Mutations at Novel Sites in pmrA/B and lpxA/D Genes and Absence of Reduced Fitness in Colistin-Resistant Acinetobacter baumannii from a Tertiary Care Hospital, India.

Background: Colistin resistance in Acinetobacter baumannii, the last resort drug for serious infections, is emerging worldwide. There has been paucity of data on this aspect from India, which is one of the largest producers of colistin. We studied colistin resistance in A. baumannii and characterized the isolates with respect to resistance mechanisms and virulence. Methods: A total of 365 A. baumannii isolates were studied. Antimicrobial susceptibility testing was performed as per standards. Colistin resistance mechanisms were studied by mutation detection in pmrA/B and lpxA/C/D genes, phenotypic loss of lipopolysaccharide, presence of mcr1-5 genes, and carbonyl cyanide 3-chlorophenylhydrazone (CCCP) effects. Biofilm formation, desiccation survival, and growth kinetics were studied and statistically analyzed for colistin-resistant and colistin-susceptible isolates. Results: All the colistin-resistant isolates (9, 2.5%) showed multiple mutations at novel sites in pmrA/B and/or lpxA/D genes with reversion of resistance with CCCP. Majority of these isolates (6, 66.6%) were from patients without prior colistin therapy. All received prior carbapenems. The resistant isolates demonstrated no significant difference in biofilm formation and desiccation survival but were slow growers. Conclusion: Mutations in pmrA/B and/or lpxA/D genes were the main resistance mechanism in these colistin-resistant isolates that showed no reduction in fitness.

Lepidium sativum: Bone healer in traditional medicine, an experimental validation study in rats.

India is a land of spices and medicinal plants. Ayurvedic medications and methods are commonly practised in India for curing several ailments. Lepidium sativum (garden cress) is an important herb that belongs to Brassicaceae family. It is believed that the plant has its origin in Ethiopia but is now cultivated throughout the world. The plant is well-known in Ayurveda for its beneficial properties it holds. The present study describes the fracture healing property of the methanolic and aqueous extract of Lepidium sativum seeds. For the study, 21 Charles foster rats were used. They were grouped into three groups each containing seven rats: control, methanolic, and aqueous group. Rats were anesthetized using ether vapors and fractures were induced in each rat from all the three groups using hand held three-point bending technique. The broken bone fragments were then stabilized using splints. The control group was administered with normal saline, along with food and water, post-fracture. The methanolic group was administered with the methanolic extract of Lepidium sativum seeds at dose of 400 mg/kg given orally, post-fracture along with food and water. The third group received aqueous extract of the seeds in doses of 550 mg/kg orally, along with daily food and water intake for a period of 8 weeks. The results were evaluated both radiologically and biochemically. X-rays were done on day 0, 2nd week, and 4th week post-fractures to look for the callus formation and serum levels for calcium, phosphorus, and alkaline phosphatases were evaluated on 0 day, 1 week, 2nd week, 4th week, 6th week, 8th week, and 10th week post-injury. It was observed at the end of the study period that the methanolic group had significant callus formation starting at the 2nd week itself post-fracture. The serum levels of calcium, phosphorus, and alkaline phosphatases at 4th, 6th, and 8th weeks had significant P values in the methanolic group rats.

Genetic Predisposition to Diabetic Nephropathy: Evidence for a Role of ACE (I/D) Gene Polymorphism in Type 2 Diabetic Population from Kutch Region.

Genetic polymorphism as described with angiotensin-converting enzyme gene has been proposed as a putative mediator of diabetic nephropathy. We substantiate the hypothesis that genetic variants of the ACE have significant impacts on diabetic nephropathy. To assess the possible association between the three ACE polymorphic variants and DN in an ethnically homogeneous type 2 diabetic population from Kutch region. A 287-bp insertion/deletion polymorphism in intron 16 of the ACE gene was examined by polymerase chain reaction using a case-control approach conducted with 309 unrelated type 2 diabetic patients of Kutch origin (159 Ahir and 150 Rabari, with >10 years duration of T2DM). Of the patients, 143 had nephropathy {AER >30 mg/day (Ahir, n:73 and Rabari, n:70)} and were considered as cases; all others {n:166 (86 Ahir and 80 Rabari)} were normoalbuminuric (AER <30 mg/day) and were treated as controls. Suitable descriptive statistics was used for different variables. Genotype frequencies in all groups were all in accordance with the Hardy-Weinberg equilibrium. Genotypic distribution was significantly different between cases and controls (Ahir: x(2) :8.87, 2 d.f. p = 0.0118; Rabari: x(2) :11.01, 2 d.f. p = 0.0041). Multivariate logistic regression analysis revealed that DD genotype was a significant and strongest independent predictor of microalbuminuria (Ahir: p = 0.0362, OR = 2.65, 95 % CI 1.89-6.36; Rabari: p = 0.024, OR = 2.81, 95 % CI 1.9-6.65). However, it did not independently change the odds of having macroalbuminuria versus microalbuminuria. Analysis of the association under various genetic models revealed that ACE I/D polymorphic variant contribute to DN susceptibility under recessive mode only. Genetic variation at the ACE locus as D/D variant in intron 16, contribute to an increased risk of nephropathy in T2DM patients but not extent of DN severity, and thus this polymorphism might be considered as genetic risk factors for DN among patients with type 2 diabetes.

