bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Valerian/Hop Mixture on Sleep Quality in Vertebrate and Invertebrate Model
Authors: Jo Kyungae , Hong Ki‐Bae , Park Yooheon , Suh Hyung Joo
Published: June 2021

Insomnia, or sleep disorder, is a common complaint in the general population and the etiologies of insomnia are complex. Treatments for insomnia can be divided into pharmacological agent and non‐ pharmacological remedies, and the most of pharmacological treatments those prescribed for insomnia bring some risks such as overdose, tolerance and addiction. Available evidence suggests that modulation of sleep can be controlled by non‐pharmacological treatments such as bioactive molecules, natural substances, precursors for serotonin and melatonin. Extracts of valerian (Valeriana officinialis) and hop (Humulus lupulus) are the most frequently used natural treatments of anxiety, depression and insomnia. The aim of this study was to investigate the effect of valerian/hop mixture in invertebrate model (Drosophila melanogaster) and vertebrate model (Rattus norvegicus). Valerian and various hops (Cascade, Haallertau, Saaz) used for the study was prepared as 70% ethanol extract. Drosophila was used to test sleep‐wake behavior and dose‐response. In addition, vertebrate model was used for electroencephalogram (EEG) analysis to acquire sleep quality, duration and latency. The results provided the evidences that valerian/hop mixture can be a plausible remedy for sleep disorder.
Vascular and Lymphatic Circulation of the American Alligator Phallus
Authors: Kelly Diane , Mascari Taylor , Hamilton Alexandra , Moore Brandon
Published: June 2021

Crocodilian phalluses are unusual because, unlike most amniote phalluses that inflate and stiffen as part of copulation, most of this organ is comprised of dense collagenous tissue that does not change size or shape during muscular‐driven phallic eversion. Despite this, phallus tissues adjacent to the sperm conducting sulcus (urethral groove) and associated with the glans do inflate before copulation. To determine how blood is supplied to both inflatory and noninflatory phallic tissues, we examined the vascularization of cloacal and phallic tissues of the American alligator (Alligator mississippiensis) using dissection, histology, digital tracing and 3D modeling of tissues in thick cloacal sections, in conjunction with CT scans of contrast‐enhanced circulatory fluid in sedated animals. We found that most vascularization in this region was limited to the cloacal walls and the tissue associated with the sulcus; no significant blood flow was observed in the densely collagenous part of the phallus. We also identified a pair of paralymphatic bodies near the proximal end of the phallus connected to lymphatic elements that extend distally into the sulcal tissues. We propose that these tissues are homologous to the paralymphatic bodies instrumental for phallus inflation in ratites; however, it is not yet known whether lymphatic fluid is involved in sulcus inflation in crocodilians. Research supported in part by an NSF EPSCoR RII award via the Louisiana Board of Regents Pfund.
Enteral feeding therapy: Characterization of patients and care in a tertiary hospital
Authors: Paiva Miriam , Okoshi Marina , Martim Vanessa , Barbin Angela , Zambom Raquel , Lima Silvana
Published: June 2021

Objective

To describe patients on enteral nutrition therapy (NET) and aspects of care held in Brazilian tertiary university hospital. Method: Retrospective and descriptive study of secondary data obtained from the electronic patient record (EPR). The study included all patients in NET by tube, admitted to the Internal Medicine (IM), Neurology (Neu) and Adult Intensive Care (ICU) units, between September and November 2014. Results: There were 222 admissions of patients with NET (33%), 44% in IM, 34% in ICU and 22% in Neu. 17% patients underwent a nutritional screening in the first 24h. 54% were men and 50% with age over 60 years. During the period were carried out nutritional assessments in 58% patients. In 184 patients who used naso/oroenteral tube, 44.6% were inadvertently removed, and in 28 patients the tube was removed 2 to 7 times. 58 patients had diarrhea, most of them (53%) for 1 day and 33% by 2‐4 days and 14% for more than 5 days. During the 2111 days of NET, 28 patients did not have the prescribed nursing care at any time and in 1531 days did not have any record of nursing evolution. 72 patients in NET died, 45 in ICU, 25 in IM and 2 in Neu.

Conclusion

EPR enabled the collection of necessary data to characterize the assisted population and aid quality aspects given in hospital units. The patient's profile and the detection of weaknesses in care indicate the need for corrective action planning and preventive and direct approach to the improvement of care.
Characterization of L‐Threonine transport in mouse embryonic stem cells
Authors: Formisano Tara , Wan Winkle Lon
Published: June 2021

Stem cells are at the forefront of current regenerative and biomedical research. There exists an imperative need to understand the mechanisms that drive stem cell function in order to exploit their use as a therapeutic tool. Amino acids are potent inducers of signaling cascades that drive stem cell proliferation and differentiation. With a focus on mouse embryonic stem (mES) cells, L‐threonine is the only amino acid required in culture medium for mES cell proliferation. Current research associates this need for L‐threonine with threonine dehydrogenase (TDH), which catabolizes L‐threonine to glycine and acetyl‐CoA in mES cells. While this may be true for mES cells, human embryonic stem (hES) cells do not express a functional tdh gene. Additionally, the concentration of 3‐hydroxynorvaline (3‐HNV), a potent inhibitor of TDH, needed to inhibit mES cell proliferation is more than an order of magnitude less than the apparent Ki for TDH inhibition. Such findings suggest a different mechanism responsible for the relationship between L‐threonine and ES cell proliferation, which may be explained by Thr transport. To explore this novel idea, we characterized the L‐threonine transport systems in mES cells by conducting radiolabelling transport studies. We found that there is a Na+‐dependent and Na+‐independent component of transport, with the Na+‐dependent component predominating. We also found that Thr, Ser, Cys, Ala, 4‐OH‐Pro, Asn, 3‐HNV, and Met were the strongest inhibitors of the Na+‐dependent component of transport. Such findings are consistent with characteristics of the ASC transport system, suggesting that this system is responsible for majority of Thr transport in mES cells. We confirmed the presence of the ASC system transporters, ASCT1 and ASCT2, in mES cells using RT‐PCR. Project supported by Midwestern University.
Glutamine Deprivation Hinders the Proliferation of Human Oligodendroglioma Cells Negative for Glutamine Synthetase
Authors: Ottaviani Laura , Chiu Martina , Bianchi Massimiliano , Bussolati Ovidio
Published: June 2021

