Abstract in English:

Bone fractures must undergo a complex healing process involving intricate cellular and molecular mechanisms. They require a suitable biological environment to restore skeletal stability and resolve inflammation. Scaffolds play a vital role in bone regeneration, thus reducing disease burden. Autologous bone graft represents the gold standard of therapy. However, its application is limited due to various reasons. Nanotechnology, in the form of nanomaterials and nano-drug delivery systems, has been proven to increase the potency of active substances in mimicking extracellular matrix (ECM), thereby providing physical support benefits and enhancing therapeutic effectiveness. Various materials, including protein, metal oxide, hydroxyapatite, and silica are modified with nanoparticle technology for the purposes of tissue regeneration therapy. Moreover, the properties of nanomaterials such as size, seta potential, and surface properties will affect their effectiveness in bone regeneration therapy. This review provides insights that deepen the knowledge of the manufacturing and application of nanomaterials as a therapeutic agent for bone regeneration.

Abstract in English:

This systematic review of inception prospective cohort studies aimed to investigate whether autoantibodies are potential prognostic factors for short- and long-term clinical outcomes of COVID-19. Searches were conducted in MEDLINE, EMBASE, AMED, GLOBAL HEALTH, and COCHRANE databases from 2019 to 2022. When possible, meta-analysis was conducted, otherwise findings from individual studies were reported using odds ratios (OR) with 95% confidence intervals (CI). Quality of evidence was summarized using the GRADE criteria. We identified 2292 references, 18 inception prospective cohort studies (3178 patients) were included in the systematic review, and 12 studies reached criteria for meta-analysis. Studies achieved, in general, low to moderate risk of bias. Moderate quality of evidence showed that anti-interferon (IFN) was associated with increased risk of severity (OR=7.75; CI=1.79-33.61) and mechanical ventilation (OR=4.19; CI=2.06-8.53), but not with COVID-19 mortality (OR=1.68; CI=0.63-4.44). Antiphospholipids were not associated with COVID-19 mortality (OR=1.42; CI=0.85-2.37; P=0.18; I2=3.21) nor with thrombosis risk (OR=1.41; CI: 0.71-2.8; P=0.33). Antinuclear antibody level was not associated with risk of mortality or severity (risk for mortality: OR=3.8; CI=0.78-18.6; P=0.1; I2: 32.3; severity: OR=1.74; CI=0.96-3.16; P=0.07). Evidence currently available is insufficient for a quantitative analysis of autoantibodies association with long COVID-19. Anti-IFN measurement should be considered in COVID-19 follow-up. In a population-based rational, optimized vaccination strategies should be considered for individuals with anti-IFN antibodies since it could represent a risk for a worse prognosis. High-quality prospective studies for short- and long-term disease effects and autoantibody evaluation are still needed.

Abstract in English:

During the climacteric period, the decline in ovarian hormones leads to changes in the lipid profile. Physical exercise is the main non-pharmacological recommendation for controlling lipid levels. However, the effects on the lipid profile in perimenopausal and postmenopausal women are incipient and inconclusive. In this context, we searched the Embase, PubMed, Scopus, and Web of Science databases for randomized clinical trials on the effects of exercise on the lipid profile of these women. We excluded studies that did not specify criteria for classifying the climacteric phase, that involved women undergoing hormone replacement therapy, or that examined combined treatments or acute effects of physical exercise. The meta-analysis indicated that general physical exercise increased high-density lipoprotein cholesterol (HDL-C) levels (mean difference [MD]=4.89; 95% confidence interval [95%CI]=0.97 to 8.81) in perimenopausal women. For obese postmenopausal women, 16 weeks of aerobic training increased HDL-C levels (MD=3.88; 95%CI=0.56 to 7.20) and reduced total cholesterol (MD=-22.36; 95%CI=-29.67 to -15.05) and low-density lipoprotein cholesterol (LDL-C) levels (MD=-17.86; 95%CI=-25.97 to -9.75), whereas 12 weeks of resistance training increased HDL-C levels (MD=4.20; 95%CI=1.16 to 7.23) and decreased triglycerides (MD=-14.86; 95%CI=-26.62 to -3.09) and LDL-C levels (MD=-16.36; 95%CI=-28.05 to -4.67). Overall, the results showed that physical exercise regulated lipid profiles in perimenopausal and postmenopausal women. Specifically, 12 weeks of resistance exercise and 16 weeks of aerobic exercise improved the lipid profile of obese postmenopausal women.

Abstract in English:

Low back pain (LBP) is a common type of pain that causes disability and impairs cognitive function. With over 80% of adults estimated to experience LBP during their lifetime, this type of pain not only has a significant impact on the individual, but also on public health systems and national economies. Unfortunately, there is no single standard of care for patients with LBP. N-acetylcysteine (NAC), which is used clinically to treat acetaminophen overdose, has recently been tested as a potential treatment for LBP. NAC is inexpensive and commercially available, and it has an established tolerance and safety profile. However, NAC's efficacy in LBP has not been established. This scoping review presents a summary of studies investigating the effects of NAC and the potential benefits in LBP treatment, and highlights its potential molecular mechanisms and side effects. A systematic literature search in Pubmed/MEDLINE, Embase, Scopus, Science Direct, Web of Science, Cinahl, and Lilacs databases was conducted. The PRISMA-ScR checklist was used to ensure integrity of the review. The scoping review protocol was registered in the Open Science Framework. No limit was set on study language and publication date. In total, 2357 articles were located, of which 16 were included. The studies show that NAC has potential for LBP treatment, but data are derived only from a few clinical trials and preclinical studies. Thus, there is much to learn and more clinical studies should be performed before NAC can be clinically recommended for the treatment of LBP.

