TBX3 transfection and nodal signal pathway inhibition promote differentiation of adipose mesenchymal stem cell to cardiac pacemaker-like cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are known as one of the best candidate cells to produce cardiac pacemaker-like cells (CPLCs). Upregulation of TBX3 transcription factor and inhibition of the nodal signal pathway have a significant role in the formation of cardiac pacemaker cells such as sinoatrial and atrioventricular nodes, which initiate the heartbeat and control the rhythm of heart contractions. This study aimed to confirm the effects of transfection of TBX3 transcription factor and inhibition of the nodal signal pathway on differentiating adipose-derived MSCs (AD-MSCs) to CPLCs. AD-MSCs were characterized using flow cytometry and three-lineage differentiation staining. METHODS: The transfection of TBX3 plasmid was carried out using lipofectamine, and inhibition of the nodal signal pathway was done using the small-molecule SB431542. The morphology of the cells was observed using a light microscope. Pacemaker-specific markers, including TBX3, Cx30, HCN4, HCN1, HCN3, and KCNN4, were evaluated using the qRT-PCR method. For protein level, TBX3 and Cx30 were evaluated using ELISA and immunofluorescence staining. The electrophysiology of cells was evaluated using a patch clamp. RESULTS: The TBX3 expression in the TBX3, SM, and TBX + SM groups significantly higher (p < 0.05) compared to the control group and cardiomyocytes. The expression of Cx40 and Cx43 genes were lower in TBX3, SM, TBX + SM groups. In contrast, Cx30 gene showed higher expression in TBX3 group. The expression HCN1, HCN3, and HCN4 genes are higher in TBX3 group. CONCLUSION: The transfection of TBX3 and inhibition of the nodal signal pathway by small-molecule SB431542 enhanced differentiation of AD-MSCs to CPLCs.

Analyze, design, develop, implement, and evaluate approach to develop a pelvic floor muscle training guidebook to treat stress urinary incontinence in women.

OBJECTIVE: Stress urinary incontinence (SUI) is a common problem that affects the quality of life of women worldwide. Pelvic floor muscle training (PFMT) is an effective conservative first-line treatment for SUI. However, low compliance with PFMT is one of the main reasons for therapeutic failure. Indirect supervision using a guidebook may improve PFMT outcomes. To develop a PFMT guidebook using the analyze, design, development, implementation, and evaluation (ADDIE) method. METHODS: A guidebook was developed from July 2020 to April 2021 using the ADDIE method. This prospective study used mixed methods, namely qualitative analysis, focus group discussions, and in-depth interviews, and involved various experts from urogynecology, urology, medical rehabilitation, and physiotherapy departments. A pilot study was conducted on patients with SUI to evaluate the effectiveness of the guidebook. RESULTS: The ADDIE method was successfully implemented to develop the PFMT guidebook. The formative evaluation of the ADDIE steps mainly focused on the PFMT technique, content clarity, illustration, design, and color choice of the book. After the pilot study, the guidebook significantly improved Incontinence Impact Questionnaire, Short Form, 1-hour pad test, and perineometer scores. However, the pilot study showed no significant improvement in Urogenital Distress Inventory, Short Form scores. CONCLUSION: The PFMT guidebook developed using the ADDIE method improved outcomes in patients with SUI.

Harnessing the power of probiotics to enhance neuroplasticity for neurodevelopment and cognitive function in stunting: a comprehensive review.

BACKGROUND: Stunting become a global concern because it's not only affecting physical stature, but also affecting on neurodevelopment and cognitive function. These impacts are resulting in long-term consequences especially for human resources, such as poor-quality labor, decreased productivity due to decreasing of health quality, including immunity and cognitive aspect. DISCUSSION: This comprehensive review found that based on many studies, there is an altered gut microbiota, or dysbiosis, in stunted children, causing the impairment of brain development through Microbiota-Gut Brain Axis (MGB Axis) mechanism. The administration of probiotics has been known affect MGBA by improving the physical and chemical gut barrier integrity, producing antimicrobial substance to inhibit pathogen, and recovering the healthy gut microbiota. Probiotics, along with healthy gut microbiota, produce SCFAs which have various positive impact on CNS, such as increase neurogenesis, support the development and function of microglia, reduce inflammatory signaling, improve the Blood Brain Barrier's (BBB's) integrity, produce neurotropic factors (e.g. BDNF, GDNF), and promote the formation of new synapse. Probiotics also could induce the production of IGF-1 by intestinal epithelial cells, which functioned as growth factor of multiple body tissues and resulted in improvement of linear growth as well as brain development. CONCLUSION: These properties of probiotics made it become the promising and feasible new treatment approach for stunting. But since most of the studies in this field are conducted in animal models, it is necessary to translate animal data into human models and do additional study to identify the numerous components in the MGB axis and the effect of probiotics on human.

