Human tolerogenic dendritic cells generated with protein kinase C inhibitor are optimal for functional regulatory T cell induction - A comparative study.

Tolerogenic dendritic cells (tDCs) are a promising therapeutic tool for specific induction of immunological tolerance. Human tDCs can be generated ex vivo using various compounds. However, the compound(s) most suitable for clinical application remain undefined. We compared the tolerogenic properties of tDCs treated with protein kinase C inhibitor (PKCI), dexamethasone, vitamin D3 (Vit D3), rapamycin (Rapa), interleukin (IL)-10, transforming growth factor (TGF)-ß, and a combination of peroxisome proliferator-activated receptor γ agonist and retinoic acid. All tDCs had a semi-mature DC phenotype. PKCI-, TGF-ß-, and Rapa-tDCs showed CCR7 expression and migration to CCL19, but other tDCs showed little or none. PKCI- and IL-10-tDCs induced functional regulatory T cells more strongly than other tDCs. The tolerogenic properties of all tDCs were stable against proinflammatory stimuli. Furthermore, PKCI-tDCs were generated from patients with rheumatoid arthritis and primary Sjögren's syndrome. Therefore, PKCI-tDCs showed the characteristics best suited for tolerance-inducing therapy.

Association between angiotensin-converting enzyme (ACE) gene I/D polymorphism with the risk of knee OA: A systematic review, meta-analysis, and meta-regression.

Background: Previous studies have linked genetics to knee osteoarthritis. Angiotensin-converting enzyme (ACE) gene I/D polymorphism may cause OA. However, evidence remains inconsistent. This study examines knee OA risk and ACE gene I/D polymorphism. Methods: We explored Europe PMC, Medline, Scopus, and Cochrane Library using keywords. Three assessment bias factors were assessed using the Newcastle-Ottawa Scale (NOS). Criteria for inclusion: (1) Split the study population into knee OA patients and healthy controls; (2) Analysed the ACE gene I/D polymorphism; (3) Case-control or cross-sectional surveys. Studies with non-knee OA, incomplete data, and no full-text were excluded. The odds ratio (OR) and 95% confidence intervals (95% CI) were calculated using random-effect models. Results: A total of 6 case-control studies consist of 1,226 patients with knee OA and 1,145 healthy subjects as controls were included. Our pooled analysis revealed that a significant association between ACE gene I/D polymorphism and risk of knee OA was only seen in the dominant (DD + ID vs. II) [OR 1.69 (95% CI 1.14 - 2.50), p = 0.009, I2 = 72%], and ID vs. II [OR 1.37 (95% CI 1.01- 1.86), p = 0.04, I2 = 43%] genotype models. Other genotype models, including recessive (DD vs. ID + II), alleles (D vs. I), DD vs. ID, and DD vs. II models did not show a significant association with knee OA risk. Further regression analysis revealed that ethnicity and sex may influence those relationships in several genotype models. Conclusions: Dominant and ID vs. II ACE gene I/D polymorphism models increased knee OA risk significantly. More research with larger samples and different ethnic groups is needed to confirm our findings. After ethnicity subgroup analysis, some genetic models in our study showed significant heterogeneities, and most studies are from Asian countries with Asian populations, with little evidence on Arabs.

Glycemic control as the main determinant factor of serum VEGF levels in type 2 diabetes mellitus patients.

