to that observed in lean animals at 12 weeks of age. These results are consistent with adrenocorticoid‐mediated insulin resistance likely due to a receptor level defect in insulin sensitivity as a contributory factor in the impairment in OGT and NST in the obese phenotype of this strain.
Mnemonics Improve Pharmacy Students' Knowledge Retention, Clinical Application, and Exam Performance in a PharmD Pharmacology Course
Authors: Munusamy Shankar , Koenigsfeld Carrie , Torry Ronald
Published: May 2022

Background

Students in healthcare professions taking pharmacology courses often feel challenged with learning and remembering crucial facts about medications, i.e., knowledge acquisition and retention. In addition, applying pharmacological knowledge into clinical settings consumes significant amounts of the student’s working memory (i.e., cognitive load). Mnemonics (memory aids) are used to reduce cognitive load and facilitate learning and knowledge retention among students. Our study assessed the impact of the mnemonics' use in a Pharmacology course offered to second‐year pharmacy students on students' perceptions of knowledge acquisition, retention, and clinical application in correlation to their exam performance.

Methods

Eighteen mnemonics were developed and used in a course covering endocrine and autonomic pharmacology topics. Students' perceptions of the impact of mnemonics' use on their knowledge acquisition, retention, clinical application, critical thinking, and confidence while answering questions on exams were collected using an anonymous survey in the fall semester of 2020. Further, students' perceptions were matched to their exam performance.

Results

Survey responses from 56 students (85% response rate) indicated that 80.4% agreed that mnemonics' use facilitated knowledge acquisition by reducing anxiety while learning, and 98.2% of students indicated that the use of mnemonics improved knowledge retention and application. Mnemonics' use was also reported to improve critical thinking while answering exam questions (89.3% agreed) and increased confidence during test‐taking (96.4% agreed). Interestingly, 64.7% of students using mnemonics answered the associated exam questions correctly compared to 35.3% of students (p<0.05) who did not use the mnemonic aids.

Conclusions

Our study suggests that mnemonics' use in pharmacology courses not only improved students' perception of knowledge acquisition, retention, application, and critical thinking but also improved their exam performance on specific questions that tested concepts taught with mnemonics. Future studies will assess students' knowledge retention and perceptions of mnemonics' use with exam performance in subsequent pharmacotherapy and pharmacy skills courses.
Does estrogen show neuroprotective effects on hypothalamic cells when induced by beta amyloid?
Authors: Henderson Kynadi , Ibragimov Angela , Gomez Roberto , Chakraborty Sanjoy , Chakraborty Tandra
Published: May 2022

Alzheimer’s disease (AD) is a neurodegenerative disorder that results in cognitive decline and eventual dementia. The two most notable pathological hallmarks of AD include beta‐amyloid plaques, formed from the extracellular aggregation and deposition of beta‐amyloid fibrils and the intracellular accumulation of hyperphosphorylated tau as neurofibrillary tangles in the brain. Estrogen has shown to have neuroprotective effects on AD in the hippocampus. The literature is unclear of the role of the hypothalamus in AD although early pathophysiology of AD, such as disturbances in sleep and emotional affect has been observed. This study is designed to replicate AD models using a control cell line, SH‐SY5Y human neuroblastoma cells and an experimental cell line, N38 murine hypothalamic cells in culture by incubating these cells with oligomerized human beta‐amyloid 42. Each cell line is then treated with estrogen in order to observe the hormone’s efficacy as a neuroprotective agent. Cell viability, cytotoxicity and proliferation were measured in order to determine estrogen’s neuroprotective potential, using a variety of bioassays such as MTT and LDH. Preliminary results show lower mitochondrial activity and higher rates of cell lysis when treated with physiologically relevant concentrations of estrogen and low beta‐amyloid concentrations in the SH‐SY5Y cell line. Treatment on N38 cells with beta‐amyloid 42 treated with estrogen at dosages similar or higher than physiologically concentrations, will be done at various time points. Cell lysis and cytotoxicity of beta‐amyloid in N38 cells will be tested in order to further elucidate the pathology of the hypothalamus in AD, as well as estrogen’s potential neuroprotective effect on this unexplored brain region.
Profiling Growth Patterns of Human Gut Microbes in Response to Preterm Human Milk and Formula
Authors: Engevik Mindy A. , Stripe Leah A. , Baatz John E. , Wagner Carol L. , Chetta Katherine E.
Published: May 2022

Background

The infant gut microbiota undergoes significant fluxes in microbial composition during the first few years of life. The intestine is colonized shortly after birth with facultative anaerobes, like as Enterococcus, Enterobacter, Streptococcus, and Staphylococcus, which reduce the intestinal oxygen content and create niches for strict anaerobes, like Bifidobacteria, Bacteroides, and Clostridia. The dominant factors which influence the infant gut composition are gestational age and diet. The gut microbiota of preterm infants is characterized by limited microbial diversity and increased levels of facultative anaerobes such as Enterococci, Staphylococci, and Enterbacteriacea (Enterobacter, Escherichia, and Klebsiellaspp.). The gut microbiota is also highly influenced by diet, specifically formula and human milk use. The intake of human milk is associated with increased Bifidobacteria, while the intake of formula correlates with increased Enterobacteriaceae. This study aimed to analyze next‐generation products including preterm formula, human milk‐oligosaccharide term formula, and preterm breastmilk using a culture‐based model.

