bluuebunny/crossref_metadata_2025 huggingface dataset:
Resting Measures of Cardiovascular Autonomic Control are not Influenced by Energy Drink Ingestion or Postural Changes in Healthy Adults
Authors: Leatherwood Matt D , Herron Robert L , Carter Stephen J
Published: November 2023

INTRODUCTION

Energy drink consumption is common among young people and linked to acute alterations in cardiovascular‐autonomic profiles. However, the magnitude and relevance of such changes is equivocal. This study evaluated resting autonomic control, as evidenced by heart rate variability (HRV), during postural changes following ingestion of a commercially‐available energy drink (ED) and zero‐kcal control drink (CD).

METHODS

Using a single‐blind, repeated measures, counter‐balanced crossover design, 14 (7 women) young (mean ± SD; 25 ± 3 y), healthy adults completed two testing sessions after an overnight fast. Participants were asked to avoid vigorous exercise and caffeine consumption the day prior to testing. Each session was separated by at least 48 h. Baseline HRV measures were recorded in the supine (Sup) and standing (St) positions prior to ingesting either an ED or control drink (CD) of equal volume. Following a 30‐min digestion period, HRV measures were repeated. The alternate drink treatment was consumed during the subsequent visit. The change in pre‐ to post‐drink HRV recordings were calculated and analyzed via repeated‐measures, two‐way ANOVA.

RESULTS

ANOVA results (Drink x Body Position) did not detect mean differences in HRV time‐ or frequency‐domains of interest (Interaction and main effects p > 0.05, RMSSD: SupED = −0.11 ± 27.13 ms, SupCD = −2.59 ± 27.18 ms, StED = 14.64 ± 15.84 ms, StCD = 10.23 ± 16.34 ms; HF nu: SupED = −1.59 ± 14.87 nu, SupCD = 5.67 ± 16.62 nu, StED = 12.02 ± 15.69 nu, StCD = 6.36 ± 18.50 nu; LF/HF: SupED = 0.06 ± 0.66 nu, SupCD = −0.14 ± 0.74 nu, StED = −2.36 ± 4.45 nu, StCD = −2.40 ± 9.41 nu).

CONCLUSION

According to these data, consumption of a commercially available ED does not alter cardiovascular autonomic control in young, healthy adults under resting conditions.
Suitability of Evasin‐3 as Molecular Imaging Marker for Early Coronary Neovascularization by Arteriogenesis
Authors: van Lier Monique GJTB , Schwarz Janina CV , de Vos Judith , de Bruin Cora M , Verberne Hein , Siebes Maria
Published: November 2023

Aim

To assess labeled molecular targets as early imaging biomarker for formation of coronary collaterals in ischemic rabbit hearts. Evasin‐3 binds specifically to CXCL1 and can provide a powerful molecular agent to visualize early arteriogenesis.

Methods

We induced regional myocardial ischemia in 10 New Zealand White rabbits by ameroid constriction of a major coronary artery. Animals were sacrificed 9 or 16 days after surgery. Fluorescently labeled (Alexa‐Fluor 488) evasin‐3 was injected by cardiac puncture and circulated for 1.5 or 3hrs prior to sacrifice in 5 rabbits. Coronary arteries of 5 excised hearts were filled with UV Blue fluorescent replica material. Using an imaging cryomicrotome, alternate cutting (14 μm slice thickness) and multispectral episcopic imaging (14 μm resolution) yielded detailed 3D representations of the heart and vasculature co‐registered with the labeled molecular marker. Vascular distribution of the fluorescent imaging biomarker was determined.

Results

Fluorescence cryomicrotome imaging clearly depicted evasin‐3‐AF488 in coronary vessels. The overlay with vascular cast images revealed presence of evasin3 along all major vessels. However, no enhanced uptake was visualized in the ischemic area.

Conclusions

The distribution of fluorescently labeled evasin‐3 in coronary vessels was not restricted to vessels in the ischemic area. It is therefore not well suited as a specific imaging marker of neovascularization by arteriogenesis.

Support or Funding Information

This research was supported by the Center for Translational Molecular Medicine (EMINENCE).
Cardiovascular risk profile in Peruvian medical students
Authors: Fernández‐Rodríguez Lissett Jeanette , Bardales‐Zuta Victor Hugo , Avalos‐Alvarado Carlos Eduardo , Hilario‐Vargas Julio Santos
Published: November 2023

We report the results of a cross‐sectional study that queries cardiovascular risk in second year medical students enrolled at Antenor Orrego Private University, Trujillo, Peru. We selected 78 students (58 women, 20 men) randomly. At the beginning of the semester, we determined medical and family history, age, adverse habits, somatometrics, blood pressure, baseline blood glucose, lipid profile and electrocardiograms of the participants over one week. The average age was 18.90 ± 1.5 for women and 18.70 ± 1.2 men. 39.7% of women and 50% men came from Trujillo, 58.6 and 75% had a family history of diabetes mellitus or arterial hypertension, 55.2 and 45% had medical history, 5.2 and 10% smoke, 16 and 35% had systolic blood pressure ≥ 130 mmHg and 7 and 20% had diastolic pressure ≥ 85 mmHg. The 38 and 55% were overweight, 3% and 10% were in obesity grade I, 2% and 5% in grade II and 5% males in grade III. The waist ‐hip index for 71% of women was > 0.80 and was > 0.95 for 30% of men. 2 and 10% had high triglycerides. 9% of women presented total cholesterol at the upper limit. 69% and 50% had low HDL, 3 and 2% of women had high and very high LDL. 46.6% had electrocardiographic abnormalities. We found that cardiovascular risk factors such as changes in lipid metabolism and ECG, overweight and obesity were common among medical students. This study underlines that universities should implement primary prevention measures to reverse or slow this trend among late‐teenage students, as earlier intervention can improve lifespan and outcome in the long term.

