Comparison of oxidant-antioxidant status in patients with vitiligo and healthy population.

BACKGROUND: Vitiligo is a well-recognized pigmentary disorder of the skin and /or mucous membrane characterized by circumscribed ivory or chalky white macules devoid of identifiable melanocytes. The pathogenesis of vitiligo is complex and still not well understood. According to autocytotoxic hypothesis, oxidative stress has been suggested to be the initial pathogenic event in melanocyte degeneration. The role of free radicals and oxidative damage in the pathophysiology of vitiligo has been documented in recent studies. OBJECTIVE: To evaluate the role of oxidative stress in patients with vitiligo and of healthy controls by measuring levels of the oxidant malondialdehyde (MDA) and antioxidants vitamin C and vitamin E in serum and catalase (CAT) in erythrocytes. METHOD: A total of 80 clinically diagnosed cases of vitiligo and 80 control subjects were included in the study to assess the activity of MDA, vitamin C and vitamin E in serum and CAT in erythrocytes of patients and controls by using the spectrophotometric assay. RESULT: There was statistically significant increase in the levels of MDA in patients with vitiligo compared to the control group (p<0.001). No significant difference was found in the levels of vitamin C (p=0.411) and vitamin E (p=0.771) between the patients with vitiligo and control group. The levels of CAT in the vitiligo patients were found to be significantly lower than those of controls (p<0.001). CONCLUSION: Increased oxidative stress and decreased catalase have been observed in vitiligo patients and the data suggesting that the free radicals may be involved in the destruction of melanocytes or dysregulation of melanogenesis.

Studies on the identification and control of pathogen Saprolegnia in selected Indian major carp fingerlings at mid hill altitude.

The Indian major carp cultured in ponds in the North Eastern hilly states of India frequently suffer from fungal disease during winter months resulting in mass mortality. This study examined the pathogenic fungi isolated from farmed raised Indian major carp fingerlings and identified as Saprolegnia. For treatment, the diseased fish were exposed to 4g salt per litre of water for 2 min followed by dip treatment with 5ppm KMnO4 for 10 min, thrice every week for a period of 6 weeks. The treatment resulted in recovery from the disease after 6 weeks from the beginning of treatment. Soon after recovery, the pond management practices such as removal of pond bottom soil, application of lime and replenishment with freshwater were followed in the infected ponds. Our study concluded that rapid decrease in pond water temperature from 22 to 8 degrees C that remains low for months together coupled with increased water pH (9) and decreas dissolved oxygen (4ppm) causes saprolegniasis to the fingerlings of Indian major carps.

The study of prevalence of Helicobacter pylori in patients with acne rosacea.

BACKGROUND: Acne rosacea is an inflammatory disease affecting the central part of the face characterized by erythema, papules, papulo pustules and telangiectasias of unknown etiology. More recently numerous studies have described an association with Helicobacter pylori (Hp) and the extra gastric symptoms of cutaneous origin. OBJECTIVES: To establish the prevalence of Helicobacter pylori (Hp) infection in the stomach in patients with rosacea based on standard Hp serological test. METHODS: All patients with the clinical staging of 2, 3 and 4 rosacea attending the Dermatology Out Patient Department from May 2009-April 2010 were included in the study. Quantitative serological test using the SERION ELISA classic Helicobacter pylori IgG was done. RESULT: A total of 26 patients were enrolled in the study with the age ranging from 26-82 years. There were 14 males and 12 females and the male: female ratio was 1.6:1. Three (11.53%) patients presented with Grade IV rosacea, a severe clinical presentation and symptoms suggestive of acid peptic diseases were found in 14/26 (53.8%) patients. A positive serology to H. pylori was found in 17/26 (65.4%) of patients. A statistically significant correlation was found when association of H. pylori positivity in patients of rosacea compared with the seropositivity in controls. However correlation in the seropositivity was not found amongst the patients having gastritis and rosacea. CONCLUSION: There still proves to find a correlation of Hp infection with patients with rosacea but it can still be hypothesised as a cutaneous manifestation of an internal peptic ulcer disease.

Hyperglycemia, glucose intolerance, hypertension and socioeconomic position in eastern Nepal.