Negativity for the astroglial marker Glutamine Synthetase (GS) is an histological hallmark of human oligodendrogliomas (OL). Given that GS accounts for the intracellular synthesis of glutamine (Gln), we hypothesized that OL cells are strictly dependent upon the availability of external Gln. To test this hypothesis, we compared the effects of several treatments on two human oligodendroglioma cell lines, HOG and Hs683, and two GS‐positive glioblastoma cells, U87 and U373 cells. Compared to HOG and Hs683 cells, the abundance of the GS‐encoding GLUL mRNA was more than 1000‐fold higher in U373 cells, while the expression of GS protein was undetectable in Western Blot in both OL lines. HOG and Hs683 cells were more sensitive than glioblastoma cells to reduced availability of extracellular Gln caused by (1) the glutaminolytic enzyme L‐Asparaginase (ASNase) from Erwinia chrysanthemi (IC50 values < 0.1 U/ml); (2) the incubation at decreasing levels of extracellular Gln; (3) the incubation in the presence of competitive inhibitors of sodium‐dependent transporters for Gln. In particular, the viability of both OL cell lines was markedly affected by MeAIB, an inhibitor of the SNAT transporters of the SLC38 gene family, while glioblastoma cells were substantially unaffected. Moreover, both OL cell lines exhibited higher levels of expression of the mRNA for the SNAT1 transporter than glioblastoma cells. Thus, OL cells are markedly dependent on extracellular Gln suggesting that GS negativity may constitute a marker of sensitivity to treatments aimed at reducing the availability of the amino acid.
A Scanning Acoustic Microscope ‐ A Novel Tool for Cytological Discrimination
Authors: Miura Katsutoshi , Tsuchida Takashi , Yamamoto Seiji
Published: June 2021

A scanning acoustic microscope (SAM) uses ultrasound to make cell images on the slide by plotting the data of cell thickness, speed of sound (SOS), and attenuation of sound (AOS) through cells without staining. We have applied this microscope to discriminate various free cells in the fluid for cytological diagnosis.

Purpose

How different are SAM images of various free cells from optical ones? Is SAM image analysis applicable to cytological diagnosis?

Materials and Methods

Residual cells of cytology specimen from body cavity were prepared to make single cell‐layer slides by liquid‐based cytology method. The slides were fixed in ethanol and kept dry until observation. The slide were soaked in distilled water and observed with SAM. Cells from squamous cell carcinomas, adenocarcinomas, reactive mesothelial cells, mesotheliomas, malignant lymphomas, and neutrophils /macrophages were compared of their SOS, AOS, and thickness.

Results

SAM could visualize not only SOS, AOS and thickness of each cell portion but also cell sizes, thicknesses, borders, contents, cell clusters and structures. Various free cells showed their own characteristic acoustic images that was equivalent to optic ones. Obtained data with SAM was statistically comparable among different cell types.

image

image

Cytological analysis

SAM
LM

Nucleus
Size
△
◎

Irregularity/groove
X
◎

chromatin density
X
◎

Cytoplasm
Size
◎
○

Border
◎
○

Content
◎
○

Irregularity
◎
△

Cell connection/Cluster

◎
◎

Structure

◎
◎

Statistical analysis

◎
○

Discussion

Each acoustic property of different cell types was distinct to make characteristic acoustic images. Statistical difference in SOS, AOS and thickness of each cell type may help discriminate cell types. SAM can be a novel diagnostic tool for cytological diagnosis.
Nigerian Medical Students at Lagos State Universtity College of Medicine (LASUCOM) Learn With Information Communication Technology
Authors: John Theresa , Agaga Luther
Published: June 2021

Introduction

Electronics and information technology are now in most spheres of educational activities. This survey examines if Nigerian students at LASUCOM have joined the global trend.

Methods

Medical students (2013/2014 session; first‐last years of MBBS Program) completed a 23‐question survey within 10min by marking one/more choices from alternatives. Instructions were: “Dear Participant, Please treat as confidential, give your true view, avoid influences, avoid crosstalk, return survey promptly.”

Results

Out of 301 students, 188 (62.46%) participated. Simple statistics gathered were: 73.7% liked e‐devices and considered them useful; 11% must always have electronics with them; they have laptops (64.4%), smart phones (62.7%), cell phones (43.6%), tablets (28.8%), desktops (2.2%), and ultrabooks (0.44%); during class they use smart phones (48%), cellphones (20.9%), tablets (19.6%), laptops (7.5%) and ultrabooks (0.89%); 5.8% started using electronics in primary school, 56% in secondary school, and 26.2%, 2.2%, 0.44%, and 0.44% in 1st, 2nd, 3rd, and 4th years of LASUCOM. Presently, to capture lectures 61.3% write in class, 58.67% collect lecturer's e‐file, actually 36% prefer to write. For schoolwork: 35.5% email classmates, 17% email lecturers; 61.8% phone classmates, 26.2% phone lecturers; 68% claimed electronics make school work easier, 29.3% claimed electronics help to enjoy work; 82% use the Internet for general information, 40.4% to add to lecture notes, 17.33% to answer exam questions.