Support or Funding Information

Antenor Orrego Private University.
β‐ENaC associated with cilia indicates the distribution of absorptive epithelial cells in the lumen of small airways
Authors: Flores‐Delgado Guillermo , Quinton Paul Marquis
Published: November 2023

Airway surface liquid (ASL) is crucial for optimal clearance of mucus, pathogens and debris from the pulmonary bronchial tree and is managed by a balance between secretory and absorptive activities along the luminal airway surfaces. Immunohistochemically, we recently demonstrated that in small airways, about half of the lining epithelium consists of cells with a secretory phenotype that are confined to the contraluminal regions (pleats) of the plicated luminal epithelium (1). However, the distribution/location of absorptive airway epithelial cells has not been demonstrated. The epithelial sodium channel (ENaC) is responsible for Na(+) absorption across the apical membranes of many salt absorbing epithelia including airways. Herein, we hypothesized that localization of β‐ENaC subunits would define the distribution of absorptive cells in airway epithelia.

We obtained small pieces of peripheral porcine lung tissue of approximately 0.5 cm3 and pieces of trachea of approximately 0.5 cm2 that were dissected and immediately snap‐frozen at −80°C. Serial tissue sections (7 μm thick) containing cross‐sections of small airways, with a diameter between 0.5 and 1 mm, were used for immunohistochemistry. We used antibodies directed to b‐subunit of ENaC (courtesy C. Fuller), as well antibodies directed to β‐Tubulin and the tight junction protein, ZO1. Western blot analysis demonstrated ENaC expression in cell extracts derived from isolated small airways and trachea. Confocal microscopy of immunostained sections of airway tissue defined the distribution of β‐ENaC subunits in the apical regions of a sub‐set of cells in the small airways and trachea. In small airways, strong staining for β‐subunit was generally restricted to ciliated cells in the region around the tips of folds in the luminal epithelium well separated from the sub‐sets of secretory cells in base of the epithelial pleats where staining was generally absent or faint, but strong for the secretory marker, NKCC1, seen previously (1). In trachea, β‐ENaC subunit staining was mainly observed at the surface of the stratified ciliated epithelium, and absent in regions where submucosal glands were present. Importantly, we observed that β‐subunit of ENaC appeared to co‐localize with the distribution β‐tubulin, a biomarker for cilia.

These findings suggest that small airway epithelium contains a subset of epithelial cells with absorptive phenotype, recognized by the high expression of β‐subunit of ENaC.

Support or Funding Information

Supported by the Cystic Fibrosis Foundation‐MCCC, CFRI and the Nancy Olmsted Trust for Pediatric Respiratory Medicine
Expression of NKCC1 and Aquaporins 4, 7 and 9 in Mouse Choroid Plexus and Ependymal Cells
Authors: Patyal Pankaj , Alvarez‐Leefmans Francisco Javier
Published: November 2023

Choroid plexus epithelial cells (CPECs) secrete cerebrospinal fluid (CSF) through poorly understood mechanisms involving Aquaporins (AQPs) and various solute transport proteins like the Na+/K+ pump and the Na+‐K+‐2Cl− cotransporter 1 (NKCC1). Net water movement across CPECs occurs from the basolateral membrane through the apical membrane. However, with the exception of AQP1 in the apical membrane, the molecular pathways for transepithelial water fluxes are either unknown, or their expression is controversial (Damkier et al. 2013, Physiol Rev 93, 1847). Because of their relatively high transcript expression (Cha et al, 2014, FASEB Journal 28 .1 Suppl 1182.3) the present work focuses on the cellular and subcellular localization of AQPs 4, 7 and 9 proteins in mouse CPECs and ependymal cells (EPCs), using immunolabeling with validated antibodies and confocal immuno‐fluorescence. Evidence for AQP4 expression in CPECs is controversial. The present results demonstrate that: 1) AQP 4 is expressed neither in CPECs nor in ependymal cells, but in radial glial endfeet surrounding the basal aspect of EPCs, where it is co‐expressed with NKCC1; the latter was also expressed in the apical membrane of both CPECs and EPCs; 2) AQP 7 expression was found to be restricted to the apical membrane of CPECs and EPCs. 3) AQP9 was expressed neither in CPECs nor in EPCs. The pathways for water influx across the basolateral membrane of CPECs remain to be elucidated. The pattern of expression of NKCC1 and AQP4 suggests a role of these proteins in K+ uptake from the CSF, both in CPECs and EPCs, thus regulating CSF and interstitial brain fluid [K+]. The expression of AQP7 in the apical membrane of CPECs suggests that this protein could work together with AQP1 in CSF secretion.

Support or Funding Information

(Supported by Wright State University Foundation FOAP 551462)
Mechanisms of Adenosine A1 Receptor‐Mediated Enhancement of Vasa Vasorum Endothelial Cell Barrier Function
Authors: Batori Robert , Kasa Anita , Shaw Mary , Sanjiv Kumar , Taraseviciene Laimute , Lapel Martin , Weston Philip , Fragoso Miguel , Moldovan Radu , Stenmark Kurt R , Verin Alexander , Gerasimovskaya Evgenia
Published: November 2023