Introduction. Diabetes mellitus (DM) is a main endocrine disorder that may cause vascular complications as the disease progresses. Vascular endothelial growth factor (VEGF) has been linked to the development of micro and macrovascular diabetic complications. This study aimed to assess several factors including blood pressure, body mass index, lipid profile, kidney function, and glycemic control that may provide the rise of serum VEGF levels in type 2 DM subjects. Methods. This cross-sectional study was carried out among 65 type 2 DM subjects. Systole, diastole, mean arterial pressure (MAP), and body mass index (BMI) were measured. The levels of serum VEGF were measured by Enzyme-linked immunosorbent assay (ELISA), Hemoglobin A1c (HbA1c) levels were measured by latex agglutination inhibition test, while serum glucose, lipid profiles, urea, and creatinine levels were tested by enzymatic photometric method. Results. The levels of serum VEGF had a significant correlation with BMI (p = 0.001, r = 0.397), fasting plasma glucose (FPG) (p = 0.001, r = 0.418), HbA1c (p < 0.001, r = 0.600), systole (p = 0.001), r = 0.397), diastole (p = 0.021, r = 0.286), and MAP (p = 0.001, r = 0.001). Further multivariate linear regression analysis revealed that HbA1c logarithm (log) was the determinant factor of VEGF levels (p < 0.001, ß = 0.631, Adjusted R2 = 38.9%) Conclusion. HbA1c is the main determinant factor of serum VEGF levels among type 2 DM patients.

A Case Series: Effect of Comorbidities on the Outcomes of Prolotherapy Injection for Frozen Shoulder Patients.

Frozen shoulder (FS) is a disease caused by an inflammatory condition that causes severe pain and decreased range of motion by loss of glenohumeral mobility. Frozen Shoulder restricts daily life's functional aspect, increasing morbidity. Hypertension and diabetes mellitus are risk factors that make an FS poor prognosis during treatment because of the diabetes glycation process and hypertension-enhanced vascularization. Prolotherapy injects an irritant solution into the tendon, joints, ligaments, and joint spaces to release growth factors and collagen deposition, reducing pain, restoring joint stability, and increasing the quality of life. We report 3 cases of patients with confirmed FS. Patient A with no comorbidity, patient B with diabetes mellitus, and patient C with hypertension, with all patient's chief complaints of shoulder pain and limited ROM, and symptoms affected the general quality of daily life. This patient was provided injection with Prolotherapy treatment combined with physical therapy intervention. Patient A had significantly improved ROM to maximum after 6 weeks with relieved pain and improved shoulder function. Patients B and C showed increased ROM, still tiny, decreased pain, and improved shoulder function. In conclusion, prolotherapy demonstrated a beneficial effect in a patient with FS with comorbidities, although not to the maximum extent in patients without comorbidity.

Dynamic interaction of obesity, age, MCP-1 Level, and ACE-1 gene with the severity of knee osteoarthritis: a cross-sectional study.

The risk factors most strongly associated with knee osteoarthritis (OA) are old age and obesity. However, few studies have evaluated the interaction between aging and obesity in conjunction with inflammatory markers and knee OA severity as part of a complete assessment of knee OA management. Therefore, this study aims to evaluate the interaction between obesity, age, inflammation [including the I/D polymorphism of angiotensin converting enzyme-1 (ACE-1)], and the severity of knee OA. Methods: A total of 80 knee OA patients were included in this cross-sectional study. The severity of knee OA was determined based on the Kellgren-Lawrence system. All patients underwent physical and radiological examination; monocyte chemoattractant protein 1 (MCP-1) markers were measured. The parameters of the ACE-1 gene were examined with sequencing DNA. Results: There was a significant relationship between age and severity of knee OA (P=0.007), with subjects aged greater than or equal to 65 having a 3.56-fold higher risk of developing moderate to severe OA than subjects aged less than 65. There was a significant difference between body weight and knee OA severity (P=0.026), in which subjects weighing greater than or equal to 60 kg had 3.14 times the risk of experiencing severe knee OA. Multivariate regression analysis indicated that age was the strongest independent variable for knee OA severity compared with body weight. MCP-1 levels were significantly higher in mild knee OA than in moderate to severe knee OA. The DD genotype of the ACE-1 gene increases the risk of severe knee OA by four times in subjects aged greater than or equal to 65 compared to subjects aged less than 65. However, the DD genotype of the ACE-1 gene does not increase the risk of severe knee OA in subjects weighing greater than or equal to 60 kg. Conclusion: While obesity and age were found to be associated with the severity of knee OA, age emerged as the independent risk factor for knee OA severity. Furthermore, MCP-1 levels were significantly higher in cases of mild knee OA compared to severe knee OA. It was observed that the DD genotype of the ACE-1 gene increases the risk of severe knee OA in individuals aged 65 years or older.

