Spotted fever group rickettsioses in Himachal Pradesh.

Rickettsiae are in many places of the world emerging or reemerging pathogens. The spotted fevers group (SFG) composes a large group of tick- and mite-borne zoonotic infections that are caused by closely related rickettsiae. The SFG rickettsiae of Southeast Asia are yet to be identified. Earlier reports have documented the endemicity of rickettsioses among adults in the Himalayan belt but no confirmed case of spotted fever have been reported from this region of India. We present two cases of SFG rickettsioses; from the northern hilly region of India that were confirmed using specific microimmunofluorescence assay.

Acute reversible hearing loss in scrub typhus.

Scrub typhus usually presents as pyrexia with or without multiple organ involvement. Acute hearing loss occurs in about one third of cases and is a useful clue toward the diagnosis. We present two cases of scrub typhus with acute reversible hearing loss from an endemic area. The diagnosis was confirmed by nested PCR.

Relevance of Weil-Felix test in diagnosis of scrub typhus in India.

OBJECTIVE: To study the relationship of Weil-Felix test and microimmunofluorescence test. METHODS: Sera of 21 patients with clinical diagnosis of scrub typhus were subjected to Weil-Felix and Microimmunofluorescence tests. RESULTS: On Weil-Felix test, 13 (62%) sera showed titers 1: > or = 40-320. 7 patients showed titers 1: > or = 320, 3 showed titers 1 : 160, 2 showed titers 1 : 80 and 1 patients showed titers 1 : 40, to Proteus OXK antigen. All 21 sera showed significant titers to O. tsutsugamushi on microimmunofluorescence. CONCLUSION: Weil-Felix test is not a very sensitive test in diagnosis of scrub typhus but due to of lack of availability of definitive tests in India it canbe a useful tool when used and interpreted in the correct clinical context.

Scrub typhus.

Scrub typhus, a dreaded disease in pre-antibiotic era, is an important military disease which caused thousands of cases in the Far East during Second World War. It is a zoonosis and is a widespread disease in Asia and Pacific Islands. Scrub typhus is an acute febrile illness which generally causes non-specific symptoms and signs. The clinical manifestations of this disease range from sub-clinical disease to organ failure to fatal disease. Deaths are attributable to late presentation, delayed diagnosis, and drug resistance. The public health importance of this disease is underestimated because of difficulties with clinical diagnosis and lack of laboratory methods in many geographical areas. Scrub typhus is known to occur all over India and physicians should be aware of this potentially serious but easily treatable disease.

Stroke at moderate altitude.

OBJECTIVE: To know the clinical profile, presence of various risk factors for stroke at moderate altitude and to study its relationship with hypertension and polycythemia at moderate altitude. METHODS: We analyzed the data of 100 patients, consecutively admitted to a hospital situated at an moderate altitude of 2200 meters MSL of Sub-Himalayan ranges and studied the age, gender, geographical distribution, clinical features and presence of various risk factors in relation to stroke. This is not only the first study conducted in Himachal Pradesh but also first study in India to be conducted at moderate altitude (2000-3000 meters MSL). RESULTS: Males outnumbered females (66% males, 34% females) with rural predominance (73% rural, 27% urban). Cerebral infarction (69%) was more common but primary intracerebral haemorrhage (26%) was more common than found in the West. Hypertension (62%) and smoking (60%) were most common risk factors present and polycythemia was not a significant risk factor at this altitude. CONCLUSION: Incidence of stroke was found to be lower than the study conducted at low altitude. incidence of various types of stroke was similar to other Indian studies. The combination of opposite effects of decreased hypertension and increased haematocrit could not be demonstrated at this altitude and smoking was more common than in other studies and other risk factors prevalent were same as that for low altitude.

Isolation of Mycoplasma bovoculi from genitally diseased bovines and its experimental pathogenicity in pregnant guinea pigs.