The vasa vasorum (VV) is a microcirculatory network that provides oxygen and nutrients to the adventitia and media of large blood vessels. Previously, we have demonstrated that angiogenic expansion of pulmonary artery (PA) VV in the chronically hypoxic calves exhibit increased permeability for circulating inflammatory cells and macromolecules. Extracellular adenosine (Ado) increased transendothelial electrical resistance (TER) of VVEC isolated from control (Denver altitude) and, in lesser extent, hypoxic (2 weeks, PB=430 mmHg) neonatal calves. Ado‐induced VVEC barrier enhancement involved activation of Ado A1 receptors (A1R), PI3K/AKT pathway, and actin remodeling. In the current study we demonstrated that inhibition of Rac1 by transduction of VVEC with dominant‐negative adenoviral Rac N17 construct almost completely abolished adenosine‐induced increase in TER supporting a critical role of Rac1 activation in Ado‐induced VVEC barrier enhancement. Inhibition of Rac1 activity also leads to the decrease of AKT phosphorylation, suggesting that Rac1 is an upstream regulator of AKT signaling in VVEC. In addition, Ado increased autophoshorylation of p21‐activated kinase 1 (PAK1), a well‐known downstream target of Rac1, reflecting its activation. Pharmacologic inhibition of PAK1 decreased basal TER and abolished Ado‐induced TER increase indicating involvement of PAK1 in VVEC barrier maintenance. Importantly, A1R expression was demonstrated in the VV of pulmonary arteries of Sprague Dawley (SD) rats, indicating that VV localization of A1R is not restricted to bovine VV. Intravenous co‐injection of Rhodamine‐conjugated Griffonia Simplicifolia lectin and FITC‐conjugated dextran in SD rats followed by two photon confocal microscopy of dissected pulmonary arteries revealed increased permeability of PA VV in chronically hypoxic SD rats. Thus, this model shows a great promise for the in vivo validation of the protective role A1R agonists in VV barrier integrity. In summary, we demonstrate for the first time that A1Rs enhance VVEC barrier function through the activation of Rac1/PAK pathway and actin cytoskeleton remodeling. We propose that A1Rs can be recognized as a vascular bed‐specific and novel therapeutic target to regulate VV barrier function and pathologic vascular remodeling in chronic hypoxia.

Support or Funding Information

Support: NIH/R01 HL086783 to EVG, PPG HL101902 to ADV
Effects of Aging on Markers of Ribosome Biogenesis in Fast‐ and Slow‐Twitch Skeletal Muscle in Rodents Aged 3 to 24 Months
Authors: Mobley Christopher Brooks , Mumford Petey W. , Kephart Wesley C. , Haun Cody T. , Holland A. Maleah , Osburn Shelby C. , Beck Darren T. , Martin Jeffrey S. , Young Kaelin C. , Kavazis Andreas N. , Lowery Ryan P. , Wilson Jacob M. , Roberts Michael D.
Published: November 2023

OBJECTIVE

Ribosome biogenesis is an essential process for regulating skeletal muscle mass by facilitating muscle protein synthesis. Aging results in numerous alterations in skeletal muscle; specifically, a decline in protein synthesis in skeletal muscle. However, less is known about the functional impact of the ribosome in aging skeletal muscle. Therefore, we sought to investigate the effects of sequential aging on various markers of ribosome biogenesis in fast‐ and slow‐twitch skeletal muscle in rodents.

METHODS

Male Fischer 344 rats (300–600 g) were sequentially aged to 3, 6, 12, 18, & 24 months (mo). Plantaris (Type II, fast‐twitch) and soleus (Type I, slow‐twitch) skeletal muscle were harvested from each respective animal and subjected to RNA isolation and analyses and Western blotting.

RESULTS

Compared to 3 mo old rats, plantaris ribosome content (total RNA) was 40% lower in 6 mo rats (p<0.05), 59% lower in 12 mo old rats (p<0.05), 69% lower in 18 mo old rats (p<0.05) and 55% lower in 24 mo old rats. There were no age‐related differences in soleus ribosome content. Plantaris c‐myc protein significantly paradoxically increased across age groups; specifically, 12 mo rats were 3.05‐fold greater (p<0.05), 18 mo rats were 5.28‐fold greater (p<0.05) and 24 mo old rats were 5.44‐fold greater (p<0.05) than 3 mo old rats. There were no age‐related differences in soleus c‐myc protein levels. Plantaris upstream binding factor (UBF) protein was 76% greater in 24 mo old rats versus 3 mo old rats (p<0.05). Soleus UBF was only different between the 18 mo old rats and 24 mo old rats (specifically ~3‐fold greater in 18 mo versus 24 mo old rats; p<0.05). Compared to 3 mo old rats, plantaris eukaryotic initiation factor (eIF)2Bɛ protein was 34% lower in 6 mo old rats (p<0.05), 49% lower in 18 mo old rats (p<0.05) and 68% lower in 24 mo old rats (p<0.05). There were no age‐related differences in soleus eIF2Bɛ protein levels. Plantaris eukaryotic elongation factor 2 (eEF2) was not different between age groups. Compared to 3 mo old rats, soleus eEF2 was 32% lower (p<0.05) in 18 mo rats and 66% lower in 24 mo old rats (p<0.05).

CONCLUSION

Aging clearly decreases ribosome content in fast‐twitch but not slow‐twitch skeletal muscle. Furthermore, there is a compensatory up‐regulation in plantaris c‐myc and UBF protein expression to potentially offset the decline in ribosome content. Identifying other intracellular mechanisms and/or co‐factors that contribute to decrements in ribosome biogenesis in skeletal muscle is warranted.