Abstract in English:

Cancer is the second leading cause of death worldwide. Cancer cachexia is a multifactorial catabolic syndrome responsible for almost one third of cancer-related deaths. Drug repurposing has been used in oncological research and drugs like clenbuterol and metformin seem to be reasonable candidates in the context of cancer cachexia, because the former is a β2-agonist that stimulates muscle gain and the latter has anti-inflammatory properties. The aim of this study was to assess the effects of a short-term treatment with metformin and clenbuterol, isolated or combined, on tumor growth and cancer cachexia parameters in Walker 256 tumor-bearing rats, a model of cancer cachexia. To this end, Wistar rats were separated into 8 groups and 4 of them were injected with Walker 256 tumor cells (W groups). Control (C) and W groups received the following treatments: metformin (M), clenbuterol (Cb), or metformin combined with clenbuterol (MCb). Body and tumor weight, metabolic parameters, and oxidative damage in the tumor were assessed. Compared to the C group, the W group showed body weight loss, hypoglycemia, hyperlactatemia, and hypertriacylglycerolemia. None of the treatments could reverse body weight loss, although they reversed the alterations of the assessed plasma metabolic parameters. Surprisingly, only clenbuterol alone reduced tumor weight. Hydrogen peroxide production and lipid peroxidation in tumor tissue was increased in this group. In conclusion, metformin and clenbuterol ameliorated metabolic cachexia parameters in Walker tumor-bearing rats, but only clenbuterol reduced the tumor weight, probably, through a lipid peroxidation-dependent cell death.

Abstract in English:

Osteosarcoma (OS) remains the most common bone tumor and the prognosis for many patients remains stagnant due to the unsatisfactory therapeutic effect of conventional treatment regimens. This research explored the effect and mechanism of a novel natural product, Eriocalyxin B (EB), in pathogenesis and immunotherapy in OS. Cell Count Kit 8 assay, colony formation assay, and wound healing assay were employed to detect the proliferative, colony-forming, and migratory abilities of human OS cells following EB treatment. Moreover, xenograft growth assay was performed to assess the effect of EB on OS in vivo. Subcutaneous OS models constructed in immunocompetent mice were employed to evaluate the effect of EB treatment in combination with immune checkpoint blockades (ICBs) PD1ab and CTLA4ab. Immunohistochemistry (IHC) staining was utilized to detect the level of CD8+ T cells infiltration and Ki67 expression. TARGET database, RNA interference technology, and qPCR assay were employed to explore the mechanism of EB on OS. EB inhibited the proliferative, colony-forming, and migratory abilities of the human OS cells MG63 and U2OS both in vitro and in vivo. TARGET data analysis demonstrated that up-regulation of TCEA3 was significantly negatively correlated with overall survival in OS patients. EB exerted anti-tumor activity via downregulation of TCEA3. EB, in conjunction with ICBs, synergistically optimized anti-tumorigenic activity against OS in immunocompetent mice. EB may promote infiltration of CD8+ T cells and down-regulate Ki67 expression. These results signaled that EB may have a role as a candidate therapeutic or preventive agent for the treatment of OS.

Abstract in English:

Necrotizing enterocolitis (NEC) is a severe intestinal disease of multifactorial origin that primarily affects premature infants. Approximately 27% of NEC babies develop short gut (SG) secondary to extensive intestinal resection, and 10% will have chronic dependence on total parenteral nutrition. We evaluated the Botox treatment in SG model rats. Twenty-day-old weanling male rats (weight range 38-70 g, n=72) were divided into four groups (n=18 each): 1) Control (fed a regular liquid diet); 2) Botox (Control submitted to laparotomy and intestinal injection of Botox®); 3) SG (short gut); and 4) SG and Botox (SG+Botox®). After seven post-operative days, samples were collected for biometrics [body weight (BW), intestine weight (IW) and IW/BW ratio (IBR), and intestine length (IL) and height (IH)], histometric analysis [villous height (VH), crypt depth (CD), muscular thickness (MT), and PCNA index)], and intestinal transit time (ITT). BW, IW, and IL decreased in SG (P<0.05). IH, VH, and PCNA index increased in Botox groups [Control = SG < Botox and SG+Botox (P<0.05)], CD increased in Botox, SG, and SG+Botox (P<0.005), and MT was higher in SG and SG+Botox. Botox groups had lower ITT (P<0.05). Botox provided dilatation and histological changes in SG. These findings suggested that Botox improved adaptation and might be applied in SG with promising results.

Abstract in English:

Frailty is a significant risk factor for adverse outcomes in elderly surgical patients. Gait speed assessment is a new tool recently used to stratify risk for these pre-operative adverse outcomes. In this prospective study of 392 frail elderly patients undergoing abdominal surgery, we investigated the predictive value of preoperative gait speed for postoperative outcomes. Patients were divided into two groups based on their 6-meter gait speed: normal (≥0.8 m/s, n=184) and slow (<0.8 m/s, n=208). The slow group was older, had more comorbidities, and higher American Society of Anesthesiologists (ASA) grades (P<0.05). They also had significantly higher rates of 30-day overall complications (38.9 vs 18.5%, P<0.01), severe complications (12.0 vs 4.3%, P<0.01), and 1-year mortality (15.4 vs 6.5%, P=0.008) compared to the normal group. Pulmonary infection, wound infection, and delirium were the most common complications. Multivariate logistic regression confirmed slow gait speed as an independent risk factor for 30-day complications (OR=2.38, 95%CI: 1.41-4.01) and 1-year mortality (OR=2.19, 95%CI: 1.07-4.48). Our findings demonstrated that preoperative 6-meter gait speed effectively predicted short-term complications and mid-term mortality in frail elderly patients undergoing abdominal surgery. This suggests the need for individualized perioperative management strategies for high-risk patients with slow gait speed to potentially improve their prognosis.

Abstract in English:

Sirtuins (SIRTs) are key regulators of cellular metabolism, involved in a wide range of physiological and pathological processes. However, there is scarce knowledge about the effect of sugar consumption and physical activity on SIRTs in kidney disorders. Here, we evaluated the impact of prolonged consumption of an isocaloric high-sugar diet (HSD) and physical training on the modulation of renal Sirts and the link between these alterations and possible obesity-associated kidney damage. Newly weaned male Wistar rats were fed a standard chow diet (STD) or HSD ad libitum and then subjected or not to regular workload swimming training for 18 weeks. Morphometric and biochemical parameters were analyzed, and the kidneys were removed for lipid quantification, histological analysis, and for Sirts1-7 expression. HSD led to the development of obesity, increased serum triglyceride levels, and glucose intolerance, regardless of higher caloric consumption. However, training was able to partially inhibit the HSD-induced obesogenic effect. No changes were identified in kidney mass, lipid content, histology, and creatinine clearance among the groups; these results were associated with a decrease in the renal expression of Sirt2-3 and Sirt7; however, training was able to reverse this modulation. The interaction between HSD and training led to an increase in Sirt4-7. However, Sirt1 remained constant among experimental groups. In conclusion, our results indicated that the transcriptional modulation of Sirts precedes HSD-induced damage and loss of kidney function, as well as a possible protective adaptive response of physical exercise on long-term Sirts expression.