The level of ratio between matrix metalloproteinase-1 (MMP-1) and tissue inhibitor matrix metalloproteinase-1 (TIMP-1) after prolotherapy intervention and the functional outcome in patient with frozen shoulder: A randomized controlled trial.

OBJECTIVE: To determine the effect of prolotherapy on functional outcome changes, along with ratio of matrix metalloproteinase-1 (MMP-1)/tissue inhibitor matrix metalloproteinase-1 (TIMP-1) as an indicator of tissue repair in the glenohumeral joint in frozen shoulder patients. DESIGN: Single-blinded randomized controlled trial. SUBJECTS/PATIENTS: Participants with frozen shoulder. METHODS: The prolotherapy group is the study group, and the normal saline (NS) group is the control group. Each group was given injections at weeks 0, 2, 4, and 6. Level of biomarker levels was measured at week 6 and week 12 after there. Functional outcomes were measured at weeks 0, 6, and 12. RESULTS: A significant difference in week 6 and week 12 was demonstrated in the ratio of MMP-1/TIMP-1 level between the prolotherapy group and the normal saline group (P value = .002). Both groups performed well regarding the Numerical Rating Scale score and functional outcome. Compared to the normal saline group, prolotherapy changed the mean range of motion in flexion and internal rotation. CONCLUSION: Prolotherapy is considered to play a role in repairing cartilage based on biomarker assessment, particularly the ratio of MMP-1/TIMP-1-prolotherapy effectiveness in improving functional outcome and Numerical Rating Scale score.

Effectiveness of Prolotherapy Combined with Physical Therapy Versus Physical Therapy Only for Frozen Shoulder: A Case Report.

BACKGROUND Frozen shoulder (FS) is a common conditions that causes significant morbidity. It is characterized by restriction of both active and passive shoulder motion (ROM) of the glenohumeral joint. The etiology, pathology, and most efficacious treatments are unclear. The purpose of FS treatment is complete elimination of pain and recovery of shoulder joint function. Prolotherapy injects certain compounds into articular spaces, ligaments, and/or tendons to relieve pain and disability around joint spaces and to stimulate a proliferation cascade to enhance tissue repair and strength. This case report aims to describe functional outcome changes in 2 patients with FS, comparing prolotherapy combined with physical therapy vs physical therapy only. CASE REPORT We report the cases of 2 patients with confirmed FS. Patient A was 66-year-old man with chief concern of right shoulder pain and limited ROM in the past 3 months, which disrupted daily life, with a visual analog scale (VAS) of 6 out of 10. Patient B was 65-year-old man with chief concern of right shoulder pain and limited ROM in the past 2 months. The symptoms affected his general quality of life, with a VAS of 5 out of 10. Patient A underwent prolotherapy combined with physical therapy and had significantly improved ROM after 2 weeks, with relieved pain and improved shoulder function. Patient B underwent physical therapy only and showed similar ROM and no significant pain improvement. CONCLUSIONS Initial treatment with prolotherapy combined with physical therapy for patients with frozen shoulder achieved fast improvement of active and passive ROM, significantly decreased pain, and improved quality of life compared to physical therapy intervention only.

Changes in levels of cartilage oligomeric proteinase and urinary C-terminal telopeptide of type II collagen in subjects with knee osteoarthritis after dextrose prolotherapy: A randomized controlled trial.