Abstract. The present study was undertaken to evaluate differences between urban and rural Nepali populations in terms of hyperglycemia, socioeconomic position (SEP) and hypertension, through a community based survey in Sunsari District, eastern Nepal. Blood glucose levels were measured in participants (N = 2,006) S30 years old from urban and rural communities and were classified according to WHO criteria (1998) into normoglycemia (NGY), impaired fasting glucose (IFG), impaired glucose tolerance (IGT) and hyperglycemia (HGY). SEP was assessed by structured health interview along with anthropometric measurements and behavioral variables. Hypertension was classified per Joint National Committee (JNC-VII) criteria. Ten point three percent and 11.9% of subjects in this survey (13.3% urban and 11.0% rural) gave a family history and personal history of diabetes mellitus, respectively. Of urban participants (n = 736) with no history of diabetes 70 (9.5%) had HGY and 143 (19.4%) had glucose intolerance (IFG and IGT). Of rural participants (n = 1,270) 114 (9.0%) had HGY and 176 (13.9%) had glucose intolerance. There was an increasing trend in numbers of cases of hyperglycemia and intolerance with increasing age (chi2 198.2, p < 0.001), body mass index (BMI) (chi2 35.1, p < 0.001), SEP (chi2 48.5, p < 0.001) and hypertension (chi2 130.6, p < 0.001). Rural participants had a lower odds ratio [0.706; 95% confidence interval (CI) 0.455-1.096] of having hyperglycemia than urban participants. Individuals with medium and higher SEP had a lower odds ratio (0.878; CI 0.543-1.868) and higher odds ratio (1.405; CI 0.798-2.474), respectively, compared to individuals with lower SEP of having HGY. Both urban and rural populations are at risk for hyperglycemia and glucose intolerance. Individuals having a medium SEP had lower risk of diabetes mellitus than individuals from lower and higher SEP.

Subclinical hypothyroidism in eastern Nepal: a hospital based study.

BACKGROUND: Subclinical hypothyroidism itself is associated with serious complications and also there is a known risk of subclinical hypothyroidism patients getting converted into overt disease. OBJECTIVES: The objective of the present study was to fi nd out the prevalence of subclinical hypothyroidism in the suspected cases i.e. amongst the cases attending the thyroid laboratory at B.P. Koirala Institute of Health Sciences, Dharan, Nepal. MATERIALS AND METHODS: It was a retrospective cross sectional study. Data of the free T3, free T4 and TSH estimations of the year 2007 of the Thyroid lab at BPKIHS, Dharan, Nepal was analyzed. ELISA based free T3, free T4 and TSH tests in the serum had been performed in all the cases. RESULTS: Total cases were 1714 including 24.446% males and 75.554% females. Cases with raised TSH levels were 26.021%, cases with normal TSH levels were 54.66% and cases with low TSH levels were 19.316%. Total 350 cases (20.42 %) had subclinical hypothyroid dysfunction which includes 84 (4.901 %) males and 266 (15.519%) females. And the maximum percentage of cases in either gender was between the age groups 20 -59 years. CONCLUSION: The prevalence of subclinical thyroid hypothyroidism amongst the suspected cases was 20.42 % which is much higher compared to the other parts of the world. The highest percentage was found in the female age group 20 - 59 years. The routine screening of the whole population is not cost effective and on the basis of the present study it is suggested that there may be routine screening of the selected populations, especially women between 20 to 59 years of age in Nepal region. The preferred screening method advised is a sensitive ELISA based TSH test.

Experimentally induced depletion of germ cells in sub-adult Patagonian pejerrey (Odontesthes hatcheri).

Germ cell (GC) transplantation (GCT) is a novel reproductive technology with application in seed production and conservation of endangered species. This study examined the suitability of treatment with Busulfan, a cytotoxic agent, and warm water, known to cause GC degeneration, for depletion of endogenous GCs in sub-adult Patagonia pejerrey Odontesthes hatcheri intended as hosts in GCT. In two experiments, fish were treated with six combinations of temperature (intermediate and high, 20 and 25 degrees C, respectively) and Busulfan (0, 20, and 40 mg/kg body weight), given intraperitoneally (ip) as a single (0 week) or repeated (0 and 4 week) dose. The effectiveness of the treatments was assessed by gonado-somatic index, histology, and (germ cell-specific) vasa gene expression after 8 weeks. Fish were allowed to recover at 17 degrees C for 4-8 weeks after the treatments to ascertain the permanency of the effects. The high temperature (25 degrees C) alone induced only incipient gonadal degeneration and germ cell loss, but was highly effective in combination with double administration of 40 mg/kg Busulfan. Males tolerated Busulfan better and were more easily depleted of germ cells than females. Animals treated for 8 weeks were severely devoid of germ cells, but were still capable of gametogenesis. Thus, the combination of Busulfan and high water temperature appeared to be efficient for depletion of GCs in adult fish; and the treated gonads retained the ability to support GC proliferation and differentiation. Furthermore, quantitative analysis of vasa transcript levels was found to be an useful to monitor the degree of gonad sterility during treatment.