Conclusion

The students' electronics culture: 73.7% like electronics, >60% possess laptops and smart phones, >50% use electronics for lectures, studying, and communication – is a predictor of effective teaching and training styles.
Quantifying Signaling Bias: A Simple Approach to Quantify Functional Selectivity and Agonist Bias
Authors: Charter Neil , Anantharaman Lakshmi
Published: June 2021

Interest in GPCR ligand bias has increased in recent years due to evidence that positive and negative aspects of drug activity can be due to differential pathway signaling. As a result it is possible to develop drugs with reduced side effects by enhancing positive effects by favoring one pathway over another. The availability of assays that can measuring signaling events such as calcium mobilization, cAMP modulation, arrestin recruitment and receptor internalization has provided to opportunity to examine specific pathways. Here we provide examples of ligand bias and how quantification of receptor potency and efficacy in different pathways can be used to generate a ligand bias index.
Calmodulin: a new partner for the α1A‐adrenergic receptor
Authors: Gebert‐Oberle Briana , VerMeer Mark , Tran Quang‐Kim
Published: June 2021

In addition to its role in vasoconstriction, the α1A‐adrenergic receptor (α1A‐AR) is increasingly recognized as a rescue mechanism under circumstances where b‐ARs are insufficient to provide stimulus in the myocardium, such as in heart failure. Mechanisms controlling α1A‐AR function therefore have profound impact on cardiovascular functions. Calmodulin (CaM) is the ubiquitous transducer of Ca2+ signals, and has recently been demonstrated to interact with a number of G protein‐coupled receptors. We have observed that in fresh ventricular tissues, α1A‐AR forms a complex with calmodulin in resting condition as well as under α1A‐AR agonism. To identify and characterize the precise interaction domains between CaM and α1A‐AR, we developed novel biosensors that span multiple fragments of the four sub‐membrane domains (SMDs) of α1A‐AR. Responses of these biosensors to Ca2+‐saturated CaM reveals that α1A‐AR directly interacts with CaM at a number of locations with disparate affinities and Ca2+ sensitivities. These data implicate CaM as a novel partner for α1A‐AR and provide background for on‐going studies on the functional impact of these interactions in the control of cardiovascular functions.
Botanical drug puerarin attenuates 6‐hydroxydopamine (6‐OHDA)‐induced neurotoxicity via upregulating mitochondrial enzyme arginase‐2
Authors: Zhao Jia , Rong Jianhui , Lao Lixing
Published: June 2021

Inhibition of nitric oxide synthases (NOS) shows promise to halt the progression of neurodegenerative diseases. The present study was designed to explore whether botanical isoflavone puerarin could attenuate nitric oxide (NO)‐mediated neurotoxicity via modulating the enzymes in the L‐arginine‐NO pathway. Neurotoxin 6‐hydroxydopamine (6‐OHDA) is well known to induce neurodegeneration via a NO‐dependent mechanism. We first validated that puerarin protected rat dopamingeric PC12 cells against 6‐OHDA‐induced neurotoxicity in a concentration‐dependent manner. We subsequently profiled the cellular responses to puerarin by a proteomic response fingerprinting approach. A total of sixteen protein spots with > 1.5‐fold change of intensity were selected and identified by mass spectrometry. As one of puerarin‐upregulated proteins, mitochondrial arginase‐2 hydrolyzes L‐arginine to L‐ornithine, thereby competing with neuronal NOS for substrate L‐arginine in mitochondria. Thus, we hypothesize that puerain may attenuate nitric oxide (NO)‐mediated mitochondrial injury via increasing arginase‐2 expression. Western blot and RT‐PCR analyses confirmed that puerarin increased arginase‐2 expression in a concentration‐ and time‐dependent manner. Accordingly, puerarin suppressed 6‐OHDA‐induced NO production and neurotoxicity in PC12 cells and primary rat midbrain neurons. Arginase inhibitor BEC diminished the effect of puerarin on 6‐OHDA‐induced NO production and neurotoxicity. The activation of arginase‐2 by puerarin represents an endogenous mechanism for specific control of NO‐mediated mitochondrial damage. Thus, puerarin is useful lead for suppressing NO‐mediated neurotoxicity in neurodegenerative diseases.
Human Argonaute‐2: For All Your RNA Slicing Needs
Authors: Sampe David , Bruss Evan , Davis T.J. , Hermsen Jack , Johnson Justin , Laughlin Robert , Lucre' Maurice , Marable Chad , Poniewaz Brett , Tuncel Virginia , Wade Gina , Weeden Michael , McNally Mark
Published: June 2021

Human cells have the remarkable capability to regulate protein production by degrading target mRNA by two pathways: RNA interference (RNAi) and micro RNA (miRNA). Central to these pathways is the protein Argonaute‐2 (Ago‐2). In the RNAi pathway, small RNAs derived from viruses are used by Ago‐2 to slice virus mRNA, protecting the cells from infection. In the miRNA pathway, Ago‐2 utilizes naturally occurring miRNA to slice cellular mRNAs to control protein production. Ago‐2 works by binding small (~22 nucleotide) regulatory RNAs (siRNA and miRNA) to target mRNA by base pairing. Ago‐2 attaches to the phosphate backbone of the regulatory RNA, that guides Ago‐2 to the target RNA. The RNase domain of Ago‐2 (containing His807, Asp669, Asp597, and Glu637 in its active site) then “slices” the target to initiate degradation. Scientists can reduce the level of disease‐causing proteins (for example, in breast cancer) using the siRNA pathway. Determining the structure of Ago‐2 allowed researchers to understand how this enzyme functions in the siRNA/miRNA pathways. The Brown Deer High School SMART (Students Modeling A Research Topic) Team has designed a model of Ago‐2 using 3D printing technology to investigate its structure‐function relationship. SMART Team programs are supported by a grant from NIH‐CTSA.
Paxillin Interaction Enhances MEKK2 Activity by Relieving Auto‐inhibition
Authors: Cuevas Bruce
Published: June 2021