Methods & Results

We cultured 8 different Bifidobacteria, 8 Lactobacilli, 8 Streptococcus, 3 Enterococci and 4 Klebsiella strains in a fully defined bacteria media and monitored growth and pH after 20 hrs of anaerobic incubation. Significant differences were identified in strain growth and culture pH between human milk and formula. Human milk significantly elevated the growth of all Bifidobacteria and Lactobacilli. In contrast, human milk supported the growth of three Streptococci and only nominally supported pathobiont Enterococci and Klebsiella strains. Preterm formula elevated the majority of Bifidobacteria and Lactobacilli, as well as several Streptococci. Interestingly, preterm and full‐term formula significantly enhanced the growth of pathobiont strains. Full‐term formula only supported the growth certain commensal microbes, which was in marked contrast to human milk and preterm formula. Using computational modeling, a correlation was observed with human milk oligosaccharide (HMO) degrading glycosyl hydrolase profiles in the genomes of commensal microbes with human milk, but not with formula; confirming the HMO preference of our representative infant commensal microbes.

Conclusions

These findings indicate a unique profile of growth in response to human milk and formula. Since alterations in the infant gut microbiota have linked to childhood health and development, understanding how bacteria differentially utilize whole milk or formula may provide insights into which microbes drive diet‐related microbiota shifts.
A Mathematical Model of the Rat Kidney IV: Whole‐Kidney Response to Hyperkalemia
Authors: Weinstein Alan M.
Published: May 2022

The renal response to hyperkalemia is mediated by increased K secretion within connecting tubule (CNT), flux that is modulated by tubular effects (e.g. aldosterone) in conjunction with increased luminal flow. It is established that peritubular K blunts Na reabsorption in proximal tubule, thick ascending Henle limb, and distal convoluted tubule (DCT). While any such reduction may augment CNT delivery, the relative contribution of each is uncertain. The kidney model of this lab was recently advanced with representation of cortical labyrinth and medullary ray. This introduced additional communication between proximal and distal nephron, in addition to tubuloglomerular feedback (TGF). Model tubules capture the impact of hyperkalemia to blunt Na reabsorption, and augment luminal flow within each upstream segment. However, this forces the question of the extent to which increased Na delivery is transmitted past macula densa and its TGF signal. Beyond the effect of high luminal Na, peritubular K is predicted to raise cytosolic Cl and depolarize macula densa cells, so that it may also activate TGF. Thus, even though upstream reduction in Na transport may be larger, it appears that the DCT effect is critical to increasing CNT delivery. Beyond the flow effect, hyperkalemia reduces ammoniagenesis and reduced ammoniagenesis can enhance K excretion. What this model provides is a possible mechanism. When cortical NH4 is taken up via peritubular Na,K(NH4)‐ATPase, it acidifies principal cells. Consequently, reduced ammoniagenesis is expected to increase principal cell pH, thereby increasing conductance of both ENaC and ROMK, further increasing K excretion.
The Magnitude of Hidden Hunger and Cognitive Deficits of Children Living in Some Selected Orphanages in Kumasi, Ghana During the COVID Pandemic
Authors: Asamoah Mary , Tandoh Marina
Published: May 2022

Many studies have assessed the magnitude of mixed micronutrient deficiencies or individual micronutrient deficiencies among children under 5 years, women of reproductive age (15‐ 49 years old) and pregnant women. This has led to various interventions for these population groups including supplementations, fortifications etc. However, the same attention has not been given to vulnerable children living in various orphanages, especially in Children’s Homes in Ghana where much is not known about their nutritional status. Socio‐ economic downturns like that induced by the current coronavirus pandemic affects food security and nutrition, thus the nutritional status of this vulnerable population could potentially be worsened.

Objective

This study assessed the magnitude of hidden hunger and cognitive deficits of 130 children (6‐ 13 years old) living in three selected orphanages in Kumasi, Ghana.

Methodology

Sociodemographic data, anthropometric assessment (BMI‐for‐age and height‐for‐age), dietary assessment (3‐day repeated 24‐ hour dietary recall), urinary iodine levels and cognitive performance (Raven’s Coloured Progressive Matrices) were assessed.

Findings

Boys formed 50.8% of the study population while girls formed 49.2%. The median age of participants was 10.50 years. About 12.3%, 7% and 10.0% of participants were stunted, thin and overweight/ obese respectively. Prevalence of mild iodine deficiency (i.e. 50‐ 99 μg/L) was 16.2%. Iodine deficiency was significantly higher (16.2%) in participants who had lived for at least 7 years in the orphanage compared to those who had lived less than 7 years (23.6%) (p = 0.047). About 17% of participants performed poorly (< 50%) in the cognition test. Mean cognitive test scores were significantly different among the orphanages (p = 0.027). Majority of participants, 89.2%, 54.6%, 76.9% and 77.7% had adequate intakes for iron, zinc, vitamin C and folate respectively whereas intake of vitamins A and B12 were inadequate for majority of participants (90.8% and 50.8% respectively). There was no significant correlation between micronutrient intake and cognitive performance. However, mean cognition test scores were significantly different between participants who had adequate and inadequate intake of iron and vitamin A (p = 0.007 and p < 0.001 respectively).