Support or Funding Information

This study is supported in part by a contract from the University of Tampa (to M.D.R. from J.M.W) as well as laboratory start‐up funds from M.D.R.
Effect of diurnal regulated‐endogenous melatonin on wound healing processes in skin injured animal model
Authors: Choi Jeonghyun , Hong Yonggeun
Published: November 2023

To elucidate the effect of diurnal regulated‐endogenous melatonin on the wound healing after skin injury, experimental animals were divided into three groups following light/dark cycle; L/D, 12/12‐hr light/dark cycle; LL, 24/0‐hr light/dark cycle; DD, 0/24‐hr light/dark cycle. We found increased mRNA expression of tumor necrosis factor alpha (TNF‐α), which is the representative pro‐inflammatory cytokine, in DD group at 15 hr after injury. However, LL group showed increased TNF‐α mRNA expression at 7 day after injury, indicating the delayed augmentation of TNF‐α. We further found that cleaved caspase‐3 expression in LL group at 15 hr and 2 day after injury. Transforming growth factor beta1 (TGF‐β1) and extracellular signal‐regulated kinase 1/2 (ERK1/2) expression of DD group was significantly increased. In histology, more fibroblast‐like cells and adipose cells were identified in acute phase of injury. Also, we found the migration of fibroblast‐like cells and accumulation of collagen‐like fibers in DD group, which are thought to indicate acceleration of re‐epithelialization in wound healing processes. Furthermore, increased wound closure rate was identified in DD group suggesting accelerated structural recovery in wound healing processes. These finding suggest that elevated endogenous melatonin concentration induced by conditional controlled photoperiodicity might contributes to the wound healing speed and quality, which may modulated by inflammation and growth factors.

Support or Funding Information

Funding: NRF‐2012R1A1A2005089, NRF‐2013R1A2A2A01067169
Critical role of Pin1 on neuronal rescue after transient ischemic brain injury
Authors: Kim Joo‐Heon , Lee Seunghoon , Hong Yonggeun
Published: November 2023

Pin1 contains two functional domains, an N‐terminal WW domain and a C‐terminal PPIase domain. We defined a critical role of Pin1 on neuronal rescue through Pin1 inhibitor treatment after ischemic brain injury (MCAo). Juglone (Jug, 1 mg/kg), Pin1 inhibitor, was administered once a day for 7 days after MCAo. Analysis in histology, protein, and gene were used to determine neural cell death and survival pathway at day 7. MCAo+Jug group lead to significant downregulation of Bcl‐2 and Bax expression compared with MCAo+Veh group (p <.01). MCAo+Jug group was shown to reduce the levels of Pin1, Tau5, and phspho‐TauThr231 in both immunoblotting and immunostaining (p<.01). On the other hands, mature‐BDNF was significantly augmented in MCAo+Jug compared to MCAo+Veh (p<.01). These results suggest that Pin1 isomerase is not only acts to accelerate an intrinsic apoptotic pathway but also interrupt to turn on the cell survival machinery after MCAo. Finally, Pin1 inhibition might alleviate the acceleration of apoptotic cell death that switch on the neuronal cell rescue signal after focal cerebral ischemic in rats.

Support or Funding Information

Funding: NRF‐2012R1A1A2005089, NRF‐2013R1A2A2A01067169
Spinal Cord Meninges Produce Attractive and Repellent Cues for Developing Axons
Authors: Suter Tracey Amelia Claire Sampath , DeLoughery Zachary , Jaworski Alexander
Published: November 2023

The meninges are connective tissue layers that envelop vertebrate brain and spinal cord. The meninges protect the central nervous system (CNS), contribute to the circulation of cerebrospinal fluid, and provide a structural framework for blood vessels entering the CNS. Recent work provides evidence that the meninges also fulfill critical functions during the development of the CNS by regulating both neural progenitor generation and migration. During development the majority of neurons within the CNS have axonal projections that remain within the CNS. However, there are unique populations, including motor and sensory neurons, which cross the boundary between the CNS and peripheral nervous system (PNS). The location of the meninges between the CNS and PNS raises the possibility that the meninges act as guidepost tissue, a source of guidance cues, for developing axons. Here, we investigate the role of meninges in axon guidance by conducting a series of co‐culture in vitro experiments to examine the response of different axonal populations to meninges. Using embryonic mouse neural tissue explants, we provide evidence that motor axons are attracted to spinal cord meninges and sensory axon growth is stimulated upon contact with the meninges. In contrast, commissural axons are repelled by a transient, secreted guidance cue produced by the meninges and results in commissural growth cone collapse. These results demonstrate that the developing meninges produce yet to be identified long‐ and short‐range axon guidance cues that regulate axonal growth in vitro. We show that each axonal population responds to meninges in a manner consistent with their in vivo trajectories relative to the meninges, suggesting that the observed meningeal guidance factors contribute to proper nervous system wiring.
Astrocytic Tissue Factor Plays An Important Role In Regulation Of CSF Circulation And Control Of Blood Spread Following Subarachnoid Hemorrhage
Authors: Golanov Eugene , Bovshik Evgueni , Foster Chase , Zhang Jonathan , Wong Kelvin , Wong Stephen , Britz Gavin
Published: November 2023

Rupture of cerebral aneurysm leads to blood accumulation within the subarachnoid space, known as a subarachnoid hemorrhage. Fatality resulting from acute subarachnoid hemorrhage (SAH) reaches 50%. Volume of blood entering subarachnoid space (SS) following aneurism rupture is one of the leading factors determining the outcome. Retardation of cerebrospinal fluid (CSF) flow emerges as one of the key mechanisms limiting spread of blood in SS. It has been established that following SAH CSF flow is significantly reduced. We hypothesize that tissue factor (TF), the key factor of extrinsic pathway of coagulation cascade expressed by brain astrocytes, plays a critical role in reduction of CSF and localization of the hemorrhage following SAH.

Perforation model of SAH was used in C57BL mice. We monitored changes of CSF flow by injecting fluorescent microspheres (0.02 um, FM) in cisterna magna (CM) from 1 hour to 30 days following SAH. We injected intracerebro ventricularly TF antibodies or IgG one hour before the perforation to explore role of TF in spread of blood across the SS and CSF flow.

