005548   Bhuvaneswari V;Rao K S;Nagini S (Biochemistry Dep, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamil Nadu) : Altered experssion of anti and proapoptotic proteins during chemoprevention of hamster buccal pouch carcinogenesis by tomato and garlic combination. Clin Chim Acta 2004, 350(1-2), 65-72.

005547   Bhaumik U;Mittal I C;Das P;Paria T (NO, Central Inland Fisheries Research Institute, Barrackpore, Kolkata-700 120) : Macrophytes and macrophyte associated fauna in a floodplain wetland system of West Bengal having both open and close characteristics. Envir Ecol 2004, 22(Spl2), 371-8.

Study of macrophytes and macrophyte associated fauna in floodplain wetlands reveals the energy being transferred through this phase and indicated tropic status of lake. Originally rich bottom coupled with a conducive physico-chemical environment encourages fast colonization by the macrophytes and macrophyte associated fauna. An investigation was carried out in a floodplain wetland having unique characteristics of both open and close system. Maintenance of a healthy aquatic environment and production or sufficient fish food organisms in the water bodies are factors of primary importance for boosting fish production. The study reveals that the seasonal fluctuations in water column were conspicuous and mostly dependent on the replenishment resources and volume. A mixed population of diversified fauna constituted the macrophytes and macrophyte associated fauna of the investigated ecosystems. The values of species diversity indicated that both the wetlands were favorable for harboring belanced population of the macrophyte associated fauna. The maximum macrophytes and macrophyte associated animal species diversity was observed in closed system. Mostly the diversity was observed to be maximum during winter seasons with coincidence of favorable temperature, dissolved oxygen and other physico-chemical parameters besides optimum solar penetration. Higher percentage of macrophytes and macrophyte associated fauna in closed system resulted in higher fish production (1,177.29 kg/ha per year) than that of open system (1,032.53 kg/ha per year).

1 illus, 5 tables, 28 ref

005546   Arockiam J P;Muruganandam M;Sudhahar A (Integrated Fish Culture Unit (IFCU), St. Xavier's College Palayamkott, , ) : Color light influence on serum cholesterol and sugar of broiler chicks. Envir Ecol 2004, 22(Spl2), 345-6.

Four groups of broiler chicks were exposed to white (control), green, blue and red light environment. At the end of the experiment serum sugar and cholesterol were estimated. The sugar and cholesterol were high in red light environment. Our previous studies proves that red light gives stress to birds so the increasing level of sugar, and cholesterol indicate birds in stress condition.

2 illus, 5 ref

005545   Agrawal N;Sachan R S (Soil Sci Dep, Coll of Agric, G B Pant Univ of Agric and Technol, Pantnagar-263 145) : Dynamics of soil microbial population influenced by wastes of sugar and yeast industries in a Mollisol. Indian J Envir Toxicol 2002, 12(1), 1-5.

The population of Azotobacter and Rhizobium was supported and that of Pesudomonas was inhibited by an increase of sugar industry effluents in the medium. The undiluted effluent of yeast industry resulted in a 323 per cent increase in Azotobacter population but had an inhibitory effect of Rhizobium (87.1% and Pseudomonas (55.9%) population. The sugar industry sludge @ 4.0% was found to be the best in creasing the population of Azotobacter and Rhizobium whereas Pseudomonas population was enhanced with the application upto 2.0% only. The pressmud application @ 2.0% caused the highest increase (29.32% over the control) in the Rhizobium population. A 20.47 per cent increase in Pseudomonas and 97.5% decrease in Azotobacter population were regrestered with 8.0% pressmud application in the soil. The Azotobacter, Rhizobium, and Pseudomonas populations were highest with 2.0, 1.0 and 0.5% yeast industry sludge application in soil, respectively over their respective control. The highest rate of sludge application @ 8.0% caused 93.2 and 24.3% reduction in Rhizobium and Pseudomonas population over their respective controls.

4 tables, 6 ref

004493   Syngenta Ltd (NO, , Syngenta European Regional Centre Priestley Road, Surrey Research Park Guildford, Surrey Gu27YH, U.K.) : Process for the preparation of an expression cassette. 0000, (NULL), (NULL).

004492   Glaxo Group Ltd (NO, , Glaxo Wellcome House, Berkeley Avenue, Greenford, Middlesex, UB6 Onn, U.K) : Assay methods. 0000, (NULL), (NULL).