The mitogen‐activated protein kinase (MAPK) regulator MEK kinase 2 (MEKK2) has been linked to control of inflammation, gene expression, cardiac hypertrophy, and breast tumor metastasis. The regulation of MEKK2 is not well understood, thus a key objective in our research is to define the mechanisms that control MEKK2 activity. Previously, we have demonstrated that attachment of invasive tumor cells to fibronectin induces MEKK2 activation and translocalization to focal adhesions, where it associates with the scaffold protein paxillin in an interaction that requires the amino‐terminal leucine‐rich motif (LD1) of paxillin. In this work, we reveal that interaction with paxillin enhances MEKK2 activity in vitro. Utilizing targeted MEKK2 mutagenesis, we demonstrate that MEKK2 kinase activity is markedly enhanced in a truncated form of the kinase that includes the kinase domain alone, suggesting that a regulatory domain exists within MEKK2 peptide outside of the kinase domain. Furthermore, we demonstrate that the observed auto‐inhibition requires the amino‐terminal region that includes the Phox and Bem1 (PB1) domain, and we show that this region is subject to post‐translational modification when transiently expressed in mammalian cells. Finally, we show that the paxillin LD1 peptide alone is sufficient to pull down MEKK2, relieve MEKK2 auto‐inhibition and enhance kinase activity. Taken together, our results reveal new mechanisms for regulation of MEKK2 kinase activity whereby MEKK2 activity is controlled by an amino‐terminal regulatory domain, and that interaction with the scaffold protein paxillin releases MEKK2 from auto‐inhibition and promotes activation.
Altered Drug and Steroid Metabolism by Mutations in Human NADPH Cytochrome P450 Reductase
Authors: Pandey Amit , Flück Christa
Published: June 2021

Cytochrome P450 oxidoreductase (POR) is required for the metabolic reactions catalyzed by cytochrome P450s in the endoplasmic reticulum. It also participates in metabolism of several other molecules like mitomycin C, ferricyanide, menadione, dichlorophenolindophenol etc either directly or as an electron donor to the actual catalyst. We had described mutations in POR from patients with disruption in sex steroid metabolism and established that alterations in POR lead to reduced activities of steroid metabolizing enzymes CYP17A1 (Flück et al. Nature Genetics 36:228 2004) and CYP19A1 (Pandey et al. Mol. Endocrinol. 21:2579 2007), heme oxygenase and CYP3A4 (Pandey & Flück, Pharmacology & Therapeutics 138:229 2013). In the patients classified as “European” or “Caucasian” most common POR mutation is A287P. We have observed variable effects of A287P variant on activities of different redox partners of POR. The CYP3A4 and 17OH‐hydroxylase and 17,20 lyase activities of CYP17A1 are reduced but CYP19A1, CYP21A2 remain unaffected. We have performed detailed enzymatic and biochemical characterization of the A287P variant of POR to study its effects on different substrates. We used purified preparations of wild type and A287P variant of human POR and performed testing with small molecule and protein substrates of POR under changing substrate, co‐factor and buffer conditions. Here we show that small molecule and protein substrates are affected in different ways by the A287P mutation. Similarly differential effects of common polymorphic variant of POR A503V have been evaluated with different drug metabolising P450s to understand the implications of POR variants on drug metabolism. In conclusion, variations in POR may lead to different activities of its redox partners that need to be evaluated individually.
Effect of Curcumin on the Production of Interleukin‐6 from MDA‐MB‐231 Breast Cancer Cells
Authors: Paciotti James , Condori‐Salas Ramiro , Rojas‐Humpire Ricardo , Pino‐Figueroa Alejandro
Published: June 2021

Cytokines such as interleukin‐6 (IL‐6) have an important role in cellular proliferation and survival. Overexpression of IL‐6 in inflammatory or degenerative diseases has been observed in patients with various types of cancer, arthritis and in the elderly; furthermore, it can be used as a marker of disease and degeneration. Natural agents such as Curcumin or its derivatives could produce their protective effects by inhibiting the production of IL‐6.

The purpose of the current study was to investigate the inhibitory effects of Curcumin and Curcumin Diethylsuccinate on the production of IL‐6 at concentrations from 1 to 10µM in MDA‐MB‐231 breast cancer cells. Cells were cultured for 3‐5 days with Dulbecco's Modified Eagle Medium (DMEM) at 37°C and 5% CO2. Media samples were used to determine the levels of IL‐6 by enzyme‐linked immunosorbant assay (ELISA) that uses antibodies to achieve a sensitive colorimetric detection at 450 nm. IL‐6 measurements were normalized to cell viability determined by MTS assay at 490 nm.

Administration of Curcumin Diethylsuccinate 1µM to MDA‐MB‐231 cells reduced their production of IL‐6 from 79±5 to 30±1 pg/mL (62%), showing that this agent has an inhibitory effect on the cytokine release from cancer cells and could be useful in the treatment of cancer and other degenerative disorders.
Mouse Models for Studying Human Disease
Authors: Beckwith Jason , Rockwood Stephen , Sasner Michael
Published: June 2021

