Unique pattern of mutations in ß-thalassemia patients in Western Uttar Pradesh.

CONTEXT: ß-thalassemia is one of the most common heterogeneous inherited single gene disorders. The disease results from one or more of 380 different mutations in the ß-globin gene. Uttar Pradesh (U.P.) is the most populous state of India, comprising various ethnic groups and Bareilly is one of the largest cities situated in Western U.P. AIMS: To examine the prevalence of five common ß-thalassemian mutations: Intervening Sequence IVS 1-5 (c. 92 + 5 G > C), codon 8/9 (c. 27_28insG), codon 41/42 (c. 124_127delTTCT), IVS 1-1 (c. 92 + 1 G > T) and codon 26 G-A (c. 79G > A) in Western U.P. SETTINGS AND DESIGN: Patients attending camps organized by the Thalassemia Society, Bareilly were selected for the study. MATERIALS AND METHODS: A total of 48 blood samples were collected from the patients of transfusion dependent ß-thalassemia from July 2011 to May 2012. All the samples were analyzed for five common mutations by using the Amplification Refractory Mutation System (ARMS)-hot start-polymerase chain reaction (PCR) technique. RESULTS: Among the five common mutations prevalent in India, we were able to detect all except codon 26 G-A (c. 79G > A), which is prevalent in northeast India. These four mutations accounted for 58% of the total number of our patients. The IVS 1-5 (G-C) was found to be the most common mutation with a frequency of 46% and the 2 (nd)most common mutation was Fr8/9 (+G) with a frequency of 21%. The frequency of other mutations was IVS1-1 (12%) and Cd 41/42 (4%). CONCLUSION: This study provides evidence that the pattern of mutations in Western U.P. is different from the rest of India and even from the neighboring states (Delhi and Punjab). To the best of our knowledge, mutation Fr8/9, the 2(nd)most common mutation in our study has never been reported to be so common from anywhere in India. Some mutations, which are prevalent in other regions are absent in our region (mutation for ε-globin). Hence, these findings can be called unique to Western U.P.

Development of a novel rapid immunodiagnostic kit based on flagellar 40 kDa antigen epitope for the detection of typhoid fever in Indian patients.

To aid the clinical diagnosis of typhoid fever in India, where most hospitals and primary health centres have no facilities for culture, we report on the development of a novel and rapid immunodiagnostic kit for the direct detection of Salmonella Typhi--specific IgG antibodies against S. Typhi flagellar H antigen. The disease often does not show a specific clinical picture, and can be confused with other febrile illness such as malaria, dengue fever and Staphylococcus aureus. To overcome the problem of cross reactivity specific epitope of the flagellar H antigen was immobilised on the testing kit strip eliminating chances of cross reactivity and false positive results thereby increasing the specificity of the test. Since the immunodiagnostic kit, uses the flagellar H antigen from bacteria present in our country, the antibodies present in the serum of patients of our country will have maximum binding affinity, enhancing the sensitivity of our test kit. The immunodiagnostic kit on analysis gave a positive result with clinically diagnosed typhoid positive patient serum and negative results were obtained with the sera of clinically diagnosed malaria, abscess of Staphylococcus aureus and Visceral leishmaniasis (Kala-azar) patients.

Antioxidant status of children with idiopathic nephrotic syndrome.

The production of free radicals can cause renal injury and play an important role in the pathogenesis of idiopathic nephrotic syndrome. Markers of reactive oxygen species (ROS) were evaluated in 48 patients with active nephrotic syndrome (ANS) and 30 age- and gender-matched healthy children. Plasma malondialdehyde (MDA), protein carbonyl, nitrite, copper, zinc, selenium, ascorbic acid, and superoxide dismutase (SOD) levels were estimated in patients with ANS and controls. Measurements were repeated in 39 cases after achievement of remission, and in 10 other children who were in remission of >6 months' duration. Plasma MDA and nitrite levels were significantly higher and selenium was lower in ANS patients compared with controls. Plasma protein carbonyl, copper ascorbic acid, zinc, and superoxide dismutase levels were comparable in ANS patients and controls. Plasma copper level was significantly higher in active cases than in the remission and long-term remission groups. Selenium value showed a rise and then normalized in long-term remission. Among different sub-groups of ANS, no significant differences were found in the levels of various parameters, except plasma selenium, which was significantly lower in first-attack nephrotic syndrome (FANS) in comparison to infrequently relapsing nephrotic syndrome (IRNS) and frequently relapsing nephrotic syndrome (FRNS) patients. Thus, we observed evidence of oxidative stress and impaired antioxidant defense during acute nephrotic syndrome. Antioxidant status recovered completely only during long-term remission.

Antioxidant status of children with acute renal failure.

The production of free radicals can cause renal injury and play a role in the pathogenesis of acute renal failure (ARF). The indirect markers of reactive oxygen species (ROS) were evaluated in children with ARF and controls. Forty patients with ARF aged 0-10 years were selected. Twenty age- and gender-matched healthy children were included as controls. Plasma malondialdehyde, protein carbonyl, nitrite, copper, ascorbic acid, zinc, and ceruloplasmin levels were estimated in patients with ARF and controls. The plasma malondialdehyde (p < 0.01), copper (p < 0.001), ascorbic acid (p < 0.05), and ceruloplasmin (p < 0.001) levels were significantly raised in ARF patients in comparison with controls. Significantly higher levels of plasma malondialdehyde (p < 0.01), nitrite (p < 0.001), copper (p < 0.001), and ceruloplasmin (p < 0.001) and lower plasma zinc (p < 0.01) were found in ARF nonsurvivors in comparison with survivors. The cutoff levels of plasma nitrite and ceruloplasmin were found to be most accurate in predicting mortality in ARF patients and had maximum sensitivity (100%) and specificity (60.7%) among the parameters studied. In conclusion, the increased levels of oxidants and antioxidants suggest the production of ROS and their possible role in ARF pathogenesis. Plasma nitrite and ceruloplasmin concentrations demonstrated predictive ability in relation to mortality.

Lymphocyte adenosine deaminase activity in children with idiopathic nephrotic syndrome.

Adenosine deaminase (ADA) activity, as a marker of cell-mediated immunity, was evaluated in the serum (S-ADA) and lymphocytes (L-ADA) of 47 children with idiopathic nephrotic syndrome, and 23 healthy controls. The mean S-ADA and L-ADA levels were significantly raised in active nephrotic syndrome (ANS) and in its sub-groups in comparison with controls. The ADA activity was significantly more elevated in relapsers than for the first attack of nephrotic patients, and the frequent relapsers had the highest enzymatic levels both in serum as well as lymphocytes. A significant positive correlation was found between serum and lymphocyte ADA levels (r =0.736, p <0.01). In remission, the S-ADA showed a significant fall in comparison with their corresponding ANS value (p <0.001) and reached the level of controls. The mean L-ADA also showed reduction but the difference was statistically insignificant and the value was significantly raised, when compared with controls. The enzyme activity in serum and lymphocytes normalized in the long-term remission group. Thus, ADA activity was abnormal in ANS cases, and L-ADA demonstrated change both in active as well as remission stage of the disease.