004491   Weightman N C;Barnham R D;Dove M (Department of Microbiology, Friarage Hospital, Northallerton & Harroga, , North Yorkshire, U K) : Streptococcus milleri group bacteraemia in North Yorkshire England (1989-2000). Indian J med Res 2004, 119(May), 164-7.

The objective of the present study was to give an account of experience with the SMG bacteraemia over a period of 12 yr in North Yorkshire. The laboratory and clinical records of all clinically significant cases of SMG bacteraemia in our district general hospital catchment (combined population 260,000) were reviewed for the 12 yr period from 1989 to 2000. Viable isolates were recovered, speciesidentified, and minimum inhibitory concentration (MIC) determined. Twenty nine episodes of infection gave an annual incidence of 0.93 cases per 100,00 population. Infections included abscess, pneumonia, septic arthritis,genital and urinary tract infections, endocarditis and diffuse septicaemia. Patient ages ranged from 18 to 90 yr but most patients were elderly, 82 per cent had evident predisposing conditions and mortality rate was 10 per cent. Species determination of the 22 isolates showed S. anginosus (64%), S. constellatus (27%) and S. intermedius (9%). Nearly all isolates were non-haemolytic and Lancefield grouping showed carriage of the F antigen (in 41%), C (14%) and no detected group (45%). Most of the isolates were susceptible to the antimicrobials tested. Bacteraemia with SMG organisms was infrequent, ofen opportunistic and featured a low overall mortality rate. Intra-abdominal sepsis was the local feature in 16 (55%) of the patients and 9 (31%) of the total patient group showed abscess or empyema. There was a low rate of antibiotic resistance in these organisms from bloodstream infection.

5 tables, 22 ref

004490   Weightman N C;Barnham R D (Department of Microbiology, Friarage Hospital, Northallerton & North Yorkshire U K) : Report of unusual clinical appearance in bacteraemia with nonhaemolytic M-type 58 Streptococcus pyogenes. Indian J med Res 2004, 119(May), 17-21.

Case notes of the two patients were reviewed. Isolates obtained from clinical specimens were recovered and identified and cultured on horse blood agar to observe pattern of haemolysis. In the first case, of a 75 year-old man with leukaemia and a retropharyngeal abscess, the isolate was consistently non-haemolytic, probably due to a failure to produce streptolysin S as has been described before in a small number of reports involving various M-types. In the second case, of an 84 year-old woman with dermatitis and septicaemia, the organism was principally beta-haemolytic but with no haemolysis on aerobic culture where the colonies were well spaced, a phenomenon though to be associated with abundant production of serum opacity factor (OF). These cases are a reminder that misleading cultural appearances can occur with S. pyogenes and that OF positive strains can produce poor haemolysis on aerobic culture, or fail even to do so at all.

28 ref

004489   Ward P N;Leigh J A (Institute for Animal Health, Compton Laboratory, Compton, Berkshire, RG20 7NN, UK) : Genetic analysis of Streptococcus uberis plasminogen activators. Indian J med Res 2004, 119(May), 136-40.

A genetic screen of streptococcal and field isolates frequently associated with mastitis was undertaken to assess the distribution, chromosomal location and sequence variation of these putative virulence factors. Southern analysis of a diverse panel of streptococci and additional bacterial isolates frequently associated with bovine mastitis was performed pauA and pauB probes. Sequence variation of PauA was assessed at the protein level following nucleotide sequence analysis of pauA alleles amplified from isolates picked from different geographical locations. Observed plasminogen activators to be universally distributed amongst S. uberis. A pauA allele was identified in all but one strain of S. uberis. This strain had a pauB allele substituted for pauA at the same locus. The remarkably low level of sequence variation demonstrated by PauA was further restricted to a limited number of residues within the molecule. The high prevalence of PauA alleles in field isolates of S.uberis supported the observation that plasminogen activators are likely to confer an advantage with respect to colonization and growth. The findings of the present study support the theory that PauA plays a critical role in the pathogenesis of S.uberis.

2 illus, 1 table, 15 ref

004488   Varma N R S;Raha A R;Ross E;Mariana N S; Yusoff K (Dep of Biotechnol, Faculty of Food Science and Biotechnology, ) : Inducible expression of green fluorescene protein in Lactococcus lactis. Curr Sci 2004, 87(9-10), 1185-7.

2 illus, 18 ref

004487   Umur H;Ozalp A A (Dep of Mec Engng, Uludag Univ, Gorukle, Bursa, Turkey) : Laminar and turbulent forced convection in accelerating and decelerating curved flows. Curr Sci 2004, 87(9-10), 1237-44.

