Impact of effluent of Pulp & Paper industry on the flora of river basin at Jaykaypur, Odisha, India and its ecological implications.

The JK Paper industry located at Rayagada discharges biologically untreated effluent more than the permissible limit prescribed by Pollution Control Board, Odisha in to the environment. The industry is seriously polluting the surrounding aquatic and terrestrial environment. No detailed intensive study was carried out by previous workers on this industry earlier. The present study aims at finding out the impact of effluent on the flora at the contaminated site. The chemically treated effluent (TE) contained significant amount of mercury and cadmium. The TE has high BOD, COD, dissolved solids and suspended solids when compared to normal river water at the site of discharge. The TE deteriorated the natural water bodies changing the physico-chemical properties of natural river water. After meeting the river water the TE was diluted after 1 km distance from the meeting point of the river. Crop plants collected from the contaminated site showed higher level of residual Hg and Cd and significant depletion in pigment was observed. Plants collected from both the sides of the treated effluent canal showed significant amount residue mercury and cadmium in the plant leaves. The plants exposed to the TE, showed variation in chlorophyll and Phaeophytin pigment content when compared to their respective control values in all terrestrial plants collected from the contaminated site. In some plant leaves little increment in the pigment level was noted but the values were not significant. The changes observed in the plant pigment might be due to heavy metal accumulation. The presence of residual Hg and Cd in crop plants and plant leaves grazed by grazing animals after absorption, accumulation and enrichment may lead to a possible biological magnification, warrants attention. Proper biological treatment, treatment of effluent by modern methods and removal of heavy metals from the effluent before discharge by the industry is suggested.

A comprehensive review on chromium induced alterations in fresh water fishes.

Chromium is considered as one of the most common ubiquitous pollutants in the aquatic environment, but the pure metallic form is absent naturally. There are three oxidation states in case of Chromium viz., Cr (II), Cr (III), Cr (VI). Among which Cr (II) is most unstable. Cr (III) and Cr (VI) are the stable oxidation state of Chromium in the environment. Being one of the commonly used metals Chromium and its particulates enter the aquatic medium through effluents discharged from different industries like textiles, tanneries, electroplating workshops, ore mining, dyeing, printing-photographic and medical industries. Among these, hexavalent chromium is considered as the most toxic form because it readily passes cellular membranes and then reduced to trivalent form. This trivalent chromium combines with several macromolecules including genetic material inside the cytosol, and is ultimately exposes the toxic and mutagenic alterations due of chromium toxicity. Chromium is taken up either through gastrointestinal tract or respiratory tract. The amount varies depending upon the medium and the form of chromium. In this review, an attempt has been made to accumulate the mammoth available data regarding impact of chromium on fresh water fishes into a systematic representation. The main objective of the review is to provide a future guideline for the scientific community and public officials involved in health risk assessment and management ensuring a better environmental condition for human health.

Congenital Bilateral Coloboma of Upper Eyelid.

Congenital coloboma of eyelid is a rare anomaly. There is partial or total absence of eyelid structures. A seven year male child had coloboma of both the upper lids lateral to lacrimal puctum affecting the medial half of lid symmetrically with symblepharon in region of defect bilaterally. The study was carried out at Maharaja Krushna Chandra Gajpati Medical College Berhampur, Odisha in year 2010. Both eyebrows were abnormal. He presented on and off diminution of vision, burning sensation, redness and watering from both the eyes on and off. On examination high refractive error was detected (visual aquity was 6/18 in righteye and 6/24 in left eye). Cornea was dry and opacities were present in both the eyes. There was limitation of ocular movement in both sides due to symblepharon. Nystagmus was present. The subject did not have any other associated anomaly. The birth and family history was normal. This case can be surgically treated and earliest management can give good fuctional as well as cosmetic results.

Correlation between thermoluminescence and mechanoluminescence of gamma-irradiated Dy activated potassium and magnesium mixed sulphate.

