Indian childhood cirrhosis (ICC) & ICC-like diseases: The changing scenario of facts versus notions.

Indian childhood cirrhosis (ICC), a disease considered to have been endemic in and unique to India has now been documented in children of non-Indian origin from other countries. More recently available findings from a large multicentre study sponsored by the Indian Council of Medical Research (ICMR) have dispelled some of the generally accepted notions and have established several new facts on different aspects of the disease. All relevant reports on ICC and ICC-like diseases, till date, were reviewed to obtain a proper perspective on the current state of our understanding on this non-Wilsonian copper overload liver disease. A primary role of exogenous copper in causing the disease was earlier debated on the basis of studies in India but investigators abroad studying some sporadic cases and a series of endemic ICC-like diseases supported a hepatotoxic injury by ingested copper in genetically susceptible infants and children in ICC- like disease and in ICC. Epidemiologic and morphologic findings in the well controlled ICMR study based on 225 cases of ICC and 426 controls, all confirmed on liver biopsy, have however, convincingly refuted this concept. Additionally, this study revealed that unlike what has been believed earlier, older children more than 3 yr age can get the disease and that in its natural course the hepatic histology can transform between the characteristic one considered diagnostic and some other patterns, any one of which can be the morphologic manifestation at first presentation of the patient. Older children and cases with milder morphologic changes at presentation had longer survival. The overall inference from critical analysis of all available data is that ICC and ICC-like diseases clinically manifest in a child of any age though common in younger ones, and a clinical diagnosis must be made in any child with so-called ‘cryptogenic cirrhosis’. Exposure to exogenous copper in food, milk and water should not be a prerequisite for this consideration. A liver biopsy whenever feasible should be mandatory for confirmation with the understanding that the morphologic changes in liver can present a few other patterns besides the characteristic one currently taken to be diagnostic. The ascribed current decline in encountering ICC is likely to be due partly to missing a diagnosis and partly to a true reduction in incidence consequent on time related economic and socio-cultural changes.

Cassia occidentalis poisoning causes fatal coma in children in western Uttar Pradesh.

We investigated cases of the annual seasonal outbreaks of acute hepato-myo-encephalopathy in young children in western Uttar Pradesh for causal association with Cassia occidentalis poisoning, by a prospective survey in 2006. During September-October homes of 10 consecutive cases were visited and history of eating Cassia beans was obtained in all. Nine children died within 4-5 days. There appears to be an etiological association between consumption of Cassia occidentalis beans and acute hepato-myo-encephalopathy.

Cassia occidentalis poisoning as the probable cause of hepatomyoencephalopathy in children in western Uttar Pradesh.

BACKGROUND & OBJECTIVE: Recurrent annual outbreaks of acute encephalopathy illness affecting young children have been reported for several years in many districts of western Uttar Pradesh (UP). Our earlier investigations over three consecutive years (2002-2005) proved that these outbreaks were due to a fatal multi-system disease (hepatomyoencephalopathy syndrome) probably caused by some phytotoxin and not due to viral encephalitis as believed so far. We conducted a case-control study to investigate the risk, if any, from various environmental factors and also to identify the putative toxic plant responsible for development of this syndrome. METHODS: Eighteen cases with acute hepatomyoencephalopathy syndrome admitted in 2005 in a secondary care paediatric hospital of Bijnor district of western UP were included in the study. Three age-matched controls were selected for each case. A semi-structured questionnaire was developed and applied to all 18 cases and 54 controls. All interviews were conducted within one week of discharge or death of each case. Quantitative data were analyzed using the relevant established statistical tests. RESULTS: Parents of 8 (44.4%) cases gave a definite history of their children eating beans of Cassia occidentalis weed before falling ill, compared with 3 (5.6% controls), the odds ratio being 12.9 (95% CI 2.6-88.8, P<0.001). History of pica was the other associated factor with the disease, odds ratio 5.20 (95% CI 1.4-19.5, P<0.01). No other factor was found significantly associated with the disease. INTERPRETATION & CONCLUSION: Consumption of C. occidentalis beans probably caused these outbreaks, described earlier as hepatomyoencephalopathy syndrome. Public education has the potential to prevent future outbreaks.

Recurrent annual outbreaks of a hepato-myo-encephalopathy syndrome in children in western Uttar Pradesh, India.