The article is concerned with accelerating and decelerating flows over straight, concave and donvex surfaces encompassing laminar and furbulent boundary layers. Flow and heat transfer experiments were carried in a blowing-type, low-speed wind tunnel and measurements indicated that the concave curvature and acceleration caused thinner boundary layers and augmented heat transfer rates, whereas the convex wall and deceleration resulted in the contrary. A new empirical equation is derived to model the combined effects of boundary layer development, surface curvature and pressure gradient on surface heat transfer values, which coincides with the experimental data within a deviation range of ±3%.

4 illus, 5 tables, 26 ref

004486   Teloni R;Von Hunolstein;Mariotti S;Donati S; Orefici G;Nisini R (Istituto Superiore di Sanita, Rome, , Italy) : Antibody classes & subclasses induced by mucosal immunization of mice with Streptococcus pyogenes M6 protein & oligodeoxynucleotides containing CpG motifs. Indian J med Res 2004, 119(May), 126-30.

Weused a recombinant M type 6 protein to immunize mice in the presence of synthetic oligodeoxynucleotides containing CpG motifs (immunostimulatory sequences: ISS) or cholera toxin (CT) to explore its possible usage in a mucosal vaccine. Mice were immunized by intranasal (in) or intradermal (id) administration with four doses at weekly intervals of M6-protein (10μg/mouse) with or without adjuvant (ISS, 10μg/mouse or CT, 0,5 μg/mouse). M6 specific antibodies were measured by enzyme linked immunosorbent assay using class and subclass specific monoclonal antibodies. The use of ISS induced in impressive anti M-protein serum IgG response but when id administered was not detectable in the absence of adjuvant. When used in, M-protein in the presence of both ISS and CT induced anti M-protein IgA in the bronchoalveolar lavage, as well as specific IgG in the serum. IgG were able to react with serotype M6 strains of S.pyogenes. The level of antibodies obtained by immunizing mice in with M-protein and CT was higher in comparison to M-protein and ISS. The analysis of anti-M protein specific IgG subclasses showed high levels of IgG1, IgG2a and IgG2b, and low levels of IgG3 when ISS were used as adjuvant. Thus, in the presenceof ISS, the ratio IgG2/IgG1 and (IgG2a+IgG3)/IgG1<62>1 indicated a type 1-like response obtained both in mucosally or systemically vaccinated mice. The study offers a reproducible model of anti-M protein vaccination that could be applied to test new antigenic formulations to induce an anti-group A Streptococcus (GAS) vaccination suitable for protection against the different diseases caused by this bacterium.

4 illus, 1 table, 16 ref

004485   Tagg J R (Department of Microbiology, University of Otago, P.O. Box 56) : Prevention of streptococcal pharyngitis by anti-streptococcus phogenes bacteriocin-like inhibitory substances (BLIS) produced by Streptococcus salivarius. Indian J med Res 2004, 119(May), 13-16.

The prototype BLIS-producing S. salivarius strains SN, 20P3, and K12 were isolated from tongue swabbings. BLIS P-typing was done using standard procedures. The BLIS molecules were purified and characterized. S. salivarius SN (P-type 226) produces a heat-laile muramidase. S. salivarius 20P3 (P-type 677) produces the 2315 Da lantibiotic salivaricin A and S salivarius K12 (P-type 777) produces two lantibiotics; salivaricin A2 (2368 Da) and salivaricin B (2733 Da). The P-type 777 S. salivarius strain produced salivaricin A2 and salivaricin B. The combined production of two anti-S. pyogenes BLIS activities by this strain indicates that it could be adopted as a colonizing strain in bacterial interference trials.

1 illus, 15 ref

004484   Suvorov A;Ustinovitch I;Meringova L; Grabovskaya K;Leontieva G;Vorobieva E;Totolian A (Institute of Experimental Medicine, Academy of Medical Sciences, Pavlova 12, St Petersburg 197376, Russia) : Construction of recombinant polypeptides based on beta antigen C (Bac) protein & their usage for protection against group B streptococcal infection. Indian J med Res 2004, 119(May), 228-32.

