Comparative evaluation of indirect and sandwich ELISA for the detection of antibodies to bovine respiratory syncytial virus (BRSV) in dairy cattle.

Seroprevalence of bovine respiratory syncytial virus (BRSV) infection in both exotic and crossbred cattle were described. A baculovirus expressed recombinant purified nucleocapsid (N) protein was used in indirect and sandwich ELISA for screening of 499 bovine sera samples from all over the state for the presence of BRSV antibodies. The seroprevalence rate of BRSV was found to be 46.09% through indirect ELISA while it would found to be 65.33% by sandwich ELISA. The result also indicated that exotic breeds were more susceptible to BRSV infection compared to crossbred cattle. A comprehensive analysis on susceptibility to BRSV as regards to various factors like age and sex was also summarized.

Clinical profile of sickle cell trait.

OBJECTIVES: Although sickle cell trait is considered a harmless condition in ordinary circumstances, a large number of pathological conditions have been attributed to it often without a scientific basis. Many patients visit this centre with various complaints and on testing are found to be sickle cell trait. Hence it was thought necessary to analyse these cases to find out the nature of their ailments. METHODS: Two hundred cases of sickle cell trait diagnosed by sickling test and hemoglobin electrophoresis on CAM, and 150 age and sex matched control cases with normal hemoglobin phenotype from a survey were studied. Hemoglobin estimation was done in all by cyanmethemoglobin method. Besides history and clinical examination other relevant investigations were done as necessary to arrive at the diagnosis. Seven cases of sickle cell trait were asymptomatic while the rest were suffering from different conditions. 51% of sickle cell trait and 86% of control cases had mild to severe anaemia which improved with iron therapy in trait cases. Hepatomegaly (11% vs 4.6%), epistaxis (5% vs 0.66%) and cholelithiasis (3% vs 0%) was seen in significantly more number in sickle cell traits than the control cases. Splenomegaly and midsystolic murmurs were present in higher percent of cases but was not statistically significant. There was one case of epilepsy with multiple small infarcts in the brain and another with focal fits with epileptogenic focus in the left cerebral hemisphere where no other cause could be found except sickle cell trait. CONCLUSION: The ailments of sickle cell trait cases are like persons with normal hemoglobin. Anaemia is not more frequent and can improve with iron therapy. However, hepatomegaly, epistaxis, cholelythiasis are seen more frequently and minor cerebral infarcts probably can occur in sickle cell trait. These require more elaborate studies to decide their pathogensis.

Splenectomy in sickle cell disease.

INTRODUCTION: Since role of splenectomy in sickle cell disease still remains controversial, this paper evaluates the same in 32 cases of splenectomy in sickle disease patients observed by the author. MATERIAL AND METHODS: After proper diagnosis by standard methods the cases were observed for 2-3 years before deciding on splenectomy. Though acute splenic sequestration (53%) and chronic hypersplenism (21.8%) were the main indication. Two cases of splenic abscess and one case with frequent pain attacks were also subjected to splenectomy. There was one immediate post-operative death. RESULTS: The cases were followed up for 1 to > 8 years. While Acute splenic sequestration disappeared, in chronic hypersplenism cases as well as others besides a sense of general well being, steady state hemoglobin rose by > 2G/dl in 81% and significant increase in height and weight was seen in 90% and 61% cases respectively. Frequency of pain attack, fever, anaemia and need for blood transfusion improved in 79% patients. CONCLUSION: Splenectomy in selected cases appears to be an acceptable procedure in sickle disease.

Cholelithiasis in sickle cell disease in India.

Eighty cases (63 Males and 17 Females) of sickle cell disease were searched for gall stone by ultrasonography, 8 (10%) cases had gall stone. Gall stones were more common in males (11.1%) than females (5.8%). It was not found below 13 years of age. Three cases had single and five cases had multiple gall stones. Three of them had thickened gall bladder wall. Typical biliary colic was uncommon (1 case). There was no case of obstructive jaundice. HbF% level was significantly lower in cases with gall stone (12.31 +/- 3.95) than without gall stones (16.73 +/- 6.30). Episodes of aggravated anaemia and total serum bilirubin was significantly higher (1.27/pt and 4.12 +/- 1.34 mg%) in cases with gall stone than without gall stone (0.31/pt and 2.74 +/- 1.47 mg%). The above findings suggest association of greater degree of haemolysis with formation of gall stone in sickle cell disease cases.

Clinical profile of sickle cell disease in Orissa.