BACKGROUND & OBJECTIVE: Outbreaks of an acute encephalopathy syndrome affecting children, with high case-fatality, have been reported in western Uttar Pradesh, India for the last many years.We investigated these cases in Bijnor district and present our findings. METHODS: Fifty five children aged 2-10 yr hospitalized from 2003 to 2005 in Bijnor, Uttar Pradesh, with features of acute encephalopathy were selected by defined clinical criteria. Various laboratory investigations were performed. RESULTS: The disease had peak incidence in early winter months. Previously healthy, 2-4 yr old rural children (mean age-3.78 yr) of very low socio-economic background were most vulnerable. Almost all had vomiting preceding unconsciousness and a majority had mild fever and abnormal behaviour/agitation. Abnormal posture of trunk and limbs were distinctive features. Fluctuation of blood pressure was seen in three-quarter cases. Serum aminotransferases, creatine phosphokinase and lactic dehydrogenase levels were found markedly raised virtually in all cases in whom the tests were performed. Serum glucose was found low (<50 mg/dl) in 47.3 per cent cases at presentation. Cerebrospinal fluid (CSF) was under normal or low pressure and without pleocytosis in all cases. No microorganism could be isolated from serum, CSF, urine and visceral specimens. Neuroimaging performed in two cases was also normal. Liver biopsy performed in 21 cases showed acute hepatotoxic injury in all with marked hydropic change and perivenular necrosis. Tibial muscle biopsy done in 8 cases showed focal necrosis while brain biopsy taken in 2 cases had mild spongiosis with focal gliosis. Forty two children succumbed to their illness (case fatality 76.4%), most within 72 h of presentation. Survivors did not show any neurological deficit. INTERPRETATION & CONCLUSION: Our findings showed that the outbreaks were due to a multi-system disease with toxic injury to liver, muscles and brain (hepato-myo-encephalopathy) and not due to viral encephalitis as believed so far. The cause remains unknown but several features suggest the possibility of phytotoxin-induced pathology.

Immunohistochemical detection of serum autoantibodies--a new technique using routine formalin fixed paraffin sections.

Detection of autoantibodies in serum is important in the diagnosis of primary immune mediated diseases. Tests of these antibodies are conventionally done by indirect immunoflourescence (IIF) on frozen sections of fresh target tissues from the murine species. More recently enzyme-linked immunoassays and IIF on cultured human epithelial cells or on fresh frozen sections of murine tissues coated onto wells or glass slides are also being used. But these tests are more expensive and generally not easily available to laboratories in developing countries. Obtaining small animals for preparing frozen sections of fresh tissue is also getting to be increasingly difficult. A simple; new and inexpensive technique was developed to perform IIF on routine paraffin sections of human tissues following antigen unmasking. This, technique offers qualitatively good, consistent, species specific and dependable results with several advantages over the conventional IIF on animal tissue frozen sections, particularly in a sausage block made from different types of tissues. Immunoperoxidase stain for autoantibodies can also be easily performed with the advantages of permanent preservation and clearer evaluation in light microscopy. Most importantly the technique is easily affordable and practicable in all histopathology laboratories.

Hepatocellular carcinoma--a model of human cancer: clinico-pathological features, etiology and pathogenesis.

Hepatocellular carcinoma (HCC), a unique human neoplasm, has interested many in several fields of biological and health sciences. This cancer is credited as the first that can be largely eliminated by a safe anti-viral vaccine and other transmission control measures. It is also the first cancer for which a reliable diagnostic tumour marker was identified and studies on this tumor in humans and animals have provided a large body of information on causation and step-wise evolution of cancers. Being a common and rapidly fatal tumour, mainly affecting males in the more populous developing countries, HCC may well be the commonest cancer of the human male. Clinical features are not specific for HCC but manifestations represent varying combinations of those due to cirrhosis which is a very frequently associated and pre-existent disease, those due to tumour and those due to malignancy. This tumour commonly takes a massive form with satellite nodules or a scattered multinodular form. A fibrolamellar variant is biologically and clinically quite different from the usual HCC and the small capsulated variant is seen in some geographic areas. Microscopically the trabecular variety is common and differentiation from metastatic cancers and benign lesions may need use of newly described immunohistochemical markers in addition to clinical evidence of cirrhosis. Hepatitis B and C viruses, dietary aflatoxin B1 and cirrhosis from any cause are common risk factors in that order of importance. Several lines of evidence including molecular biology and animal studies support these etiological linkages. Studies in experimental animals using chemical carcinogens, different hepatotropic viruses and aflatoxin have greatly helped our understanding of carcinogenesis in general and hepatocarcinogenesis in particular. Individual HCC risk factors may contribute to summation of mutations subsequent to repeated and continued liver cell injury act as carcinogen/co-carcinogen or be involved in both capacities.

Multicentric clinical trials for safety and efficacy evaluation of alpha;beta arteether in complicated P. falciparum malaria.

