bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impact of Krill Oil Supplementation on Neuroinflammation and Spatial Memory in Aged Rats
Authors: Andraka John M. , Graham Eldon , Szczesny David , Marchalant Yannick , Sharma Naveen 
Published: May 2022

ObjectiveInvestigate the potential of dietary krill oil (KO), a marine‐based, long chain, omega‐3 (n‐3) polyunsaturated fatty acid (PUFA), to negate the deleterious effects of high fat diets and aging on neuroinflammation and spatial learning in a rat model.Background/HypothesisKO exhibits anti‐inflammatory properties comparable to other sources of n‐3 PUFA, but with several biochemical advantages. Phospholipid‐bound fatty acids in KO exhibit improved bioavailability when compared to triacylglyceride bound n‐3 PUFA such as fish oil. Moreover, KO contains astaxanthin, a powerful antioxidant that counters harmful oxidative stress in the brain, as well as choline which is an essential neurotransmitter precursor with neuroprotective properties. Hence, we hypothesized that KO supplementation would improve deficiencies in spatial learning and elevated neuroinflammatory protein levels induced by high fat diet and normal aging in rats.MethodsThirty‐two male, Sprague Dawley rats, aged 15 months were divided into four dietary groups consisting of varying amounts of saturated fats, with and without krill oil supplementation: Control (CON), Control with krill oil (CONKO), High fat (HF), High fat with krill oil (HFKO). HF and CON experimental diets consisted of 60% and 23% calories from saturated fats, respectively. Food intake and body weight were assessed weekly. After 12‐weeks of dietary supplementation, Morris Water Maze (MWM) testing was conducted to compare spatial learning abilities between groups. Moreover, inflammatory cytokine concentrations, including interleukin (IL)‐1β, IL‐4, IL‐6, IL‐10 and tumor necrosing factor‐α (TNF‐α), were analyzed using multiplex bead‐based protein assays.ResultsDespite evidence of learning by each group over the course of MWM testing, no significant difference between dietary groups in reaching the platform was identified. Furthermore, no statistically significant difference in inflammatory cytokines was identified. Total food consumption was significantly less in the HF group when compared to CON (p=.047) and CONKO (p=.030), whereas body weight was significantly higher in the HFKO group when compared to CON (p=.024) and CONKO (p=.030). Otherwise, no significant difference in food consumption or body weight was identified.ConclusionsKO supplementation did not significantly improve the effects of aging and high fat diet consumption on spatial memory or neuroinflammatory signaling proteins. Some evidence suggests that the unique properties of KO, a more novel source n‐3 PUFA, have the potential to decrease neuroinflammation and improve cognition. Indeed, considering an aging population, the proliferation of western‐style diets, and the associated increase in neurodegenerative diseases such as Alzheimer’s disease, there is a need for preventive interventions. Further investigations are still needed to weigh the potential benefits of n‐3 dietary supplementation, aging, obesity, and neurodegenerative disease.
Cyclosporine A but not tacrolimus induces pro‐apoptotic endoplasmic reticulum stress in renal tubular cells
Authors: Yilmaz Duygu E. , Kirschner Karin , Demirci Hasan , Himmerkus Nina , Bachmann Sebastian , Mutig Kerim 
Published: May 2022

