bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypometabolism in Endotoxic Shock Reflects a Thermoregulatory Response and Occurs Independently of Circulatory Hypoxia
Authors: Steiner Alexandre , Corrigan Joshua , Fonseca Monique , Flatow Elizabeth , Lewis Kevin , Brito Camila
Published: June 2021

Oxygen utilization (VO2) is thought to fall in the late stages of septic shock because of mitochondrial dysfunction, circulatory hypoxia or both. However, the applicability of this conceptual framework to early septic shock is unclear. We investigated this matter in a rat model of lipopolysaccharide (LPS)‐induced shock (endotoxic shock). A novel experimental preparation was devised to simultaneously monitor oxygen delivery (DO2) and VO2 in rats whose thermoregulation had not been disrupted by anesthetics. Using a β3‐adrenergic agonist known to activate brown adipocytes, we found no evidence of metabolic impairment between 20 and 150 min post‐LPS. In spite of that, VO2 fell early in the LPS‐challenged rats, and such a fall in VO2 seemed to be driving a decrease in body temperature. The fall in VO2 was not preceded by any fall in cardiac output or DO2. Furthermore, no imbalance between VO2 and DO2 were encountered, and an analysis of NAD+/NADH ratios provided no indication of tissue hypoxia. Only when hypometabolism and hypothermia were prevented by exposure to a warm environment did an imbalance in the DO2/VO2 ratio become evident, and such an imbalance was associated with tissue hypoxia (reduction in NAD+/NADH ratio). It is concluded that reduced VO2 in early endotoxic shock (i) results from neither metabolic failure nor circulatory hypoxia, (ii) reflects a thermoregulatory adjustment, and (iii) serves as preemptive strategy to avoid hypoxia. Clinical data and conduct should be reexamined in light of these new findings.

Funding

FAPESP, AHA & CNPq.
Fluorescent Microspheres Injected into Skeletal Muscle to Observe Fluid Movement in Interstitial Space
Authors: Evertz Loribeth , Greising Sarah , Morrow Duane , Sieck Gary , Kaufman Kenton
Published: June 2021

The force within a muscle may be assessed using intramuscular pressure measurements (IMP). However, a limiting factor in implementing this technology in the clinic is the observed variability of IMP measurements. The variability may be due in part to the flow of interstitial fluid. The objective of this study was to develop a technique for tracking interstitial fluid flow during muscle lengthening using fluorescent microspheres. Additionally, the effect of microsphere diameter on dispersion was evaluated. Fluorescent microspheres were injected into the rat tibialis anterior (TA) either before (right limb) or after (left limb) lengthening to 10% strain (n = 5). To help discriminate the effects of microsphere size, 10 µl of microspheres (0.25% by volume) were injected simultaneously: 0.1 µm in sky blue and 0.2 µm in yellow green. Once frozen, 20 µm muscle cross‐sections were stained and imaged using confocal microscopy to evaluate the microsphere's axial dispersion as a percent of muscle length. The microsphere dispersion was normalized to the muscle length. The fluorescent microspheres flowed with the interstitial fluid and did not penetrate the muscle fiber. There was significantly greater microsphere dispersion with passive strain (p‐value > 0.01) for lengthened samples (31±7%) compared to static samples (17±9%). Dispersion did not differ for the two bead sizes. In conclusion, this illustrates the microspheres track the interstitial fluid and passive lengthening results in additional dispersion.

Supported by NIH under award number R01HD31476 and T32HL105355.
A Role for CD34 in the Response of Skeletal Muscle to Hypoxic Conditions
Authors: Pagé Mélissa , Langlois Anick , Dubé Annie , Patoine Dany , Maltais François , Morissette Mathieu , Debigaré Richard , Blanchet Marie‐Renée
Published: June 2021

CD34, a satellite cell (SC) marker participates in skeletal muscle (SM) regeneration, facilitating the migration of SC. A fraction of patients with chronic obstructive pulmonary disease (COPD) develop SM atrophy, in part due to increased SC senescence. COPD patients experience transient hypoxemia, and it was suggested these hypoxic episodes may participate in SM wasting. Recently, we have observed that in vitro exposure of SC to hypoxic conditions increased CD34 expression and retained SC in an undifferentiated state. The goal of the present study was to evaluate the importance of CD34 expression in the muscle response to hypoxic conditions in vivo. Total weight and lean body mass were measured in WT and Cd34 knockout (Cd34‐/‐) mice exposed to hypoxic conditions (10% O2). The gastrocnemius expression of CD34 and genes involved in muscle regeneration were detected by qRT‐PCR. Results show an important loss of total weight and lean body mass at 2 days, which is significantly worse in Cd34‐/‐ mice. Alterations in Mrf4 and MHC expression were observed in Cd34‐/‐ mice vs. WT, suggesting a role for this protein in the response of muscle to hypoxic conditions. All in all, our results suggest that CD34 expression is important for the response of SM to hypoxic conditions, leading to a possible role in SM wasting in COPD.
Estradiol enhances neutrophil infiltration into traumatically‐injured skeletal muscle
Authors: Le Gengyun , Jergenson Matthew , Lowe Dawn , Warren Gordon
Published: June 2021

Purpose

To probe how estradiol modulates neutrophil infiltration and function following traumatic muscle injury.

Methods

Adult, female C57BL/6 mice were ovariectomized and randomly assigned to no treatment or implantation of a 17‐b estradiol slow‐release pellet. Four weeks later, freeze injury was induced to tibialis anterior (TA) muscles and mice were allowed to recover for 1, 2, 3, or 4 days before sacrifice, blood collection, and TA muscle excision. Myeloperoxidase (MPO) activity, an indicator of neutrophil function, was examined in the muscles. H&E staining of TA cross‐sections were used to address neutrophil localization and FACS analysis was performed to quantify neutrophil infiltration. Neutrophils in the blood were counted by automated hematological analysis.