In naive and sham (no perforation) mice FM injected into CM filled ventral cisterns and spread along the perivascular space of Willis circle and middle cerebral arteries (MCA) reaching the olfactory bulbs within 1 hr. Following perforation of Willis circle, blood spread along the paravascular space, ventral cisterns and proximal trunk of ipsilateral MCA. FM injected into CM following SAH, were confined to the caudal ventral cisterns limited by the rostral pons border. CSF flow remained impeded for up to 30 days. The spread of hemorrhage did not correlate with the degree of CSF flow retardation. Block of TF by specific antibodies lead to significant (p<0.05) expansion of hemorrhage by 95%. In non‐SAH animals, intraventricular administration of TF antibodies significantly increased spread of CM injected FM compared to their spread following IgG administration (p<0.05). Subsequent immunohistochemistry demonstrated substantial association of TF with glia limitans, especially on the ventral brain surface.

We conclude that TF plays an important role in localization of blood spread following arterial perforation. Moreover, block of TF by specific antibodies significantly facilitates spread of FM injected into CM allowing speculating that TF may play a regulatory role in CSF flow. It was suggested that astrocytes are the only source of TF in brain. We propose that astrocytes may exercise their CSF flow regulatory function and limit the blood spread through TF‐dependent mechanism.

Support or Funding Information

Oden Foundation
A Study of the Influence of Single Nucleotide Gene Polymorphisms on the Risk of Hypertension
Authors: Soliman Naglaa Raafat Abdraboh , Hemimi Neveen Salah
Published: November 2023

Background

Several allele variants of human genes are proved to be linked with parthenogenesis of hypertension. The aim of this study is to identify the association of hypertension with single nucleotide polymorphism in four genes including, endothelial nitric oxide synthase (eNOS rs 1799983), angiotensinogen (AGT rs 699), lipoprotein lipase (LPL rs 320), and methyl tetrahydrofolate reductase (MTHFR rs 180113338, MTHFR rs 1801131 ).

Material and Method

The SNPs were genotyped in 180 hypertensive and 198 age and gender matched normotensive individuals using polymerase chain reaction‐ restriction fragment length polymorphism.

Results

eNOS rs 1799983 and AGT rs 699 were found to be significantly associated with hypertension OR (95% CI) 2.7(1.27–5.60); and 2.3 (1.00–5.10) respectively, p<0.05. The other 3 SNPs have shown a tendency to increase the risk with OR between 1.5 and 2.2 with no statistical significance. The risk of hypertension was increased with simultaneous presence of risky alleles of eNOS rs 1799983 with other SNPs as MTHFR rs 1801131 OR 2.2 (0.70–2.69), AGT rs 699 OR 2.6 (0.3–22.99), LPL rs 320 OR 4.2 (1.88–9.41) and rs 180113338 OR 4.8 (1.19–19.27).

Conclusion

We concluded that coincident appearance of some genetic alleles may increase the risk of hypertension and could be a predictive genetic profile for the disease.

Support or Funding Information

The research was funded by Dubai Medical College.
The role of conditional controlled photoperiodicity with propionic acid in induction of autism‐mimetic animal model during prenatal phase
Authors: Lee Seunghoon , Hong Yonggeun
Published: November 2023

We determined whether prenatal pups were affected the brain and physiological development by exposure of propionic acid (PPA) as well as conditional photoperiods, which are could be modulate the secretion of endogenous melatonin secretion. Indirectly PPA‐treated rats were significantly increased body weight at 3‐ and 4‐week‐old after partus. All of PPA‐treated rats brain was organized the neuronal cell more than control groups following controlled photoperiodicity. All of experimental groups were not significantly altered organization of neural cells components. In behavioral test, constant dark condition (DD) revealed to delayed organization of neural architecture in cerebral cortex and hippocampus compared to constant light condition (LL). There were not significantly altered an exploring activity. Conversely, PPA+DD was similarly increased an inner square exploring activity but not in the PPA+LL group. We were not clearly defined for autism‐mimetic animal model induction during prenatal period via PPA exposure. Conditional controlled photoperiodicity might act on environmental stress for embryonic neural development in both control and PPA exposure. In future, we will process the embryonic stage for PPA exposure via placenta. Thus, we will clarify the effect of prenatal PPA exposure during pregnancy with conditional photoperiod.

Support or Funding Information

Funding: NRF‐2012R1A1A2005089, NRF‐2013R1A2A2A01067169
Effects of endogenous melatonin regulated by conditional controlled photoperiodicity on neural reorganization through ER stress response and autophagy pathway after spinal cord injury
Authors: Jin Yunho , Hong Yonggeun
Published: November 2023

Spinal cord injury (SCI) causes serious neurological deficits. The purpose of this study was to elucidate the effect of endogenous melatonin on neural reorganization and locomotor recovery through regulation of autophagy and endoplasmic reticulum (ER) stress response after SCI. To diversify the concentration of endogenous melatonin, subjected animals were divided into 5 groups; group1 (12‐hr light/12‐hr dark, L/D), group2 (6‐hr light/18‐hr dark, 6L/18D), group3 (24‐hr dark, DD), group4 (18‐hr light/6‐hr dark, 18L/6D), group5 (24‐hr light, LL). At 7 days after injury, locomotor test based on BBB (Basso, Beattie, Bresnahan) scale was conducted to identify functional recovery and the animals were sacrificed for further molecular and histological analyses. The rats under LL condition showed great recovery of motor function after SCI. Moreover, the expression of neuronal marker and autophagy marker was also upregulated in DD group. However, the protective role of ER stress response seems to be ambiguous. In conclusion, autophagy rather than ER stress response in SCI might play a critical role in neuroprotection, which is thought to be intensified by endogenous melatonin induced by conditional controlled photoperiodicity.