Calcineurin inhibitors (CNI), cyclosporine A (CsA) or tacrolimus (Tac), belong to the first‐line immunosuppressive strategies in organ‐transplanted patients. Both CNI may exert renal side effects, which are typically stronger in patients receiving CsA. Continued clinical demand for CsA requires improved understanding of its nephrotoxicity. Calcineurin inhibition by CsA occurs via complexes with cyclophilins, whereas Tac recruits FKBP12 instead. We hypothesized that impaired chaperone function of cyclophilins may aggravate CsA cytotoxicity due to induction of endoplasmic reticulum (ER) stress and pro‐apoptotic unfolded protein response (UPR). To address this hypothesis, effects of CsA vs. Tac (10 µM for 6 h) on the UPR signaling were evaluated in cultured human embryonic kidney cells (HEK293), human renal proximal tubular (PT) epithelial cells (HRPTEpC), and isolated rat PTs using quantitative PCR, immunoblotting, and immunofluorescence staining. An established ER stress inducer, thapsigargin (Tg) served as a positive control. CsA and Tg, but not Tac, induced ER stress and pro‐apoptotic UPR in HEK293 cells, which was reflected by increased levels of key UPR products (BiP, CHOP, spliced XBP1, and phosphorylated IRE1a) and cleaved caspase 3 (cCas3). Similar effects were observed in HRPTEpC cells and isolated rat PTs. Knockdown of cyclophilin A or B isoforms in HEK293 cells using siRNA augmented CHOP and cCas3 to an extent comparable with CsA treatment suggesting relevance of cyclophilin activity for intact proteostasis. Application of chemical chaperones, TUDCA or 4‐PBA, alleviated the CsA‐induced UPR suggesting that boosting the protein folding may have protective effects against CsA cytotoxicity. Along the same line, genetic suppression of UPR in HEK293 cells by CRISPR/Cas9‐mediated deletion of the key ER stress sensors, PERK or ATF6, blunted the pro‐apoptotic UPR in response to CsA.In summary, these results suggest that nephrotoxic effects of CsA are partially mediated by suppression of cyclophilins, ER stress, and pro‐apoptotic UPR. Pharmacological modulation of UPR bears the potential to alleviate CsA nephrotoxicity.
Obeticholic Acid Prevents Fibrosis in a Model of Tubulointerstitial Kidney Disease
Authors: Davidson Shania R. , Adapa Sharmila , Lopez‐Santiago Isabel , Dial Katelyn , Rowland Emma , Mykala Komuraiah , Wang Xiaoxin , Rosenberg Avi , Levi Moshe , Jones Bryce A. 
Published: May 2022

ObjectiveThis study investigates the protective properties of farsenoid X receptor (FXR) agonism by the drug Obeticholic Acid (OCA) in the adenine‐induced tubulointerstitial fibrosis model of kidney disease male and female mice.HypothesisOCA treatment will decrease the expression of SMAD3 and phosphorylated‐SMAD3, and therefore will be nephroprotective in the adenine model of kidney disease in male and female mice.MethodsMale and female C57BL/6J mice (12 weeks old) were fed chow (Research Diets D1912040li) or chow admixed with adenine (0.2% w/w) ad lib for 7 weeks. Mice were treated with either vehicle (corn oil) or obeticholic acid (10 mg/kg BW) by gavage 5 days per week. Plasma and organs were collected and processed for bio‐chemical analysis.ResultsOCA treatment reduced BUN and plasma creatinine in diseased mice of both sexes. FXR agonism reduced SMAD3 expression in both male (P<0.05) and female (P<0.05) mice. Importantly, FXR agonism also reduced SMAD3 phosphorylation in both male (P<0.05) and female (P<0.001) mice. On histology, OCA reduced renal fibrosis in male (P<0.05) but not female mice.ConclusionsFXR agonism by OCA is anti‐fibrotic in the adenine model of tubulointerstitial fibrotic crystalline nephropathy in male and female mice. Mechanistically, OCA prevents renal fibrosis by reducing SMAD3 expression and phosphorylation.
Harnessing antibacterial properties of biomaterials for fabricating wound healing patch
Authors: Masoud Abdul , Mills David K. 
Published: May 2022

IntroductionStudies aimed at elucidating the role of infections caused by bacterial colonization in wound healing are gaining increasing recognition. Proper management of wound infections purposely aimed at keeping low levels of bacteria on wound surfaces is pivotal in enhancing the wound healing process and preventing local and systematic infections. In this research, biomaterials with inherent antimicrobial properties were exploited for fabricating a wound healing composite with an improved potential of reducing or completely inhibiting bacterial growth. The use of biomaterials with inherent antibacterial properties was necessitated by the desire to reduce the incorporated antibiotic load, thus reducing the risk of development of resistance to antibiotics by these bacteria.MethodsCarboxymethyl cellulose (CMC), one of the most common derivatives of cellulose, chitosan (CTS), a natural, biodegradable, non‐toxic polymer that demonstrates diverse biological properties, metalized and non‐metalized halloysites (HNT & mHNT) as well as variants loaded with antibiotics (LHNT, LmHNT) were tested for antimicrobial properties against E.coli at concentrations of 2mg/ml and 0.5mg/ml in 3 repeats. CTS and CMC were then physically cross‐linked and impregnated with halloysites and metal coated halloysites. A Fourier‐transform infrared spectroscopy (FTIR) characterization test was then carried out to validate the cross‐linking of the polymers. The biocompatibility of the fabricated composite was assessed by studying the proliferation rate of human dermal fibroblasts treated with the test samples.Results2mg/ml and 0.5mg/ml of the biomaterials tested all inhibited growth of E. coli with metal‐coated halloysites loaded with antibiotics exhibiting the most antimicrobial effect. CTS and CMC both exhibited characteristic absorption peaks on the FTIR spectra which conforms with data reported in other studies, with cross‐linking between the two polymers characterized by broadening of the bands at 3400–3301 cm−1. The proliferation assay showed that all the samples did not inhibit fibroblast cell growth with the exception of CTS‐CMC composites incorporated with 5% HNT and 5% metalized HNT.DiscussionCTS and CMC composites incorporated with HNTs have the potential to effectively suppress microbial growth when used to fabricate wound dressings. The dressing is easy to fabricate, and its components are tunable from a personalized medicine perspective.
Temporal Development of Cardiometabolic Complications in A Hyperandrogenemic Model of Polycystic Ovary Syndrome
Authors: Pruett Jacob E. , Everman Steven , Romero Damian G. , Yanes Cardozo Licy L. 
Published: May 2022

