Circadian time series proteomics reveals daily dynamics in cartilage physiology.

OBJECTIVE: Cartilage in joints such as the hip and knee experiences repeated phases of heavy loading and low load recovery during the 24-h day/night cycle. Our previous work has shown 24 h rhythmic changes in gene expression at transcript level between night and day in wild type mouse cartilage which is lost in a circadian clock knock-out mouse model. However, it remains unknown to what extent circadian rhythms also regulate protein level gene expression in this matrix rich tissue. METHODS: We investigated daily changes of protein abundance in mouse femoral head articular cartilage by performing a 48-h time-series LC-MS/MS analysis. RESULTS: Out of the 1,177 proteins we identified across all time points, 145 proteins showed rhythmic changes in their abundance within the femoral head cartilage. Among these were molecules that have been implicated in key cartilage functions, including CTGF, MATN1, PAI-1 and PLOD1 & 2. Pathway analysis revealed that protein synthesis, cytoskeleton and glucose metabolism exhibited time-of-day dependent functions. Analysis of published cartilage proteomics datasets revealed that a significant portion of rhythmic proteins were dysregulated in osteoarthritis and/or ageing. CONCLUSIONS: Our circadian proteomics study reveals that articular cartilage is a much more dynamic tissue than previously thought, with chondrocytes driving circadian rhythms not only in gene transcription but also in protein abundance. Our results clearly call for the consideration of circadian timing mechanisms not only in cartilage biology, but also in the pathogenesis, treatment strategies and biomarker detection in osteoarthritis.

Thiamine-Responsive Megaloblastic Anemia-Related Diabetes: Long-Term Clinical Outcomes in 23 Pediatric Patients From the DPV and SWEET Registries.

OBJECTIVES: To describe clinical presentation and long-term outcomes in a large cohort of children diagnosed with thiamine-responsive megaloblastic anemia (TRMA)-related diabetes. METHODS: Data from the Diabetes Patienten Verlaufsdokumentation (DPV) and Better control in Pediatric and Adolescent diabeteS: Working to crEate CEnTers of Reference (SWEET) registries were used to identify cases. Complementary information was collected through a chart review of each case. Descriptive analyses with medians and interquartile ranges and numbers (proportions) were tabulated. RESULTS: We identified 23 cases (52% male) in the 2 registries. Eighteen (78%) had genetic confirmation of TRMA. Median age at diabetes onset was 1.4 (quartiles 0.8 to 3.6) years and median age at initiation of thiamine treatment was 5.9 (2.4 to 12.4) years. At their most recent visit, patients' median age was 14.3 (8.1 to 17.5) years, glycated hemoglobin level was 6.9% (6.1% to 7.9%), insulin dose was 0.9 (0.4 to 1.2) units/kg per day and thiamine dose was 200 (100 to 300) mg/day. Three patients were not treated with insulin or antidiabetic drugs. There was no difference in diabetes outcomes in patients with initiation of thiamine ≤1 year after diabetes onset compared to patients with initiation of thiamine >1 year after diabetes onset. CONCLUSIONS: This is the longest case series of pediatric TRMA-related diabetes reported to date. Diabetes onset often occurs several years before initiation of thiamine supplementation. Early initiation of thiamine (within 1 year of diabetes onset) was not linked to improved diabetes outcome. However, the role of thiamine in pancreatic function needs further assessment. Patients with TRMA-related diabetes maintained good glycemic control even after 9 years (median) of follow up.

Bidirectional screening of tuberculosis patients for diabetes mellitus and diabetes patients for tuberculosis.

To assess the feasibility and results of screening diabetes mellitus (DM) patients for tuberculosis (TB) and TB patients for DM within the routine health care setting. Prospective observational study carried out within the Diabetes Centre and Pulmonary Medicine Department from February 2012 to September 2012. The screening for active TB in DM and DM in TB patients is followed as per the guidelines of the Revised National Tuberculosis Control Programme and national programmes in India. Total of 307 patients diagnosed with TB during the study period. Among the TB patients 9.77% were smokers, 19.54% were known cases diabetes, and 15.96% were newly diagnosed cases of diabetes. Total of 4,118 diabetes patients were screened for TB in which 111 patients found to have TB. The strengths of this study are that we implemented screening within the routine health system. It is feasible to screen DM patients for TB resulting in high rates of TB detection.