Results

MPO activity was 25‐125% greater in the injured muscles of estradiol‐treated mice (P<0.0001). Infiltration of neutrophils appeared in the first 24 hours and peaked at 3‐days post‐injury in muscles of mice with and without estradiol. FACS analysis revealed that compared to injured muscles from untreated mice, infiltrated neutrophils were 6‐61% greater in injured muscles from estradiol‐treated mice, with the peak difference at day 4 (24±6 vs 15±3 % of hematopoietic cells). The concentration of neutrophils in blood of estradiol‐treated mice was 40‐60% lower than that in untreated mice at 1, 2, and 3‐days post‐injury (P<0.001).

Conclusion

Estradiol increases neutrophil infiltration into muscle following traumatic injury. Supported by NIH grant R01‐AG031743.
Astaxanthin Supplementation Improves Decreased Mitochondrial Enzyme Activity and Capillary Regression during Recovery from Muscle Atrophy
Authors: Maezawa Toshiyuki , Kanazashi Miho , Tanaka Masayuki , Maeshige Noriaki , Kondo Hiroyo , Fujino Hidemi
Published: June 2021

Reactive oxygen species (ROS) impairs mitochondrial respiratory chain and capillary structure in atrophied muscle. Impaired mitochondria and capillary are also associated with a decrease of exercise tolerance. Reloading promotes recovery from atrophy, whereas reloading induces the overexpression of ROS. Therefore, we investigated the effects of reloading and anti‐oxidant supplementation for the mitochondria enzyme activity, capillary number and oxidative stress marker during recovery from disuse‐induced muscle atrophy. Hindlimb unloading resulted in a reduction of citrate synthase (CS) activity, capillary‐to‐fiber (C/F) ratio and expression of PGC‐1α protein, and an increase of superoxide dismutase (SOD)‐2. Reloading after unloading improved the decreased CS activity and PGC‐1α expression level, but C/F ratio could not be recovered. Astaxanthin, anti‐oxidant, supplementation maintained all of them to control level through the unloading and reloading conditions. In addition, astaxanthin supplementation attenuated the overexpression of SOD‐2. These results suggest that astaxanthin supplementation approach may be effective as a therapeutic strategy under recovery conditions involving muscular debilitation.

Supported by Grants‐in‐Aid for Science Research from the Japanese Ministry Education, Culture, Sports, Science and Technology, and the internal funds of Fuji Chemical Industry Co. Ltd.
Muscle‐Specific RING Finger 1 (MuRF1) Dependent Activation of Dual‐specificity Phosphatase 4 (Dusp4) Under Neurogenic Atrophy Conditions
Authors: Haddock Ashley , Waddell David
Published: June 2021

Skeletal muscle atrophy is the result of many physiological conditions, including denervation, corticosteroid treatment, and aging. The E3 ubiquitin ligase, MuRF1, is induced under atrophic conditions and is believed to play a role in protein degradation in wasting muscle. However, the data described in this study provides evidence that MuRF1 may also act as a transcriptional modulator of atrophy‐induced gene activity, including the regulation of Dusp4 expression. In order to characterize the transcriptional regulation of Dusp4, reporter gene constructs with fragments of the proximal promoter region of the gene were developed, transfected into C2C12 cells with or without a MuRF1 expression plasmid and analyzed for differences in reporter gene activity. The Dusp4 reporters showed repressed activity in cells ectopically expressing MuRF1 compared to cells that did not overexpress MuRF1. Interestingly, overexpression of the myogenic regulatory factors (MRFs), MyoD and myogenin, also caused repression of the Dusp4 reporter constructs, while co‐overexpression of MuRF1 with MyoD or Myogenin resulted in cooperative repression of reporter gene activity. Mutagenesis of the single conserved E‐box in the Dusp4 proximal promoter resulted in a 20‐fold repression in reporter gene activity, but the trend in cooperative repression was still observed. To further characterize the role of MuRF1 in the repression of Dusp4, a MuRF1 RING domain mutant was created. The MuRF1 RING mutant failed to cooperate with the MRFs to repress the Dusp4 reporter gene activity, suggesting that catalytic activity is required for transcriptional regulation by MuRF1. This data offers evidence that MuRF1 may act as a transcriptional modulator of atrophy‐induced gene expression.
TRPV4 Induces Warm‐Defense Responses in Non‐Genetically Modified Rats
Authors: Scarpellini Carolina , Correa Gabriela , Almeida Maria Camila , Gargaglioni Luciane , Bícego Kenia
Published: June 2021

TRPV4 is a warmth sensitive channel, suggested to act as a thermosensor but with no thermoregulatory role described in vivo (sensitive to temperatures in vitro at the range of ~24 ‐ 34ºC). Thus, we aimed at investigating which thermoeffectors are affected by TRPV4 activation in rats. To this end, we measured core body temperature (Tc), oxygen consumption and heat loss index (HLI) in male rats (290‐320g) under different ambient temperatures (Ta) after the intravenous blockade of this channel with HC‐067047. Moreover the preferred ambient temperature was assessed: i) after thermal stimulation combined with the intravenous blockage of TRPV4 with HC‐067047; and ii) after topical application of agonist RN‐1747. HC‐067047 caused an increase on Tc at Ta of 26 and 30°C (~0.65°C; p<0.05), but not at 22 and 32°C. At 26°C, HC‐067047‐induced hyperthermia was accompanied by increase in oxygen consumption (1.24 ± 0.2 mL O2 kg‐¹ min‐¹ more than the controls; p< 0.05), but no difference on HLI was observed. Furthermore, rats chemically stimulated with TRPV4 agonist leaned towards colder Tas (29°C for controls and 21°C for agonist‐treated; p< 0.05) and the cold‐seeking behaviour after thermal stimulation (28‐31°C) was inhibited by TRPV4 antagonist (24°C for controls and 28°C for antagonist‐treated; p< 0.05). Our results suggest, for the first time, that TRPV4 channel is involved in the recruitment of behavioural and autonomic warmth defense responses in order to regulate Tc. Financial Support: FAPESP
Rapid Onset Pressor Response to Static Handgrip in Young Normotensive Women with a Family History of Hypertension
Authors: Matthews Evan , Greaney Jody , Wenner Megan
Published: June 2021