IntroductionPolycystic Ovary Syndrome (PCOS) is the most common endocrine disorder in young women, being characterized by androgen excess and irregular ovulation. Insulin resistance (IR), dyslipidemia, obesity, and hypertension are highly prevalent in this population. The etiology of PCOS is unknown. Most patients begin developing symptoms around puberty. However, there is a delay in the diagnosis of PCOS, so the metabolic characteristics of PCOS early on are unclear. We have demonstrated in a hyperandrogenemic PCOS rat model that obesity, increases in blood pressure, and IR are present after 3 months of androgen excess. Using this model, we tested the hypothesis that androgen‐induced IR, obesity, and dyslipidemia would develop in a time‐dependent manner.MethodsFour‐week‐old Sprague Dawley female rats were randomly selected for sham or dihydrotestosterone pellet (DHT, 7.5mg/90 days) implantation (n=13‐17/group). Body composition was measured by EchoMRI; 24‐hour urine was collected with metabolic cages; and fasting plasma glucose, insulin, and lipid panel were collected at 1 (early), 2 (intermediate), and 3 (late) months of DHT treatment. IR was assessed by homeostatic model assessment for IR (HOMA‐IR).ResultsAfter 1 month of DHT, PCOS rats had decreased fasting glycemia (122.0 ± 2.8 vs 133.6 ± 3.1 mg/dL, P<0.01) and insulinemia, leading to decreased HOMA‐IR (4.3 ± 0.4 vs 7.4 ± 0.5, P<0.001). However, there were no differences between the groups in IR at 2 months of DHT, while at 3 months both fasting insulin (21.0 ± 1.6 vs 14.2 ± 1.4 µU/mL, P<0.01) and HOMA‐IR (6.3 ± 0.5 vs 4.3 ± 0.5, P<0.05) increased in PCOS compared to controls. Low‐density lipoprotein cholesterol (LDL) was increased in PCOS compared to controls at all time points, while triglycerides were only elevated at 2 months (84.9 ± 5.4 vs 65.1 ± 2.9 mg/dL, P<0.01) and 3 months (70.7 ± 5.3 vs 54.5 ± 4.5 mg/dL, P<0.05) of DHT. Meanwhile, there were no differences in high‐density lipoprotein cholesterol throughout the study. Lean mass was elevated at all time points in PCOS compared to controls, though fat mass was only elevated after 2 and 3 months (21.6 ± 1.9 vs 16.7 ± 0.8 g, P<0.05) of DHT. Proteinuria, a marker of renal injury, was elevated in PCOS at all time points.ConclusionsIn this PCOS model, early cardiometabolic complications include increases in LDL and renal injury. Intermediate cardiometabolic complications include increases in fat mass and triglyceridemia. However, DHT had a differential effect on fasting insulin and HOMA‐IR by lowering them initially at 1 month of treatment and then gradually increasing them above control levels by 3 months of DHT. These data indicate that androgen excess has a differential effect in cardiometabolic complications with time in female rats. In women with PCOS, these data may lead to a better characterization, and more effective treatments, for the evolving metabolic profile in women with PCOS.
Peripheral Chemoreflex Sensitivity is Augmented in Human Type 2 Diabetes
Authors: Lis Eric C. , Ott Elizabeth P. , Harper Jennifer L. , Manrique‐Acevedo Camila , Limberg Jacqueline K. 
Published: May 2022