Bidirectional Screening of Tuberculosis Patients for Diabetes Mellitus and Diabetes Patients for Tuberculosis

To assess the feasibility and results of screening diabetes mellitus (DM) patients for tuberculosis (TB) and TB patients for DM within the routine health care setting. Prospective observational study carried out within the Diabetes Centre and Pulmonary Medicine Department from February 2012 to September 2012. The screening for active TB in DM and DM in TB patients is followed as per the guidelines of the Revised National Tuberculosis Control Programme and national programmes in India. Total of 307 patients diagnosed with TB during the study period. Among the TB patients 9.77% were smokers, 19.54% were known cases diabetes, and 15.96% were newly diagnosed cases of diabetes. Total of 4,118 diabetes patients were screened for TB in which 111 patients found to have TB. The strengths of this study are that we implemented screening within the routine health system. It is feasible to screen DM patients for TB resulting in high rates of TB detection.

Type 1 diabetes mellitus with ketoacidosis in infancy.

Type 1 diabetes mellitus is considered a common form of diabetes mellitus in young people. Type 1 diabetes in infants is rare. However, the condition is rare in infants. Type 1 diabetes has not been reported in the literature in 45 days old child of an Indian population. Type 1 diabetes typically begins between the ages of 7 and 13 years, but 1-3% of patients are under 1 year of age. This communication describes a case of type 1 diabetes in a 45 days old male child which presented as diabetic ketoacidosis. It was effectively managed with continuous intravenous regular insulin infusion. The present report is made because of the rarity of the condition in the early age group of Indian children.

Familial early onset of type-2 diabetes mellitus and its complications.

BACKGROUND: Globally, the prevalence of chronic, non-communicable diseases is increasing at an alarming rate. Furthermore, approximately 197 million people worldwide have impaired glucose tolerance. Consequently, diabetes is rapidly emerging as a global health problem that threatens to assume a pandemic level by 2030. In Indian population, genetic predisposition to trigger diabetes at an early age as compared to western counterpart has been focused very much. AIM: To gain further insight into the positive correlation between the diabetes and family history was the objective of this study. MATERIALS AND METHODS: Patients attending the Diabetes Centre, K.L.E.S Dr. Prabhakar. Kore Hospital and Medical Research Centre; J. N. Medical College; KLE University Belgaum, Karnataka- India, were recruited, diagnosed and analyzed as per WHO criteria. RESULTS: The prevalence of diabetes was higher among patients with diabetic mother (25.6%) compared to patients with diabetic father (21.2%) and there was early onset of type -2 diabetes among patients having both parents with diabetic when compared to other patients. CONCLUSION: Based on the present observation, it would be appropriate to emphasize again that a strong family history for diabetes, would signal at an early age, the onset of diabetes perhaps with its complications.

Erythrocyte glutathione peroxidase, glutathione reductase activities and blood glutathione content in leprosy.