Support or Funding Information

Funding : NRF‐2012R1A1A2005089, NRF‐2013R1A2A2A01067169
An Empirical Test of Metabolic Effects and Adaptive Potential in Response to Global Ocean Warming in a Model Fish Species, the Threespine Stickleback
Authors: Morozov Sergey , McCairns R.J. Scott , Leinonen Tuomas , Merilä Juha
Published: November 2023

Climate change has had a significant influence on the world's oceans, which have shown constant increases in mean water temperature over the last 50 years. The biota of Arctic environments, ectotherms in particular, may be particularly threatened by this change: whilst southern animals have the possibility of migrating north, those adapted to colder northern climes have nowhere left to move and may be faced with the ‘choice’ of adapt or die. A key factor in adaptation to the changing thermal environment is the physiological response. Metabolic scope (the difference between minimum and maximum oxygen consumption rate) is likely a key fitness related trait because it defines energy available for locomotion, growth and reproduction. The unifying aim of our research is to develop a deeper understanding of the constraints and/or adaptive potential in metabolic scope with respect to thermal changes predicted due to climate change.

As obligate ectotherms, threespine stickleback (Gasterosteus aculeatus) populations adapted to localized thermal conditions provide an excellent and logistically tractable model for study of thermal and metabolic evolution in a changing environment. We collected broodstock from anadromous Barents Sea fish and bred them under controlled conditions for two generations to standardize environmental & maternal sources of variance. The multi‐generational pedigree and large numbers of families (N=180, 40 families) permits estimation of additive genetic variance, whereas repeated measurements in multiple environments allows us to estimate direct environmental variance on metabolism and swimming performance. Maximum and minimum metabolic rates, in addition to critical swimming performance, were measured with the same individuals at three different temperatures: (i) contemporary summer temperature for Barents Sea coastal waters (12°C), (ii) extreme predictions under global warming (17°C), and (iii) stressful temperature (80% of CTmax). Individual critical temperature in observed population varied between 27.2 and 32.1°C. The preliminary results showed that minimum and maximum metabolic rates increased with the temperature rise: 61.6 ± 10.1 and 126.2 ± 36.1 mg O2/kg/hour at 12°C; 70.19 ± 11.8 and 144.9 ± 38.8 mg O2/kg/hour at 17°C; 114.0 ± 15.7 and 183.7 ± 32.0 mg O2/kg/hour at individual stressful temperature, respectively. However, the highest aerobic metabolic scope and swimming performance were observed at 17°C. Quantitative genetic analyses are ongoing, but point to the broad potential for adaptation to environmental change.

Support or Funding Information

SM: Alfred Cordelin Foundation, Otto A. Malm Foundation, Oscar Öflund Foundation, LUOVA doctoral programme; RJSM, JM, TM: Academy of Finland
Effects of the antibiotics succinylsulfathiazole and sulfaquinoxaline on development and viability of the fruit fly Drosophila melanogaster
Authors: Singh Nimisha , Blatch Sydella A
Published: November 2023

Sulfa‐drugs and other antibiotics are often used in nutritional studies to reduce the synthesis of vitamins by microbes in the gastrointestinal tract of animals. Succinylsulfathiazole (SS) has been used in this way to study the effects of dietary folate on mice. Observations of mice consuming 1% SS in nutritional studies raise the possibility that this antibiotic has direct and/or interactive effects on the growth and development of mice. It is important to clarify whether this is the case, so effects caused by SS are not entirely attributed to diet or nutrient status alone. Towards this, we sought to use fruit flies as a model system to determine whether SS has effects on the development and growth of animals. We hypothesized the 1% SS dosage used in mice would be fatal to fruit flies, so we conducted a dose‐response experiment to determine tolerable doses. Fruit flies were reared from egg to adulthood on one of six doses of SS (twenty eggs per dose), spanning eight orders of magnitude. One group was reared on a different sulfa‐drug, sulfaquinoxaline (SQ), for which a tolerable dose for flies is already known. Stage‐specific viability and time to develop, as well as larval mass were determined for each group, and for flies receiving no antibiotics. As predicted, 1% SS was fatal: no larvae survived this dose. The other five doses were tolerable and the flies responded generally similar across the groups. Numbers of larvae developing, larval mass, and time spent the larval stage were not significantly different in the remaining SS groups. However, more pupae were present by day 20 for flies on 0.05% SS (the second highest dose), compared to some of the other SS doses. Interestingly, larvae fed a low dose of SQ developed faster than all of the SS flies, including SS flies that were at the same or lower doses. These results indicate that Drosophila cannot survive doses of SS comparable to mice, and that lower doses should be used in fruit fly studies. Future experiments can use these lower but tolerable levels of SS to help determine direct and interactive effects of the sulfa drug on fruit fly development.
Hypermetric Tracheal Scaling as a Factor in Insect Gigantism
Authors: Wagner Julian , Duell Meghan , Harrison Jon
Published: November 2023

Why are insects small? In the Paleozoic Era, atmospheric oxygen levels peaked significantly higher than the current level. Along with this oxygen peak came giant insects, many times larger than those currently observed. This suggests that oxygen transport may inhibit insects from becoming giants. If this were the case, then one would expect to observe an increase of investment in gas exchange structures in larger insects, i.e. hypermetric scaling. This has been observed in tenebrionid beetle trachea. In this study, we look at scarab beetles, since they have a large range of sizes including the largest extant insect species. Using sets of micro‐CT images of freshly dead scarabs taken at 180 degrees around the beetle, 3d reconstructions of the specimens were made. Using Aviso Fire, the eight spiracles, coxal‐trochantal areas, and the mesothoracic femoral tracheal volumes were measured. The spiracle areas showed isometric scaling with a slope of 0.70. However, spiracle area over spiracle depth showed a steeper slope in the proximal abdominal and thoracic spiracles as compared to the distal abdominal spiracles, suggesting that larger beetles may exhibit different dimensions in their spiracles to promote the spiracle's diffusive abilities (larger area compared to depth allows more air to flow a shorter distance via diffusion). The scaling of the area of the coxal‐trochantal trachea showed a larger slope than the scaling of the area of the coxa with a p value of 0.09 in a t test. The femur structure showed isometric scaling, whereas the femoral trachea showed hypermetry with a slope of 1.46, which differed from a slope of 1 with a p value of 0.0012 in a t test. These indicate that structures far from oxygen intake at the spiracles scale hypermetrically in scarabs.