Older hypertensive adults display exaggerated blood pressure (BP) and muscle sympathetic nerve activity (MSNA) responses at the onset of static muscle contraction. Young normotensive adults with a family history of hypertension (+FH) are at a greater risk for developing future hypertension. We tested the hypothesis that young women with a +FH wouldhave exaggerated increases in BP and MSNA at the onset of static muscle contraction compared to those without a family history of hypertension (‐FH). We retrospectively examined beat‐by‐beat BP, heart rate (HR), and MSNA during the initial 30 seconds of isometric handgrip exercise at 30% of maximum voluntary contraction in 14 +FH (22±1 yrs, 21±1 kg·m‐2) and 14 –FH (22±1 yrs, 22±1 kg·m‐2) women. Resting mean arterial pressure (MAP;+FH 80±2 vs. –FH 79±3 mmHg), HR (+FH 62±3 vs. –FH 66±2 bpm), and MSNA (+FH 7±1 vs. –FH 10±2 bursts·min‐1) were similar between groups (all P>0.05). At exercise onset, the increases in MAP (+FH Δ11±2 vs. –FH Δ4±1 mmHg, P<0.05) was greater in +FH compared to –FH women. The increase in HR was also greater in +FH (+FH Δ 13±2 vs. –FH Δ 4±1 bpm P<0.05). The MSNA responses were not different between groups (+FH Δ3±1 vs. –FH Δ22±1 bursts·30s‐1, P=0.29). These data demonstrate that there is an exaggerated pressor response at the onset of static muscle contraction in young normotensive women with a +FH and further suggest that this may be due to greater increases in HR at exercise onset.

Research supported by NIH Grant U54‐GM104941 (PI: Binder‐Macleod)
Nighttime blood pressure dipping under controlled sodium conditions: effect of sex and race
Authors: Brian Michael , Matthews Evan , Muth Bryce , Ramick Meghan , Lennon‐Edwards Shannon , Wenner Megan , Edwards David , Farquhar William
Published: June 2021

Blunted nighttime blood pressure (BP) dipping is related to increased cardiovascular risk. Altered dietary sodium can influence short‐term and long‐term BP, but studies that have assessed nighttime BP dipping have not controlled for sodium intake in normotensives. We therefore measured nighttime BP dipping during a controlled sodium diet in normotensive adults (n=96; age= 38±1 yrs, BP= 119±1 / 75±1 mmHg), and examined sex (women n=48, age= 41±2 yrs, BP= 115±2 / 73±2 mmHg vs. men n=48, age= 35±2 yrs, BP= 123±1 / 76±1 mmHg) and racial differences (Caucasians n=75, age=39±2 yrs, BP= 120±1 / 75±1 mmHg; African American n=13, age= 35±3 yrs, BP= 116±3 / 71±2 mmHg) in subgroup analyses. All subjects consumed a recommended sodium diet (100 mmol/day) for 7 days and 24‐hour ambulatory BP was measured on the 7th day of the diet. BP was assessed every 20 minutes during daytime (wake) and every 30 minutes during nighttime (sleep). Nighttime BP dipping was assessed by measuring the percent (%) difference between mean systolic daytime BP and mean systolic nighttime BP. Twenty‐four‐hour urine sodium excretion was 82.9±4.2 mmol/24 hrs for all subjects; no difference was observed between sex and race (p>0.05). Nighttime BP dipping under a controlled sodium condition was 10.9±0.6% for all subjects. No differences were observed in nighttime BP dipping between sexes (women= 10.9±0.8% vs. men= 10.8±0.9%, p>0.05) or between races (Caucasian= 10.7±0.7% vs. African American= 9.6±1.3% p>0.05). These data suggest that under controlled sodium conditions, there are no sex or race differences in nighttime BP dipping.
Identification of Pin1 Isomerase on Neuronal Cell Rescue in Ischemic Brain Injury
Authors: Lee Seunghoon , Won Jinyoung , Hong Yonggeun
Published: June 2021

Pin1 contains two functional domains, an N‐terminal WW domain and a C‐terminal PPIase domain. The WW domain is a phosphorylation specific protein interaction module that directs Pin1 to its substrate‐proteins phosphorylated at certain serine or threonine residues followed by proline. Upon this binding, the PPIase domain catalyzes conformational change of the Pin1 substrates by isomerizing specific pSer/Thr‐Pro bonds; thus, we hypothesized that Pin1 can regulate neuronal cell death and neurogenesis through Pin1 inhibitor treatment after MCAo. To test the hypothesis, we elucidated roles of Pin1 following treatment of the Pin1 inhibitor at the sub‐acute phase after MCAo in rats. Juglone (1 mg/kg) was administered once a day for 7 days after MCAo. Histological and molecular analyses were used to determine apoptosis and neurodegeneration. MCAo+Jug leds to significant downregulation of Bcl‐2 and Bax the protein level compared with MCAo+veh (**p<.01). Ischemic brain injury promoted phosphorylation of Thr231 motif on tau as shown in Western blot. MCAo+Jug was shown to reduce the levels of Pin1 and tau5 in both Western blot and immunostaining (**p<.01). These results suggest that Pin1 isomerase acts as a pro‐apoptotic molecule in ischemic‐induced neuronal cell death after MCAo. Additionally, we suggest that Pin1 inhibition might alleviate the acceleration of apoptotic cell death from acute to sub‐acute after MCAo in rats.