Reverse micelles for the removal of dyes from aqueous solutions.

The ability of reverse micelles to solvate organic dyes in the aqueous core was investigated with methyl orange (MO) and methylene blue (MB) using hexadecyl trimethyl ammonium bromide (HTAB) and sodium dodecyl benzene sulphonate (SDBS) surfactants in a polar amyl alcohol medium. The removal trend of the dyes from water was studied with different concentrations of the dyes. The effects of NaCl and CaCl2 salts on removal efficiency of the surfactants were investigated and results were compared. It was observed that the separation of dyes from the aqueous phase to the organic phase depends on the electrostatic interaction between the dye molecule and surfactant head groups. In the case of NaCl, with increasing salt concentration, the removal (%) of dye decreases. For CaCl2, removal of methyl orange shows a gradual increase with increasing dye concentration, whereas, for methylene blue, its removal decreases with increasing dye concentration.

Erythrocyte glutathione status in human visceral leishmaniasis.

A glutathione redox cycle is a major antioxidant defense system for the detoxification of reactive oxygen species within erythrocytes. Reactive oxygen species such as superoxide anions, hydrogen peroxide and hydroxyl radicals are generated as a host defense mechanism for killing of engulfed Leishmania donovani, a causative agent of visceral leishmaniasis, are capable of damaging lipids and other biomolecules when produced in excess. Erythrocytes are most vulnerable to Reactive oxygen species. In present study we aimed to evaluate erythrocyte reduced glutathione (GSH) levels as an antioxidant and erythrocyte malondialdehyde (MDA) as a marker of lipid peroxidation. The study included twenty-five Visceral leishmaniasis patients and they were followed up after their complete chemotherapy with antileishmanial drugs (sodium stibogluconate) for 30 days. Forty six age and sex matched healthy individuals were taken as controls. GSH levels in erythrocytes of visceral leishmaniasis patients were increased in spite of significant increased erythrocyte MDA as compared to controls. Whereas erythrocyte GSH and MDA levels of follow up patients were decreased as compared to patients before treatment groups. We concluded that visceral leishmaniasis patients are in oxidative stress which most likely induces the endogenous antioxidant such as GSH or its poor utilization by cells.

Antibacterial properties of human beta defensin-3 derivative: CHRG01.

Antibiotic resistance in bacteria is a major health concern. Antimicrobial peptides (AMPs) are a class of peptides that are efficient in killing most microbes yet development of resistance to AMPs is rare. However, complex secondary and tertiary structures and difficulties in isolating AMPs have limited their use as antibiotics. It has been demonstrated earlier that small peptides derived from human ß defensin-3 (HBD-3) also show antibacterial activity. Here, we perform a detailed characterization of the antibacterial activity of one such derivative: CHRG01. While HBD-3 has 45 amino acids with three disulphide bonds and a ß-sheet folded structure, CHRG01 has only 14 amino acids with the cysteine residues replaced by serine. The antibacterial nature of CHRG01 was studied using scanning electron microscopy (SEM), confocal microscopy, circular dichroism (CD) and small-angle X-ray scattering (SAXS). CD data show that CHRG01 is random coiled in solution. SEM and confocal studies show that the mode of action of CHRG01 is pore forming. SAXS studies show that CHRG01 induces a negative Gaussian curvature, the type of curvature needed for pore formation. The above results show that CHRG01, a small peptide without any complex structure, is capable of killing bacteria by permeabilizing their outer membranes.

Incidence of megaloblastic anaemia and its correction in leishmaniasis--a prospective study at BPKIHS hospital, Nepal.