Abstract in English:

It has been confirmed that the expression of miR-501-3p is closely related to the behavior of several cancers. This study aimed to elucidate the effects of miR-501-3p/SPC24 axis on the behavior of renal cancer cells and to identify its prognostic value in renal cancer. First, the expression of miR-501-3p in the renal cell carcinoma (RCC) cell line was detected using real-time quantitative polymerase chain reaction (RT-qPCR). Second, cell function identification experiments were performed, including CCK-8, scratch, transwell invasion, and flow cytometry assays. Several databases were applied to explore the possible mechanism of miR-501-3p tumor suppressor effect in RCC. To explore the value of miR-501-3p/SPC24 axis in predicting renal cancer patient overall survival (OS), GEPIA (http://gepia.cancer-pku.cn/index.html) was used. Finally, western blot was performed to detect the expression level of SPC24 in renal cancer cells predicted by bioinformatics analysis. Dual-Luciferase Reporter Assay was used to verify if SPC24 is a target of mir-501-3p. MiR-501-3p was found to be down-regulated in cancer cells and tissues and to play a role in suppressing tumor cell proliferation, cell viability, cell migration, and cell invasion, while promoting apoptosis. We also found that high expression levels of SPC24 were associated with shorter OS time in patients diagnosed with renal cell carcinoma. In addition, the results of TCGA data analysis and western blot showed that the tumor suppressor effect of miR-501-3p may be achieved by targeting SPC24. The MiR-501-3p/SPC24 axis affects cell proliferation, migration, invasion, apoptosis, and prognosis in renal cell carcinoma.

Abstract in English:

Malaria is a parasitic disease of great relevance in global public health. The development of new sensitive and specific diagnostic high-throughput methods remains a challenge in the eradication of this disease. In this study, we developed a flow cytometry test using latex microbeads and polyclonal antibodies obtained from rabbits and mice for the detection of the P. vivax lactate dehydrogenase (PvLDH) antigen. We processed 50 samples from Brazilian patients diagnosed with malaria caused by P. vivax and 40 samples from healthy individuals. The assay presented sensitivity of 64%, specificity of 97%, a positive predictive value of 97%, and a negative predictive value of 57% when analyzed using the fluorescent labeling method. Using the mean fluorescence intensity (MFI) analysis method, the sensitivity was 53%, specificity was 89%, the positive predictive value was 95%, and the negative predictive value was 33%. In both methods of analysis, we observed significant statistical differences between the analyzed groups (P-value <0.0001). A high correlation (0.60) between the two methods and a low correlation between PvLDH concentration and parasite density was found. The test was able to detect the PvLDH protein with high specificity, but its sensitivity should be improved. More promising results were observed when the samples were analyzed according to the percentage of fluorescent labeling. Improvement of this assay would enable its application as a serological test for the detection of asymptomatic patients and for the validation of rapid diagnostic tests.

Abstract in English:

The aim of the present study was to describe the use of tapering, carbohydrate (CHO) supercompensation, and supplementation strategies self-reported by athletes in the Olympic triathlon category. A total of 72 triathletes (61 males and 11 females) answered an online questionnaire about their training and performance, supercompensation strategies, carbohydrate supplementation, and use of supplements and other ergogenic substances. The information was summarized and subjected to descriptive analysis. Shapiro-Wilk test was applied to check data normality. The t-test was used to investigate differences in the analyzed variables between sexes. Almost all triathletes reported to have performed tapering (93.05%) and approximately half of them adopted a CHO supercompensation strategy (48.61%); updated CHO supercompensation was the most used strategy (27.77%). Most participants (86.11%) used CHO supplementation during competitions, but in amounts below the 60 g/h recommended for most athletes (96.77%). Thus, since few triathletes performed supercompensation, in addition to the insufficient amount of supplemented carbohydrate taken by them, it could be concluded that triathletes were not sufficiently aware of nutritional recommendations or did not adopt them.

Abstract in English:

The stiffening of the conductance arteries is a hallmark of ageing and increases drastically after menopause. Therefore, the augmentation index (AIx), a surrogate for arterial stiffness, could be related to the decline in baroreflex sensitivity. We sought to investigate the relationship between resting AIx and spontaneous cardiac baroreflex sensitivity (cBRS) during handgrip exercise in ageing women. Thirteen young women (YW: 24±5 years; 24±2 kg/m2) and nine postmenopausal women (PMW: 60±5 years; 26±3 kg/m2) underwent the protocol, which consisted of 10 min of supine resting followed by 3 min of static handgrip exercise at 40% of the maximal voluntary force. The AIx was provided by the aortic pressure waveform and cBRS was calculated using the sequence technique, and vagal activity was accessed via heart rate variability using the root mean square of successive differences (RMSSD) index. Resting AIx was higher in PMW compared to YW (YW: 8±10%; PMW: 23±8%; P<0.01), while the cBRS (YW: 16±12 ms/mmHg; PMW: 10±5 ms/mmHg; P=0.08) and RMSSD (YW: 46±35 ms; PMW: 34±12 ms; P=0.26) were similar in YW and PMW. At rest, there was no significant (P>0.05) relationship between the AIx and cBRS in YW and PMW. However, in PMW, a negative (slope=-0.22) and strong (r=-0.70; P=0.03) relationship was observed between AIx and cBRS for the increment of blood pressure during the handgrip exercise. The stiffening of the arterial tree is one possible mechanism to explain the decrease of spontaneous cardiac baroreflex sensitivity during exercise in postmenopausal women.