Funding: 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512
Improvement of nephrotic syndrome by Banggihwanggitang.
Authors: Lee So min , Chung Sun‐Ku , Lee Seo‐young , Kang Dae Gill , Lee Ho Sub
Published: June 2021

Nephrotic syndrome is associated with altered renal handling of sodium along with changes of renal epithelial Na channel (ENaC), leading to the development of edema and ascites.

The dried sclerotia of Banggihwanggitang (BHT) has been used for the treatment of chronic kidney disease. This study was conducted to evaluate the effects of BHT on puromycin aminonucleoside (PAN)‐induced renal functional derangement and the change of ENaC expression. The nephrotic syndrome animal models were constructed by PAN 75 mg/kg injection and then were treated with losartan (30 mg/kg/day) or BHT (200 mg/kg/day) for 7 days. Consequently, BHT group was significantly improved the proteinuria and ascites. In addition, the plasma level of triglyceride, total cholesterol, and low density lipoprotein (LDL)‐cholesterol were significantly decreased in BHT. Besides, BHT group attenuated the PAN‐induced increase in ENaC protein expression. BHT was significantly suppressed PAN‐induced organic osmolytes regulator such as serum‐ and glucocorticoid‐inducible protein kinase (Sgk1), and mineralocorticoid receptor (MR) mRNA expression.

Taken together, BHT improve nephrotic syndrome including proteinuria and ascites, through inhibition of ENaC. Thus, Banggihwanggitang is involved in the body‐fluid regulation via inhibition of water and sodium channels against renal disorder such as edema or nephrosis.
Functional implications of a novel point‐mutation at an ACTN4 phosphorylation site
Authors: Feng Di , Krishnan Ramaswamy , Steinke Julia , Pollak Martin
Published: June 2021

Focal segmental glomerulosclerosis (FSGS) is a significant cause of human kidney disease. Mutations in the actin‐crosslinking protein alpha‐actinin‐4 (ACTN4) constitute a rare cause of glomerular podocyte damage and consequent FSGS. To this end, we report here a novel ACTN4 mutation that occurs at the tyrosine phosphorylation site, Y265H. This mutation was identified in a sporadic FSGS patient through exome sequencing and validated through Sanger sequencing. Using purified protein, we found that compared to WT ACTN4, Y265H ACTN4 mutant protein increased binding affinity to F‐actin. In immortalized human podocytes, transfection with mutant Y265H ACTN4 resulted in abnormal ACTN4 aggregation in the cytosol. These structural changes were associated with a two‐fold enhancement in traction forces that the podocytes exerted on their underlying substrate measured by traction force microscopy. Taken together, we demonstrate that a point mutation at an ACTN4 phosphorylation site can lead to aberrant biochemical and biophysical changes in the podocyte. Moreover, the Y265H mutation could be a novel ACTN4 mutation leading to FSGS in humans.
Anti‐Fibrotic Effect of Ajulemic Acid in Bleomycin‐Induced Lung Fibrosis
Authors: Lucattelli Monica , Fineschi Silvia , Selvi Enrico , Garcia Gonzales Estrella , Balistrieri Epifania , Lorenzini Sauro , Galeazzi Mauro , Distler Jorg , Lungarella Giuseppe
Published: June 2021

Ajulemic acid (AjA) a synthetic analogue of tetrahydrocannabinol can prevent progression of skin fibrosis in experimental systemic sclerosis. We aimed to investigate if AjA modulate lung fibrogenesis induced in mice by bleomycin (BLM).

The efficacy of AjA was evaluated in inflammatory (within 7 days after BLM) or in early fibrogenic phase (from 7 days onward). DBA/2 mice were i.tr. instilled with a single dose of BLM (0.1 μg/50μl saline) and AjA given orally, once a day,starting at day 1 (Groups 3, 4) or day 8 from BLM (Groups 5, 6). AjA in safflower oil was given at a dose of 1mg/kg (Groups 3, 5) and 5mg/kg (Groups 4,6). Group 1 and Group 2 received safflower oil or BLM, respectively. Mice were sacrificed at day 14 and 21 after BLM and lungs were processed for histology and morphometry, examined for HO‐proline and expression of TGF‐β, CTGF, α‐SMA, and PPAR‐ γ.

BLM induces lung fibrosis with significant increase of HO‐proline (Group 1: 174.32 ± 30 μg/lung; Group 2: 280± 35 μg/lung; p<0.01). AjA treatment decreases significantly the fibrotic response as revealed by morphometry and HO‐proline content (Group 3: 172±29; Group 4: 209± 38; Group 5: 189±32; Group 6: 186±20). Marked decrease of TGF‐β and CTGF expression, significant reduction of α‐SMA positive cells and significant increase of PPAR‐γ expression is observed in AjA groups.