Fifty (50) cases of Leishmaniasis were included in a prospective study after making the diagnosis based on the demonstration of L D Bodies on bone marrow aspirate. Hemoglobin (Hb) and total leucocyte count (TLC) were less than 10 gm/dl and 4000 per/ml respectively in 22 out of 50 cases. Among 50 cases, 30 showed macrocytic red cell and hypersegmented neutrophils on peripheral blood smear as features of megaloblastic anemia. Out of 30 cases of leishmaniasis with megaloblastic anemia, 10 cases showed decreased reticulocyte count. Bone marrow aspiration cytology showed megaloblastic maturation in erythroid series and giant metamyelocytes in myeloid series in all 30 cases of leishmaniasis with megaloblastic changes. Bone marrow iron was also increased in these cases. However megakaryocyte was normal in all. Among 30 cases of leishmaniasis with megaloblastic changes, 15 cases were treated with sodium antimony gluconate (SAG) only and other 15 with SAG, folic acid and Vit. B12. Rest 20 cases of leishmaniasis without megaloblastic changes were also treated with SAG only. After treatment with SAG alone as well as with SAG, folic acid and vit. B12, hemoglobin and TLC improved in all 50 cases. Of 15 cases of leishmaniasis with megaloblastic changes, who were treated with SAG, folic acid and vit. B12, the reticulocyte count reached at peak on the 7th day and normalized at the end of 4th week. Red blood cells (RBC) also became normocytic normochromic and neutrophils became normal on peripheral blood smear. But no change was observed in reticulocyte count, RBC and white blood cell (WBC) morphology on peripheral blood smear in cases of leishmaniasis without megaloblastic changes and other 15 cases of leishmaniasis with megaloblastic changes, who received SAG only.

Diagnosis of multiple myeloma by demonstration of M protein in bone marrow aspirate by agar gel electrophoresis: a case report.

A number of laboratory tests are used to confirm the diagnosis of multiple myeloma, including M protein in the serum. Since M protein in the serum originate from tumour cells in the bone marrow before circulating in the serum, demonstration of M protein in bone marrow aspirate can be added to the batteries of diagnostic parameters.

Association of obesity and physical activity in adult males of Dharan, Nepal.

OBJECTIVES: Obesity is one of the most apparent-yet most neglected public health problems. Physical activity is a crucial element in the development of obesity but still its importance in the occurrence of obesity varies in different populations. This paper aims to study the burden of obesity and its association with physical activity was carried out in a rapidly urbanizing town. METHODS: A cross-sectional study to investigate the prevalence of obesity and its association with physical activity was carried out in one thousand adult males of Dharan municipality. Tools of data collection included interview and physical measurements such as blood pressure, height and weight measurements, and waist and hip circumferences. Odds ratios (ORs) and their 95% confidence intervals for obesity were computed across various demographic and other variables without adjusting and then adjusting for physical activity. RESULTS: The prevalence of overweight and obesity in the population was 32.9% and 7.2% respectively. The study showed that physical inactivity is more importantly associated with obesity in the older population. The trend of young being more obese is reversed after adjusting for physical activity so that those in the older age were more obese than the younger ones. Similarly, those in to the business, vocational and clerical works, those who were more literate and those in the higher socio-economic status were significantly associated with obesity even after correcting for physical activity. CONCLUSION: The prevalence of overweight and obesity is high in the males of Dharan. The value and effect of physical activity seem to vary across different age-groups and socio-economic status and occupations. The young, the technical persons or businessmen and the more prosperous ones probably need to bring down their calorie intake along with emphasis on physical activity in order to bring down their weight and cardiovascular risk.

Effects of microgravity on microcirculation.

The blood behaviour in human circulatory system observed during space flights reveals that the microgravity environment reduces the flow rate and increases the haematocrit compared with the situation on the earth surface. The present study gives a theoretical analysis of the above phenomena by considering a two layer Newtonian model for blood flow in capillaries. The effect of gravity on plasma layer as well as on the blood viscosity is taken into account for calculating the percentage change in flow rate of blood. The corresponding apparent viscosities on the earth surface and in space are estimated. The results are analyzed and presented graphically. The findings agree reasonably well with the data observed in spaceflight experiments.

Status of iron, oxidant and antioxidants in chronic type 2 diabetes mellitus patients.

