The selection of proper reference genes and materials is critical in the design of PCR experiments, especially for differential expression studies. In this study, we propose a method to identify robust endogenous control miRNAs in the visceral adipose tissue of C57BL/6J mice with non-alcoholic fatty liver disease induced by alternating Western and control diets. This study outlines a comprehensive methodology for the analysis of microRNA endogenous controls using microfluidic cards in conjunction with miRNA profiling through small RNA sequencing and subsequent validation by quantitative PCR and the RefFinder algorithm. Criteria included were fold change, p-value, reads per million, and gene stability assessment. A set of six putative endogenous microRNAs was identified (miR-331-3p, let-7a-5p, miR-1839-5p, miR-151a-5p, let-7d-5p, and let-7c-5p). Subsequent validation and analysis using the RefFinder algorithm assessed the stability of the selected genes, and a combination of the three most stable endogenous miRNA controls (miR-331-3p, let-7a- 5p, and miR-1839-5p) exhibiting consistent expression patterns with minimal variability was set. Given the absence of universal endogenous controls, individual evaluation of normalizers for each experiment is imperative for accurate miRNA expression measurements. This approach, which combines multiple techniques and assessments, provides a reliable strategy for identifying and validating endogenous controls in miRNA studies.
- MeSH
- algoritmy MeSH
- mikro RNA * genetika metabolismus MeSH
- modely nemocí na zvířatech * MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nealkoholová steatóza jater * genetika metabolismus patologie MeSH
- nitrobřišní tuk * metabolismus MeSH
- regulace genové exprese MeSH
- stanovení celkové genové exprese metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
INTRODUCTION: A comparison of body composition assessments using military circumferences to bioelectrical impedance analysis (BIA) and the reference standard dual-energy X-ray absorptiometry (DEXA) can gauge effectiveness of assessments. High-frequency (500 KHz) direct segmental multifrequency bioelectrical impedance analysis (DSM-BIA) accurately calculates total water mass and body fat% (BF%), but it is unknown whether higher frequencies (1,000 KHz) increase measurement accuracy. The purpose was to compare DSM-BIA 500, DSM-BIA 1000, the DoD Circumference Method (CM), and the reference-standard DEXA. MATERIALS AND METHODS: Design: Cross sectional, observational study. Participants/Setting: A total of 62 participants from the military healthcare system (n = 25 males, 38.8 ± 11.4 years, n = 37 females 43.7 ± 15.95 years) were measured in an outpatient clinic setting. Statistical Analysis: BF% was estimated via DEXA, DSM-BIA 500, DSM-BIA 1000, and CM to identify the relationship between methods using Pearson correlation, intraclass correlation coefficients (ICCs), and Bland-Altman plots. The study was approved by the IRB from Walter Reed National Military Medical Center at Bethesda and Concordia University Chicago. RESULTS: Circumference Method BF% was moderately correlated with DSM-BIA 500 (males r = 0.63, ICC = 0.76; females r = 0.77, ICC = 0.85), DSM-BIA 1000 (males r = 0.59, ICC = 0.74; females r = 0.77, ICC = 0.85), and DEXA (males r = 0.62, ICC = 0.62; females r = 0.73, ICC = 0.82). DSM-BIA 500 BF% was strongly correlated with DSM-BIA 1000 (males r = 0.99, ICC = 0.99; females r = 0.99, ICC = 0.99) and DEXA (males r = 0.93, ICC = 0.94; females r = 0.89, ICC = 0.89). Lastly, DSM-BIA 1000 BF% was also strongly correlated with DEXA (males r = 0.93, ICC = 0.94; females r = 0.84, ICC = 0.90) (P for each reported r < 0.01). Bland-Altman analysis confirmed an overall mean bias of -1.72% CM vs. DEXA in females, indicating the tendency of CM to underestimate BF% compared to DEXA limits of agreement from -14.24 to 10.8. There was an upward slope of the linear relationship between the bias and mean of the measures (Beta = 0.34, P = 0.01). In the full cohort, there was an overall mean bias of 1.14% of CM vs. DSM BIA 1000, with CM tending to overestimate BF% compared to DSM BIA 1000 with limits of agreement -11.13 to 13.41%. There is an upward slope line of the linear relationship between the bias and the mean of the measures (Beta = 0.17, P = .03). CONCLUSION: This study found that CM BF% was moderately correlated with DSM-BIA 500 kHz, DSM-BIA 1,000 kHz BIA, and DEXA. Both DSM-BIA 500 and DSM-BIA 1,000 kHz strongly correlated well with DEXA implying that there was no further increase in correlation with increased frequency. Additionally, there was proportional bias in BF% in the female group between CM and DEXA and in the total group between CM and DSM BIA 1000.