Access to model organisms is key to advancing the research objectives of the biomedical research community. To facilitate this access, The Jackson Laboratory (JAX) Mouse Repository has served as a centralized resource for the distribution, cryopreservation and development of high health status mouse models for over 50 years. Hundreds of new strains are added annually to one of the largest collections of characterized mouse strains available.
DREADDs (designer receptors exclusively activated by designer drugs) are mutant G‐protein coupled receptors activated by the pharmacologically‐inert molecule clozapine‐N‐oxide. Several pharmacogenetic tool strains have Cre‐ or Tet‐inducible expression of the DREADDs hM3Dq, hM4Di or rM3Ds. Newly available Cre‐lox, Tet‐On/‐Off, fluorescent, optogenetic, CRISPR/Cas9 and other tool strains will also be described.
Search JAX Mouse Repository holdings using the newly designed JAXMice database (jaxmice.jax.org/query). In addition, we show other web resources useful to identify and obtain mouse models for human disease research. The International Mouse Strain Registry (IMSR; findmice.org) displays mouse strains and mutant ES cell lines available worldwide. The Human/Mouse Disease Connection (HMDC; diseasemodel.org) is a human‐to‐mouse data traversal that enables clinical and translational researchers to access mouse model data/annotations, and allows mouse researchers to connect their findings directly to genetic associations reported in human disease.
Donating a strain to JAX Mouse Repository fulfills the NIH requirement for sharing mice; researchers wishing to do so may submit them via our online submission form (jax.org/donate‐a‐mouse). JAX Mouse Repository is supported by NIH, The Howard Hughes Medical Institute and several private charitable foundations.
Aqueous extract of Gumiganghwal‐tang, traditional herbal medicine, reduces pulmonary fibrosis through transforming growth factor‐β1/Smad signaling pathway in murine model of chronic asthma
Authors: Jeon Woo‐Young , Shin In‐Sik , Shin Hyeun‐Kyoo , Lee Mee‐Young
Published: June 2021

Gumiganghwal‐tang is a traditional herbal prescription that is widely used for the treatment of the cold and inflammatory diseases in Korea and other Asian countries. In this study, we investigated the protective effects of aqueous extract of Gumiganghwal‐tang (GGTW) against airway inflammation and pulmonary fibrosis using a mouse model of chronic asthma. Chronic asthma generated by sensitizing and inhalation BALB/c mice with intraperitoneal injection and nebulized 1% ovalbumin (OVA) for 4 weeks. GGTW (100 mg/kg or 200 mg/kg) was also administered by oral gavage once a day for 4 weeks. We investigated the production of Th2 cytokines, chemokine and transforming growth factor (TGF)‐β 1 in bronchoalveolar lavage fluid (BALF), the levels of immunoglobulin (Ig)E in plasma, and the expression of TGF‐β 1, Smad‐3 and collagen in lung tissue. Our results revealed that GGTW decreased the production of Th2 cytokines, chemokine and TGF‐β 1 in BALF, and attenuated levels of total and OVA‐specific IgE in plasma. Moreover, GGTW significantly reduced the expression of TGF‐β 1, Smad‐3 and collagen deposition in lung tissue. These results indicate that GGTW reduces the airway inflammation and pulmonary fibrosis via regulation of Th2 cytokines and TGF‐β1/Smad‐3 pathway, providing a potential treatment for chronic asthma.
Intravital Video Microscopy of Forelimb Capillary Blood Flow in Rat Skeletal Muscle
Authors: Fraser Graham
Published: June 2021

Objective

The present study describes the development of a novel in vivo microvascular preparation for the investigation of capillary blood flow in the extensor carpi radialis longus (ECRL) muscle of rat. Blood flow measurements in the forelimb provide an alternative to other contemporary vascular preparations and is particularly useful for investigating capillary blood flow in disease models where hind limb flow is impaired (e.g. hind limb unloading, reduced uterine perfusion pressure etc.).

Methods

Two male Sprague‐Dawley rats, 174 and 161g were anaesthetized with pentobarbital, tracheotomized, ventilated, and maintained in a normotensive state via minimal saline infusion in the jugular vein. The skin on the lateral side of the right forelimb was removed exposing the muscle capsule surrounding the extensor carpi radialis longus and brevis muscles. The distal tendon of the ECRL was secured with a silk ligature and the muscle was carefully isolated and reflected onto an inverted microscope stage. The muscle was bathed in warm saline and isolated from the air via polyvinylidene chloride film and a glass coverslip. Ten – twenty, 1 minute video sequences of capillary blood flow at 20X magnification were recorded in each animal. Videos were analyzed offline using custom software written in MATLAB yielding frame‐by‐frame measurements of capillary hemodynamics and oxygen saturation.

Results

Capillary measurements were made in a total of 69 vessels (31 ‐ 38 in each animal), with erythrocyte velocities of 196.8±40.2 μm/s; tube hematocrit 19.9±2.9 %; 10.7±0.7 cells/s; and entrance and exit oxygen saturation of 70.4±5.6 % and 55.8±4.6% respectively.

Conclusions

We demonstrate direct measurement of microvascular hemodynamics and capillary oxygen saturation in a novel intravital preparation using rat ECRL muscle.
Effect of Gastric Bypass Surgery on Endothelial function and Heart Rate Variability
Authors: Lee Kyuwan , DeVallance Evan , Grogg Kristen , Mason Rebecca , Hull Robert , Rosati Kristen , Tabone Lawrence , Frisbee Jefferson , Chantler Paul
Published: June 2021

The obesity epidemic in the US has reached 155 million people. Obesity is associated with increased incidence of cardiovascular (CV) and metabolic diseases. The use of gastric bypass surgery has been shown to decrease excess body weight and reduce CV risk. Although, more than 100,000 procedures were performed during the past decade, it is unclear how it contributes to endothelial and autonomic function. The purpose of this study was to examine the outcomes of gastric bypass surgery on endothelial function and heart rate variability (HRV).

We assessed 12 gastric bypass patient's peripheral arterial and endothelial function along with HRV using EndoPAT device, pre and 1 week post‐surgery.