Localizes the region of IgA binding on Bac protein, to evaluate the role of one of the Bac protein regions MLKKIE in IgA binding, and to investigate the ability of Bac based recombinant proteins to generate protective antibodies against GBS infection. Recombinant proteins based on beta antigen C were generated after PCR amplification of the fractions of bac gene with the following cloning of the PCR products into expression plasmids. Recombinant peptides were tested for IgA binding by immunoprecipitation and Western blot. One of the recombinant proteins expressing IgA binding was used as an antigen for immunization of mice and for GBS protection studies. Several bac gene constructs were generated. Their ability to bind IgA varied dramatically depending on the size of the construct and location of the fragmenton the bac gene map. The smallest peptide expressing IgA binding was 14 kD in size. Amino acid substitutions in MLKKIE region facilitated IgA binding ability. Immunization of mice with recombinant Bac based peptide induced the appearance of anti-GBS antibody with high affinity level providing protection against GBS infection. Size dependence of Bac based recombinant peptides proved that the effective IgA binding required specific folding of the protein binding IgA. Region MLKKIE could not be considered as region, responsible for IgA binding. Generation of antibodies against Bac based recombinant peptides with high titre and affinity makes these proteins a potent candidates for generating a vaccine against GBS infection.

2 illus, 1 table, 16 ref

004483   Strakova L;Motlova J (National Reference Laboratory for Streptococci & Enterococci, National Institute of Public Health, Prague, Czech Republic) : Active surveillance of early onset disease due to group B streptococci in newborns. Indian J med Res 2004, 119(May), 205-7.

Early onset disease (EOD) due to group B streptococci (GBS) poses a serious threat in many countries. In the Czech Republic neither summarized data on the EOD incidence are available nor a nationwide prevention program has been initiated. The present surveillance was initiated to establish the incidence of EOD due to GBS in newborns in the Czech Republic, distribution of GBS serotypes and GBS susceptibility to antimicrobials. Both invasive and carrier GBS isolates from newborns and the data on teh newborns' clinical status and maternal colonization and intrapartum prophylaxis were colected from 30 microbiological and clinical centres all over the Czech Republic within prospective active surveillance. HCI extracts of the GBS strains were precipited with rabbit polysaccharide (I-VIII) and protein (c,R) antisera. Between January 2001 and Septembe 2002, GBS isolates from 239 full-term and 46 preterm newborns were collected. Of the 285 GBS positive newborns, 105 had invasive EOD, 42 showed suspected EOD, and in 56 clinical diagnosis was not specified. Eighty two GBS isolates were obtained from healthy colonized infants. The isolates obtained from newborns with confirmed invasive EOD were mostly of serotype III (42%), followed by serotypes V a Ia (13% each). Types Ia (26%), III (22%) and II (20%) were most frequent among the isolates from colonized inidividuals. Protein antigens (c protein, R protein) either coupled with polysaccharide or alone were found in 70 per cent (30 and 40%, respectively) of the study isolates. The incidence of EOD due to GBS found in the Czech Republic 0.7-1.0 per 1000 live births was comparable witht eh rates reported in the countries where the prevention programme has been implemented nation wide. Serotypes III, V and Ia prevailing among the isolates from Czech newborns with EOD belonged to those most frequently identified in the USA and Western European countries.

2 illus, 9 ref

004482   Smith B L;Flores A;Dechaine J;Krepela J; Bergdall A;Ferrieri P (Department of Laboratory Medicine & Pathology & Pediatrics, University of Minnesota Medical School, USA) : Gene encoding the group B streptococcal protein R4, its presence in clinical reference laboratory isolates & R4 protein pepsin sensitivity. Indian J med Res 2004, 119(May), 213-20.

Reports the gene encoding the R4 protein from a type III group B streptococcal isolate (76-043) well characterized in the laboratory. Trypsin extracted GBS protein were assayed for protease sensitivities by double-diffusion Ouchterlony using varying conditions for the enzyem pepsin. Standard haemoglobin assay was ued to examine pepsin enzymatic activity. Thirty clinical isolates of varying protein profiles identified by double-diffusion from our reference strain laboratory were screened by PCR and Southern technique. SDS-PAGE gel purified R4 amino acid sequences were determined and used to design oligonucleotide primers for screening a 76-043 genomic library. R4 was sensitive to pepsin at pH2 but appeared resistant at pH4, the reported pH used for Rib. By standard haemoglobin assay and trypsin extract studies of R4 protein, pepsin was shown to be active at pH2, yet easily inactivated; assays of GBS surface proteins are critical at pH2. Of the amino acids initially sequenced from R4, 88 per cent (61/69) showed identity to Rib; the r4 nucleotide sequence was identical to that of rib. All isolates with strong positive protein reactions for R4 were positive in both PCR and Southern technique, whereas isolates expressing alpha, beta, R1/R4, and R5 (BPS) protein profiles were not. Sequenced PCR products aligned with identity to the R4 and Rib nucleotide sequences and confirmed the identity of these proteins and their molecular sequences.