Abstract in English:

Head and neck squamous cell carcinoma (HNSCC) is a common malignant tumor that poses a major hazard to people's health. ZC3H12D, which belongs to the family of CCCH-type zinc finger-containing proteins, is a negative regulator with a key function in immune modulation. However, it is still unclear how ZC3H12D affects the immune infiltration and prognosis of HNSCC. In this study, the data obtained from various databases were used to assess ZC3H12D expression in HNSCC and in various tumors under the HNSCC classification. The association between clinical features and ZC3H12D expression in HNSCC was evaluated using the UALCAN database. Additionally, a ROC curve was employed to analyze the diagnostic value of ZC3H12D. The effect of ZC3H12D on prognosis was assessed using Kaplan-Meier curves, Cox analysis, and the nomogram model. Gene Set Enrichment Analysis, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were employed to investigate the underlying role of ZC3H12D in HNSCC. The association between ZC3H12D expression and the tumor microenvironment and immune checkpoints were investigated by TIMER2 and Tumor Immune Single Cell Hub 2 databases and various packages in R. The findings demonstrated a significant up-regulation of ZC3H12D expression in HNSCC, while ZC3H12D expression was found to be associated with clinical parameters. Our study also demonstrated that ZC3H12D could act as a potential prognostic biomarker for HNSCC, especially oral squamous cell carcinoma. Additional analyses have shown that ZC3H12D was associated with common immune checkpoint genes and may be related to immune infiltration in HNSCC.

Abstract in English:

Alpinia zerumbet, a plant native to East Asia, is widely found on the Brazilian coast, where it is used in folk medicine as an antihypertensive, diuretic, and anxiolytic. This study investigated the effects of the hydroalcoholic extract obtained from Alpinia zerumbet leaves (AZE) on cardiovascular changes and oxidative status in spontaneously hypertensive rats (SHR). SHR and Wistar-Kyoto male rats, 90 days old, treated or not with AZE (50 mg/kg/day in drinking water) for six weeks, were used in this study. Blood pressure (BP) was assessed weekly by tail plethysmography. At the end of treatment, the animals were anesthetized with thiopental (70 mg/kg, ip), blood was collected through abdominal aorta puncture, the thoracic aorta and left ventricle were isolated for morphometric analysis and immunostaining of NOX-4, SOD-2, 8-isoprostane, and angiotensin II AT1 receptors (AT1R), and the mesenteric arterial bed (MAB) was isolated for the assessment of vascular function. Oxidative damage in lipids and proteins and the enzymatic antioxidant activity were evaluated in plasma samples by spectrophotometry. AZE normalized BP in SHR. Although the treatment did not improve the MAB vascular dysfunction, it reversed the cardiovascular remodeling in the aorta and left ventricle. In addition, AZE improved antioxidant activity in plasma and SOD-2 immunostaining in the thoracic aorta and left ventricle, decreased protein carbonylation in plasma, and reduced 8-isoprostane, NOX-4, and AT1R immunostaining in the cardiovascular system. The results suggested that AZE reversed hypertension and cardiovascular remodeling in SHR, which was associated with lower oxidative stress and AT1R.

Abstract in English:

Nasopharyngeal carcinoma (NPC) is a malignant tumor predominantly influenced by Epstein-Barr virus infection and genetic factors. The transforming growth factor-beta (TGF-β) superfamily is implicated in various cellular processes, including tumorigenesis. This study aimed to detect the role of one TGF-β superfamily member activin receptor type IIB (ACTRIIB) in NPC. This study analyzed NPC datasets, including GSE12452, GSE102349, and GSE53819. ACTRIIB expression and N-glycosylation levels were assessed by western blot, real-time PCR, immunofluorescence, and immunohistochemistry in NPC cells and tissues. As indicated by the datasets, ACTRIIB was significantly upregulated in NPC tissues, and the up-regulation was associated with poor prognosis. This study confirmed the N-glycosylation of ACTRIIB primarily at the forty-second amino acid, an asparagine. The N-glycosylation of ACTRIIB promoted the localization of ACTRIIB to the cell membrane and prevented the degradation of the protein by lysosomes, through which ACTRIIB activated the downstream Smard1/2 to promote tumor cell proliferation and invasion. Inhibition of N-glycosylation or knockdown of ACTRIIB resulted in reduced cell proliferation and invasion and increased the cell sensitivity to docetaxel. In conclusion, N-glycosylation of ACTRIIB was a critical post-translational modification that enhanced protein stability and induced membrane localization, which facilitates the functions of ACTRIIB in cell proliferation and invasion in NPC. Inhibition of ACTRIIB N-glycosylation could potentially serve as a therapeutic strategy to improve the efficacy of chemotherapy in NPC.

Abstract in English:

Hepatic ischemia reperfusion injury (HIRI) is a pathophysiological and complex systemic process involving multiple tissues and organs. Gastrodin (GSTD), a natural compound from Gastrodia elata, displays a variety of interesting pharmacological activities. Heme oxygenase-1 (HO-1), a stress-responsive protein, has a cytoprotective defense response against oxidative and inflammatory injuries. The aim of this investigation was to elucidate whether GSTD plays a protective role against HIRI by regulating HO-1 expression. GSTD (100 mg/kg) or zinc protoporphyrin (15 mg/kg; an HO-1 inhibitor) was administered to HIRI C57 male mice. GSTD decreased glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels in HIRI mice. Inflammatory (TNF-α and IL-6) and oxidative-stress (malondialdehyde, MDA) markers of HIRI mice were decreased by GSTD. GSTD up-regulated HO-1 protein and mRNA expression in HIRI mice but decreased caspase-3 and -9 protein expression. GSTD lowered mRNA expression of apoptosis-related genes (caspase-3, -9, -12, and Bax) in the liver of HIRI mice but enhanced mRNA level of the anti-apoptotic Bcl-2 gene. Consistent with in vivo results, GSTD displayed a similar regulatory effect on the expression of mRNA (HO-1, caspase-3, -9, -12, Bax, and Bcl-2) and protein (HO-1, caspase-3 and -9) as well as inflammatory (TNF-α and IL-6) and on oxidative stress factors (superoxide dismutase and MDA) in BRL-3A cells transfected with small interfering HO-1 RNA in a hypoxia-reperfusion model. In conclusion, GSTD up-regulated HO-1 expression to play a protective role in HIRI by anti-apoptotic, anti-inflammatory, and antioxidant effects. GSTD is a promising natural compound that alleviated HIRI in liver surgery.