OBJECTIVE: To assess the effects of dextrose prolotherapy in patients with knee osteoarthritis on the levels of serum cartilage oligomeric proteinase and urinary C-terminal telopeptide of type II collagen, and on the Western Ontario McMaster Universities Index and numerical rating scale score for pain. METHODS: A randomized controlled trial, in which participants were randomly allocated into 2 groups, receiving injections of either hyaluronic acid or dextrose prolotherapy. The hyaluronic acid group received 5 injections, 1 each on weeks 1, 2, 3, 4 and 5, and the dextrose prolotherapy group received 3 injections, 1 each on weeks 1, 5 and 9. Serum cartilage oligomeric proteinase, urinary C-terminal telopeptide of type II collagen, Western Ontario McMaster Universities Index score, and numerical rating scale score for pain were measured at baseline and 3 weeks after the last injection. Comparative analysis was conducted using Wilcoxon test within groups and analysis of covariance (ANCOVA) test between groups. RESULTS: A total of 47 participants (21 allocated to hyaluronic acid, 26 allocated to dextrose prolotherapy) completed the protocol. Both interventions resulted in significant improvements in numerical rating scale scores for pain, total Western Ontario McMaster Universities Index scores, and its subscales score. However, the dextrose prolotherapy outperformed hyaluronic acid in numerical rating scale score for pain and level of urinary C-terminal telopeptide of type II collagen, with score changes differences of 0.93 (p = 0.042) and 0.34 (p = 0.048), respectively. No significant changes in level of serum cartilage oligomeric proteinase were found in either group. CONCLUSION: Dextrose prolotherapy is an alternative injection therapy for knee osteoarthritis, which was found to be associated with a significant reduction in urinary C-terminal telopeptide of type II collagen compared with hyaluronic acid injection. Neither injection method resulted in reduced serum cartilage oligomeric proteinase.

Insulin resistance and testosterone level in Indonesian young adult males.

INTRODUCTION: Central obesity is characterized by the accumulation of abdominal fat which may lead to several diseases including insulin resistance. The prevalence of central obesity is higher in male and the incidence in young adult males is increased. Central obesity is also related to low testosterone levels. The research aimed to assess the relationship between the testosterone levels and insulin resistance of young adult males with central obesity. METHODS: This was a cross-sectional study, the subjects were young adult males of 18 to 25 years old. The central obesity consisted of 50 samples and non-central obesity comprised 70 samples. The examination of testosterone and insulin was performed by the ECLIA method, glucose used the enzymatic method, the insulin resistance was calculated by using the HOMA-IR index. RESULTS: The mean of the testosterone level in central obesity was lower than non-central obesity (5.24 + 1.17 vs 7.18 + 1.54 ng/mL, p < 0.001). HOMA-IR index in central obesity was higher than non-central obesity (4.29 + 2.23 vs 2.46 + 1.72, p < 0.001). Testosterone levels had negative correlation with HOMA-IR (r = -0.470, p < 0.001). There was significant difference in HOMA-IR among the quartiles of testosterone levels. CONCLUSION: There is negative correlation between testosterone level with HOMA-IR, the lower the testosterone level the higher the insulin resistance in young adult males.

Anthropometric features in predicting insulin resistance among non-menopausal Indonesian adult females.

INTRODUCTION: The prevalence of obesity is increasing worldwide in high, low, and middle-income countries such as Indonesia. Obesity rate is higher in females in Indonesia. Obesity has important contribution in the occurrence of insulin resistance (IR) and type 2 diabetes mellitus. Several anthropometric measurements such as waist circumference (WC), body mass index (BMI), body mass (BM), total body fat percentage (Fat%) and visceral fat (VF) are related to IR. This study aimed to investigate which of those measurements could be used as a better predictor of IR in non-menopausal Indonesian adult females. METHODS: Total of 80 non-menopausal Indonesian adult females ranging from 21 to 40 years were recruited in this study. Insulin resistance was measured by using Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) equation. Subjects with HOMA-IR index >75th percentile with cut-off 2.74 were defined as IR. Waist circumference, BMI and BM were measured, while TF and VF were measured by bioelectrical impedance analysis (BIA). RESULTS: HOMA-IR had significant correlation with WC (r = 0.563, p < 0.001), BMI (r = 0.537, p < 0.001), BM (r = 0.515, p < 0.001), VF (r = 0.515, p < 0.001), Fat% (r = 0.490, p < 0.001). The area under curve of VF (0.809), BMI (0.807), WC (0.805), and BM (0.799) are slightly larger than and Fat% (0.766). CONCLUSION: Insulin resistance had strong correlation with all anthropometric measurements, but the correlation was less significant with Fat%.