1 illus, 21 ref

004481   Skjaervold N K;Bergh K;Bevanger L (Norwegian University of Science & Technology, Department of Laboratory Medicine, Trondheim, Norway) : Distribution of PFGE types of invasive norwegian group B streptococci in relation to serotypes. Indian J med Res 2004, 119(May), 201-4.

A total of 78 epidemiological unrelated invasive GBS strains were examined by PFGE using Smal digested chromosomal DNA. Of these, 11 were nontypeable (NT) with regard to capsular polysaccharide (CPS) serotype. PFGE patterns were analyzed and classified in a dendrogram. 75 Strains were typeable by PFGE, and a total of 62 restriction profiles were identified. At an 85 per cent similarity level, 53 different PFGE patterns were identified. Within each serotype. PFGE patterns differed considerably, the largest degree of heterogeneity observed among type IV, Ia and II strains. Serotype Ib, III and V strains were more homogeneous. Strains with identical macrorestriction profiles belonged to the same CPS type, but varied with regard to serosubtypes. Any strain among the ones investigated showing a greater than 88 per cent similarity to a restriction profile in the detabase, could correctly be ascribed to a particular CPS type. Of the 11 NT strains 10 restriction profiles were found, two of which were identical to teh PFGE profile for a cluster of type V strains, and one profile were identical to the profile showed by a cluster of 5 type Ib strains. PFGE is a useful technique for classifying strains that are non-typeable by conventional serotyping.

2 illus, 9 ref

004480   Sinha S;Ashok KUmar;Gupta V;Kumar S;Singh V P;Raman R (BPS Lab, Centre for Diagnostic Haematology, , Sankatmochan, Varanasi 221 005) : Haemoglobinopathies-thalassaemias and abnormal haemoglobins in eastern Uttar Pradesh and adjoining districts of neighbouring states. Curr Sci 2004, 87(6), 775-80.

The haemoglobinopathies-thalassaemias and abnormal haemoglobins-constitute a major burden of genetic diseases in India. The study, based on index cases from 120 families detected between May 1999 and May 2003, highlights the ethnic distribution of haemoglobinopathies in regions in and around Varanasi comprising 8-10 districts of easter Uttar Pradesh and

3 tables, 17 ref

004479   Shet A;Kaplan E;Johnson D;Cleary P P (Department of Pediatrics, World Health Organization Collaborating Center for Reference & Researc, Minneapolis, MN 55455, USA) : Human immunogenicity studies on group A streptococcal C5a peptidase (SCPA) as a potential vaccine against group A streptococcal infections. Indian J med Res 2004, 119(May), 95-8.

Group A streptococcal C5a peptidase (SCPA) is a major virulence surface factor. Its highly conserved nature among all tested serotypes of group A streptococci (GAS) as well as animal protection studies make SCPA a prime vaccine candidate. The present study was undertaken to explore the human immunogenicity to SCPA usingan indirect enzyme-linked immunosorbent assay. Children (n=72) who had signs and symptoms of acute pharyngitis and had GAS isolated from the throat at initial visit were included. Acue and convalescent sera were collected 4 weeks apart. ELISA was performed using recombinant SCPA peptide as antigen. The mean convalescent anti-SCPA level was twice the level of mean acute anti-SCPA and the difference was statistically significant (P<60>0.0001). There was a rise in convalescent anti-SCPA in all children aged 2-12 yr. Our observations confirmed that SCPA was highly immunogenic in children infected with group A streptococcal pharyngitis. Further studies need to be done to characterize the immune response including antibody subclass.

20 ref

004478   Shakleina E;Drnitriev A;Tkacikova L;Suvorov A;Mikula I;Totolian A (Department of Microbiology, Institute of Experimental Medicine, acad Pavlov str., 12, st-Petersburg 197376, Russia) : Presence of insertion sequences (IS elements) in group B streptococci of bovine origin. Indian J med Res 2004, 119(May), 242-6.