Abstract in English:

Although correlated to peak oxygen uptake (VO2), the six-minute walk test (6MWT) alone cannot provide precise physiological insight regarding the specific cause(s) of exercise limitations. We aimed to analyze whether 6MWT is able to properly detect differences in the cardiorespiratory responses between patients with stable coronary artery disease (SCAD) and those with ischemic cardiomyopathy (IC) and determine whether the degree of abnormality in ejection fraction is related with impaired submaximal exercise capacity. Twenty-two subjects with SCAD and 19 subjects with IC underwent a 6MWT while simultaneously using a mobile telemetric cardiopulmonary monitor to assess cardiorespiratory responses. VO2 response at exercise onset was used to obtain VO2 on-kinetics, and the slope of ventilation vs carbon dioxide output (VE/VCO2) was calculated. IC subjects exhibited significantly delayed VO2 on-kinetics compared with the SCAD group (P<0.01) and higher VE/VCO2 slope (IC=40.45 [95%CI: 39.76 to 41.1] vs SCAD=34.36 [95%CI: 34.03 to 34.69], P=0.001). The left ventricular ejection fraction (LVEF) was moderately correlated with VO2 on-kinetics in the SCAD group, but no relationship was found in the IC group. Pulmonary function was correlated with the VE/VCO2 slope only in the IC group. Subjects with IC presented slower VO2 on-kinetics during the 6MWT than those with SCAD. Once reduction in left ventricular function is achieved, LVEF had no association with exercise capacity. Pulmonary function could help identify IC patients at risk of ventilatory inefficiency and may add diagnostic power to the 6MWT.

Abstract in English:

Sleep duration is associated to various health impairments, while its comprehensive association with tinnitus is rarely investigated. The current study aimed to explore the relationship between sleep duration and tinnitus incidence, and to determine the optimal sleep duration relating to the lowest tinnitus risk. Data of participants from the National Health and Nutrition Examination Survey (NHANES) from 2005 to 2012 and 2015 to 2018 were retrieved. A total of 13,871 participants were eligible and included in the analysis. Generally, sleep duration was lower in participants with tinnitus compared to those without (7.15±1.76 vs 7.30±1.51 h, P<0.001). After adjustment by demographics, lifestyle, and chronic diseases, a U-shaped relationship between sleep duration and tinnitus incidence was observed, with the inflection point at 8.5 h. Interestingly, in participants with sleep duration <8.5 h, sleep duration exhibited an independent negative correlation with tinnitus risk [OR=0.88 (95%CI: 0.84-0.93), P<0.001], while in participants with sleep duration ≥8.5 h, sleep duration had an independent positive association with tinnitus risk [OR=1.16 (95%CI: 1.04-1.28), P=0.006]. In conclusion, a U-shaped relationship was found between sleep duration and tinnitus incidence, with a sleep duration of about 8.5 h being associated with the lowest tinnitus risk.

Abstract in English:

Neutrophil extracellular traps (NETs) are a novel regulatory mechanism of neutrophils, which can promote endothelial cell inflammation through direct or indirect pathways and play a crucial role in the occurrence and development of atherosclerosis (AS). This study aimed to explore the mechanism of NETs in AS progression using bioinformatics methods. We acquired datasets from Gene Expression Omnibus (GEO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) and used Weighted Gene Co-expression Network Analysis (WGCNA) to identify communal genes shared by NET-related genes. Gene Ontology (GO) and KEGG enrichment analyses were conducted. Machine learning algorithms were used to identify hub genes, then protein-protein interaction (PPI), CO-expression network construction, nomogram model building and validation, and immune infiltration analysis were performed. Data were verified by qPCR. Four datasets related to AS progression were included. Module genes shared 27 genes with NRGs. Pathways related to immune regulation, leukocyte migration, and others were identified. Machine learning revealed SLC25A4 and C5AR1 as hub genes. SLC25A4 and C5AR1 were confirmed to have predictive value for intraplaque hemorrhage (IPH), advanced AS plaques, ruptured plaques, and unstable plaques. These pathologic changes are closely related to AS progression and are the main contents of AS progression. Immune infiltration analysis revealed 4 immune cells associated with IPH, among them resting dendritic cells, which were closely related to SLC25A4. In qPCR validation, SLC25A4 and C5AR1 were shown to be consistent with the bioinformatic analysis results. These findings provided novel insights into the molecular characteristics of NRGs and potential therapies for AS progression.

Abstract in English:

1α,25-Dihydroxyvitamin D3 (VD3), the active form of vitamin D, plays a crucial role in wound healing. In this study, we aimed to investigate the effect of VD3 on the proliferation and differentiation of epidermal stem cells (EpSCs) and monitor its impact on re-epithelialization. We established a murine full-thickness skin defect model and applied four doses of VD3 (0, 5, 50, and 250 ng/mouse/day) to the wounds topically for three days. Immunostaining and flow cytometry confirmed the effect of VD3 on the proliferation and differentiation of EpSCs in wounds. This effect of VD3 (0, 1, 10, and 50 nM) on EpSCs and its possible mechanism were further confirmed in vitro by CCK8, westen blot, immunostaining, and flow cytometry. We found that on day five post-wounding, the means±SD length of the neo-epidermis was 195.88±11.57, 231.84±16.45, 385.80±17.50, and 268.00±8.22 μm in the control, 5, 50, and 250 ng groups, respectively, with a significant difference from the control (all P<0.05). Immunostaining and flow cytometry showed that VD3 improved the proliferation and differentiation of K15+ EpSC (vs control, all P<0.05), K14+ epidermal progenitor cells (vs control, all P<0.05), and K10+ epidermal terminal cells (vs control, all P<0.05) in vivo and in vitro. The PI3K signaling pathway appeared to underlie this response because significant inhibition of the response was found when inhibitors were used to inhibit PI3K. Our study demonstrated that VD3 is a potent promoter of cutaneous wound healing by stimulating EpSC proliferation and differentiation through PI3K activation.

Abstract in English:

During the COVID-19 pandemic in Shanghai, medical workers were more vulnerable to psychological problems. This two-phase cross-sectional survey was conducted by online questionnaires to investigate the symptoms of depression, anxiety, stress, post-traumatic stress disorder (PTSD), and fatigue in healthcare workers during the outbreak of COVID-19 and after the resumption of work and production in Shanghai. The questionnaire included the Depression Anxiety Stress Scale-21 (DASS-21), the Impact of Event Scale-Revised (IES-R), and the Fatigue Assessment Instrument (FAI). In Phase I (n=2192), the prevalence of depression, anxiety, stress, and PTSD symptoms among medical staff was 45.48, 41.93, 20.35, and 75.55%. In Phase II (n=1031), after work resumed in Shanghai, the prevalence was 19.79, 21.44, 28.23, and 12.22%, respectively. Fatigue had a mean score of 121.23±45.776 in Phase I and 144.73±44.141 in Phase II. Binary logistic regression identified risk factors associated with this psychological status: personal and familial chronic disease history; occupation, including doctor, nurse, or administrative staff; working in the fever clinic, infectious disease department, emergency or intensive care unit, hemodialysis room, or clinical laboratory; work experience of 3-6 years or 7-10 years; and involvement in nucleic acid sampling team. Medical staff self-reported comparatively high rates of depression, anxiety, stress, and, especially, PTSD symptoms during the COVID-19 pandemic in Shanghai. Our study indicated that after work resumption in Shanghai, it appeared that the overall mental health of medical staff improved somewhat. Nevertheless, the high level of fatigue exhibited still cannot be ignored.