Obesity and Its Relation to Excessive Daytime Sleepiness in Civilian Pilots.

INTRODUCTION: Excessive daytime sleepiness (EDS) is often associated with decreased work performance and fatigue in civil pilots. However, aeromedical recommendations for the evaluation of EDS are associated with suspicion of obstructive sleep apnea (OSA). Currently, many studies have found an association between obesity and EDS, regardless of OSA. This study aims to determine whether there is a relationship between obesity and EDS in Indonesian civilian pilots, as well as its risks for developing OSA.METHODS: This study used a cross-sectional design and was carried out at the Directorate General Civil Aviation Medical. Subjects were asked to fill out questionnaires, including the Epworth Sleepiness Scale to measure EDS and STOP-Bang to assess OSA risk, followed by anthropometric measurements for body mass index (BMI) and waist circumference as obesity indicators.RESULTS: A total of 156 subjects were obtained, with an EDS prevalence of 16.7%. There was no significant relationship between obesity and EDS, but the prevalence of EDS was higher in obese subjects based on waist circumference than based on BMI (17.8% vs. 15.6%). Most obese pilots with EDS had a low risk of OSA (83.3% and 80%).CONCLUSION: The prevalence of EDS was found to be higher in pilots with central obesity compared to BMI-categorized obesity. The incidence of EDS was not correlated with the risk of OSA.Brahmanti RS, Sampurna B, Ibrahim N, Adi NP, Siagian M, Werdhani RA. Obesity and its relation to excessive daytime sleepiness in civilian pilots. Aerosp Med Hum Perform. 2023; 94(11):815-820.

Prediction of Translational Regulation by Network Interaction in Synaptic Plasticity Induced with Centella asiatica.

Background: Recently, human life expectancy, aging, and age-related health disorders, especially neurodegenerative diseases such as Alzheimer's disease (AD), have increased. The increasing number of AD patients causes a heavy social and economic burden on society. Since there is no treatment for AD, utilization of natural products is currently accepted as an alternative or integrative treatment agent against AD. Methods: Selection of protein databases related to synaptic plasticity was obtained from a gene bank. The protein-protein interaction (PPI) analysis was performed using Cytoscape 3.9.1. Prediction of Centella asiatica target constituents and their relationship with target synaptic plasticity was performed using STITCH, followed by GO and KEGG pathway enrichment analysis and molecular binding of ligands to presynaptic and postsynaptic receptors afterwards. Results: From the protein database, 446 protein coding genes related to synaptic plasticity were found. PPI and KEGG pathway analysis showed potentiality to inhibit AKT and mTORC1 pathways. The targeted proteins were TSC1, Rheb, and FMRP. Conclusion: This study showed potentiality of Centella asiatica in AD through its binding to several proteins such as TSC1, Rheb, and FMRP. This compound in Centella asiatica was able to bind to the AKT1 and mTOR signaling pathways. Centella asiatica may behold greater potency in AD therapy.

Analysis of the infant's acute pain assessment using developed conductance skin electric instrument compared to the behavioural and faces pain scale in painful injected vaccine.

This study compared the scale of infant pain during vaccinated injection using conductance skin electric (Skin Conductance), the Wong-Baker Faces Scale (WBFS) instrument, and Face Leg Activity Cry and Consolability (FLACC) instruments. It was observational cohort study with pre-experimental design using vaccinated injection as pain stimuli. This study investigated 121 infants (59 boys, 62 girls), age/PNA 4.37 ± 2.97 months, and current body weight 6522 ± 1378.65 grams). Most infants had adequate birth weight 71 (89.9%) about 2985.74 ± 405.83 kg and mature infants as 67 (84.4%), about 38,52 ± 2,09 weeks. Pain measurement of all three instruments was do simultaneously using a developed Skin Conductance (SC) apparatus and video recording (to assess behaviour and face). WBFS, FLACC and SC have the same significance in measuring infants' pain scale during vaccination injection. Statistical analysis showed a significant difference in the three pain measurement instruments between before and during injection with a p value of <0.001. So that SC can be recommended for pain measurement.

Effect of altitude zone exposure on visuospatial function in military aircrew member.