IntroductionThe peripheral chemoreceptors have been implicated in the pathogenesis of type 2 diabetes. Our main objective was to examine peripheral chemosensitivity in individuals with type 2 diabetes, as well as the contribution of the peripheral chemoreceptors to basal cardiovascular control. We hypothesized individuals with type 2 diabetes exhibit exaggerated peripheral chemoreflex sensitivity to hypoxia compared to healthy, non‐diabetic controls. We further hypothesized the peripheral chemoreceptors contribute to cardiovascular dysfunction in type 2 diabetes.MethodsEight adults with diagnosed type 2 diabetes (3M/5F, 54±3 yrs, HbA1c 8.1±0.6%) and nineteen healthy controls (8M/11F, 50±3 yrs, HbA1c 5.3±0.1%) participated (IRB #2010295 and #2018486). Ventilatory and hemodynamic responses to acute hypoxia were assessed to determine peripheral chemosensitivity [i.e., hypoxic ventilatory (HVR), heart rate (HHRR) and blood pressure (HBPR) response]. Separately, heart rate, blood pressure and forearm blood flow (Doppler ultrasound) were measured while breathing normoxic or hyperoxic air (100% O2; Modified Dejours test) to acutely attenuate peripheral chemoreceptor activity. Forearm blood flow was normalized for mean arterial blood pressure for measures of vascular resistance.ResultsPeripheral chemosensitivity was augmented in adults with type 2 diabetes compared to control as assessed via the HVR (‐1.10±0.16 vs ‐0.50±0.10 L/min/%, p<0.01) and HBPR (‐0.92±0.17 vs ‐0.42±0.19 mmHg/%, p=0.06), with no differences in HHRR (‐0.68±0.22 vs ‐0.56±0.12 beats/min/%, p=0.64). Higher HVR and HBPR were associated with higher fasting insulin (R=‐0.58, p<0.01 and R=‐0.38, p=0.06) and HbA1c (R=‐0.60, p<0.01 and R=‐0.39, p=0.05). Attenuation of peripheral chemoreceptor activity with acute hyperoxia decreased heart rate (p<0.01) and blood pressure (p=0.02), with no effect on forearm vascular resistance (p=0.55); any effect of hyperoxia did not differ between adults with type 2 diabetes and controls (p>0.05).ConclusionIndividuals with type 2 diabetes exhibit exaggerated peripheral chemoreflex sensitivity which is associated with disease severity (i.e., HbA1c). In contrast, the contribution of the peripheral chemoreceptors to basal cardiovascular function in type 2 diabetes does not differ from non‐diabetic controls. Results advance our understanding of the pathogenesis of type 2 diabetes and related complications.
Quantifying flow‐induced transport of membrane‐linked proteins in model and cell membranes
Authors: Pash Samuel , Ankrom Emily , Anthony Autumn , Ratajczak Amanda , Honerkamp Smith Aurelia , Thevenin Damien 
Published: May 2022

Shear flow sensing is crucial in defining fundamental cell properties, including cell fate, self‐renewal, motility, and homing behaviors. For instance, endothelial cells lining blood vessels translate blood flow into responses that in turn regulate the tension in blood vessels, and, in tumors, flow can regulate genes that promote cancer cell migration and metastasis.Recent studies suggest that glypican‐1 (a lipid‐anchored heparan sulfate proteoglycan at the surface of endothelial cells) may play a key role in the flow signaling pathway: its flow‐mediated redistribution at the membrane appears to mediate the production of nitric oxide (NO), a key signaling molecule produced by the NO synthase eNOS. However, the molecular mechanisms that endothelial cells use to sense changes in flow and translate this mechanical stimulus into intracellular molecular responses (e.g., activation of eNOS by glypican‐1) are currently unknown. We hypothesize that protein concentration gradients in the plasma membrane are formed by physical interactions with fluid flow. This is based on the premise that extracellular lipid‐anchored proteoglycans like glypican‐1 can be transported along the membrane by external flow, with the aqueous part of the protein acting as a molecular sail.To investigate the effect of flow on membrane‐associated proteins and cell signaling, it is crucial to visualize and quantify the motility of membrane bound proteins in response to flow. Here, we present results in both reductionist model membrane systems and cell membranes aimed at determining the relationship between drift velocity and protein size, membrane lipid composition, and lipid anchor structure: (1) Our first approach measures the flow coupling and frictional coefficients of lipid‐anchored model proteins (monomeric streptavidin fusions) of varying size reconstituted in glass‐supported bilayers. (2) We also present the development of a unique labeling approach allowing the visualization of glypican‐1 expressed at the membrane of living endothelial cells under flow.While glypican‐1 is specific to endothelial cells, the principles of fluid mechanics are universal. We, therefore, anticipate that our methodology and results will apply to multiple cell lines and flow conditions.
Modelling the Applications of CRISPR‐Cas9 for Treating Sickle Cell Disease
Authors: Cooper Grace , Akteke Elfin , Dalvi Viggo , Delage Laura , Feberova Natalie , Fujikawa Kento , Klousseh Edem , Manoharan Harish , Mcdaniel Kieran , Powter Morgan , Seyoum Michael , Greig James , MacGillivray Brendan 
Published: May 2022