Children comprised 52% of patients with Sickle Cell Disease (SCD). Types of Sickle Cell Disease encountered were SS (92.7%). SB thalassaemia (6.7%) and SD disease (0.7%). The disease was widespread in almost all castes and communities in the society; largest number of patients (20%) belonging to scheduled castes and only 1.4% were from scheduled tribes. Maximum number of cases were in the age group 2-4 and 4-6 years, many of whom died around this age. Besides attacks of pain, jaundice and anemia, frequent attacks of fever with anemia or only anemia in childhood were a predominant presenting feature. Splenic sequestration was frequent (10.1%). The patients usually had a steady state hemoglobin level of 6-10 g/dl, with which they thrived well. Fetal hemoglobin was 5-30%. Blood transfusion was not a frequent requirement, but prophylactic long acting penicillin was helpful in preventing frequency of crisis.

Serum immunoglobulins in sickle cell disease.

Immunoglobulins IgG, IgA and IgM were estimated in 20 cases of sickle cell disease in steady state, eleven cases of sickle cell disease in crisis and/or infection, 20 cases of sickle cell trait and 14 normal healthy controls. Immunoglobulin deficiency has not been observed in sickle cell disease. Significantly raised IgG but normal IgA and IgM found in sickle cell disease cases was probably due to chronic stimulation of reticuloendothelial cells secondary to chronic hemolysis. IgM levels were significantly higher in patients with splenic enlargement > 3 cm. There was no alteration in immunoglobulin levels in sickle cell trait.

Sickle cell disease in India.

A screening programme involving 9,822 hospitalised patients revealed the frequency of individuals with S gene to be 11.1 per cent. A population survey of 1,000 randomised subjects from amongst about 70,000 people in one block of the area showed the frequency to be 15.1%. The gene is not confined to tribal peoples, but is prevalent throughout the society, being more frequent in scheduled castes and some caste Hindus. With the available Indian data a sickle cell belt can be mapped out in the country. Analysis of clinical data on the first 700 cases of sickle cell disease seen in the Sickle Cell Research Centre (ICMR) at Burla shows patients of all ages, even beyond 40 years, though many patients tend to die by 20 years of age. Genetically, while most patients are SS and 8.1% are S-beta thalassaemia, cases of SD disease and SE disease were also encountered. A frequency of 0.32% of alpha thalassaemia gene was noted in SS patients against 0.28% in sickle cell trait and 0.12% in AA controls. The disease was found to manifest as early as 3 months or may remain asymptomatic till adult life. Though generally running a milder course, moderate to severe anaemia, vaso-occlusive attacks (86.5-89.36%), splenic sequestration (8.43%-12.76%), crippling avascular bone necrosis (5.7%-35.08%), osteomyelitis (5/700), and epistaxis (28.92%-35.08%) remain a few clinical events deserving competent and urgent medical management.(ABSTRACT TRUNCATED AT 250 WORDS)

Sickle cell-beta+ thalassaemia in Orissa State, India.

The clinical, haematological, and some molecular genetic features of 17 Orissan Indian patients with sickle cell-beta+ thalassaemia (S beta+ thal) are described and compared with those in 131 Indian patients with homozygous sickle cell (SS) disease. Patients with S beta+ thal had higher Hb A2 levels, and lower mean cell volume (MCV) and mean cell haemoglobin (MCH) compared to SS disease but no other haematological difference of statistical significance. High levels of Hb F occurred in both genotypes and the alpha+ thalassaemia gene frequency reached 0.47 in S beta+ thal and 0.32 in SS disease. Clinically there were no significant differences between the genotypes indicating that the low levels of HbA (3-5%) in this condition were insufficient to modify the clinical features. The thalassaemic beta globin gene is inactivated by a G----C mutation at position 5 of the first intron of the beta globin gene (IVS1-5 G----C) in all cases. This finding should facilitate the introduction of a prenatal diagnosis programme aimed at the prevention of beta thalassaemia or S beta+ thalassaemia in that population.

Sickle haemoglobin, G-6PD deficiency and malaria in western Orissa.

Sixty cases of malaria were screened for sickle haemoglobin and G-6PD deficiency. Plasmodium vivax was detected in 40 (66%) and Plasmodium falciparum in 21 (35%) cases, with six of the latter having cerebral manifestation. Sickle Hb was found in 7 (11.5%) patients and G-6PD deficiency in 3 (5%) cases. Both patients with SS disease had vivax malaria, while of 5 with sickle cell trait 3 had only vivax, one only falciparum and one mixed infection. Amongst G-6PD deficient patients one had vivax and two falciparum malaria. One of the latter had both SC trait and G-6PD deficiency. Thus, adult persons with SS disease or SC trait were not found to be resistant to either vivax or falciparum malaria. A high frequency (5%) of G-6PD deficiency amongst malaria patients warrants a caution against indiscriminate use of 8-aminoquinoline drugs.

Yellow oleander (cerbera thevetia) poisoning with jaundice and renal failure.