Thermoluminescene (TL) and mechanoluminescence (ML) properties of gamma-irradiated Dy activated potassium and magnesium mixed sulphate have been studied. (K2Mg2:Dy) (SO4)3 samples having different concentrations of Dy were prepared by solid-state diffusion method. Two distinct peaks, the first approximately 130 degrees C and the second approximately 273 degrees C, are observed in the TL glow curve. It is also observed that TL intensities of both peaks decrease when TL glow curves were recorded after deforming the irradiated samples. Only one peak is observed in the ML intensity vs. time curve and the ML intensity decreases markedly with the post-irradiation annealing (to remove 130 degrees C TL peak) of the sample. Both ML and TL intensities have been observed optimum for 1 mol% of Dy in the mixed sulphate system. It is suggested that the recombination of electrons with the free radicals (anion radicals produced by gamma irradiation) released from the traps during the thermal or mechanical excitation is responsible for luminescence in this system.

Composition of the editing complex of Trypanosoma brucei.

The RNA editing that produces most functional mRNAs in trypanosomes is catalysed by a multiprotein complex. This complex catalyses the endoribonucleolytic cleavage, uridylate addition and removal, and RNA ligation steps of the editing process. Enzymatic and in vitro editing analyses reveal that each catalytic step contributes to the specificity of the editing and, together with the interaction between gRNA and the mRNA, results in precisely edited mRNAs. Tandem mass spectrometric analysis was used to identify the genes for several components of biochemically purified editing complexes. Their identity and presence in the editing complex were confirmed using immunochemical analyses utilizing mAbs specific to the editing complex components. The genes for two RNA ligases were identified. Genetic studies show that some, but not all, of the components of the complex are essential for editing. The TbMP52 RNA ligase is essential for editing while the TbMP48 RNA ligase is not. Editing was found to be essential in bloodstream form trypanosomes. This is surprising because mutants devoid of genes encoding RNAs that become edited survive as bloodstream forms but encouraging since editing complex components may be targets for chemotherapy.

Four related proteins of the Trypanosoma brucei RNA editing complex.

RNA editing in kinetoplastid mitochondria occurs by a series of enzymatic steps that is catalyzed by a macromolecular complex. Four novel proteins and their corresponding genes were identified by mass spectrometric analysis of purified editing complexes from Trypanosoma brucei. These four proteins, TbMP81, TbMP63, TbMP42, and TbMP18, contain conserved sequences to various degrees. All four proteins have sequence similarity in the C terminus; TbMP18 has considerable sequence similarity to the C-terminal region of TbMP42, and TbMP81, TbMP63, and TbMP42 contain zinc finger motif(s). Monoclonal antibodies that are specific for TbMP63 and TbMP42 immunoprecipitate in vitro RNA editing activities. The proteins are present in the immunoprecipitates and sediment at 20S along with the in vitro editing, and RNA editing ligases TbMP52 and TbMP48. Recombinant TbMP63 and TbMP52 coimmunoprecipitate. These results indicate that these four proteins are components of the RNA editing complex and that TbMP63 and TbMP52 can interact.

Mitochondrial ribonuclease P activity of Trypanosoma brucei.

Ribonuclease P (RNase P) is an essential enzyme that cleaves the 5' leader sequences of precursor tRNAs (pre-tRNAs) to generate mature tRNAs. The RNase P-like activity from Trypanosoma brucei mitochondria (mtRNase P) was purified over 10000-fold by sequential column chromatography. This is the first demonstration of such activity from mitochondria of parasitic protozoa. Its apparent molecular weight is approximately 70 kDa, considerably less than bacterial RNase P. Preliminary characterizations revealed no RNA component that is essential for this activity. Like other RNase P activities, the cleavage generates mature tRNAs with a terminal 5'-phosphate at the cleavage site and the 5' leader sequence with a 3'-hydroxyl. Disruption of the pre-tRNA tertiary structure inhibits the cleavage of the substrates. These data suggest that although all mitochondrial tRNAs are encoded in nuclear DNA in T. brucei, these cells contain an RNase P in the mitochondrion that cleaves the 5' terminal leader sequences of pre-tRNAs to generate mature tRNAs. Cleavage by mtRNase P of a pre-tRNA substrate that was divided into two fragments was demonstrated. This shows the feasibility of artificial regulation of gene expression that can be achieved by creating a complex made of target mRNA and a complementary small oligonucleotide that resembles natural substrates for RNase P.