AjA exerts potent anti‐fibrotic effects in BLM‐lung fibrosis also when given during the fibrogenic phase. The reduced ECM deposition is associated with decreased expression of the profibrotic cytokines TGF‐β and CTGF and increase of PPAR‐γ positive cells.
Ventilatory and Neurochemical Effects of Microdialysis of a µ‐opioid Receptor Agonist (DAMGO) into the Region of the Ventral Respiratory Column in Awake Goats
Authors: Langer Thomas , Neumueller Suzanne , Crumley Emma , Muere Clarissa , Hodges Matthew , Pan Lawrence , Forster Hubert
Published: June 2021

Prior studies (J.appl Phisiol. 114, 694‐647, 2012., Respir Phisiol 205: 7‐15 2014.) have shown that dialysis of antagonists of excitatory neuromodulator receptors in the ventral respiratory column (VRC) resulted in changes in the release of other excitatory neuromodulators with no net change in ventilation. In the present study, we tested the hypothesis that agonists of inhibitory receptors in the VRC will also increase excitatory neuromodulator release by dialyzing DAMGO ([D‐Ala2, N‐MePhe4, Gly‐ol]‐enkephalin), a µ‐opioid receptor agonist, in chronically cannulated awake goats. In five goats we dialyzed mock cerebral spinal fluid (mCSF) with or without DAMGO (10, 50, or 100µM concentrations). Dialysis of 100µM DAMGO resulted in a decrease in tidal volume which was followed by an increase in frequency to maintain ventilation. The concentrations of serotonin, substance P, GABA and glycine in the effluent mCSF were not altered by DAMGO dialysis. We conclude that increased inhibition of VRC respiratory neurons is not compensated for by the same mechanism as during decreased excitation. This work was funded by the National Heart, Lung, and Blood Institute grants HL‐25739, HL‐112996, HL‐007852, and by the Department of Veterans Affairs.
Eye Can Learn About Physiology through PhUn Week
Authors: Alarcon Fortepiani Maria , Ibarra Jessica
Published: June 2021

Physiology Understanding (PhUn) Week, a national science outreach program of the American Physiological Society provided a great opportunity to deliver innovations in teaching into the classroom in San Antonio. The city has a high prevalence of vision problems, some of them which may be preventable with proper nutrition. Most PhUn Week activities engage students on the physiology of the heart, circulation, and the lungs, but we decided to introduce and enhance middle school student's understanding of the eye during PhUn Week. We developed a collection of hands‐on vision physiology activities specifically for middle school students.

Method

In 2014, over 500 middle school students from Clara Driscoll and E.T. Wrenn Middle Schools in San Antonio, Texas participated in several 10 minute activities that we developed during PhUn Week. The activities included examining the parts of the eye looking up close at cow eye specimens and a human eye anatomical model to make functional correlates for each structure examined. We further evaluated the integrity of this functional system by demonstration of pupils responding to light, finding their blind spot, showing them x‐ray vision, and shifting images. During the 45 minute session the students were also given a presentation describing the eye safety, common diseases of the eye, and contained a discussion on eye nutrition.

Result

Many students gained a greater understanding of the eye, how it works, its importance, and the link between good nutrition and healthy eyesight. These activities also highlighted for students the impact of physiology in their daily lives – to sense the world around them.

Conclusion

Because of PhUn week, we were able to deliver innovative vision physiology activities at middle school in our community.
Repetition of Key Words During Phun Week Activities Increases Correct Answers in a Population of Junior Students in Puerto Rico
Authors: Martínez‐Orengo Neysha , Marrero‐Criado Marangelie , Ramos Sugeily , Cora Evelyn , Santiago Cariluz , Appleyard Caroline , Chompre Gladys
Published: June 2021

Puerto Rico Physiological Society has been conducting PhUn Week activities in schools located in the south of the Island. This year junior students received talks and participated in hands‐on activities related to several topics in physiology.

Aim

The main was to provide information on basic physiological concepts, what scientists do, and how to pursue a scientific career. We hypothesized that combining conventional talks with hands‐on activities emphasizing key words will increase the students' attention and retention, leading to correct answers on a written evaluation.

Methods

Students from eleventh grade (22 males and 8 females) were given a pre‐test which consisted of 5 multiple choice questions related to cardiovascular, respiratory, nervous, and gastrointestinal systems. Oral presentations were given followed by hands‐on activities using repetition of key words during each activity. A post‐test was given to measure learning outcomes.

Results

In the pre‐test, 59.3% of the overall questions were answered correctly, and this increased to 98.7% in the post‐test. Repetition during the talks and hands‐on activities increases the selection of correct answers in the written evaluation.

Conclusions

We were able to call the students attention by sharing our personal experiences as scientists and explaining physiology via a variety of assessments, which includes the repetition of key words in each activity. This outreach endeavor provides the student with alternative ways of learning physiology and a better understanding of a career in the sciences.
Student Reflections from an Upper Level Problem‐Based Learning Biochemistry Course
Authors: Lescallette Adam , Knecht Vincent , Dries Daniel
Published: June 2021

Problem‐based learning (PBL) is a pedagogical method structured around the idea of a student recognizing his/her own barriers in learning a particular topic. Here, we as students reflect upon our experiences in an upper level PBL course in biochemistry that uses the primary literature as the framework for the class. The theme of our class was the native South American poison known as curare. Students were given articles to read outside of class and were asked to come to the next class with specific “learning issues” identified. (A learning issue is any question that impedes the student from fully understanding the information given in the article.) In class, these learning issues were addressed in small groups. This method is particularly appealing to some students, as it is logical and systematic ‐ as a student is forced to question his/her understanding, the student more naturally probes deeper into the problem being experimentally addressed. After a few weeks, we students saw that there was one major problem: students were not able to focus on the most important aspects of each article without a little direction from the professor. When direction was given, it was much easier to work through each article from the general to the specific. Moreover, retention of information became easier as the method relied less on memorization and more on deeper understanding. PBL allows for the systematic solving of learning issues in a way that is easiest for the student, allowing learning to happen organically rather than rigidly. An inherent criterion for the success of PBL, however, is the students' willingness to recognize his/her own ignorance, which may be the biggest struggle for some.
Superresolution Imaging Reveals the Internal Organization of Syntaxin Clusters in Hippocampal Processes
Authors: Lowenstein Sarah , Shaver Daniel , Thomson Holly , Lochner Janis
Published: June 2021