Relationship between insulin resistance, metabolic syndrome components and serum uric acid.

BACKGROUND: Metabolic syndrome is cluster of abnormality related with increasing cardiovascular events. Hyperuricemia is level of uric acid more than 7 mg/dL for men. Some research have reported relation between metabolic syndrome mediated by insulin resistance with increasing of serum uric acid level. OBJECTIVE: Assess relationship between insulin resistance and metabolic syndrome components with the level of serum uric acid. METHOD: Observational study with cross sectional approach conducted on 102 outpatient subjects at Dr. RSUP Wahidin Sudirohusodo (RSWS) hospital and Hasanuddin University Hospital in the period of July-September 2018. RESULTS: Subjects with IR were found to be significantly higher for having MetS (88.23% vs. 11.77% p = 0,000). In subjects with IR, the average serum uric acid level was higher compared to non-IR subjects, but this difference was not significant (6.63 vs 6.42 mg/dL; P = 0.325). In subjects with MetS, the average serum uric acid level was higher compared to subjects with non-MetS but this difference was not significant (6.62 vs. 6.28 mg/dL; P = 0.556). No significant relationship was found between IR and MetS with serum uric acid level. CONCLUSION: Insulin resistance is related to the incidence of MetS and in both of these circumstances an independent tendency is found to increase uric acid levels. The role of insulin resistance in the relationship between metabolic syndrome and uric acid levels was not proven in this study.

Novel Autoantigens Associated with Lupus Nephritis.

Systemic lupus erythematosus (SLE) is characterized by production of a variety of autoantibodies. Although anti-double-stranded DNA (anti-dsDNA) antibodies contribute to the pathogenesis of lupus nephritis (LN), they are not sufficient for diagnosis and evaluation of disease activity. To obtain other autoantibodies associated with LN, we screened autoantigens reacting with the sera of LN patients by using an N-terminal biotinylated protein library created from a wheat cell-free protein production system. We screened 17 proteins that showed higher positive signals in the active phase than in the inactive phase of SLE, and higher positive signals in the serum of SLE patient with nephritis than in that of patient without nephritis. Of these, two LN-associated autoantigens, ribosomal RNA-processing protein 8 (RRP8) and spermatid nuclear transition protein 1 (TNP1) were identified by immunoprecipitation and immunofluorescence of renal tissues. Circulating anti-RRP8 and anti-TNP1 autoantibodies were recognized and deposited as an immune complex (IC) in glomeruli. IC was deposited preferentially in glomeruli rather than in other organs in C57BL/6 mice injected with RRP8 or TNP1. ELISA analysis of sera from patients with various rheumatic diseases demonstrated reactivity for RRP8 and TNP1 in 20% and 14.7% of SLE patients, respectively, whereas there was little or no reactivity in patients with other rheumatic diseases. Among SLE patients, 63.6% and 45.5% of those with LN were positive for anti-RRP8 and anti-TNP1 antibodies, compared with 12.5% and 9.4% of SLE patients without nephritis, respectively. Both proteins are cationic, and their respective antibodies did not cross-react with dsDNA. These proteins released from apoptotic cells form ICs with each autoantibody, and their ICs may become trapped at anionic sites in the glomerular basement membrane, leading to deposition in glomeruli. These autoantibodies may be useful for prediction of LN in subsets of SLE patients who are negative for anti-dsDNA antibodies.