Support or Funding Information

This research was supported in part by funds from the School of Life Sciences Undergraduate Research (SOLUR) Program through the School of Life Sciences at Arizona State University, Tempe Campus, and by NSF 1122157
Cytochrome c Release from Dynamic Mitochondria in Endothelial Cells under Flow
Authors: Hu Ying‐Li , Hur Sung Sik , Chien Shu
Published: November 2023

Endothelial mitochondria are essential to the functional integrity of endothelial cells (ECs) as they integrate a wide range of cellular processes. Cytochrome c (Cyt c) is located in the mitochondrial inner membrane space (IMS), where it functions in the electron transport chain. The morphology of mitochondria changes dynamically due to their frequent fusion and fission in response to cellular conditions. Mitochondrial fission is an early event during apoptosis and it occurs very close in time to Cyt c release, and fragmented mitochondria can readily be seen retaining Cyt c. ECs were subjected to oscillatory and laminar flows, with shear stresses of 0.4±4 and 12 dynes/cm2, respectively. Cyt c staining of the mitochondria was detected in ECs under both types of flow. In comparison to static control, Cyt c was significantly reduced in the mitochondria of ECs exposed to both flow patterns for 4 h, 8 h, and 24 h. The reduction of Cyt c in endothelial mitochondria was greater under oscillatory flow for 8 h and 24 h.

4 h
8 h
24 h

Oscillatory Flow/Static
0.16±0.05
0.25±0.06*
0.59±0.05*

Laminar Flow/Static
0.19±0.03
0.44±0.03
0.90±0.03

Values are mean±SEM.

P<0.05 for Oscillatory vs. Laminar Flow.

After laminar flow for 24 h, Cyt c rose to almost the level of the static ECs. We used time‐lapse imaging to monitor the spatial‐temporal dynamics of mitochondria in live ECs transfected with RFP‐mito. During live ECs shearing process mitochondria networks are dramatically reorganized from long filamentous, interconnected tubules into small punctate spheres in ECs under both flow patterns. Under oscillatory flow, more small fragmentations of endothelial mitochondria were observed than those under laminar flow. The results suggest that mechanical force affects the mitochondria dynamics in ECs under flow to cause Cyt c release, which was greater under oscillatory than laminar flow. Understanding of the endothelial mitochondria responses to different flow patterns will help to elucidate the mechanism of the region‐specific localization of atherosclerosis in the arterial system.

Support or Funding Information

This work was supported by NHLBI Research Grants HL‐104402 and HL‐106579 (S.C.)
Gene Polymorphism of Matrix Metalloproteinases 3, and 9 in Breast Cancer
Authors: Soliman Naglaa Raafat Abdraboh , Bayoumi Fatehia Ali
Published: November 2023

Background

Genetic factors have a considerable influence on development and progress of malignancy. Matrix metallo‐proteinases (MMPs) are a group of proteolytic enzymes which are proved to be involved in pathogenesis of breast cancer. Some functional single nucleotide polymorphisms (SNPs) affect the transcriptional activity of genes of MMPs. We aimed in the present study to investigate the possible association of 2 SNPs, C/T polymorphism of MMP9 gene, and 5A/6A polymorphism of MMP3 gene, with the occurrence and clinico‐pathological criteria of breast cancer.

Subject and method

The present case control study included 59 breast cancer female patients, and 77 tumor free age matched control females. The SNPs were genotyped using polymerase chain reaction ‐ restriction fragment length polymorphism assay.

Results

Data analysis have shown higher frequency of T allele of C/T MMP9 and A5 of A5/A6 MMP3 in breast cancer patients as compared to control subjects with odd ratio (OR), 95%confidence interval (95% CI) 1.8 (1.02 – 3.26) and 1.2 (0.71 – 1.89) respectively. When the frequency of both alleles was analyzed in comparison to prognostic clinico‐pathological criteria of the disease, there was a significant association of T allele of C/T MMP9 with advanced tumor size and histological grade with OR (95%CI) 2.25 (1.12 – 5.64), 1.8 (0.78 – 4.17) respectively.

Conclusion

the results of the current study suggested that T allele of C/T MMP9 gene polymorphism is strongly linked to the incidence, growth and differentiation of breast cancer.