N‐ethylmaleimide sensitive factor attachment protein receptor (SNARE) proteins are the universal molecular machinery used to mediate membrane fusion events. Syntaxins are SNARE proteins that reside in the target membrane and determine the allowable sites for membrane fusion. Clusters of syntaxin molecules release reactive syntaxin molecules for interactions with their partners in SNARE formation. In hippocampal neurons, syntaxin‐1a (Stx1a) mediates neurotransmitter release from synaptic vesicles whereas syntaxin‐3 (Stx3) facilitates activity‐dependent α‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionic acid(AMPA) receptor exocytosis. To explore the single‐molecule distribution of these syntaxin variants in hippocampal neurons, we implemented photoactivated localization microscopy (PALM), a superresolution fluorescence microscopy technique. PALM revealed single Stx1a molecules as well as clusters of Stx1a distributed throughout the hippocampal processes. The clusters of Stx1a varied markedly in size and the density of Stx1a molecules within the clusters was nonuniform with the greatest density of Stx1a molecules observed in the core of the clusters. The largest Stx1a clusters contained multiple density gradients of Stx1a within the cluster. PALM images revealed that Stx3 also forms clusters in hippocampal processes, and these Stx3 clusters likewise contained a density gradient of syntaxin molecules with Stx3 density most pronounced in the cluster core. This work is supported by National Institutes of Health grant 2 R15 NS40425‐03.
Phenolics Constituents and In‐vitro α‐amylase, α‐glucosidase and Ecto‐ 5'nucleotidase Inhibitory Properties of Selected Solanum spp
Authors: Nwanna Esther , Shodeinde Sidiqat , Adeyemo Adenike
Published: June 2021

Control of postprandial plasma glucose levels is critical in the early management of diabetes mellitus,and in reducing chronic vascular complications such as obesity.This study investigate the phenolics, antioxidant properties, and effect of extracts from selected Solanum spp (S. melongena, S. macrocarpon, S.anguivi, S. aethiopicum, S. kumba and S.gilo) on α‐amylase, α‐glucosidase and 5'nucleotidase related to diabetes in‐vitro. HPLC‐DAD was used to characterize the phenolic constituents. Aqueous extracts were prepared and its effect on α‐amylase, α‐glucosidase, and ecto‐5'nucleotidase activities as well as antioxidant [Fe2+ induced lipid peroxidation, hydroxyl radical, 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging, Fe2+ chelating ability and ferric reducing antioxidant properties were determined. The predominant phenolics detected were quercetin, quercitrin, kaempferol, chlorogenic acid, caffeic acid and rutin. The results showed that all the Solanum species significantly (P<0.05) inhibited α‐amylase, α‐glucosidase, in a concentration dependent manner (0‐2µg/ml) with S.aethiopicum having the highest α‐glucosidase inhibitory ability while S.melongena had the highest α‐amylase inhibitory activity. Extract from these species S.melongena, S.macrocarpon, S.anguivi,S. kumba) had inhibitory effect on ecto‐5'nucleotidase activity while others had stimulatory effect. The enzyme inhibitory and antioxidant properties exhibited these species could be attributed to their individual /synergistic phenolic constituents and Vitamin C content, thus explaining possible biochemical rationale behind their use in the management of diabetes.
Efficacy and Safety of Intraventricular Antibiotic Administration
Authors: Llave Nathaniel , Young Nicholas , Abernathy Casey , Reinert Justin P.
Published: June 2021

The administration of antibiotics via intraventricular administration for treatment of central nervous system infections is an understudied topic that lacks data for specific patient outcomes regarding dosing and adverse events. A comprehensive review of the literature was performed on PubMed, MEDLINE, Scopus, and CINAHL using the following terminology: “intraventricular” OR “intraventricular drug administration” AND “antibiotic” OR “antimicrobial” AND “infection” AND “CNS” NOT “Intrathecal.” Studies that described adult human patients who received an antibiotic via intraventricular administration with complete dosing, efficacy, and safety data were included. A total of 8 studies met inclusion and were used in this review and detailed the use of antibiotics targeting resistant Gram-positive and Gram-negative bacteria. Minimal adverse effects were reported, although the determination of appropriate dosing and markers of efficacy remains elusive due to the common concomitant administration of intravenous antibiotics. Further clinical evaluation is necessary.
Topical Therapy Failure in Chronic Suppurative Otitis Media is Due to Persister Cells in Biofilms
Authors: Santa Maria Peter L. , Kaufman Adam C. , Bacacao Brian , Thai Anthony , Chen Xiaohua , Xia Anping , Cao Zhixin , Fouad Ayman , Bekale Laurent A.
Published: June 2021

Objective:

Chronic suppurative otitis media (CSOM) is characterized by a chronically draining middle ear. CSOM is typically treated with multiple courses of antibiotics or antiseptics which are successful in achieving quiescence; however, the disease is prone to relapse. Understanding why these treatment failures occur is essential.

Study Design:

The minimum inhibitory concentration (MIC), minimal biofilm eradication concentration, and the inhibitory zone were determined for ototopicals and ofloxacin for the laboratory strains and CSOM-derived isolates. The percentage of persister cells and bacterial biofilm formation were measured. Disease eradication was tested in a validated in-vivo model of CSOM after treatment with ofloxacin.