Clustered Regularly Interspaced Short Palindromic Repeats or CRISPR‐associated protein 9 (CRISPR‐Cas9) is a genome editing tool that can remove, modify, or add sections of DNA. It is made up of two main molecules: the Cas9 protein and the guide‐RNA (gRNA), which is used to match a target gene to the CRISPR‐Cas9 molecule. The desired sequence of DNA must be between 2 and 5 nucleotides and must be followed by the protospacer adjacent motif (PAM) at the 3’ end of the gRNA. This enables the target gene to be found and cut by the Cas9 endonuclease. Once cut, the DNA can be repaired using non‐homologous end joining (where a random insertion or deletion of DNA occurs) or homology directed repair (where a homologous piece of DNA is used as a template to repair the DNA). The latter is used for genome editing, as the target sequence can be changed with the CRISPR‐Cas9 system.Sickle Cell Disease (SCD) is an autosomal recessive hereditary disorder that affects erythrocytes and is caused by a single base substitution of adenine to thymine on the hemoglobin beta (HbB) gene. Currently two different groups are examining two different techniques to treat SCD. One group has shown that by deleting the BCL11A gene, fetal hemoglobin production can occur, which greatly diminishes the amount of Sickle hemoglobin (HbS). An example of this application of CRISPR‐Cas9 for treating SCD is the story of Victoria Gray, the first person to have received genome editing treatment for SCD.
Cardiovascular and core temperature responses to acute organophosphate intoxication in rats
Authors: Pan Shiyue , Bruun Donald A. , Lein Pamela J. , Chen Chao 
Published: May 2022

IntroductionOrganophosphates (OP) are cholinesterase inhibitors that are used as pesticides in agriculture and as nerve agents in chemical warfare. Surprisingly, we have limited information on the time course of cardiovascular responses to acute OP intoxication at doses that cause cholinergic crisis in humans. OP‐induced QT prolongation has been shown to lead to fatal ventricular arrhythmia and delayed death, however, not all cardiac failures are associated with QT prolongation.MethodsRats were implanted with a BP/ECG telemetry device and 14 d later, acutely intoxicated with the OP diisopropylfluorophosphate (DFP, 4 mg/kg, sc) followed 1 min later by atropine sulfate (2 mg/kg, im) and pralidoxime (25 mg/kg, im) to block peripheral cholinergic toxicity. BP/ECG data were grouped based on the rat’s survival time: <15 min (n=11, group A), >15 min and <17 hours (n=9, group B), and >17 hours (n=4, group C).ResultsIn conscious, freely moving rats, DFP injection significantly increased BP (+87±10 mmHg), HR (+177±32 bpm), core temperature (+4.0±0.7 °C), and occurrences of premature ventricular complexes (PVC). These increases were similar in all three groups. Around the peak pressor response, group A had abrupt drops in BP and HR (cardiovascular collapse) that was triggered by supra‐ventricular events (sinus bradycardia and/or AV blocks). A similar pattern of cardiovascular collapse was observed in group B, hours after DFP injections. Furthermore, after the initial pressor response, half of the rats in group B had significant lower pulse pressure and BP despite a maintained HR.ConclusionOur data suggest that DFP‐induced cardiovascular collapse may be centrally mediated and that compromised cardiac function may also contribute to the cardiovascular collapse in delayed death.