- MeSH
- absorpční fotometrie * metody statistika a číselné údaje MeSH
- dospělí MeSH
- elektrická impedance * MeSH
- index tělesné hmotnosti MeSH
- lidé středního věku MeSH
- lidé MeSH
- průřezové studie MeSH
- složení těla * fyziologie MeSH
- tuková tkáň diagnostické zobrazování fyziologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- srovnávací studie MeSH
Fibroblast growth factor 21 (FGF21), a metabolic hormone with pleiotropic effects, is beneficial for various cardiac disorders. However, FGF21's role in heart failure with preserved ejection fraction (HFpEF) remains unclear. Here, we show that elevated circulating FGF21 levels are negatively associated with cardiac diastolic function in patients with HFpEF. Global or adipose FGF21 deficiency exacerbates cardiac diastolic dysfunction and damage in high-fat diet (HFD) plus N[w]-nitro-L-arginine methyl ester (L-NAME)-induced HFpEF mice, whereas these effects are notably reversed by FGF21 replenishment. Mechanistically, FGF21 enhances the production of adiponectin (APN), which in turn indirectly acts on cardiomyocytes, or FGF21 directly targets cardiomyocytes, to negatively regulate pyruvate dehydrogenase kinase 4 (PDK4) production by activating PI3K/AKT signals, then promoting mitochondrial bioenergetics. Additionally, APN deletion strikingly abrogates FGF21's protective effects against HFpEF, while genetic PDK4 inactivation markedly mitigates HFpEF in mice. Thus, FGF21 protects against HFpEF via fine-tuning the multiorgan crosstalk among the adipose, liver, and heart.
- MeSH
- adiponektin * metabolismus genetika MeSH
- dieta s vysokým obsahem tuků * škodlivé účinky MeSH
- energetický metabolismus * účinky léků MeSH
- fibroblastové růstové faktory * metabolismus genetika MeSH
- fosfatidylinositol-3-kinasy metabolismus MeSH
- kardiomyocyty * metabolismus účinky léků MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- PDH-kinasa * metabolismus genetika MeSH
- protoonkogenní proteiny c-akt metabolismus MeSH
- signální transdukce MeSH
- srdeční mitochondrie * metabolismus účinky léků MeSH
- srdeční selhání * metabolismus prevence a kontrola genetika MeSH
- tepový objem účinky léků MeSH
- tuková tkáň metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Vaccines represent an essential tool for the prevention of infectious diseases. Upon administration, a complex interaction occurs between the vaccine formulation and the recipient's immune system, ultimately resulting in protection against disease. Significant variability exists in individual and population responses to vaccination, and these differences remain the focus of the ongoing research. Notably, well-documented factors, such as age, gender, and genetic predisposition, influence immune responses. In contrast, the effects of overweight and obesity have not been as thoroughly investigated. The evidence indicates that a high body mass index (BMI) constitutes a significant risk factor for infections in general, with adipose tissue playing a crucial role in modulating the immune response. Furthermore, suboptimal levels of vaccine seroconversion have been observed among individuals with obesity. This review provides a plausible examination of the immunity and protection conferred by various vaccines in individuals with an overweight status, offering a comprehensive analysis of the mechanisms to enhance vaccination efficiency.