An RNA ligase essential for RNA editing and survival of the bloodstream form of Trypanosoma brucei.

RNA editing in trypanosomes occurs by a series of enzymatic steps that are catalyzed by a macromolecular complex. The TbMP52 protein is shown to be a component of this complex, to have RNA ligase activity, and to be one of two adenylatable proteins in the complex. Regulated repression of TbMP52 blocks editing, which shows that it is a functional component of the editing complex. This repression is lethal in bloodforms of the parasite, indicating that editing is essential in the mammalian stage of the life cycle. The editing complex, which is present in all kinetoplastid parasites, may thus be a chemotherapeutic target.

Association of two novel proteins, TbMP52 and TbMP48, with the Trypanosoma brucei RNA editing complex.

RNA editing in kinetoplastid mitochondria inserts and deletes uridylates at multiple sites in pre-mRNAs as directed by guide RNAs. This occurs by a series of steps that are catalyzed by endoribonuclease, 3'-terminal uridylyl transferase, 3'-exouridylylase, and RNA ligase activities. A multiprotein complex that contains these activities and catalyzes deletion editing in vitro was enriched from Trypanosoma brucei mitochondria by sequential ion-exchange and gel filtration chromatography, followed by glycerol gradient sedimentation. The complex size is approximately 1,600 kDa, and the purified fraction contains 20 major polypeptides. A monoclonal antibody that was generated against the enriched complex reacts with an approximately 49-kDa protein and specifically immunoprecipitates in vitro deletion RNA editing activity. The protein recognized by the antibody was identified by mass spectrometry, and the corresponding gene, designated TbMP52, was cloned. Recombinant TbMP52 reacts with the monoclonal antibody. Another novel protein, TbMP48, which is similar to TbMP52, and its gene were also identified in the enriched complex. These results suggest that TbMP52 and TbMP48 are components of the RNA editing complex.

Uridylate addition and RNA ligation contribute to the specificity of kinetoplastid insertion RNA editing.

RNA editing in Trypanosoma brucei inserts and deletes uridylates (U's) in mitochondrial pre-mRNAs under the direction of guide RNAs (gRNAs). We report here the development of a novel in vitro precleaved editing assay and its use to study the gRNA specificity of the U addition and RNA ligation steps in insertion RNA editing. The 5' fragment of substrate RNA accumulated with the number of added U's specified by gRNA, and U addition products with more than the specified number of U's were rare. U addition up to the number specified occurred in the absence of ligation, but accumulation of U addition products was slowed. The 5' fragments with the correct number of added U's were preferentially ligated, apparently by adenylylated RNA ligase since exogenously added ATP was not required and since ligation was eliminated by treatment with pyrophosphate. gRNA-specified U addition was apparent in the absence of ligation when the pre-mRNA immediately upstream of the editing site was single stranded and more so when it was base paired with gRNA. These results suggest that both the U addition and RNA ligation steps contributed to the precision of RNA editing.

Evaluation of third generation anti-HCV enzyme immunoassays.

The Hepatitis C Virus (HCV) is a major cause of post transfusion hepatitis. The introduction of HCV antibody screening has reduced the risk of post transfusion hepatitis significantly. However, the test is yet to be used routinely in blood banks of several developing countries with limited resources. We have developed an Enzyme immunoassay using synthetic peptides. The test was compared to seven commercial tests available in the Indian market. The test was evaluated using a panel of 90 sera which were chosen from an earlier panel based on detection of HCV RNA by Reverse Transcription Polymerase Chain Reaction RT-PCR. In case of any discrepancy the sera were further analysed by Line immunoassay (LIA). The sensitivity of the in house EIA was 90%. The specificity of the commercial EIAs varied.