Thirteen strains of M. bovoculi, 6 from frozen bull-semen (3.5% of 168), 3 from neat bull-semen (3.0% of 100), one each from heart blood and stomach contents of aborted foetus of 85 (1.18%) bovine-abortions, one each from stomach contents and pooled internal organs of 9 (11.1%) stillborn calves, were isolated. All the isolates were resistant to ampicillin and sensitive to spiramycin, vibramycin, demeclocyclin, oxytetracycline, lincomycin and tylosin. However, variation in resistance to tetracycline, erythromycin, neomycin, kanamycin and streptomycin was observed. The gross lesions like congestion of lungs, liver, kidney and spleen were noted only in stillborn calf. However, significant microscopic lesions were encountered in internal tissues of both the aborted bovine fetuses and stillborn calf. Thickened alveolar wall, congestion of blood vessels, mesenchymal cell proliferation along with infiltration of lymphocytes and macrophages were observed in lungs. The liver showed mild infiltration of lymphocytes, macrophages in hepatic triad and necrosis of hepatic cells. The kidney tissues had focal lymphocytic infiltration in the interstitium. One strain of M. bovoculi (isolate # SBC-7/84,IO) isolated from a stillborn calf was found abortigenic upon experimental inoculation in pregnant guinea pigs.

Neurocysticercosis presenting with psychosis.

Cysticercosis consists of infection with the small bladder-like larvae of the pork tapeworm Taenia solium. The life cycle of parasite is maintained between man and pig infected with cysticerci. Epilepsy is the most common presentation of neurocysticercosis; focal signs, headache, involuntary movements and global mental deterioration are other symptoms. Psychosis is a rare presentation and may be seen in up to 5% of patients. We present a 25 years old male, who had been under treatment from psychiatry OPD for psychosis for one year, developed generalized tonic-clonic seizures. CT scan of brain revealed multiple calcified and hypodense lesions with surrounding oedema. Histopathological examination of subcutaneous nodule confirmed the diagnosis of neurocysticercosis.

Experimental pathogenicity evaluation of Mycoplasma canadense from bovine mastitis in vitro and in vivo laboratory models.

Mycoplasma canadense, a clinical isolate from milk of a mastitic buffalo, was experimentally tested for its pathogenic potential in hamster tracheal ring and rabbit fallopian tube explant organ cultures (in vitro) and rat and rabbit mammary gland (in vivo) models. The activity percentage reduction in M. canadense infected hamster tracheal rings was 99.1% in comparison to 16.4% in control rings. Mycoplasma canadense, also induced complete ciliostasis at 11-day post-infection in rabbit fallopian tube explants. Histopathological lesions in these infected organ cultures were loss of cilia, desquamation or denudation of epithelium, infiltration of inflammatory cells and proliferation of macrophages as well as oedema in lamina propria. At the end of the experiments, M. canadense organisms were reisolated in pure colonies from the infected but not the control organ cultures. In the rat and rabbit mammary glands, M. canadense organisms persisted upto 6-day and 7-day postinfection, respectively and caused histopathological changes suggestive of subacute to chronic mastitis during the experimental period. The results indicate that the tested M. canadense clinical isolate was virulent.

Duodenal diverticular haemorrhage in a patient taking NSAID.

We present a 55 years old male with severe anemia with history of pain abdomen for 2 weeks and malena of 10 days duration. He was taking NSAID (Diclofenac sodium). Upper GI endoscopy done twice did not reveal any abnormality. Upper gastrointestinal (UGI) barium series with small bowel follow through revealed a diverticulum on medial wall of second part of duodenum and there was evidence of ulcer in diverticulum. He underwent diverticulectomy. On follow up after 6 months patient was asymptomatic.

Effects of angiotensin converting enzyme inhibitor on renal function in patients of membranoproliferative glomerulonephritis with mild to moderate renal insufficiency.