Perivascular adipose tissue (PVAT) envelops the majority of systemic vessels, providing crucial mechanical support and vessel protection. In physiological conditions, PVAT releases various bioactive molecules, contributing to the anti-inflammatory environment around neighboring vessels. However, in conditions like obesity, PVAT can exacerbate cardiovascular issues such as atherosclerosis. Communication between PVAT and nearby vessels is bidirectional, with PVAT responding dynamically to signals from the vasculature. This responsiveness positions PVAT as a promising indicator of vascular inflammation. Recently, the role of PVAT in the CVD risk prediction is also greatly discussed. The objective of this review is to summarize the current state of knowledge about the PVAT function, its role in physiologic and pathophysiologic processes and its potential in CVD risk prediction. Keywords: Perivascular adipose tissue, inflammation, atherogenesis, Fat attenuation index.
- MeSH
- ateroskleróza * metabolismus patologie patofyziologie MeSH
- lidé MeSH
- obezita metabolismus patofyziologie patologie MeSH
- tuková tkáň * metabolismus patologie patofyziologie MeSH
- zánět * metabolismus patologie patofyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
We investigated the sex-dependent effects of inflammatory responses in visceral adipose tissue (VAT) and perivascular adipose tissue (PVAT), as well as hematological status, in relation to cardiovascular disorders associated with prediabetes. Using male and female hereditary hypertriglyceridemic (HHTg) rats-a nonobese prediabetic model featuring dyslipidemia, hepatic steatosis, and insulin resistance-we found that HHTg females exhibited more pronounced hypertriglyceridemia than males, while HHTg males had higher non-fasting glucose levels. Additionally, HHTg females had higher platelet counts, larger platelet volumes, and lower antithrombin inhibitory activity. Regarding low-grade chronic inflammation, HHTg males exhibited increased serum leptin and leukocyte levels, while females had increased serum interleukin-6 (IL-6). Both sexes had increased circulating plasminogen activator inhibitor-1 (PAI-1), higher PAI-1 gene expression in VAT and PVAT, and elevated intercellular adhesion molecule-1 (ICAM-1) gene expression in the aorta, contributing to endothelial dysfunction in the HHTg strain. However, HHTg females had lower tumor necrosis factor alpha (TNFα) gene expression in the aorta. Severe dyslipidemia in this prediabetic model was associated with hypercoagulation and low-grade chronic inflammation. The increase in PAI-1 expression in both VAT and PVAT seems to indicate a link between inflammation and vascular dysfunction. Despite the more pronounced dyslipidemia and procoagulation status in females, their milder inflammatory response may reflect an association between reduced cardiovascular damage and prediabetes.
- MeSH
- dyslipidemie * metabolismus patologie genetika MeSH
- inhibitor aktivátoru plazminogenu 1 * metabolismus genetika MeSH
- krysa rodu rattus MeSH
- modely nemocí na zvířatech * MeSH
- nemoci cév metabolismus patologie etiologie genetika MeSH
- nitrobřišní tuk * metabolismus patologie MeSH
- pohlavní dimorfismus MeSH
- zánět * metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: The main objective of the present cross-sectional cohort study was to determine whether there is an association between cardiac autonomic regulation, as expressed through heart rate variability (HRV), and cardiorespiratory fitness (CRF), visceral adipose tissue (VAT), and over the long-term living in areas with low or high air pollution. METHODS: The study sample included 1036 (487 females) healthy runners (603) and inactive participants (age 18-65 years) who had lived for at least 5 years in an area with high (Moravian-Silesian; MS) or low (South Bohemian; SB) air pollution in the Czech Republic. A multivariable regression analysis was used to evaluate the associations between multiple independent variables (CRF (peak oxygen consumption), VAT, sex, socioeconomic status (education level), and region (MS region vs. SB region) with dependent variable HRV. The root mean square of successive RR interval differences (rMSSD) was employed for the evaluation of HRV. RESULTS: The multivariable linear regression model revealed that cardiac autonomic regulation (rMSSD) was significantly associated with CRF level (p < .001) and age (p < .001). There were no associations between rMSSD and region (high or low air-pollution), sex, education level or VAT (p > 0.050). CONCLUSIONS: We showed that living in an area with low or high air pollution is not associated with cardiac autonomic modulation in healthy runners and inactive individuals. CRF and age significantly directly and inversely, respectively, associated with HRV. There were no other significant associations.