Abstract in English:

Maximum oxygen uptake (V̇O2max) equations from developed countries are inaccurate for developing countries. Accordingly, we aimed to develop equations to predict treadmill V̇O2max over time based on variables other than exercise test in adults from the USA and Brazil undergoing cardiopulmonary exercise testing (CPET). We analyzed data from 2,170 adults who underwent two CPETs (1,307 men; 20-85 years) from the USA (n=1,880) and Brazil (n=290) with a second test after 2.0±1.7 years on average. We fit linear mixed-effects models to develop equations using 90% of the sample, randomly selected. In the remaining 10% of the cohort, we used the coefficient of variation, intraclass correlation coefficient, and the Bland and Altman plots to cross-validate the optimal equation. Our best linear mixed model equation was as follows: V̇O2max (mLO2·kg-1·min-1) = 62.01 - (0.23×Ageyears) - (0.001×Age×Age) - (0.65×Body mass indexkg/m 2) + (5.47×Sexfemales=0; males=1) + (2.78×CountryBrazil=0; USA=1) - (0.68×Arterial hypertensionno=0; yes=1) - (0.45×Hyperlipidemiano=0; yes=1) - (2.02×Smokingno=0; yes=1) - (4.36×Insufficiently activeno=0; yes=1) - (1.67×Beta-blockersno=0; yes=1); R2=0.566. Our main equation was reliable at baseline according to Bland and Altman plot results (mean difference, 0.01 mLO2·kg-1·min-1: 95%CI, -13.94 to 13.98; P=0.966) and over time (0.44 mLO2·kg-1·min-1: 95%CI, -13.5 to 12.4; P=0.439). Demographic and anthropometric attributes, cardiovascular risk, and beta-blockers are valuable for predicting V̇O2max at baseline and over time. The developed equations may apply to countries with socioeconomic and demographic characteristics such as Brazil and the USA.

Abstract in English:

As limb muscle function is age- and sex-related, both elbow and knee isokinetic muscle functions and their main predictors, such as physical activity level and cardiovascular risk factors, should be determined. We aimed to describe the percentiles of normality of the isokinetic muscle function of the knee and elbow joints. Secondarily, we developed equations to predict muscle function in apparently healthy adults aged 20-80 years, including cardiovascular risk factors. We conducted a cross-sectional study with 1,334 adults. We collected sociodemographic data, self-reported cardiovascular risk, anthropometry, body composition (bioelectrical impedance), moderate-to-vigorous physical activity (MVPA) (triaxial accelerometry), and isokinetic muscle function. Multiple regression analysis was used to develop equations to predict isokinetic muscle function. Percentiles of normality for muscle function were described by sex and age (20-39, 40-59, and >60 years). The models accounted for 49.6-70.9% of the total variability of muscle function, but MVPA and cardiovascular risk slightly influenced the coefficient of determination (additional ΔR2=0.003-0.006). Demographic and anthropometric variables were more relevant predictors of isokinetic muscle function (R2=0.50-0.70) than MVPA and cardiovascular risk. Even though they correlated with muscle function, cardiovascular risk and MVPA failed to explain the variability of muscle function largely determined by anthropometric and sociodemographic data. The percentile values and equations developed will help in interpreting the isokinetic muscle function and improve its clinical use.

Abstract in English:

Age-related macular degeneration (AMD), particularly the wet form characterized by choroidal neovascularization, is a leading cause of vision loss. Dysregulation of regulatory T cells (Tregs), key modulators of inflammatory responses, may contribute to wet AMD pathogenesis. This study explored the involvement of Tregs and the Rac1 signaling pathway in modulating Treg-derived cytokine expression and their role in choroidal neovascularization during wet AMD progression. Peripheral blood samples from healthy controls, dry AMD patients, and wet AMD patients were collected. An in vitro transmembrane co-culture system of Tregs and human choroidal endothelial cells (HCECs) was employed to investigate the impact of Tregs (with or without Rac1 silencing) on the angiogenic phenotype of HCECs. A mouse model of AMD was established to evaluate the effects of a Rac1 inhibitor and IL-10/TGF-β neutralization on Tregs and choroidal neovascularization. An increased Treg percentage in the CD4+ T lymphocyte population was found in the peripheral blood samples of wet AMD patients. Tregs from wet AMD patients showed an increased expression of Rac1 and an elevated production of IL-10 and TGF-β1. Rac1 silencing suppressed Treg stability and differentiation, and impaired the pro-angiogenic effect of Tregs on HCECs. In the animal model of AMD, the administration of a Rac1 inhibitor or neutralizing antibodies against IL-10/TGF-β1 reduced Treg abundance and attenuated choroidal neovascularization. Rac1 upregulation in Tregs promoted IL-10 and TGF-β1 production to mediate choroidal neovascularization in wet AMD. Targeting Rac1 and Treg-derived IL-10/TGF-β1 production in Tregs may serve as a strategy to ameliorate AMD progression.