Setting:

Microbiology Laboratory.

Methods:

Basic science experiments were performed to measure the effectiveness of a number of compounds against CSOM bacteria in a number of distinct settings.

Results:

The minimal biofilm eradication concentration is higher than is physiologically achievable with commercial preparations, except for povo-iodine. Clincial isolates of CSOM have equivalent biofilm-forming ability but increased proportions of persister cells. Ofloxacin can convert to inactive disease temporarily but fails to eradicate disease in an in-vivo model.

Conclusions:

Higher percentages of persister cells in clinical CSOM isolates are associated with resistance to ototopicals. Current ototopicals, except povo-iodine, have limited clinical effectiveness; however, it is unknown what the maximum achievable concentration is and there are ototoxicity concerns. Fluoroquinolones, while successful in producing inactive disease in the short term, have the potential to encourage antimicrobial resistance and disease recalcitrance and do not achieve a permanent remission. Given these limitations, clinicians should consider surgery earlier or use of clinically safe concentrations of povo-iodine earlier into the treatment algorithm.
Outcomes of Initial Observation Versus Upfront Microsurgical Resection for Small to Medium-sized Vestibular Schwannomas
Authors: Patro Ankita , Totten Douglas J. , Sherry Alexander D. , Manzoor Nauman F. , Cass Nathan D. , Tawfik Kareem , Bennett Marc L. , O’Malley Matthew R. , Haynes David S. , Perkins Elizabeth L.
Published: June 2021

Objective:

To assess postoperative outcomes and predictive factors of patients observed prior to microsurgery and those undergoing upfront resection for small and medium-sized VS.

Study design:

Retrospective cohort.

Setting:

Tertiary referral center.

Patients:

VS patients who had microsurgery from 2003 to 2018 for tumors up to 2.5 cm.

Main outcome measures:

Postoperative outcomes including facial nerve function and interventions, complications, extent of resection, and salvage therapy.

Results:

Of 220 patients, 120 were initially observed, and 100 pursued upfront microsurgery. There was no significant association between initial observation and upfront microsurgery for postoperative facial nerve function at 2 to 3 weeks (p = 0.18) or 12 months (p = 0.5), facial nerve intervention (p = 0.5), major/minor complications (p = 0.48/0.63), recurrence (p = 0.8), subtotal resection (p = 0.6), or salvage therapy (p = 0.9). Time from initial consultation to surgery did not significantly impact outcomes. Intrameatal tumors were more likely to be observed (odds ratios [OR] 2.93; 95% CI 1.53–5.63; p = 0.001). Patients with larger tumor volume (OR 0.52; 95% CI 0.37–0.72; p < 0.0001), brainstem compression (OR 0.28; 95% CI 0.09–0.91; p = 0.03), or higher PTA were less likely to undergo observation (OR 0.99; 95% CI 0.97–0.997; p = 0.02). On multivariable analysis, predictive factors for observation were smaller tumor volume (OR 0.53; 95% CI 0.38–0.75; p < 0.001), lower PTA (OR 0.99; 95% CI 0.98–0.999; p = 0.04), and diabetes (OR 2.54; 95% CI 0.95–6.83; p = 0.06).

Conclusions:

Patients with worse hearing, larger tumor volume, and brainstem compression were more likely to pursue upfront microsurgery. A watchful waiting period does not appear to worsen outcomes and can be considered for patients with better hearing and smaller tumors without brainstem compression.
Next generation plasma proteome profiling of COVID-19 patients with mild to moderate symptoms
Authors: Zhong Wen , Altay Ozlem , Arif Muhammad , Edfors Fredrik , Mardinoglu Adil , Uhlén Mathias , Fagerberg Linn
Published: June 2021

COVID-19 has caused millions of deaths globally, yet the cellular mechanisms underlying the various effects of the disease remain poorly understood. Recently, a new analytical platform for comprehensive analysis of plasma protein profiles using proximity extension assays combined with next generation sequencing has been developed. Here, we describe the analysis of the plasma profiles of COVID-19 patients with mild and moderate symptoms by comparing the protein levels in newly diagnosed patients with the plasma profiles in the same individuals after recovery 14 days later. The study has identified more than 200 proteins that are significantly elevated at infection and many of these are related to cytokine response and other immune-related functions. In addition, several other proteins are shown to be elevated, including SCARB2, a host cell receptor protein involved in virus entry. A comparison with the plasma protein response in patients with severe symptoms shows a highly similar pattern, but with some interesting differences. In conclusion, the results will facilitate further studies to understand the molecular mechanism of the immune-related response of the SARS-CoV-2 virus.
Self-collection and pooling of samples as resources-saving strategies for RT-PCR-based SARS-CoV-2 surveillance, the example of travelers in French Polynesia
Authors: Aubry Maite , Teiti Iotefa , Teissier Anita , Richard Vaea , Mariteragi-Helle Teheipuaura , Chung Kiyojiken , Deen Farah , Paoaafaite Tuterarii , Cao-Lormeau Van-Mai
Published: June 2021