Perform IS analysis of a large number of bovine GBS and to use the IS elements for classification and molecular epidemiology of GBS strains. A total of 101 GBS isolates obtained from the dairy cows were tested. These were analyzed by PCR and multiplex PCR. Southern hybridization was accomplished with the Enzo^TM DNA Labeling and Detection Kit. The computer techniques were used for selection of the specific primers and for analysis of the sizes of PCR products. GBS isolates collected at three diferent dairy farms were studied for the presence of IS elements. Multiplex PCR was used for the fast screening. It was found that IS861 presented in 29 GBS isolates (28.7%), IS1548 in 9 (8.9%), ISSa4 in 48 (47.5%) and IS1381 in 26 isolates (25.7%). A total of 28 bovine GBS isolates (27.7%) did not posses any of the IS elements, 36 (35.6%) possessed, 35 (34.7%) possessed two and 2 (1.9%) possessed three different IS elements. The GBS with four different IS elements were not found. Taken together, 10 different variants of GBS strains were discovered. Two out of 10 variants veing specific for 51 isolates (50.5%) were predominant in bovine GBS. The results of the study demonstrated that the presence of IS elements significantly varied in bovine GBS. The present data demonstrated that variants of IS elements present in GBS genome could be used as effective criteria for molecular epidemiology. In future this approach could probably by used as an additional tool for the epidemiological control and prevention of other bacterial infections.

2 illus, 2 tables, 13 ref

004477   Selvaraj P;Raja A;Kurian S M;Uma Devi K R; Narayanan P R (Tuberculosis Research Centre (ICMR), , Mayor V R Ramanathan Road, Chetput, Chennai 600 031) : HLA-DR phenotypes and IgG, IgA and IgM antibody responses to Mycobacterium tuberculosis culture filtrate and 30 kDa antigens in pulmonary tuberculosis. Curr Sci 2004, 87(6), 771-5.

The role of HLA-DR genetic make-up on the IgG, IgA and IgM antibody response to Mycobacterium tuberculosis culture filtrate and 30 kDa antigens was studied in pulmonary tuberculosis. The study was carried out in HLA-DR typed active pulmonary tuberculosis (ATB) patients (n=37), inactive (cured) pulmonary tuberculosis (ITB) patients (n=79) and normal healthy subjects (NHS;n=46). In ATB and ITB (cured) patients, IgG antibody (optical density at 490 nm for 1:3200 dilution) as measured by enzyme-linked immunosorbent assay was the predominant one than IgA and IgM antibodies. Increased IgG antibody titre to culture filtrate (P=0.03) and decreased titre to 30 kDa antigen were observed with HLA-DRI-positive ATB patients than non-DR1 (ATB) patients. Moreover, HLA-DR4- and HLA-DR6-positive ATB patients showed trends toward an increased IgG antibody response to 30kDa antigen than HLA-DR4-and HLA-DR6-negative (ATB) patients respectively. Significantly increased IgA antibody to 30kDa antigen was observed with HLA-DR1-positive ATB patients than non-DR1 patients (P=0.03). The study suggests that multiple HLA-DR molecules may regulate the IgG and IgA antibody responses to various proteins of M.tuberculosis. Moreover, HLA-DR phenotypes and increased IgG and IgA antibody titres may be useful to differentiate M.tuberculosis-infected subjects from normal subjects and cured patients with the same HLA-DR phenotypes or genetic make-up.

1 illus, 2 tables, 20 ref

004476   Sehlegel L;Grimont F;Grimont P A;Bouvet A (Centre National de Reference des Streptocoues, Service of Microbiologie, Hotel Dieu AP-HP, University Paris V U389, Institut Pasteur, Paris, France) : New group D streptococcal species. Indian J med Res 2004, 119(May), 252-6.

Improve the phenotypic delineation between species and to clarify their respective phylogenetic position. Physiological and genomic analyses were carried out in 84 representative strains of the group D streptococci. Biotypes were determined with the API 20 strep and rapid ID 32 STREP systems of identification. Quantitative DNA-DNA hybridization under stringent conditions and values of the ΔT_m allowed to delineate species and subspecies. The phylogenic position of the different genomic groups was determined by comparing the sequences of their 16S rDNA. Four DNA-clusters, including seven species or subspecies, were characterized. Differential associations of biochemical characters allowed their identification. S. equinus and the type strain of S. bovis belonged to a single species. S. gallolyticus, S. bovis biotype II.2, and S. macedonicus formed a single DNA-cluster including three different subspecies. These were designated as S. gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, and S. gallolyticus subsp. macedonicus. The two other DNA-clusters corresponded to the two subspeciesof S. infantarius, and to S. alactolyticus. The study presented a new classification associated with an identification scheme of group D streptococci. The changes in this classification demonstrate the interest of a polyphagic approach ot the bacterial identification.