AIM: To determine the effect of enalapril, angiotensin-converting-enzyme inhibitor (ACE-I) on progression of renal insufficiency in primary membranoproliferative glomerulonephritis with mild to moderate renal insufficiency. METHOD: Thirty patients with histopathologically proved MPGN having hypertension (grade I and II of JNC-VI criteria of hypertension) and mild to moderate impairment of renal function (creatinine clearance varying from 30-80 ml/min, significant albuminuria and serum creatinine 1.2-3.0 mg/dl) were initially treated with diuretics and 3-blockers to bring down BP < 140/90 mm Hg. These patients were then randomly divided into three groups of 10 each, group I--Control; group II--Nifedipine and group III--Enalapril. In group II and III Nifidepine 30 mg/day and in group III Enalapril 10 mg/day respectively were added in addition and treatment was continued for nine months. These patients were followed up monthly for drug efficacy, side effects and any adverse drug reaction. RESULTS: Out of 30, 28 patients completed the study. At the end of nine months of treatment the patients of control group revealed significant increase in serum creatinine (1.65 +/- 0.38 to 2.17 +/- 0.31 mg/dl), blood urea (34.0 +/- 3.9 to 40.0 +/- 3.1 mg/dl), and 24 hours albuminuria (3.6 +/- 0.6 to 4.2 +/- 0.6 gm) and decrease in creatinine clearance (60.3 +/- 13.3 to 37.5 +/- 11.8 m/min); however, in enalapril group there was decrease in serum creatinine (1.72 +/- 0.45 to 1.24 +/- 0.58 mg/dl), blood urea (34.6 +/- 4.7 to 28.1 +/- 6.7 mg/dl) and 24 hours albuminuria (3.3 +/- 1.0 to 1.6 +/- 1.1 gm) and increase in creatinine clearance (56A +/- 15.8 to 77.1 +/- 23.5 ml/min). The patients on nifedipine showed statistically nonsignificant changes in creatinine clearance, blood urea and serum creatinine; while albuminuria increased from 3.0 +/- 1.3 to 3.9 +/- 0.4 gm/24 hours (p < 0.01). The blood pressure was well controlled in all patients. None of the patient had side effects leading to withdrawal of drugs. No adverse drug reaction was noted. CONCLUSION: ACE-I (enalapril) provided protection against the progression of renal insufficiency in patients of MPGN having hypertension with mild to moderate renal impairment. The renoprotective effects of ACE inhibitor (enalapril) is associated with substantial decrease in albuminuria.

Cloning and characterization of an insecticidal crystal protein gene from Bacillus thuringiensis subspecies kenyae.

A sporulating culture of Bacillus thuringiensis subsp. kenyae strain HD549 is toxic to larvae of lepidopteran insect species such as Spodoptera litura, Helicoverpa armigera and Phthorimaea operculella, and a dipteran insect, Culex fatigans. A 1.9-kb DNA fragment, PCR-amplified from HD549 using cryII-gene-specific primers, was cloned and expressed in E. coli. The recombinant protein produced 92% mortality in first-instar larvae of Spodoptera litura and 86% inhibition of adult emergence in Phthorimaea operculella, but showed very low toxicity against Helicoverpa armigera, and lower mortality against third-instar larvae of dipteran insects Culex fatigans, Anopheles stephensi and Aedes aegypti. The sequence of the cloned crystal protein gene showed almost complete homology with a mosquitocidal toxin gene from Bacillus thuringiensis var. kurstaki, with only five mutations scattered in different regions. Amino acid alignment with different insecticidal crystal proteins using the MUTALIN program suggested presence of the conserved block 3 region in the sequence of this protein. A mutation in codon 409 of this gene that changes a highly conserved phenylalanine residue to serine lies in this block.

Comparison of aminophylline and insulin-dextrose infusions in acute therapy of hyperkalemia in end-stage renal disease patients.