The significance of the research is to provide data for relevant policy makers and stakeholders involved in the care of orphans and vulnerable children in Ghana like the Social Welfare Department. It is also expected that this research will stimulate further studies in the occurrence of multiple micronutrient deficiencies in vulnerable children since nutrition is not only necessary for physical development, but also for cognitive development and life‐long health (WHO, 2016).

Conclusion

Findings of this study warrant a closer look at nutritional intakes in orphanages to improve on hidden hunger.
Evidence for widespread cytoplasmic structuring into mesoscopic condensates
Authors: Wühr Martin , Keber Felix , Nguyen Thao , Brangwynne Cliff
Published: May 2022

Eukaryotic cytoplasm organizes itself via both membrane‐bound organelles and membrane‐less biomolecular condensates (BMCs). Known BMCs exhibit liquid‐like properties and are typically visualized on the scale of ~1 µm. They have been studied mostly by microscopy, examining select individual proteins. Here, we investigate the global organization of native cytoplasm with quantitative proteomics, using differential pressure filtration, size exclusion, and dilution experiments. These assays reveal that BMCs form throughout the cytosplasm, predominantly at the mesoscale of ~100 nm. Our data indicate that at least 18% of the proteome is organized via such mesoscale BMCs, suggesting that cells widely employ dynamic liquid‐like clustering to organize their cytoplasm, at surprisingly small length scales.
Brain Serotonin Deficiency Increases Susceptibility to Stress‐Induced Increases in Binge Drinking in Females, but not Males
Authors: Sachs Benjamin , Dimitratos Elisabeth , Waltrip Leah H. , Bavley Charlotte , Karbalaei Reza , Wimmer Mathieu
Published: May 2022

Approximately 30 million Americans currently suffer from alcohol use disorders (AUDs), which cost the US economy over $249 billion annually. Men are more likely than women to have an AUD, but rates of AUDs in women are rising rapidly, and women tend to experience more negative health consequences from excessive alcohol consumption than men. The causes of gender disparities in AUDs and alcohol responses remain unknown, but stress has been associated with the development of AUDs, particularly in women, and genetic factors are thought to increase AUD risk. Although the specific genetic factors that confer AUD risk have not been conclusively established, dysfunction of the brain serotonin (5‐HT) system has been hypothesized to play a role. Based on our published work demonstrating that brain 5‐HT deficiency increases susceptibility to stress, we hypothesized that low brain 5‐HT would predispose animals to excessive alcohol consumption following stress. Our unpublished findings demonstrate that chronic restraint stress increases binge‐like alcohol consumption in the drinking in the dark paradigm in 5‐HT‐deficient females but not in WT females or in males of either genotype. In contrast, in the absence of stress, low 5‐HT increased binge‐like drinking in males only. The precise molecular mechanisms underlying this enhanced susceptibility of 5‐HT‐deficient females to stress‐induced binge drinking have not been conclusively established, but RNA sequencing studies followed by gene ontology enrichment analysis reveals that chronic stress leads to dysregulation of several gene sets involved in serotonin signaling and alcoholism in 5‐HT‐deficient females, but not in WT females. These data provide new insight into the mechanisms through which low 5‐HT impacts stress‐sensitive behaviors in a sex‐dependent manner.
Intentional Innovation in Educational Technology and Media to Promote Students’ Holistic Development
Authors: Lee Michael , Witchel Harry
Published: May 2022

We consider how educational technology might promote students’ holistic development. Based on a new edited book in the Methods in Physiology series, we contextualize how far technology has taken bioscience education, and also speculate on where technology needs to go. Technology, in conjunction with innovations in pedagogy, has profoundly influenced the experience of teaching and learning biology, and examples are provided, some familiar (e.g., video lectures in flipped classroom), and some emerging. Compared to the personal changes technology has wrought in students' experiences, past innovations in technology and pedagogy have led to group outcomes more weighted toward inclusivity, opportunities, and participation rates. This is welcome progress. For the future, we propose an intentional development of educational technology and media that supports holistic development of students as individuals. This goes beyond design‐centered or market‐driven research and focuses on student competencies and skills known to play a role in learning and professional maturity: social, metacognitive, and non‐cognitive competencies such as team work skills, social behaviors, empathy, emotional regulation, and professionalism. The challenge is to use intentional innovation to achieve student learning of competencies, by leveraging what computers are particularly good at: timing, repetition, and vigilance. The expected result will probably involve learning with repetition, deeper learning with models, collaborative and cooperative learning, and self‐regulated learning. We engage in serious discussion on the challenges and obstacles inherent in re‐focusing media creation and educational technology innovation. We propose that intention should be the guiding approach toward a cohesive narrative of future developments.