- MeSH
- autonomní nervový systém fyziologie MeSH
- dospělí MeSH
- kardiorespirační zdatnost * fyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- nitrobřišní tuk * MeSH
- průřezové studie MeSH
- senioři MeSH
- srdce fyziologie MeSH
- srdeční frekvence * fyziologie MeSH
- znečištění ovzduší * škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
AIM: To propose a standardized, high-resolution ultrasound (US) protocol to assess the patellar tendon-Hoffa fat pad interface (PTHFPI) in patients with (proximal) patellar tendinopathy (PPT). METHODS: Using a high-frequency transducer and a high-level machine, we matched the cadaveric and histological microarchitecture of the PTHFPI with multiple sonographic patterns of patients with PPT. Likewise, high-sensitive color/power Doppler assessments were also performed to evaluate the microcirculation of the soft tissues beneath the patellar tendon. RESULTS: Modern US equipment allows for detailed assessment of the potential pain generators located inside the PTHFPI in patients with PPT. They include anterosuperior portion of the Hoffa body and the loose connective tissue of the deep paratenon with its microvascular plexus. CONCLUSIONS: In patients with PPT, accurate sonographic assessment of the PTHFPI can be performed using adequate technological equipment. Accordingly, tailored ultrasound-guided interventions can also be planned if/when clinically indicated.
- MeSH
- kolenní kloub diagnostické zobrazování MeSH
- lidé středního věku MeSH
- lidé MeSH
- ligamentum patellae * diagnostické zobrazování MeSH
- mrtvola MeSH
- senioři MeSH
- tendinopatie * diagnostické zobrazování patologie MeSH
- tuková tkáň * diagnostické zobrazování MeSH
- ultrasonografie metody MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVES: Submental fullness has been associated with being perceived as unattractive. Technology combining radiofrequency and muscle stimulation offers submental contouring through fat reduction, muscle stimulation, and skin tightening. This study aims to demonstrate the effectiveness and safety of fat reduction aspect with a novel submentum applicator delivering HIFES and synchronized radiofrequency+ (RF+) energies. MATERIALS AND METHODS: Six white pigs (sus scrofa domesticus, n = 6, 60-80 kg) were recruited for this study, five in the active group (n = 5) received four treatments on the abdominal area, one sow served as a control (n = 1). Ultrasound, histological, and RT-qPCR methods were used as evaluation methods. RESULTS: Fat thickness decreased at 1 month by -17.35% and at 2 month by 31.40%. Proapoptotic caspase-9 gene expression increased (at 1 h, 6 h, 24 h to +43.45%, +21.22%, -8.36%), as well as caspase-3 (+15.28%, +21.77%, -6.71%), while bcl2l1 activity decreased (-11.46% at 1 h, -17.02% at 6 h, -3.9% at 24 h). While the AI in the control animal had minimal change (at 1 h -0.08%, at 6 h -0.09%, and at 24 h -0.025%), the active group's AI increased from the baseline of 9.14 to 44.85 at 1 h (+391%), peaked at 6 h to 53.50 (+485%), and at 24 h to 38.17 (+318%). CONCLUSION: The study results indicate the efficacy and safety of subcutaneous fat reduction following the novel technology combining HIFES and RF+ energies, designed to target small localized areas.