OBJECTIVE: To evaluate efficacy of alpha;beta arteether in patients of P. falciparum malaria presenting with complications was undertaken in a multicentric clinical trial. METHOD: Each patient who consented to undergo clinical trial with parenteral Arteether was treated with a fixed dose schedule of Arteether given intramuscularly in a dose of 150 mg once a day on three consecutive days. Every patient was followed upto 28 days with clinical, haematological and parasitological monitoring every day upto one week and thereafter at 14, 21 and 28 days. The response was assessed in terms of fever clearance time, parasite clearance time, cure rate and parasite reappearance rate. RESULTS: A total of 211 patients of P. falciparum malaria were included in the study from four centres (Bhilai, Guwahati, Jamshedpur and Rourkela). Results of this study showed that fever clearance time ranged between 24-168 hours, parasite clearance time ranged between 24-120 hours and overall mortality ranged between 4-8.5%. Out of 211, only 14 patients expired during the study, of these, 10 patients expired within first two days i.e. before completing the three day schedule of arteether therapy. Tolerability to arteether injection was good in all these patients and no untoward effects were experienced or reported during the study. Overall cure rate observed in these studies was 93%. CONCLUSION: This study shows a rapid parasite and fever clearance in patients of complicated P. falciparum malaria.

A multicentric study with arteether in patients of uncomplicated falciparum malaria.

Two hundred and sixty seven patients of uncomplicated P. falciparum malaria completed study in a multicentric phase III clinical trial of Arteether. Arteether was given intramuscularly in a dose of 150 mg daily for three consecutive days. Each patient was followed upto 28 days of alpha, beta arteether therapy. The cure rate was 97% with fever clearance time between 1-7 days (24-168 hours) and parasite clearance time between 1-3 days (24-72 hours). Parasite reappearance rate was found to be 3% and reported at only three of the centres. Following the treatment no adverse effect was observed on haematological, biochemical and vital clinical parameters.

Aflatoxin B1 induced hepatocarcinogenesis in neonatal rats.

Role of cell replication on aflatoxin B1 (AFB1) induced hepatocarcinogenesis was investigated in neonatal rats showing persistence of cell replication in the liver for 21 days of post natal life. Adult (8-10 weeks old) rats displaying no hepatocytic proliferation served as controls. Three doses of AFB1 were administered to both the groups at intervals of 48 hr with the doses starting on 10th day of age in the neonatal group. Appearance of phenotypically altered preneoplastic hepatocytes was quantitated in both the groups. A significantly higher incidence of preneoplastic foci was recorded in neonatal rats as compared to adult animals. The results suggest that presence of cell replication in neonatal rats at the time of AFB1 administration enhances the process of hepatocarcinogenesis.

A light & electron microscopic study on apoptotic bodies in acute aflatoxin B1 induced injury in the rat liver.

A subnecrogenic dose of aflatoxin B1 (AFB1) was injected intraperitoneally into 8-10 wk old male rats of the Holzman strain. Cytoplasmic and sinusoidal eosinophilic bodies were seen in the liver which appeared at 30 h and reached a maximum at 48 h. Electron microscopically some of the cytoplasmic structures were seen to be of mitochondrial origin whereas others resembled apoptotic bodies. The sinusoidal bodies were similar to apoptotic bodies.

Synovial fluid changes in mycoplasma induced septicaemic polyarthritis of goat kids.

Synovial fluid samples of goat kids inoculated (ip) with 5 ml of 48 hr log phase culture of Mycoplasma mycoides sub sp. mycoides (large colony type) containing 10(7) cfu/ml were analysed for physical, cytological and biochemical properties. The synovial effusions were exudative in nature with increased volume. Gross appearances were serofibrinous, haemorrhagic and turbid containing flocculent materials with immediate clot formation. Mucinous precipitate quality was very poor having friable precipitates with cloudy supernatant. There were high total leucocytic and erythrocytic counts with significant high numbers of both neutrophils and lymphocytes. Synovial fluid sugar contents were significantly reduced, whereas total protein contents were significantly increased with concomitant reduction in albumin:globulin ratio. The alkaline phosphatase and transaminase values were also markedly increased in the synovial fluids of mycoplasma induced polyarthritic goat kids. The results may provide a clinical guideline for diagnosis, chemotherapy and prognosis of different joint diseases in domesticated animals.

Etiologic spectrum of acute sporadic viral hepatitis in children in India.

The relative magnitude by hepatitis A virus (HAV), hepatitis B virus (HBV) and hepatitis Non-A, Non-B virus (HNANBV) was determined in 496 children from three different parts of India suffering from acute viral hepatitis by tests for specific IgM class anti-HAV and anti-HBV antibodies in the serum. HAV, HBV and NANB infections accounted for 55.8 per cent, 20.2 per cent and 23.2 per cent of cases respectively. Hepatitis A largely (59.5%) affected younger children of 1-5 yr. Nearly a third of children affected by NANB hepatitis were additionally positive for HBsAg. The proportions of HAV and HBV infected cases respectively decreased and increased with increasing age whereas the incidence of HNANBV infection remained almost constant throughout childhood. Acute NANB hepatitis, a major health problem in the adults of India is also common throughout childhood. This study suggests that this infection does not impart long lasting protective immunity.