The mean of augmentation index (AI) was lower (p<0.05) 1 week post‐surgery (1.7±8.0 to ‐7.2±4.1) with an increase in mean of AI pulse (178±12 to 193±46, p<0.05). LF/HF ratio was increased (1.03±0.26 to 1.34±0.28, p<0.05) after gastric bypass surgery suggesting that it may contribute to improved autonomic modulation of CV function. In contrast, no changes were found in endothelial function. These findings show that gastric bypass surgery improves the parasympathetic to sympathetic input on the heart and peripheral arterial stiffness despite no change in endothelial function.
Manipulation of Induced Pluripotent Stem Cell Differentiation to Endothelial Cells
Authors: DiVincenzo Lola , Kolz Chris , Chilian William , Yin Liya
Published: June 2021

With the potential use of induced pluripotent stem cells (iPSCs) in regenerative medicine, strategies must be implemented to guide iPS cell differentiation. Without guidance, the pluripotent nature may yield tumorigenic and/or undesired cell types. Previously we observed that the endothelial cell secretome (EC‐S) derived from conditioned media (EC‐CM) guided iPS cell differentiation to an endothelial cell phenotype. Specifically, the EC‐S induced iPS cell differentiation as evidenced by LDL scavenging and tube formation in vitro. Of keen interest is the question of what product(s) of the EC‐S induce the differentiation of iPS cells. To determine this, we first confirmed that treatment of IPS cells with conditioned media from normoxic or hypoxic (1‐2% O2) endothelial cells produced differentiation of iPS cells into endothelial cells. To determine if a protein factor is responsible for the differentiation, we subjected the conditioned media to heat denaturation (10 min @ 70 C). In media obtained from normoxic endothelial cells, heat denaturation greatly reduced the differentiation of iPS cells into endothelial‐like morphology. In fact the vast majority of cells in culture retained an IPS cell phenotype. Conversely, the heat denatured conditioned media from hypoxic endothelial cells did reduce the differentiation of IPS cells into an endothelial phenotype. These results suggest that the products of the secretome that induce differentiation of iPS cells into endothelial cells are changed by hypoxia. The results also suggest that during normoxia, the active component of the secretome are proteins (the activity is lost with heat denaturation), but this changes during hypoxia to a factor (or factors) that is resistant to heat denaturation.
Nicotinamide Phosphoribosyltransferase (NAMPT) Inhibition Yields Promising Future Implications
Authors: Kaiser Carl , Klestinski Keith , Tsuji J , Sabatino T , Yorke N , Johnson R , Arnhold K , Otten J , Ahn P , Cephus K , Dittlof A , Ortega L , Foster H , Hernandez M , Rivera M , Strom D , Strom J , Yang N , Dittrich N , Waas Matthew
Published: June 2021

Human embryonic stems cells (hESC) and induced pluripotent stem cells (hiPSC), collectively termed human pluripotent stem cells (hPSC), differentiate into any cell type. The creation of hPSC free from potentially tumorigenic pluripotent stems is advantageous for research strategies and necessary for future hPSC‐based clinical therapies. The STF‐31 molecule inhibits an important metabolic enzyme, NAMPT, providing selective toxicity of hPSC in diverse cell culture conditions. This strategy effectively eliminates potentially tumorigenic cells but spares differentiated progeny. The MUHS SMART (Students Modeling A Research Topic) Team has designed a model of NAMPT bound with STF‐31 using 3D printing technology. What makes STF‐31 unique as a NAMPT inhibitor is its ability to occupy the protein's active site and act as a substrate for the enzyme. The pyridine ring of STF‐31 is situated between the F193 and Y188 sidechains of NAMPT. The central phenyl ring of STF‐31 occupies the tunnel region of NAMPT. Other binding sites between STF‐31 and NAMPT include H191, R196, S241, V242, A244, S275, I309, and R311. NAMPT inhibition research will lead towards the development of clinically safe hPSC progeny for human stem cell based therapies, drug development, and toxicity testing. This is supported by a grant from NIH‐CTSA.
Aging and Atherosclerosis Impair Hematopoietic Stem Cell Function
Authors: Porto Marcella , Rodrigues Bianca , Bezerra Geisiane , Campagnaro Bianca , Menezes Thiago , Pereira Thiago , Vasquez Elisardo , Meyrelles Silvana
Published: June 2021

Background

Stem cell function is compromised with aging, especially when a cardiovascular disease is associated. The aim of this study was to evaluate how aging and atherosclerosis affect bone marrow hematopoietic stem cell (HSC) function in mice.

Methods: Male mice were divided into C57 and apoE‐/‐ (2, 12 and 24‐month‐old) groups. Oxidative stress was assessed by 2,7‐dichlorofluorescein (DCF) by flow cytometer. For apoptosis detection, HSC were incubated with Annexin V‐FITC and propidium iodide (PI). Plasma inflammation markers, IL‐6 and MCP‐1, were measured. Also, senescence was evaluated by acridine orange protocol. Lipid profile and aortic lipid deposition were also analyzed. Data are mean±SEM. Statistical analysis was performed with two way ANOVA test and Tukey pos hoc.

Results: Aging and atherosclerosis increased ROS levels and apoptosis in HSC. Senescence was also increased with aging and atherosclerosis, especially in 24‐month‐old mice. Atherosclerotic adult and aged animals showed higher levels of IL‐6 and MCP‐1. As expected, atherosclerosis augmented total serum cholesterol levels and aortic lipid deposition.

Conclusion

The present study provides evidence that aging and atherosclerosis can cause oxidant stress and impair HSC function. Such findings could lead to new insights into the mechanisms by which aging and atherosclerosis can affect immunity and inflammation.
PKD Regulates Mitochondrial Morphology and Function via Phosphorylation of DLP1 in Cardiac Myocytes.
Authors: Jhun Bong Sook , O‐Uchi Jin , Mishra Jyotsna , Xu Xiaole , Hurst Stephen , Mende Ulrike , Sheu Shey‐Shing
Published: June 2021