Hepatitis G virus in multitransfused thalassaemics from India.

Hepatitis G virus (HGV)/GB virus-C (GBV-C) has been identified as a blood-borne agent with disputed pathogenicity. This virus belongs to the flaviviridae with a distant relationship to hepatitis C virus (HCV). Genetically divergent HGV isolates have been reported from different parts of the world. This study describes the prevalence of HGV in multitransfused thalassaemic children in India and genomic sequence variations in 11 HGV isolates from the same geographical location. Hepatitis G virus RNA was detected in 39.7% multitransfused thalassaemic children. The seroprevalence of hepatitis B virus (HBV) and HCV was 23.8% and 17.1%, respectively, and 11.4% had dual infection. The nucleotide sequence of a 166 bp HGV genomic segment from the putative capsid-envelope region (nucleotide; nt 578-743) from 11 Indian isolates was compared to the sequences available in the nucleotide databases. The isolates from India were 81.3-94.5% homologous to the isolates from other parts of the world. On phylogenetic analysis, it was observed that HGV isolates from India may belong to two genetically divergent types.

Magnitude of hepatitis C virus infection in India: prevalence in healthy blood donors, acute and chronic liver diseases.

An enzyme immunoassay (EIA) was developed in-house for the detection of anti-hepatitis C virus (HCV) antibody against the prevailing genotypes in India. The specific reactivity of the test was compared with commercial second and third-generation EIAs and reverse transcription nested polymerase chain reaction (RT-nested PCR). Fifteen thousand nine hundred twenty-two healthy blood donors at the All India Institute of Medical Sciences (AIIMS), New Delhi, India, were screened for anti-HCV antibody. Two hundred ninety-five (1.85%) of these donors were positive. The screening was also used to determine how many patients with acute hepatitis and chronic liver diseases were positive for anti-HCV antibody. Five hundred sixty-four chronic liver disease patients were screened for anti-HCV antibody and 78 (13.83%) were found positive. Two hundred forty-seven sporadic acute viral hepatitis patients were screened for viral infection markers. Hepatitis B and E viruses (HBV and HEV) were the major etiologic agents. HCV was associated with 9% of the acute cases. Anti-HCV core IgM with HCV RNA detection were found to be helpful for the diagnosis of acute HCV infection.

Genotype determination of hepatitis C virus from northern India: identification of a new subtype.

Hepatitis C virus (HCV) shows substantial nucleotide sequence diversity distributed throughout the viral genome, with many variants showing only 68-79% overall sequence homology. This has led to problems in diagnosis of HCV using commercial immunoassays. Based on clustering of homologous sequences, various genotypes and subtypes of HCV have been described from different geographical regions. In the present study, 11 isolates from India were genotyped using sequence comparison for part of the non-structural (NS5) and structural (core) regions. Parts of the genome covering 451 bp (nt 9-459) of the core gene and a 249 bp fragment (nt 7959-8207) of the NS5 gene were reverse transcribed and amplified using nested polymerase chain reaction (RT-PCR). The amplified fragments were cloned and sequenced. The classification into genotypes was done on the basis of phylogenetic analysis. Four isolates showed sequence homology to type 1b. Two of the isolates were classified as type 3a. One isolate was classified as type 3b and the remaining four isolates were found to be variants of type 3 but did not belong to any designated subtype. On the basis of phylogenetic analysis two of the unclassified isolates were put into a new subtype of 3 named as 3g. In one of these variants, parts of a 5'-noncoding (5' NCR; 204 bp), envelope-E1 (435 bp), and NS3 (502 bp) regions were also amplified, cloned, and sequenced. This study demonstrates the type 3 variants including a new subtype (3g) to be the major cause of HCV infection in India.

Diagnosis of hepatitis C virus-associated chronic liver disease in India: comparison of HCV antibody assay with a polymerase chain reaction for the 5' noncoding region.