Diabetes mellitus is a common health problem of the world. Iron may be a part of the cause of the disease and its complications. Iron is a trace element which produces reactive oxygen species (ROS) participating through Fenton reaction and that ROS may be a cause to produce oxidative stress and further diabetic complications. The study aims to access the iron and its effect in producing oxidative stress in type 2 diabetic patients. Serum iron, total iron binding capacity (TIBC) and percentage transferrin saturation are calculated as the index of iron. Malondialdehyde (MDA) is estimated as index of oxidant and vitamin C, vitamin E are measured as index of antioxidants. This is a case control study conducted in the department of Biochemistry in collaboration with department of Medicine at B P Koirala Institute of Health Sciences (BPKIHS), Dharan, Nepal. 52 chronic type 2 diabetes mellitus patients and 52 age and sex matched normal healthy controls were included in the study. Plasma iron, TIBC, percentage transferrin saturation were found (89.14±30.50 µg/dL), (266.78±48.80 µg/dL), (36.61±14.31%) in diabetic cases as compared to (83.98±24.19 µg/dL), (279.08±40.23 µg/dL), (31.05±10.98%) of healthy controls. A significant increase in MDA level (6.35±1.52 nmol/ml in cases and 4.18±1.12 nmol/ml in controls, p<0.001) and significant decrease in vitamin C (0.85±0.19 mg/dL in cases and 1.28±0.21 mg/dL in controls, p<0.001) and vitamin E (0.85±0.25 mg/dL in cases and 1.34±0.38 mg/dL in controls, p<0.001) were observed.

Oral zinc supplementation for reducing mortality in probable neonatal sepsis: a double blind randomized placebo controlled trial.

OBJECTIVE: To study the role of Zinc in the treatment of neonatal sepsis. DESIGN: Double blind, randomized, placebo controlled trial. SETTING: Tertiary Care Hospital. PARTICIPANTS: 614 neonates with probable neonatal sepsis. INTERVENTION: The drug group (n=307) received 1mg/kg/day of elemental zinc, and placebo group (n=307) received the placebo, in addition to antibiotic therapy and supportive care, till the final outcome (discharge/death). OUTCOME MEASURES: Decrease in mortality rates (primary outcome), duration of hospital stay and need of higher lines of antibiotic therapy (secondary outcomes) were tested. RESULTS: Baseline characteristics of the two groups were similar. No statistically significant differences between drug and placebo group were found in mortality rate (9.77% vs 7.81%; P=0.393), mean duration of hospital stay (142.85 ± 69.41 hrs, vs. 147.99 ± 73.13 hrs; P=0.841), and requirement of higher lines of antibiotic therapy (13.35% vs 12.05%, P=0.628) after supplementation. CONCLUSIONS: This study does not report decrease in mortality rates, duration of hospital stay and requirement of higher lines of antibiotic therapy following zinc supplementation in neonatal sepsis.

Distribution and ethnic variation of â-thalassemia mutations in Nepal.

This is the first study characterizing spectrum of beta-thalassemia mutations in Nepalese population. Mutations were analyzed in 22 patients using 10 sets of allele-specific primers. Five of the mutations, namely F.S 41/42 (--TCTT), IVS1 nt5 (G-->C), IVS1 nt1 (G-->T), 619 bp deletion and F.S 8/9 (+G), were found to constitute 87.82% of total alleles studied. F.S 41/42 (--TCTT) was the commonest mutation. -88 (C-->T), Codon 16 (--C) and Codon 15 (G-->A), had a combined frequency of 12.18%. Distribution of mutations causing beta-thalassemia in different ethnic Nepalese groups was analyzed. The mutational profile in Nepal share several similarities with that from the two neighboring countries, India and China. Detection of more than one mutation in three cases of thalassemia trait raises the likelihood of existence of multiple mutations in cis in Nepalese thalassemic carriers. Such possibility has to be carefully considered while developing prenatal screening program for Nepalese population.

Adenosine deaminase in CSF and pleural fluid for diagnosis of tubercular meningitis and pulmonary tuberculosis.