OBJECTIVES: Leprosy is a chronic granulomatous infection caused by Mycobacterium leprae involving cutaneous tissue and peripheral nerves producing skin lesions, nerve degeneration, anaesthesia and deformities. In leprosy, the activated phagocytes produce reactive oxygen species (ROS) as a part of their microbicidal function. Such ROS are capable of damaging the host tissue by lipid peroxidation. Increased lipid peroxidation has been reported in leprosy. The glutathione antioxidant system with glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione (GSH) as components protect the cells from ROS toxicity and lipid peroxidation. The objective of the present study was to assess blood glutathione content and erythrocyte antioxidant enzyme activities of glutathione peroxidase and glutathione reductase in leprosy. DESIGN: The parameters were studied in 100 leprosy patients and 50 normal healthy controls. The data was analysed by grouping the patients into Ridley-Jopling (RJ) types [tuberculoid leprosy (TT), borderline tuberculoid leprosy (BT), borderline leprosy (BB), borderline lepromatous leprosy (BL), lepromatous leprosy (LL)] and into different levels of Bacteriological Index (BI) [bacteriologically negative (BI=0), BI=0.1-1, BI=1.1-2, BI=2.1-3, BI=3.1-4, BI=4.1-6]. METHODS: Venous blood sample was used for the study. The GSH level was estimated in the blood by DTNB [5,5'-dithiobis(2-nitrobenzoic acid)] reduction method. The enzyme activities were measured in the red blood cell haemolysate by kinetic methods using NADPH. RESULTS: GSH, GSH-Px and GR were significantly low in leprosy (total patients) as compared to the control group (p<0.001). A progressive decrease in GSH level and enzyme activities was noted along the leprosy spectrum from TT to LL. A significant decline of GSH in BB (p<0.05), BL (p<0.005) and LL (p<0.001); and of GSH-Px and GR in BT (p<0.05, p<0.02), BB (p<0.02), BL (p<0.005) and LL (p<0.001) was noted as compared to controls. A significant lowering of GSH-Px in LL (p<0.005); the GR in BB (p<0.02), BL (p<0.05) and LL (p<0.05); and the GSH in BL (p<0.01) and LL (p<0.001) was noted in comparison to the TT group. The GSH and GSH-Px were significantly low in LL (p<0.05) as compared to BT. A progressive decreasing trend in GSH level and enzyme activities was also noticed along the leprosy groups with advancing level of BI. The GSH, GSH-Px and GR were significantly low in BI levels 1.1-2 (p<0.005, p<0.05, p<0.02), 2.1-3 (p<0.005, p<0.001, p<0.005), 3.1-4 (p<0.005) and 4.1-6 (p<0.01, p<0.005, p<0.05) as compared to controls. A significant decline in GSH was noted in BI levels 1.1-2 (p<0.005), 2.1-3 (p<0.005), 3.1-4 (p<0.005) and 4.1-6 (p<0.01) as compared to the bacteriologically negative group. The GSH-Px (p<0.05) and GR (p<0.05) were significantly low in BI levels 2.1-3, 3.1-4 and 4.1-6 as compared to the bacteriologically negative group. CONCLUSION: The findings suggest oxidative stress associated with diminished antioxidant defence potential in leprosy. The study identifies association of diminished antioxidant potential with bacterial load and type of leprosy.

Erythrocyte superoxide dismutase, catalase activities and hydrogen peroxide induced lipid peroxidation in leprosy.

OBJECTIVES: To assess erythrocyte superoxide dismutase (SOD) and catalase (CAT) activities and hydrogen peroxide induced lipid peroxidation in leprosy. DESIGN: One hundred leprosy patients and 50 normal healthy controls were studied for the parameters. The data was analysed by grouping the patients into Ridley-Jopling (RJ) types [Tuberculoid leprosy (TT, n = 22), Borderline tuberculoid leprosy (BT, n = 28), Borderline leprosy (BB, n = 13), Borderline lepromatous leprosy (BL, n = 16) and Lepromatous leprosy (LL, n = 21)] and into different levels of Bacteriological Index (BI) [bacteriologically negative (n = 32), BI = 0.1-1 (n = 22), BI = 1.1-2 (n = 16), BI = 2.1-3 (n = 14), BI = 3.1-4 (n = 10) and BI = 4.1-6 (n = 06)]. RESULTS: The induced peroxidation was significantly high and the enzyme activities were significantly low in leprosy (total patients) as compared to controls. A progressive increase in peroxidation was detected along the leprosy spectrum from TT to LL and the increase was significant in BB, BL and LL groups as compared to controls. Induced peroxidation in LL group as compared to TT, BT and BB and in the BL group as compared to TT and BT were significantly different. A concomitant progressive decline in enzyme activity was detected along the leprosy spectrum from TT to LL. The SOD activity in BB, BL and LL and the CAT activity in BL and LL were significantly low as compared to controls. SOD activity in BB, BL and LL groups as compared to TT and in the LL group as compared to BT were significantly different. A progressive trend of increasing peroxidation and decreasing SOD and CAT activity were also detected along the leprosy groups with advancing level of BI. Induced peroxidation and SOD activity were significantly different in bacteriologically positive groups as compared to controls and in the BI levels 1.1-2, 2.1-3, 3.1-4 and 4.1-6 as compared to bacteriologically negative group. The peroxidation was significantly different in BI levels 2.1-3, 3.1-4 and 4.1-6 as compared to BI level 0.1-1. The CAT activity was significantly different in BI levels 2.1-3, 3.1-4 and 4.1-6 as compared to controls. CONCLUSION: The study findings suggest oxidative stressful state associated with reduced antioxidant defence potential in erythrocytes of leprosy patients. The study implicates association of erythrocyte oxidative stress with bacterial load and type of leprosy.