Introduction: visuospatial is a type of high-level visual perception necessary for identification, information integration, analysing of visual and spatial forms, details, structures and spatial relation. These functions are required in order to successfully complete aviation-related tasks, such as analysing movement, distance perception, and spatial navigation. The aim of this study is to examine whether hypobaric hypoxia can significantly influence changes in visuospatial function, thus increasing the risk of accident or serious incident during flight operation. Methods: this study is a quasi-experiment of pre-post study including before and after hypobaric hypoxia simulated through an altitude chamber. In this study, 42 military aircrews were exposed to different altitude zones at ground level, 10,000 ft (ft) and 25,000 ft respectively, for five minutes. At each altitude zone, the participants were instructed to complete a clock drawing test as a measurement for visuospatial function. The results were analysed using the McNemar non-parametric test. Results: among the 42 subjects, six show impaired visuospatial function at 10,000 ft and 26 participants show it at 25,000 ft. There were significant increased on the proportion of impaired visuospatial function between the ground level to 10,000 ft (p=0.031), 10,000 to 25,000 ft (p=0.0001) and ground level to 25,000 ft (p=0.0001). Conclusion: hypobaric hypoxia may have a significant influence on visuospatial function, starting from as early as 10,000 ft to 25,000 ft. This decrease of visuospatial function could affect human cognitive performance when flying and increase the risk of aviation accidents.

Anti-inflammatory Effects of Hibiscus Sabdariffa Linn. on the IL-1ß/IL-1ra Ratio in Plasma and Hippocampus of Overtrained Rats and Correlation with Spatial Memory.

Overtraining leads to an increase in IL-1ß systemically due to muscle microtrauma, which affects the hippocampus, an important structure in spatial memory consolidation. The administration of Hibiscus sabdariffa Linn is expected to decrease IL-1ß and increase IL-1ra, thereby potentially preventing impairments in spatial memory consolidation. This research was an experimental study using 20 male Wistar rats. The overtraining of Wistar rats altered the ratio of IL-1ß/IL-1ra in the plasma and hippocampus. Moreover, this overtraining impaired spatial memory consolidation. The methanol extract of H. sabdariffa improved spatial memory consolidation in Wistar rats and prevented impairment in spatial memory consolidation by maintaining the ratio of IL-1ß/IL-1ra in the plasma and hippocampus of Wistar rats who experienced overtraining. H. sabdariffa is a potent anti-inflammatory substance that prevents impairments in spatial memory consolidation in overtrained Wistar rats.

Correcting of pronated feet reduce skeletal muscle injury in young women with biomechanical abnormalities.

Biomechanical abnormalities of pronated feet accompanied by functional leg length disparity may increase the risk of skeletal muscle injury. Objective of the study is to prove that correction of pronated feet by the foot orthoses will reduce the creatine kinase-MM (CK-MM) concentrations as the muscle injury indicator. The design study was double blind randomized clinical trials with control. Research subjects were divided into two groups, group 1 used the foot orthoses while group 2 did not used the foot orthoses. The whole subject examined the concentrations of the CK-MM enzyme before, and 24-72 hours after the walking test. The walking test was conducted 15 minutes with maximum speed. The concentration of the CK-MM enzyme before walking test on treatment group was 70.07±15.33 International Unit (IU), similar with the control group was 69.85±17.03 IU (P=0.971). The increased in CK-MM enzyme concentrations 45 hours after the walking test was lower in the treatment group (7.8±9 IU) than the control group (22.0±11.5 IU) (P=0.001). The CK-MM enzyme concentrations continued to decline in the treatment group after the second walking test (77.21±17.47 IU), and after the third walking test (69.86±11.88 IU) (P=0.018). The foot orthoses for correcting the pronated feet on the young women with biomechanical abnormalities is able to reduce the degree of the skeletal muscle injury after walking activity.

The role of curcumin as an inhibitor of oxidative stress caused by ischaemia re-perfusion injury in tetralogy of Fallot patients undergoing corrective surgery.