1 tables, 22 ref

004475   Ray S K (Dep of Molecular Bio and Biotechnol, Tezpur Univ, Tejpur 784 028) : Riboswitch: a new mechanism of gene regulation in bacteria. Curr Sci 2004, 87(9-10), 1168-69.

^ssc2 illus, 8 ref

004474   Rastogi G;Dharne M;Bharde A;Pandav V S; Ghumatkar S V;Krishnamurthy R;Patole M S;Shouche Y S (Molecular Biology Unit, National Centre for Cell Science, Ganeshkhind, Pune 411 007) : Species determination and authentication of meat samples by mitochondrial 12S rRNA gene sequence analysis and conformation sensitive gel electrophoresis. Curr Sci 2004, 87(9-10), 1278-81.

Partial fragment of 450 bp of mitochondrial (mt) 12S rRNA gene was PCR-amplified and sequenced from cooked and uncooked meat samples of various known and unknown origin, viscera, and biological fluids like blood and semen of suspected origin, using universal primers. Homology search for obtained sequences was carried out using BLAST and NCBI. The technique was found to identify the meat tissue, semen, blood and viscera up to species level, even when the tissues were preserved at ambient temperature for prolonged periods of time. Further, to detect the adulteration in meat conformation-sensitive gel electrophoresis (CSGE) was used, which successfully detected adulteration in meat tissue even when the level of adulteration was as low as 10%. Overall, the simplicity of CSGE patterns could make this technique suitable for meat authentication in routine food forensic analysis.

4 illus, 2 tables, 12 ref

004473   Ranin L;Opavski N;Djukic S;Mijac V (Institute of Microbiology & Immunology, School of Medicine, University of Belgrade, Dr Subotioea I, 11000 Belg, Serbia) : Epidemiology of diseases caused by Streptococcus pyogenes in Serbia during a nine-year period (1991-1999). Indian J med Res 2004, 119(May), 155-9.

Carry out an epidemiological analysis of GAS infections in Serbia in a 9-yr period (1991-1999) and evaluation of susceptibility of GAS isolates obtained during the same period to penicillin and erythromycin. Occurrence of tonsillopharyngitis, scarlatina and rheumatic fever was analyzed and GAS carrier status in healthy children was examined over 1 9-yr period from 1991 to 1999. Susceptibility to penicillin and erythromycin was determined for 1657 GAS isolates obtained from patients diagnosed with pharyngitis or scarlet fever and 512 isolates from healthy carriers. M-type antigen was also determined in these isolates. The average incidences of tonsillopharyngitis and scarlet fever were 76.2 and 30.8 per cent respectively. A total of 166 cases of rheumatic fever registered. Per cent of carriers varied from 5.5 to 11.4 per cent over the study period. Predominating M serotpes among GAS isolates tested were M1, M3, M4, M6, M11, M12 and M18, depending on the source of clinical material and period of isolation. Antimicrobial susceptibility testing showed susceptibility to penicillin in all isolates tested and resistance to erythromycin in 2.41 per cent of the isolates. Although the fluctuations in incidence were noted during the nine-year period, the incidence of streptococcal tonsillopharyngitis is low but with a steady raise in Serbia. No significant changes in the incidence of scarlet fever and rheumatic fever were noted. Susceptibility to penicillin remained unchanged, but the number or erythromycin resistant strains have increased.

3 tables, 18 ref

004472   Randhawa V S;Kapoor L;Singh V;Mehta G (Department of Microbiology, Lady Hardinge Medical College, New Delhi) : Aminoglycoside resistance in enterococci isolated from paediatric septicaemia in a tertiary care hospital in north India. Indian J med Res 2004, 119(May), 77-9.

The study was carried out to find out the occurrence of high level aminoglycoside resistant isolates of enterococci in paediatric septicaemia cases in a tertiary care set up in north India. Blood of paediatric cases with a clinical diagnosis of septicaemia was cultured to isolate and identify enterococci. Agar screen method was uded to detect high level streptomycin and gentamicin resistance in these isolates. Vancomycin susceptibility of these isolates were determined as per the National Committee for Clinical Laboratory Standards (NCCLS) recommendations. Fifty one enterococal strains were isolated from 21 neonates, nine infants and 21 children with a clinical diagnosis of septicaemia. Sixty eight per cent of these isolates had high level gentamicin resistance and forty three per cent had high level streptomycin resistance. All the isolates with high level streptomycin resitance also had high level gentamicin resistance. More than ninety five per cent of these isolates were sensitive to vancomycin. The occurrence of high level gentamicin and high level streptomycin resistance in enterococcal isolates in our set up was high. This would require routine testing of the enterococcal isolates for high levelaminoglycoside resistance. Alternative treatment regimes need to be sought, besides prudent use of antibiotics.