OBJECTIVE: This study was performed to compare the efficacy of aminophylline and insulin-dextrose infusion as acute treatment modality of hyperkalemia in patients with end-stage renal disease (ESRD). METHODS: This study was conducted on 30 ESRD patients with serum potassium > 6.0 mEq/L. These patients were divided in two groups of 15 each. Group A patients were treated with aminophylline infusion, whereas group B patients were treated with insulin-dextrose infusion. Serum potassium and other biochemical parameters such as blood sugar were measured at beginning of treatment followed by at 60 minutes, 180 minutes, and 360 minutes after treatment. RESULTS: Intervenous infusion of aminophylline lowered plasma potassium from 6.48 +/- 0.39 mEq/L to 5.92 +/- 0.40 mEq/L at 180 minutes (p < 0.001 Vs basal) and 6.05 +/- 0.53 mEq/L at 360 minutes (p < 0.01 Vs basal). Whereas, intravenous infusion of insulin-dextrose decreased plasma potassium from 6.59 +/- 0.31 mEq/L to 5.76 +/- 0.32 mEq/L (p < 0.001 Vs basal) and 5.84 +/- 0.21 mEq/L (p < 0.001 Vs basal). Thus in both groups, plasma potassium levels were significantly less than basal levels throughout the study. The decrease in plasma potassium was significantly more in group B patients (p value is < 0.001 after 60 minutes, < 0.05 after 180 minutes and < 0.05 after 360 minutes) when compared to group A patients. There was one episode of hypoglycemia (blood sugar < 60 mg%) in insulin-dextrose infusion group. No other side effects were observed throughout the study. CONCLUSION: Aminophylline is an effective modality for acute treatment of hyperkalemia, though it is less effective than insulin-dextrose infusion. However, more studies are required to confirm these results.

Dissimilar Deletion In DMD Gene Of Two Brothers : A Case Study.

Two brothers who are affected by DMD were studied with respect to CPK assay, muscle biopsy and deletion analysis of their DNA. The severity of phenotype in these two brothers is comparable. Onset of the disease in both the cases was at 4 years of age. CPK values obtained at the age of 10 years for the elder brother and 6years for the younger one were 2550 IU and 1650 IU respectively. Muscle biopsy of the patients indicated dystrophy. PCR analysis of the patient's DNA was carried out for deletion detection in DMD gene. The deletion pattern observed for these two cases was found to be quite different from each other. In case of the elder brother, deletion was found to be expanding exons 8-13 whereas, for the younger brother exons 8-13 as well as exons 43-45 were found to be deleted.

Human carbonic anhydrase I: Effect of specific site mutations on its function.

Carbonic anhydrase I (CAI) is one out of ten CA isoenzymes that have been identified in humans. X-ray crystallographic and inhibitor complex studies of human carbonic anhydrase I (HCAI) and related studies in other CA isoenzymes identified several residues, in particular Thr199, GlulO6, Tyr7, Glull7, HislO7, with likely involvement in the catalytic activity of HCAI. To further study the role of these residues, we undertook, site-directed mutagenesis of HCAI. Using a polymerase chain reaction based strategy and altered oligonucleotide primers, we modified a cloned wild type hCAI gene so as to produce mutant genes encoding proteins with single amino acid substitutions. Thrl99Val, Thrl99Cys, Thr199Ser, GlulO6Ile, GlulO6Gln, Tyr7Trp, Glu.117Gln, and His I 07Val mutations were thus generated and the activity of each measured by ester hydrolysis. Overproduction of the Glu 117Gln and His I 07Val mutant proteins in Escherichia coli resulted in a large proportion of the enzyme forming aggregates probably due to folding defect. The mutations Thr199Val, GlulO6Ile and GlulO6Gln gave soluble protein with drastically reduced enzyme activity, while the Tyr7Trp mutation had only marginal effect on the activity, thus s.uggesting important roles for Thr199 and GlulO6 but not for Tyr7 in the catalytic function of HCAI.

Subcapsular tubercular liver abscess.

A young female married for the last 2 years but without any issue presented with lump in the right upper abdomen. This was proved to be a subcapsular liver abscess on USG and CT scan and proved to be tubercular on needle aspiration cytology. She was given four drugs antitubercular treatment (ATT). After four months of ATT she conceived and it was decided by both parents to continue the pregnancy. The three drug ATT was continued throughout the pregnancy and she delivered a perfectly healthy baby. Upto three months follow up the mother and baby were perfectly healthy. The subcapsular tubercular liver abscess is extremely rare and conception during treatment may be the first case in literature.