Abstract in English:

Osteonecrosis of the femoral head (ONFH) is a debilitating condition characterized by the death of bone cells in the hip joint, resulting in profound disability. This condition has a significant global prevalence. Glucocorticoid (GC)-induced apoptosis of bone cells serves as a crucial cellular mechanism underlying ONFH. The protein kinase C zeta (PKCζ) and c-Jun N-terminal kinase (JNK)/c-Jun cascades have been implicated in the progression of ONFH, yet their interrelationship and contributions to disease development remain unclear. The objective of this study was to investigate the combined impact of PKCζ and JNK/c-Jun signaling on dexamethasone (Dex)-induced apoptosis in osteoblasts in vitro and in GC-induced ONFH rat models in vivo. In vitro experiments were conducted using hFOB1.19 osteoblastic cells to scrutinize the effects of Dex-induced apoptosis. The role of the PKCζ/JNK/c-Jun signaling pathway in this process was examined using naringenin-7-O-β-D-Glucuronide (N7G), a PKC inhibitor, and anisomycin, a JNK activator. The findings were further validated using a rat model of ONFH in vivo. Our results revealed that PKCζ activation augmented JNK/c-Jun signaling and facilitated Dex-induced osteoblast apoptosis. Inhibition of PKCζ with N7G mitigated these effects, while JNK activation with anisomycin intensified them. Similar regulatory effects on osteoblast apoptosis and ONFH progression were observed in the in vivo rat models. Glucocorticoids can induce osteoblast apoptosis and contribute to the development of ONFH by activating the PKCζ/JNK/c-Jun signaling pathway. This study provides compelling evidence supporting the potential therapeutic value of comprehending the pathogenesis of ONFH and developing targeted treatments for this debilitating condition.

Abstract in English:

Prenatal care is of fundamental importance and must be carried out by a multidisciplinary healthcare team, including dental care, as several changes and complications affecting the oral cavity may occur during pregnancy. This was a cross-sectional study that aimed to analyze the hematological profile of pregnant women with and without periodontal disease (PD). Data were obtained by consulting medical and dental records, which were stratified into two subgroups: pregnant women with PD (n=107) and pregnant women without PD (n=42). Study variables were related to PD, sociodemographic, clinical, and laboratory characteristics. Data were collected from the complete blood count and the following indices were calculated: neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), derived NLR (dNLR), systemic inflammation response index (SIRI), aggregate index of systemic inflammation (AISI), and systemic immune-inflammation index (SII). The mean age in both subgroups was 27 years. Pregnant women with less education had more PD. Mean corpuscular volume was significantly higher in pregnant women with PD, probably a reflection of folate deficiency. White blood cell and lymphocyte counts were significantly higher in pregnant women with periodontitis, possibly reflecting an inflammatory process caused by bacterial invasion of the periodontium with systemic repercussions. This study reinforces the need for a multidisciplinary team, including a dentist, in prenatal care, to lower the risk of complications for the mother and child.

Abstract in English:

Dysfunction of the intestinal epithelium barrier (DIEB) is frequent and can lead to serious complications in early childhood when diagnosis and clinical intervention are limited, especially in children with environmental enteric disease and malnutrition. The use of refined analytical techniques is increasingly necessary in this context. This study aimed to validate the high-performance liquid chromatography method coupled with tandem mass spectrometry (LC-MS/MS) to measure DIEB by lactulose:mannitol ratio detection (LM test) in samples of children with different social profiles from Fortaleza, Ceará. The first experimental set was conducted to validate the method through laboratory parameters, such as limit of detection (LD), limit of quantification (LQ), specificity/selectivity, linearity, accuracy, and precision. All validation parameters achieved detection and recovery standards within an acceptable coefficient of variation. Community samples (human development index (HDI) from 0.000 to ≤0.499) were obtained from children from the cohort study Malnutrition-Enteric Diseases, Fortaleza-CE (environmental enteric disease; EED group). The control group samples came from a school located in a region with a high HDI (>0.8). Mannitol excretion was lower in the EED group than in the control group (P<0.0001). On the other hand, LM was higher in this group compared to the control group (P<0.0001). For the first time, a robust analytical approach was used to detect biomarkers of environmental enteropathy (LM) in community samples, confirming with high-sensitivity the damage to the intestinal epithelial barrier function in populations living in low socio-economic conditions.

Abstract in English:

Coptisine (COP), a naturally occurring alkaloid, is recognized for its varied pharmacological impacts and its supportive function in intestinal well-being. However, the role of COP to protect the colonic epithelium in colitis has not been extensively investigated. The objective of this study was to assess the efficacy of COP in ameliorating colitis by investigating intestinal histopathology, mucosal barrier function, and transient receptor potential (TRP) signaling pathways in mice with colon disease compared to a control group, thereby elucidating the underlying mechanisms of its action. The results demonstrated a marked improvement in diarrhea and bleeding, an improvement in general behavioral competencies of the mice, and a decrease in disease activity index (DAI) scores. Histopathological analysis indicated a reduction in intestinal inflammation and an enhancement of intestinal mucosal barrier function. Our research identified that the protein expressions of the TRP family including transient receptor potential cation subfamily M member 8 (TRPM8), transient receptor potential vanilloid 1 (TRPV1), and transient receptor potential ankyrin 1 (TRPA1) were significantly upregulated with COP treatment. Compared with the model, COP markedly downregulated cytosolic phospholipase A2 (cPLA2) levels, while upregulating calcitonin gene-related peptide-1 (CGRP-1) protein expressions. Our study revealed that COP enhanced intestinal barrier function by modulating the cPLA2/TRPM8/CGRP-1 signaling pathway, thus shedding light on the mechanism by which COP mitigates inflammation in the intestinal mucosa. These findings provided new insights on COP as a therapeutic agent in ulcerative colitis (UC).

Abstract in English:

Rapid dissemination of Klebsiella pneumoniae carbapenemase (KPC) is a leading cause of treatment failure, significantly increasing morbidity and mortality rates among inpatients, particularly in the intensive care unit (ICU). This study aimed to detect the occurrence of carbapenemase- and carbapenem-resistant-encoding genes in K. pneumoniae isolates from COVID-19 positive and negative patients, and to assess their impact on patient outcomes. A prospective analysis was conducted at a tertiary care hospital in Saudi Arabia, collecting 97 carbapenem-resistant K. pneumoniae (CRKP) isolates from patients with COVID-19 during 2020-2021. Isolates were obtained from various clinical specimens. Antimicrobial susceptibility assays were performed using the Automated Vitek-2 system, and data were analyzed using IBM SPSS Statistics. The predominant carbapenemases identified were Oxacillinase-48 (OXA-48), followed by KPC and New Delhi metallo-β-lactamase (NDM), with Imipenemase (IMP) and Verona integron-encoded metallo-β-lactamase (VIM) being the least prevalent. COVID-19 did not significantly affect the distribution of these genes (P>0.05); however, COVID-19 status and age over 60 years significantly impacted the outcomes of CRKP patients. Other factors such as gender, total ICU or ward stay, and comorbidities did not significantly affect CRKP infection outcomes. The most common carbapenem-resistant genes identified were blaKPC, blaNDM, and blaOXA-48; however, they were not significantly associated with increased mortality.