Tuberculosis (TB) is one of the most common infectious diseases in developing countries including Nepal. Delay in diagnosis and treatment of tuberculosis results in poor prognosis of the disease. This study was conducted to estimate diagnostic cut off values of Adenosine Deaminase (ADA) in cerebrospinal fluid (CSF) and pleural fluid and to evaluate the sensitivity, specificity, positive and negative predictive values ofADA in pleural fluid and CSF from patients with tuberculous and non-tuberculous disease. A total of 98 body fluid (CSF: 24, Pleural fluid: 74) specimens were received for the estimation of ADA. ADA activity was measured at 37 degrees C by spectrophotometric method of Guisti and Galanti, 1984 at 625nm wavelength. Among the patients enrolled for the study subjects for which CSF were received (n = 24) included 8 tuberculous meningitis (TBM), and 16 non-tubercular meningitis (NTM). Pleural fluid samples (n = 74) were received from 19 pulmonary TB with pleural effusion, 17 PTB without pleural effusion and 37 of non-tuberculous disease patients. CSF ADA activity were (11. 1 +/- 2.03 IU/L) and (5.3 +/- +1.89 IU/L) (p <00001) in TM and non-NTM groups and Pleural fluid ADA activity were (10 +/- 22.18 IU/L) and (23.79 +/- 11.62 IU/L) (p < 0.001) in PTB and non-TB groups respectively. ADA test in body fluids, which is simple, cost-effective and sensitive, specific for the tubercular disease is recommended to perform before forwarding the cumbersome and expensive procedures like culture and PCR for TB diagnosis.

Efficiency of osmotic and chemical treatments to improve the permeation of the cryoprotectant dimethyl sulfoxide to Japanese whiting (Sillago japonica) embryos.

Insufficient cryoprotectant permeation is one of the major obstacles for successful fish embryo cryopreservation. The purpose of this study was to test the effectiveness of osmotic and chemical treatments to enhance cryoprotectant uptake by fish embryos. Japanese whiting Sillago japonica embryos at the somites and tail elongation stages were treated with hyperosmotic sugar solutions (1 M trehalose and sucrose) for 2-6 min, or a permeating agent (2-6 mg/mL pronase) for 30-120 min, and then impregnated with 10-15% DMSO in artificial sea water or aqueous solutions containing inorganic salts (0.125-0.25 M MgCl(2) and CaCl(2)). The viability of the embryos after the treatments was estimated from hatching rates and the internal DMSO concentration was measured by HPLC. Treatment with trehalose for 3 min prior to impregnation with DMSO enhanced the uptake of the cryoprotectant by 45% without significantly affecting embryo viability, whereas pronase had no noticeable effect on cryoprotectant permeation. Incorporation of DMSO into the embryos was enhanced by 143-170% in the presence of 0.25 M MgCl(2) and 0.125 M CaCl(2) compared to sea water. A combination of treatments with trehalose and MgCl(2) was even more effective in promoting DMSO permeation (191% compared to untreated embryos). Tail elongation embryos were less tolerant of the treatments, but had higher DMSO impregnation. In conclusion, the use of trehalose (as dehydrating agent) and MgCl(2)/CaCl(2) (as a vehicle during impregnation) greatly promoted cryoprotectant uptake and may be a promising aid for the successful cryopreservation of fish embryos.

Glycemic status, lipid profile and proteinuria in diabetic nephropathy.

INTRODUCTION: Diabetic nephropathy is one of the major complications of Diabetes Mellitus characterized by persistent albuminuria, elevated arterial blood pressure, a relentless decline in glomerular filtration rate (GFR) and a high risk of cardiovascular morbidity and mortality. METHODS: In this study, urinary micro-albumin estimation was done in 177 diabetic patients. This study aims to ascertain association of glycemic status, lipid profile and proteinuria in Type 2 Diabetes Mellitus with nephropathy. RESULTS: Among 177 patients, 26 had frank proteinuria, 79 had micro-albuminuria and 72 were without proteinuria. Increased frequency of proteinuria was seen in male than female. Micro-albuminuria and frank proteinuria was seen more in older age group. The multiple comparisons showed the significantly increased levels of urea, creatinine, fasting blood glucose in micro-albuminuria and overt proteinuria patients in comparison to without proteinuria. Glycated hemoglobin level was increased with the increasing age group particularly in overt proteinuric patients. CONCLUSIONS: The glycemic control, monitoring of lipid profile and early urinary protein estimation with better management may delay diabetic nephropathy or its further complications in diabetes mellitus.