BACKGROUND: Cardiopulmonary bypass during tetralogy of Fallot corrective surgery is associated with oxidative stress, and contributes to peri-operative problems. Curcumin has been known as a potent scavenger of reactive oxygen species, which enhances the activity of antioxidants and suppresses phosphorylation of transcription factors involved in inflamation and apoptosis. OBJECTIVES: To evaluate the effects of curcumin as an antioxidant by evaluating the concentrations of malondialdehyde and glutathione, activity of nuclear factor-kappa B, c-Jun N-terminal kinase, caspase-3, and post-operative clinical outcomes. METHODS: Tetralogy of Fallot patients for corrective surgery were randomised to receive curcumin (45 mg/day) or placebo orally for 14 days before surgery. Malondialdehyde and glutathione concentrations were evaluated during the pre-ischaemia, ischaemia, re-perfusion phases, and 6 hours after aortic clamping-off. Nuclear factor-kappa B, c-Jun N-terminal kinase, and caspase-3, taken from the infundibulum, were assessed during the pre-ischaemia, ischaemia, and re-perfusion phases. Haemodynamic parameters were monitored until day 5 after surgery. RESULTS: In all the observation phases, malondialdehyde and glutathione concentrations were similar between groups. There was no significant difference in nuclear factor-kappa B activity between the groups for three observations; however, in the curcumin group, c-Jun N-terminal kinase significantly decreased from the pre-ischaemia to the re-perfusion phases, and caspase-3 expression was lower in the ischaemia phase. Patients in the curcumin group had lower temperature and better ventricular functions, but no significant differences were found in mechanical ventilation day or length of hospital stay in the two groups. CONCLUSION: Cardioprotective effects of curcumin may include inhibition of the c-Jun N-terminal kinase pathway and caspase-3 in cardiomyocytes, particularly in the ischaemia phase.

Hypothalamic proopiomelanocortin neurons are glucose responsive and express K(ATP) channels.

Hypothalamic proopiomelanocortin (POMC) neurons are critical for controlling homeostatic functions in the mammal. We used a transgenic mouse model in which the POMC neurons were labeled with enhanced green fluorescent protein to perform visualized, whole-cell patch recordings from prepubertal female hypothalamic slices. The mouse POMC-enhanced green fluorescent protein neurons expressed the same endogenous conductances (a transient outward K(+) current and a hyperpolarization-activated, cation current) that have been described for guinea pig POMC neurons. In addition, the selective micro -opioid receptor agonist DAMGO induced an outward current (maximum of 12.8 +/- 1.2 pA), which reversed at K(+) equilibrium potential (E(K+)), in the majority (85%) of POMC neurons with an EC(50) of 102 nM. This response was blocked by the opioid receptor antagonist naloxone with an inhibition constant of 3.1 nM. In addition, the gamma-aminobutyric acid(B) receptor agonist baclofen (40 micro M) caused an outward current (21.6 +/- 4.0 pA) that reversed at E(K+) in these same neurons. The ATP-sensitive potassium channel opener diazoxide also induced an outward K(+) current (maximum of 18.7 +/- 2.2 pA) in the majority (92%) of POMC neurons with an EC(50) of 61 micro M. The response to diazoxide was blocked by the sulfonylurea tolbutamide, indicating that the POMC neurons express both Kir6.2 and sulfonylurea receptor 1 channel subunits, which was verified using single cell RT-PCR. This pharmacological and molecular profile suggested that POMC neurons might be sensitive to metabolic inhibition, and indeed, we found that their firing rate varied with changes in glucose concentrations. Therefore, it appears that POMC neurons may function as an integrator of metabolic cues and synaptic input for controlling homeostasis in the mammal.

Estrogen modulation of K(+) channel activity in hypothalamic neurons involved in the control of the reproductive axis.

Here we report on the progress we have made in elucidating the mechanisms through which estrogen alters synaptic responses in hypothalamic neurons. We examined the modulation by estrogen of the coupling of various receptor systems to inwardly rectifying and small conductance, Ca(2+)-activated K(+) (SK) channels. We used intracellular sharp-electrode and whole-cell recordings in hypothalamic slices from ovariectomized female guinea pigs. Estrogen rapidly uncouples mu-opioid receptors from G protein-gated inwardly rectifying K(+) (GIRK) channels in beta-endorphin neurons, manifest by a reduction in the potency of mu-opioid receptor agonists to hyperpolarize these cells. This effect is blocked by inhibitors of protein kinase A and protein kinase C. Estrogen also uncouples gamma-aminobutyric acid (GABA)(B) receptors from the same population of GIRK channels coupled to mu-opioid receptors. At 24 h after steroid administration, the GABA(B)/GIRK channel uncoupling observed in GABAergic neurons of the preoptic area (POA) is associated with reduced agonist efficacy. Conversely, estrogen enhances the efficacy of alpha(1)-adrenergic receptor agonists to inhibit apamin-sensitive SK currents in these POA GABAergic neurons, and does so in both a rapid and sustained fashion. Finally, we observed a direct, steroid-induced hyperpolarization of both arcuate and POA neurons, among which gonadotropin-releasing hormone (GnRH) neurons are particularly sensitive. These findings indicate a richly complex yet coordinated steroid modulation of K(+) channel activity that serves to control the excitability of hypothalamic neurons involved in regulating the reproductive axis.