In French Polynesia, the first case of SARS-CoV-2 infection was detected on March 10th, 2020, in a resident returning from France. Between March 28th and July 14th, international air traffic was interrupted and local transmission of SARS-CoV-2 was brought under control, with only 62 cases recorded. The main challenge for reopening the air border without requiring travelers to quarantine on arrival was to limit the risk of re-introducing SARS-CoV-2. Specific measures were implemented, including the obligation for all travelers to have a negative RT-PCR test for SARS-CoV-2 carried out within 3 days before departure, and to perform another RT-PCR testing 4 days after arrival. Because of limitation in available medical staff, travelers were provided a kit allowing self-collection of oral and nasal swabs. In addition to increase our testing capacity, self-collected samples from up to 10 travelers were pooled before RNA extraction and RT-PCR testing. When a pool tested positive, RNA extraction and RT-PCR were performed on each individual sample. We report here the results of COVID-19 surveillance (COV-CHECK PORINETIA) conducted between July 15th, 2020, and February 15th, 2021, in travelers using self-collection and pooling approaches. We tested 5,982 pools comprising 59,490 individual samples, and detected 273 (0.46%) travelers positive for SARS-CoV-2. A mean difference of 1.17 Ct (CI 95% 0.93 – 1.41) was found between positive individual samples and pools (N=50), probably related to the volume of samples used for RNA extraction (200 µL versus 50 µL, respectively). Retrospective testing of positive samples self-collected from October 20th, 2020, using variants-specific amplification kit and spike gene sequencing, found at least 6 residents infected by the B1.1.7 UK variant. Self-collection and pooling approaches allowed large-scale screening for SARS-CoV-2 using less human, material and financial resources. Moreover, this strategy allowed detecting the introduction of SARS-CoV-2 variants in French Polynesia.
Clinical Pharmacy Services Provided in Public Sector Hospitals in Nigeria: A National Survey
Authors: Udoh Arit , Akpan Mary , Ibrahim Umar Idris , Lawal Basira Kankia , Labaran Kamilu Sarki , Ndem Ekpedeme , Amorha Kosisochi , Matuluko Ayodeji , Tikare Olubukola , Ohabunwa Unoma , Kpokiri Eneyi
Published: June 2021

Background

Studies show that clinical pharmacy services are effective in optimizing medicines use and patients’ outcomes. This study aimed to determine the clinical pharmacy services provided in public sector hospitals in Nigeria.

Methods

This was an online survey of 296 primary, secondary and tertiary care hospitals sampled purposively across the 36 States and Federal Capital Territory in Nigeria. Data analysis was conducted descriptively, and via Chi-square test and multivariate analysis of variance (MANOVA).

Key findings

Responses were obtained from 272 hospitals in the country with a survey completion rate of 88%. This included 55 tertiary, 72 secondary, and 145 primary healthcare centres (PHCs). Pharmacists provided pharmaceutical care services in all the tertiary care hospitals, 94% of the secondary, and in only 6% of the PHCs surveyed. The composition of the pharmacy department per level of care was similar across the six geopolitical regions (V = 0.383, F = 1.453, P = 0.06) with more pharmacy staff employed in the tertiary care hospitals compared to the secondary care facilities. The majority (≥75%) of the tertiary and secondary care hospitals provided medicines information, patient education and counselling, alongside in- and outpatient dispensing services. However, fewer than 30% reported routine pharmacists’ involvement in multidisciplinary ward rounds, medication chart review, therapeutic guidelines development, antibiotic stewardship programmes, and drug therapy monitoring. Pharmacists routinely provided medication error reporting services in only about half of the hospitals in the sample, and this was not associated with the level of care (p > 0.05).

Conclusions

The findings of this study demonstrate disparity in clinical pharmacy service availability across the various levels of care in Nigeria. It also highlights the need to scale up and prioritize their integration within the primary care sector.
Emergence of border-ownership by large-scale consistency and long-range interactions: Neuro-computational model to reflect global configurations
Authors: Kogo Naoki , Froyen Vicky
Published: June 2021

The visual system performs remarkably well to perceive depth order of surfaces without stereo disparity, indicating the importance of figure-ground organization based on pictorial cues. To understand how figure-ground organization emerges, it is essential to investigate how the global configuration of an image is reflected. In the past, many neuro- computational models developed to reproduce figure-ground organization implemented algorithms to give a bias to convex areas. However, in certain conditions, a convex area can be perceived as a hole and a non-convex area as figural. This occurs when the surface properties of the convex area are consistent with the background and, hence, are grouped together in our perception. We argue that large-scale consistency of surface properties is reflected in the border-ownership computation. We developed a model, called DISC2, that first analyzes relationships between two border-ownership signals of all possible combinations in the image. It then enhances signals if they satisfy the following conditions: 1. the two signals fit to a convex configuration, and 2. the surface properties at the locations of the two signals are consistent. The strength of the enhancement decays with distance between the signals. The model gives extremely robust responses to various images with complexities both in shape and depth order. Furthermore, we developed an advanced version of the model (“augmented model”) where the global computation above interacts with local computation of curvilinearity, which further enhanced the robust nature of the model. The results suggest the involvement of similar computational processes in the brain for figure-ground organization.
Comparative performance of between-population vaccine allocation strategies with applications for emerging pandemics
Authors: Joshi Keya , Rumpler Eva , Kennedy-Shaffer Lee , Bosan Rafia , Lipsitch Marc
Published: June 2021

2

Vaccine allocation decisions during emerging pandemics have proven to be challenging due to competing ethical, practical, and political considerations. Complicating decision making, policy makers need to consider vaccine allocation strategies that balance needs both within and between populations. Due to limited vaccine stockpiles, vaccine doses should be allocated in locations where their impact will be maximized. Using a susceptible-exposed-infectious-recovered (SEIR) model we examine optimal vaccine allocation decisions across two populations considering the impact of population size, underlying immunity, continuous vaccine roll-out, heterogeneous population risk structure, and differences in disease transmissibility. We find that in the context of an emerging pathogen where many epidemiologic characteristics might not be known, equal vaccine allocation between populations performs optimally in most scenarios. In the specific case considering heterogeneous population risk structure, first targeting individuals at higher risk of transmission or death due to infection leads to equal resource allocation across populations.