Support or Funding Information

Fund supplied by Dubai Medical College
Unlocking a more comprehensive assessment of diet with continuous, chronic blood glucose and complete hemodynamic monitoring in rodents
Authors: Main Bradley , Holliday‐White Kimberly , Bogie Heather , Lillegard Kathryn E , Lindquist Ryan
Published: November 2023

New technology was employed in a novel way to monitor the effect of diet change on continuous blood glucose in addition to standard hemodynamic parameters. A new telemetry implant (HD‐S11‐F2, DSI) enables researchers to collect and transmit data at a different radio‐frequency (18MHz) than the traditional 455KHz implants. While a second frequency allows for social housing and tighter density of research subjects, in this experiment the 18MHz device was paired with a continuous glucose telemetry implant (HD‐XG, DSI) in the same animal. Implantation of two devices in a single animal allowed us to measure blood pressure, ECG, arterial blood glucose, body temperature and activity in a single rat. Six male Sprague Dawley rats were implanted with 2 telemetry implants that each required an arterial site for sensor placement. The HD‐XG sensor was placed in the descending abdominal aorta and the device body was located IP. The HD‐S11‐F2 was inserted in the femoral artery and the catheter tip was advanced to the abdominal aorta (just cranial to the iliac bifurcation), not overlapping the glucose sensor. The HD‐S11‐F2 implant body was placed subcutaneously on the flank. After recovery from surgery, animals were enrolled in a cross‐over designed study to compare the physiologic effects of offering high carbohydrate or low carbohydrate treats after a 12 hour fast. Blood pressure, heart rate and blood glucose values are presented. Marshmallows and mangos were offered as high carbohydrate treats while cheese cubes were offered as the low carbohydrate treat. Approximately 1 hour after the treat offering, the standard rat chow was restored to the cage feeders. The excitement from the technician opening the cage was evident in HR and BP changes associated with that activity. Blood glucose levels from 9–12 hours post‐fast were approximately 92 mg/dl in both groups. Eating marshmallows and mango caused a rapid rise to ~103 mg/dl in ~30 minutes. Eating cheese resulted in a modest rise in blood glucose (~96 mg/dl). These experiments demonstrate a successful surgical approach using two implants to get a more comprehensive picture of whole animal cardiovascular and metabolic physiology.
Estrogen Receptor α induces microRNA‐365 expression to attenuate classical macrophage activation
Authors: Chang Richard Cheng‐An , Tseng Alexander Mike , Wang Haiqing , Lin Yu‐Lieh , Zhou Beiyan
Published: November 2023

The sex steroid hormone estrogen, besides its role in reproduction, has been reported as a potential regulator for immune responses. Albeit the increasing co‐morbidity associated with obesity, the pandemic threat marked with chronic inflammation and insulin resistance, only a handful of studies suggest estrogen may exert immune suppressive effect in this context. In this study, we report the first set of evidence to support that estrogen receptor alpha (ERα) signaling can selectively suppress classical activation of macrophages, which is the primary contributor to obesity‐induced chronic inflammation.

First, our microRNA array analysis from macrophages subjected to polarized activation conditions revealed a set of microRNAs were actively regulated in macrophage classical activation. Bone marrow‐derived macrophages (BMDMs) stimulated with lipopolysaccharide/interferon gamma (LPS/INFγ) inflammatory responses (M1) and alternative activation (M2) by IL‐4/IL‐13 treatment. There were 33 miRNAs detected in naïve macrophage (M0), 156 miRNAs detected in M1, and 159 miRNAs detected in M2. Furthermore, there were 33 miRNAs displayed significant differential expression patterns in M1 or M2. Genomic analysis by surveying the promoters of these miRNAs identified a cluster of 10 miRNAs bearing ERα binding sites within 10 upstream kilobases. Interestingly, BMDMs subjected to 17β‐estradiol treatment exhibited suppressed M1 responses accompanied by significantly higher levels of ERa and miR‐365; whereas no detectable changes in ERa level in M2 activation. BMDMs treated with LPS/INFγ in the presence of 17β‐estradiol exhibited significantly lower levels of IL‐6, TNFα, and IL‐1β. Of note, miR‐365 level was significantly lower in M1 compared to M0, which was reversed by 17β‐estradiol treatment. Validation of the regulatory axis of ERa‐miR365 in macrophage classic activation will provide crucial evidence to support a novel paradigm shift in understanding the function of estrogen in obesity‐induced tissue inflammation and subsequent systemic disorders.

Support or Funding Information

American Diabetes Association (1‐13‐JF‐59 to B. Zhou) and National Institute of Health/National Institute of Diabetes and Digestive and Kidney Diseases (NIH/NIDDK 1R01DK098662 to B. Zhou)
Physiological And Transcriptomic Analyses In Escherischia coli Reveal Effects On Stress Associated Metabolism In The Presence of Oxidized Graphene
Authors: RAMPAL SAKSHI , Barabote Ravi Damodar
Published: November 2023

Graphene is a carbon nanomaterial that has a great potential to be used in wide variety of fields especially in biomedical applications. However, the environmental and health effects of graphene‐derived nanomaterials are not fully understood. Oxidized graphene is widely used in the synthesis of graphene‐derived nanomaterials. We assessed the biological effects of oxidized graphene using Escherichia coli as a model bacterium. Growth of E. coli was monitored in the presence of pristine graphene and oxidized graphene using dilution plating and spot test analysis. Growth analysis revealed that addition of oxidized graphene at concentrations greater than 80 μg/ml inhibited the growth of E. coli. To understand the molecular response of E. coli to graphene, RNA‐seq technology (Illumina platform) was used. Transcriptomic analysis of E. coli exposed to pristine graphene and oxidized graphene revealed several genes that were substantially differentially expressed. The genes that showed higher expression in the presence of oxidized graphene encode for stress response proteins, membrane proteins, transporters, acid resistance proteins, and proteins involved in sulfur metabolism. These data suggests that oxidized graphene likely elicits oxidative stress in E. coli. Using gene deletion knockouts, we studied the physiological role of a few of the genes involved in above‐mentioned functions. Deletion mutants of an inner membrane protein and a sulfite reductase were grown in the absence or presence of oxidized graphene. Growth data showed that both genes are critical for the bacteria to survive under stress conditions induced by oxidized graphene. In addition, we are studying the expression responses of select genes using GFP reporter constructs to identify biomarker genes specific to oxidized graphene.

Support or Funding Information

Department of Biological Sciences, University of Arkansas; Cell and Molecular Biology Program, University of Arkansas; Arkansas Research Alliance