Regulation of mitochondrial morphology and dynamics is essential for maintaining cardiomyocyte function. Abnormal mitochondrial morphology and mitochondrial dysfunction are frequently observed in both human heart failure (HF) and animal HF models. However, it is still unclear which cardiac signaling pathway(s) regulate(s) mitochondrial morphology/function under pathophysiological conditions. Since Gq‐protein coupled receptor (GqPCR) signaling is critical for the development and progression of HF, we tested the hypothesis that GqPCR stimulation induces pathological changes in mitochondrial morphology/function in cardiomyocytes. We found that protein kinase D (PKD) was activated and translocated to the outer mitochondrial membrane by GqPCR stimulation. GqPCR‐mediated PKD activation induced mitochondrial fragmentation, increased reactive oxygen species generation and mitochondrial permeability transition pore opening followed by caspase‐3 activation. These morphological and functional changes in cardiac mitochondria were mediated via PKD‐dependent phosphorylation of Dynamin‐Like Protein 1 (DLP1) at S637. Importantly, PKD‐dependent DLP1 phosphorylation at S637 concurrent with abnormal mitochondrial morphology and apoptotic signaling were also observed in ventricular tissue from transgenic mice with cardiac‐specific overexpression of constitutively active Gq. In conclusion, we demonstrate that GqPCR stimulation induces mitochondrial fragmentation and dysfunction via PKD‐dependent phosphorylation of DLP1 at S637, which likely contributes to cardiomyocyte dysfunction during HF.
Regulation of the Tight Junction Protein Cingulin in Ovarian Theca Cells of Cattle
Authors: Zhang Lingna , Schutz Luis , Robinson Cheyenne , Totty Morgan , Spicer Leon
Published: June 2021

Recently, we discovered using microarray analysis that expression of cingulin (CGN), a cytoplasmic tight junction protein, is significantly down‐regulated by fibroblast growth factor 9 (FGF9) in cultured ovarian theca cells (TC). FGF9 is known to increase cell proliferation and decrease steroidogenesis in bovine TC. The objective of the present studies was to investigate the hormonal regulation of CGN gene expression in bovine TC. Ovaries of cattle were collected at a local slaughterhouse, and TC isolated from large follicles (8‐22 mm). TC were cultured for 2 days in 10% fetal bovine serum followed by 24 h in serum‐free medium containingvarious treatments in three experiments, and TC RNA was isolated to measure abundance of CGN mRNA by real‐time PCR. Exp. 1: TC were treated with 30 ng/ml of FGF9 for either 0, 4, 12, or 24 h, and CGN mRNA measured; data were analyzed via 3×2 factorial ANOVA. CGN mRNA abundance averaged 60% less (P < 0.05) in FGF9‐treated than in control TC at both 12 h and 24 h. Exp. 2: TC were treated with 30 ng/ml of either FGF9, insulin‐like growth factor 1 (IGF1), tumor necrosis factor (TNFα), IGF1 plus FGF9, or dexamethasone (300 ng/ml) for 24 h, and CGN mRNA measured; data were analyzed via one‐way ANOVA. CGN mRNA abundance was 64% less (P < 0.01) in TNFα‐treated TC and 49% greater (P < 0.05) in dexamethasone‐treated TC than in controls. IGF1 tended (P < 0.10) to increase CGN mRNA abundance while FGF9 attenuated (P < 0.01) the IGF1 effect. Exp. 3: Estrogenic and androgenic compounds had no significant effect on CGN mRNA expression. These results indicate that FGF9 and TNFα inhibit whereas glucocorticoids stimulate CGN mRNA expression in TC, and suggests a role of inflammatory and stress hormones in regulating the tight junction protein CGN.
Progressive mechanisms on naïve wound healing process in an animal model with skin injury
Authors: Choi Jeonghyun , Hong Yunkyung , Hong Yonggeun
Published: June 2021

Cutaneous wound healing is a complex and coordinated series of events. A wide variety of growth factor and cytokines are involved in the regulation of all phased of wound healing. Inflammatory phase of the healing process develops during 24 hours from the moment when an injury occurred and last for up to 48 hours or above that. The early inflammatory phage of wound healing cascade is characterized by subsiding of the vessel contraction followed by widening their lumen, which promotes leaking of plasma to the wound site. The changes are supported by reactive oxygen species (ROS). Low levels of ROS are important in wound repair by protecting the injured area against microbes along with enhancing angiogenesis. Normal ROS levels promote the collagenase activity MMPs and other signalings which help wound reepithelilization through maintaining normal keratinocyte migration and proliferation. Normal ROS also aid in the production of collagenI,III and their subsequent cross linking. High levels of ROS increase apoptosis of keratinocytes and damages fibroblasts, causing them to become senescent and lose the ability to produce extracellular matrix. ROS stimulate apoptosis through the C‐junN‐terminal kinase (JNK) pathway and stimulates translocation of JNK to the mitochondria. The aim of this study was that to provide inflammatory phage of normal wound healing process without intervention.

Funding: 2012R1A1A2005089, 2013R1A2A2A01067169, KGM4611512
Comparing the effects of rapid and gradual cooling following acute hyperthermia on body temperature and inflammatory response
Authors: Johnson Jay , Sapkota Avi , Lay Donald
Published: June 2021