Impact of lifestyle modification on glycemic control in patients with type 2 diabetes mellitus.

BACKGROUND: Current treatment guidelines support the role of lifestyle modification, in terms of increasing the quantity and quality of physical activity to achieve target glycemia in patients with type 2 diabetes mellitus. OBJECTIVE: To assess the effect of structured exercise training and unstructured physical activity interventions on glycemic control. MATERIALS AND METHODS: This was a randomized six-month exercise intervention study conducted with previously inactive 279 patients of type 2 diabetes mellitus. Before randomization, all enrolled T2DM participants (n: 300; 30 to 60 year old, having diabetes for more than a year with HbA1c levels of 6.5% or higher) entered a one-month run-in phase to reduce dropout and maintain adherence. RESULTS: A recommendation to increase physical activity was beneficial (0.14% HbA1c reduction; P = 0.12), but was not bringing significantly declines in HbA1c, whereas, structured exercise training is associated with a significant HbA1c decline of 0.59%. (P = 0.030). In a subgroup analysis limited to participants with a baseline HbA1c value > 7%, both the unstructured (0. 48%; P = 0.04) and structured exercise training (0.77%; P < 0.01) groups experienced significant decline in HbA1c Vs the control, whereas among participants with baseline hemoglobin A1c values less than 7%, significant reduction occurred only in the structured exercise training group. Changes in blood pressure; total cholesterol, HDL-cholesterol (high-density lipoprotein), LDL-cholesterol (low-density lipoprotein) and the atherogenic index factors did not statistically significantly differ within (baseline to follow-up) and among groups. CONCLUSION: Supervised structured training was more efficacious than unstructured activity in achieving declines in HbA1c. Although both structured and unstructured training provide benefits, only the former was associated with significant reductions in HbA1c levels. Therefore, T2DM patients should be stimulated to participate in specifically designed exercise intervention programs.

Diagnosis of Sanfilippo disease correlating clinical, radiological and biochemical findings-a case report.

Mucopolysaccharidoses (MPS) are a group of genetic diseases and its diagnosis is a challenging task due to multiple differential diagnosis. We had combined clinical findings, radiological and ophthalmological features. Biochemical test for urine glycosaminoglycans (GAG) was done for confirmation of diagnosis in the patient. The case of Sanfilippo disease was characterized by slowly progressive, severe CNS involvement with mild somatic disease. Radiological features were suggestive of Sanfilippo disease and urine GAG test for MPS was positive in the case. With the clinical features we had multiple differential diagnoses. The radiological investigations minimized the list and the biochemical test confirmed GAG in urine. In this case the combination of clinical, radiological and biochemical findings confirmed the diagnosis of Sanfilippo disease.

Estimation of serum malondialdehyde in oral cancer and precancer and its association with healthy individuals, gender, alcohol, and tobacco abuse.

BACKGROUND: Tobacco and alcohol induces generation of free radicals and reactive oxygen species, which are responsible for high rate of lipid peroxidation. Malondialdehyde is the most widely used marker of lipid peroxidation. The aim of the study was to estimate serum malondialdehyde level in oral precancer, oral cancer, and normal individuals. MATERIALS AND METHODS: In this study serum malondialdehyde was measured according to the method of Ohkawa et al in 30 normal individuals and 30 patients each with histopathologically diagnosed oral precancer, and oral cancer. RESULTS: The mean serum malondialdehyde level in the control group was found to be 5.107 ± 2.32 ηmol/ml, whereas it was 9.33 ± 4.89 ηmol/ml and 14.34 ± 1.43 ηmol/ml in oral precancer and oral cancer, respectively. There was statistically significant increase in serum malondialdehyde levels in the oral precancer and oral cancer patients compared with the control group. CONCLUSION: Increased serum malondialdehyde in oral cancer and oral precancer would serve as a valuable marker for both preventive and clinical intervention, and may deserve further investigation for the early diagnosis, treatment, and prognosis.