Abstract in English:

Axons of dopaminergic neurons projecting from substantia nigra to striatum are severely affected in the early stage of Parkinson's disease (PD), with axonal degeneration preceding the loss of cell bodies. Our previous study indicated that the dysfunctional retrograde axonal transport could lead to the death of dopaminergic neurons resulting in PD (10.1111/j.1471-4159.2008.05526.x). However, dynein, as the main molecule involved in retrograde axonal transport, was not affected. This study aimed to verify the hypothesis that dynactin rather than dynein may be one of the key factors in the retrograde degeneration of dopaminergic neurons in the early stage of PD. Dynactin morpholino was used to inhibit the expression of dynactin in transgenic (Vmat2:GFP) zebrafish, resulting in a significant decrease of diencephalon dopamine neurons and synuclein aggregation in the basal plate region. In the dopaminergic SH-SY5Y cell line, dynactin-siRNA knockdown resulted in the expression of dynein shifting from dispersed distribution to concentration in synapses and cytoplasm near axons, and the fusion rate of dynein to dynactin was decreased, especially in axons, which blocked the retrograde axonal transport of α-synuclein and autophagy flow. Our results linked the knockdown of dynactin gene to the dysfunction of axonal microtubule transport system, suggesting that dynactin may be one of the key factors contributing to the retrograde degeneration of dopaminergic neurons in the early stage of PD.

Abstract in English:

Sprint interval training (SIT), which consists of vigorous-intensity exercise interspersed with periods of rest or low-intensity exercise, can improve human anaerobic performance. Probiotic strains, including yeasts (e.g. Saccharomyces boulardii; Sb), have beneficial effects on human health; however, evidence regarding the effects of probiotics on anaerobic performance is unavailable. The current study investigated whether Sb supplementation influences the SIT-induced changes to the following performance variables: peak (PPO) and mean (MPO) power output. Fifteen healthy individuals (twelve men and three women) were randomly divided into two groups: placebo (PLA; n=8) and Sb (n=7). The individuals performed six SIT sessions on a cycle ergometer (four to seven 30-s all-out sprints thrice weekly). During the training period, participants ingested a capsule containing PLA or at least 1×109 Sb cells daily for 14 days. Performance-related variables were compared between the first and last training sessions. Sb supplementation did not influence the changes in PPO and MPO across the two weeks of training (P>0.05); therefore, the data from both groups were analyzed collectively to assess performance changes induced by SIT. Training increased PPO, an index of anaerobic power, in the sixth session compared to the first session (by 8±11% in the first sprint; +1.0±1.2 W/kg; P=0.008) but did not change MPO. In conclusion, short-term SIT improved the participants' anaerobic performance (power), as evidenced by increased PPO. Sb supplementation did not affect the improved anaerobic power caused by SIT.

Abstract in English:

The COVID-19 pandemic has caused a global crisis, overwhelming hospitals and intensive care units (ICU) and leading to an increase in nosocomial infections due to prolonged hospitalization and other risk factors. The present study evaluated the prevalence of secondary fungal infections in critically ill patients with COVID-19. This is a retrospective, single-center study conducted in a hospital in northeastern Brazil, which evaluated 1,364 medical records of patients admitted to a COVID-19 ICU during 2020 and 2021. A total of 327 pathogenic yeasts were isolated from 132 (40.4%) respiratory, 70 (21.4%) blood, 124 (37.9%) urine, and one (0.3%) surgical wound samples. Fungal infections were diagnosed in the intermediate (5 to 12 days) or late (≥12 days) stage of hospitalization. The most frequent yeast isolated from critically ill COVID-19 patients was Candida albicans [126 (67.7%) and 60 (42.6%)], followed by Candida tropicalis [25 (13.4%) and 39 (27.7%)]. Candida parapsilosis isolates increased 5.7-fold in 2021 [40 (28.4%)] compared to 2020 [7 (3.8%)]. The least frequently isolated in 2020 and 2021 were Nakaseomyces glabratus [4 (2.2%) and 1 (0.7%)], and Pichia kudriavzevii, which was isolated only in 2021 (1 (0.7%)). During the study period, a decrease in susceptibility to antifungals was observed: susceptibility to voriconazole reduced from 100 to 77.2%, to flucytosine from 99.4 to 78.8%, and to micafungin from 99.4 to 83.6%. The changes in the frequency of species causing secondary infections in critically ill COVID-19 patients and susceptibility to the antifungals indicate the need for early and adequate diagnosis to minimize negative outcomes.

Abstract in English:

Cellular senescence is an important cause of age-related degenerative diseases, including osteoarthritis (OA). Chondrocyte senescence is crucial in OA onset and progression. As a non-invasive, safe, and widely used physical rehabilitation factor, the effect and mechanism of low intensity pulsed ultrasound (LIPUS) on chondrocyte senescence remain unclear. This study evaluated the inhibitory effect of LIPUS on OA chondrocyte senescence in vitro and in vivo. The effect of LIPUS on chondrocyte senescence was examined by RT-qPCR, enzyme-linked immunosorbent assay (ELISA), and western blotting. Changes in levels of reactive oxygen species (ROS) and γ-h2ax foci in senescent chondrocytes were detected using fluorescent staining. Chondrocyte senescence was evaluated by senescence-associated β-galactosidase (SA-β-gal) staining. The PI3K inhibitor LY294002 and the PI3K agonist 740Y-P were used to investigate whether PI3K/AKT/mTOR signalling was involved in the effect of LIPUS in senescent chondrocytes. Chondrocyte senescence and cartilage degeneration were analyzed in a destabilization of the medial meniscal (DMM) mouse model by immunohistochemistry, hematoxylin and eosin staining, and safranin-O/fast green staining. LIPUS inhibited the expression of the senescence-associated secretory phenotype (SASP) factors CCL4 and CCL2 and the senescence phenotype in doxorubicin-treated chondrocytes by inhibiting the PI3K/AKT/mTOR pathway. LIPUS alleviated chondrocyte senescence and attenuated OA progression in the DMM mice. These results demonstrated a novel role for LIPUS in inhibiting chondrocyte senescence and the SASP by modulating PI3K/AKT/mTOR signalling. Our findings expanded the clinical application of LIPUS and provide a new, non-invasive, and safe treatment approach to prevent and treat age-related degenerative joint disorders.