Hyperthermia negatively impacts human and animal health and may cause morbidity and mortality if recovery is not properly managed. The study objective was to evaluate the effects of two cooling methods on body temperature and the inflammatory response after acute heat stress (HS) exposure in a pig model for human hyperthermia. Male pigs (88.7 ± 1.6 kg BW) were exposed to thermoneutral conditions (TN; n = 12; 19.5 ± 0.1°C) for 6 hrs, or HS (36.4 ± 0.1°C) for 3 hrs, followed by a 3 hr recovery period of either rapid cooling (HSRC; n = 12; immediate TN exposure and ice water dousing) or gradual cooling (HSGC; n = 12; gradual decrease to TN temperature). Rectal (TR) and gastrointestinal temperatures (TGI) were obtained every 15 min. Blood samples were collected from 6 pigs per treatment at 60, 180, 210 and 240 min to determine TNFα and IL‐1ß concentrations. TR was increased (p < 0.01) in HSRC and HSGC pigs compared to TN controls during HS (1.2°C) and recovery (0.6°C), and was reduced (p < 0.01; 0.4°C) in HSRC versus HSGC pigs during the recovery period. TGI was increased (p < 0.01) in HSRC and HSGC compared to TN pigs during HS (1.6°C and 1.5°C, respectively) and recovery (1.2°C and 1.3°C, respectively), but was similar (40.8°C) in HSRC and HSGC pigs during both periods. During HS, HSRC and HSGC pigs had increased TNFα concentrations (p < 0.04; 27.5%), but IL‐1ß concentrations were similar (p > 0.60; 10.2 pg/mL) to TN controls. In the recovery period, HSRC pigs tended to have increased TNFα concentrations (p < 0.07; 41.2%) compared to HSGC pigs. In summary, although rapid cooling was more effective than gradual cooling in reducing TR during HS recovery it had no effect on TGI and this differential may be linked to the increased inflammatory response observed in HSRC pigs.
Cerebellar HSP70 Remains Elevated During Heat Stroke Recovery despite Return to Baseline Core Temperature
Authors: Fulco Cameron , Dineen Shauna , Audet Gerald , Leon Lisa
Published: June 2021

Heat Stroke (HS) remains a significant health threat for civilians and U.S. Armed Forces. Unknown is whether return to normal core temperature (Tc; ±0.1°C, radiotelemetry) within 2 days of HS is an accurate indicator of full recovery. Cerebellar damage is implicated in HS sequelae, but inflammatory responses in this brain region during prolonged recovery have not been investigated. We hypothesized that the cerebellum would show a differential time course of HS recovery compared to Tc during 7 days of HS recovery. C57BL/6J mice (N=3‐7/group) were heated to Tc of 42.4ºC and the cerebellum was extracted at 4 time points (3 hours, 1, 2, and 7 days) of recovery. Twelve common HS biomarkers were examined using rtPCR, ELISA, and Western Blot. There were no changes in any genes at 7 days, indicating normalization of expression. HSP70 protein peaked at day 1 and remained elevated through 7 days. At 3h, five cytokine genes (IL6, HSP70, TNFα, HSF‐1, IL‐1β) and three chemokine genes (Ccl2, Ccl3, Ccl4) were elevated, while one chemokine (Ccl5) was down‐regulated. At 1 day, only Ccl2 was still significantly elevated, yet returned to baseline levels by 2 days. Elevated HSP70 protein at 7 days following HS suggests persistent protein denaturation, which could indicate incomplete recovery even though other inflammatory markers and Tc have returned to normal. These data suggest that a return to normal Tc should not be the only indicator for HS recovery and persistent inflammatory changes may be occurring in the brain through 7 days. Author views not official US Army or DoD policy.
Reliability of Skin Blood Flow Measures by Laser‐Doppler Flowmetry
Authors: Herron Robert , Carter Stephen , Collins Ann , Leatherwood Matthew
Published: June 2021

Laser‐Doppler flowmetry (LDF) is a non‐invasive technique often used in combination with local heaters to examine changes in skin blood flow (SkBF). Though customary to normalize cutaneous vascular conductance (CVC) to maximum, CVC normalized to baseline is less common. Accordingly, we sought to evaluate between site measures of SkBF to test the reliability of such practices. Nineteen participants (12 males, 7 females, age 23 ± 3 yr) completed two trials (T1 and T2). Participants rested supine as the dorsal aspect of their dominant forearm was instrumented with two LDF probes and local heaters. Baseline SkBF (in arbitrary units; au) and blood pressure were monitored during which local skin temperature was clamped at 34 °C. Each site was then heated to 42 °C at a rate of 0.2 °C·10 s‐1 for 30 min to attain a maximal SkBF response (i.e., plateau). Blood pressure was measured by automated oscillometry at 10 min intervals. Mean arterial pressure (MAP) was used to calculate CVC (au/MAP). The same procedures were conducted during the second visit. Data were averaged from the two sites and analyzed in raw au's and CVC at baseline and max. Intraclass correlations (ICC) of raw au's at baseline and max were moderately reliable, 0.68 and 0.62, respectively. Additionally, baseline and max CVC were moderately reliable, 0.65 and 0.52, respectively. However, when CVC was expressed as percent change (%Δ) from baseline to max ICC was lower (0.39). These data suggest that indices of SkBF normalized to baseline offer moderate reliability.
Screening for early detection of stomach cancer in Kazakhstan: first results
Authors: Orazova Galiya , Karp Leonid , Dossakhanov A.
Published: June 2021

Introduction

Over the last 15 years (1999‐2013) more than 42 thousand patients have been registered with stomach cancer (SC) identified for the first time.

Materials and methods

In 2013, screening was introduced in Kazakhstan with the aim of early detection of stomach and esophageal cancers. The target population is persons aged 50‐60 years. Screening by endoscopic examination was conducted in a pilot mode in 4 regions and in Astana and Almaty cities.

Results

150,546 persons were examined in the targeted age groups.1,649 patients were identified with disorder of gastro‐intestinal tract mucosa (0.9% from examined patients), of whom 1,589 people (96.4%) were registered for medical checkup. Statistically significant differences of endoscopic esophageal and stomach disorders frequency were identified between max (2.8%) and min (0.4%) parameters (p<0.05). The frequency of the disorder was the highest in the age group of 60 years. According to the data of primary medical care the diagnosis of SC was achieved in 87 cases (0.05%). According to the endoscopic examination 79 cases of SC were identified (including 39 morphologically verified). According to the cancer registry, SC detectability is 0.04% from the number of examined patients. Complete agreement between primary medical care and cancer‐register data was observed only in one region. SC is identified more frequently at stage II (47.0%).

Conclusions

According to the primary medical care and cancer registry data, there is mismatch in the results: almost in all regions both cases of stomach cancer over diagnosis in some regions and misreporting in others.