The relative value of an anti-hepatitis C virus (HCV) serological assay and reverse transcriptase-nested polymerase chain reaction assays (RT-PCR) were investigated for the constant 5' putative noncoding region of HCV for the diagnosis of HCV-associated chronic liver diseases in India. One hundred fifteen patients with biopsy proven chronic active hepatitis and 140 cases of cirrhosis of the liver were investigated for anti-HCV antibody using a second generation commercial enzyme-linked immunosorbent assay (ELISA). A proportion of these patients: 42 with chronic hepatitis and 27 with cirrhosis of the liver were analysed further for HCV RNA in the serum using RT-nested PCR assay. Thirty-three (12.9%) of the 255 patients were positive for anti-HCV antibody and 23 of 69 (33.3%) patients were positive for HCV RNA in serum. Fifteen of the 33 (45.5%) anti-HCV positive patients had HCV RNA in the serum. Eight of 36 (22.2%) HCV seronegative patients tested were found with HCV RNA. This indicates that the diagnosis of HCV infection is not possible if it is based solely on the available serodiagnostic tests. Inclusion of both assays improved the diagnostic efficiency, 18.8% (13/69) were negative for all virological markers associated with HBV and HCV infection. Since a majority of the chronic liver disease patients (143/255 [56%]) were seronegative for either HBV or HCV infection, it is significant that HCV RNA was detected in 38% (8/21) of a randomly selected group from these patients. The antibody assay and PCR were compared using interclass correlation (kappa statistics).(ABSTRACT TRUNCATED AT 250 WORDS)

Etiological role of hepatitis E virus in sporadic fulminant hepatitis.

Non-A, non-B hepatitis viruses have been implicated as the etiological agent(s) in up to 60% of patients with fulminant hepatitis. These agents are reported to induce a higher mortality than other causes of fulminant hepatitis. Hepatitis E virus (HEV) and hepatitis C virus (HCV) at present constitute the major identifiable non-A, non-B hepatitis agents. Of these, HEV has been established as the sole cause of epidemic hepatitis in Afro-Asian countries, and fulminant hepatitis has been recorded during such epidemics. However, in sporadic cases, the etiological role of HEV in fulminant hepatitis has remained uncertain. The role of HCV in acute liver disease and fulminant hepatitis remains unclear. The present study was undertaken to investigate the association of HEV and HCV in patients with fulminant hepatitis by direct detection of the viral genome using reverse transcription-polymerase chain reaction (RT-PCR). Serum samples from 50 serologically identified non-A, non-B fulminant hepatitis cases negative for cryptic hepatitis B virus (HBV) infection examined via PCR were tested for HEV and HCV RNA using RT-PCR. For HEV primers from the nonstructural region (ORF-1) were used, and for HCV primers from the highly conserved 5' untranslated regions were used. The products were analysed using agarose gel electrophoresis and confirmed by hybridisation with radiolabelled internal oligonucleotide probes. HEV was detected in 31 (62%) of the 50 fulminant non-A, non-B hepatitis cases. In 18 (36%) cases, HCV RNA was detected. In 11 (22%) of the HCV cases, the HEV genome was also amplified. In 20 (40%) cases, HEV was detected alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of methyl mercuric chloride treatment on haematological characteristics and erythrocyte morphology of Swiss mice.

Effects of methyl mercuric chloride (MMC) on the blood parameters of Swiss mice (Mus musculus) were studied. The mice received an initial dose of MMC (24 mg kg(-1) body wt) as intraperitoneal injection followed by a second similar dose on the 14th day of the first dose administration. Significant (p < or = 0.001) decreases in haemoglobin content, red blood cell (RBC) count and haematocrit value were observed in the MMC injected mice when compared to the control mice. The effect of the second dose was severe, after which no significant recovery in the values of these parameters was observed. The result also showed a high degree of mercury accumulation in the blood of the MMC exposed mice. Interesting features were marked in the erythrocyte morphology of the exposed mice. An initial shrinkage followed by swelling of the cells was observed after each injection. The outline of the exposed cells was irregular with beak like or small finger like projections. Rupturing and disintegration of the erythrocyte membrane, leading to erythrolysis, were also noticed.