Jaundice and renal failure in yellow oleander poisoning have not been reported previously. Similarly no post-mortem report has shown renal tabular necrosis and glomerular pathology, liver and brain changes in this poisoning. Four cases of yellow oleander poisoning with jaundice and renal failure and the postmortem findings in another three cases are reported.

The molecular basis of alpha thalassemia in India. Its interaction with the sickle cell gene.

The alpha globin genotype of a total of 282 Indians from Orissa state has been analyzed. The overall alpha thalassemia gene frequency is 0.29, most frequently caused by the -alpha 3.7 and -alpha 4.2 deletions. In one family a novel -alpha 3.5 deletion removing the alpha 1 globin gene with some of its flanking sequences has been found, suggesting further sequence homology of the alpha globin gene cluster 3' to the alpha 1 globin gene. Patients with sickle cell disease and alpha thalassemia had higher hemoglobin (Hb) levels, RBC counts, and Hb A2 levels, and lower reticulocyte counts, MCV, MCH, and Hb F levels than those with a normal alpha genotype. The frequency of splenomegaly was not influenced by the alpha globin genotype. A higher prevalence of alpha thalassemia was found in patients greater than or equal to 10 years of age than in the younger group, suggesting a possible advantageous effect of alpha thalassemia on the survival of patients with sickle cell disease.

Fetal hemoglobin levels and beta (s) globin haplotypes in an Indian populations with sickle cell disease.

To further explore the cause for variation in hemoglobin F (Hb F) levels in sickle cell disease, the beta globin restriction-fragment length polymorphism haplotypes were determined in a total of 303 (126 SS, 141 AS, 17 S beta(0), 7 A beta, (0) and 12 AA) Indians from the state of Orissa. The beta(s) globin gene was found to be linked almost exclusively to a beta(S) haplotype ( -++-), which is also common in Saudi Arabian patients from the Eastern Province (referred to as the Asian beta(s) haplotype). By contrast, the majority of beta A and beta(0) thalassemia globin genes are linked to haplotypes common in all European and Asian populations (+-----[+/-]; --++-++). Family studies showed that there is a genetic factor elevating Hb F levels dominantly in homozygotes (SS). This factor appears to be related to the Asian beta(s) globin haplotype, and a mechanism for its action is discussed. There is also a high prevalence of an independent Swiss type hereditary persistence of fetal hemoglobin (HPFH) determinant active in both the sickle cell trait and in sickle cell disease.

The sickle cell gene is widespread in India.

The sickle cell gene was first described in India in a tribal population in the south, leading to the belief that it was confined to tribal groups. The present study confirms that it is widespread in the state of Orissa and spreads throughout Hindu society, being more common in upper and scheduled castes than in tribal groups.

Raised Hb F levels in sickle cell disease are caused by a determinant linked to the beta globin gene cluster.

A haplotype of restriction fragment length polymorphisms in the beta S globin gene cluster was determined in six different African and Asian ethnic groups with sickle cell disease. The geographical distribution of a particular haplotype, designated Asian beta S haplotype, coincides with the occurrence of raised Hb F levels in sickle cell disease. Detailed hematologic and genetic studies of a large geographically isolated group of Orissan Indian patients with sickle cell disease and a mixed Asian Indian group with homozygous beta thalassemia provided evidence that the determinant responsible for raised Hb F levels is linked to the beta globin gene cluster. The possible nature of this determinant is discussed.

Sickle cell disease in Orissa State, India.

A study of 131 patients with homozygous sickle cell (SS) disease in Orissa State, India, indicated that, compared with Jamaican patients, Indian patients have higher frequencies of alpha thalassaemia, higher fetal haemoglobin, total haemoglobin, and red cell counts, and lower mean cell volume, mean cell haemoglobin concentration, and reticulocyte counts. Indian patients have a greater frequency and later peak incidence of splenomegaly, and hypersplenism is common. Painful crises and dactylitis are not uncommon in Indian patients but chronic leg ulceration is rare. Homozygous sickle cell disease in Orissa is similar to that in the Eastern Province of Saudi Arabia and is very different from that in populations of West African origin.

Geographical survey of beta S-globin gene haplotypes: evidence for an independent Asian origin of the sickle-cell mutation.

The haplotypes of 152 beta S-chromosomes were characterized in six different population groups. The chromosomes of individuals from Nigeria and from the southwest of the Arabian peninsula have the haplotype - - - - + + - + previously found in west African, Jamaican, and U.S. American blacks, whereas those from the eastern oases of Saudi Arabia and from the west and the east coast of India showed a different haplotype not found in Africa (+ + - + + + + -). These data are most consistent with an independent Asian origin of the sickle-cell mutation and provide further information about the geographic distribution of beta S-haplotypes in the Old World. The distribution of the Asian beta S-haplotype corresponds to the reported geographic distribution of a mild clinical phenotype of homozygous SS disease.