1 table, 12 ref

004471   Ramoliya P J;Patel H M;Pandey A N (Biosciences Dep, Saurashtra University, Rajkot-360 005) : Effect of salinization of soil on growth and macro- and micro-nutrient accumulation in seedlings of Salvadaora persica (Salvadoraceae). For Eco Magmt 2004, 202(1-3), 181-93.

004470   Purushothaman S S;Park H S;Cleary P P (Department of Microbiology, University of Minnesota, MN 55455, USA) : Promotion of fibronectin independent invasion by C5a peptidase into epithelial cells in group A Streptococcus. Indian J med Res 2004, 119(May), 44-7.

Group A Streptococcus, causative agent of several clinical manifestations codes for multiple protein invasins which help the bacterium to enter non-phagocytic cells. C5a peptidase (SCPA) is a surface protein conserved among different serotypes of M1 strain. The present study was taken up to study SCPA promoted fibronectin independent entry of GAS into epithelial cells. An isogenic 90226 emm1∧AB (M1^-mutant was constructed with thermosensitive pGhost vector. This isogenic M1^- strain exhibited 54 per cent decreased invasion as compared to the bacteria incubated with control serum. Also, purified recombinant SCPA proteins blocked internalization of M1^- streptococci into HEp-2 cells. The M1^- strain invaded at the same efficiency in the presence or absence of fibronectin. These results suggest that SCPA acted as a potential invasin of group A streptococcus and promoted invasion independent of fibronectin.

2 illus, 11 ref

004469   Prymula R;Motlova J;Kriz P (Purkyne Military Medical Academy, Hradec Kralove & National Institute of Public Health, Prague, Czech Republic) : Comparison of Streptococcus pneumoniae serotypes causing acute otitis media & invasive disease in young children in the Czech Republic. Indian J med Res 2004, 119(May), 168-70.

Analyses the prevalence of Streptococcus pneumoniae serotypes in children suffering from acute otitis media or invasive pneumococcal disease and to compare a coverage of serotypes by individual pneumococcal vaccines. Children suffering from acute otitis media and invasive pneumococcal disease were analysed in the Czech Republic from October 1999 to Noember 2000. Serotyping was performed by the quellung technique using antisera from Statens Serum Institute (Denmark). The most frequent serotypes in patients with acute otitis media were 3, 19F, 23F, 14, 9V, 1, 6B, IIA and 28F. Vaccine coverage for the identified serotypes in acute otitis media patients was 52.1 per cent for the 7 valent vaccine, 57.8 per cent for the 9-valent vaccine and 75.7 per cent for the 11-valent form of the vaccine. In 108 patients with invasive pneumococcal disease, the most frequent serotypes were 6B, 9V, 14, 19F, 3 and 23F. Vaccine coverage for the identified serotypes in patients with invasive pneumococcal disease was 62 per cent for the 7 valent vaccine, 66.4 per cent for the 9-valent vacine and 7.5 per cent for the 11-valent form of the vaccine. Vaccine coverage for the identified serotypes for the 11-valent pneumococcal vaccine was better than the other two vaccines.

1 tables, 5 ref

004468   Proft T;Webb P D;Handley V;Fraser J D (Department of Molecular Medicine & Pathology, Faculty of Medical & Health Sciences University of Auckland, Private Bag 92019, Auckland, New Zealand) : Identification & characterization of the two novel streptococcal pyrogenic exotoxings SPE-L & SPE-M. Indian J med Res 2004, 119(May), 37-43.

The aim of this study was to search unfinished streptococcal genomes for novel superantigens, to generate recombinant proteins from potential open reading frames (ORFs) and to analyse them for superantigen activity. The microbial genome detabase was searched using a TBLASTN search programme. Genotyping of S. equi and S. pyogenes isolates was done using the specific primer pairs. The spe-l and spe-m genes were amplified by PCR. Two novel streptococcal superantigen genes (sepe-1 and sepe-m) were identified from the Streptococcus equi genomic database at the Sanger Centre. Genotyping of S. pyogenes isolates resulted in the detection of the orthologous genes spe-1 and spe-m in a restricted number of S. pyogenes isolates and revealed a link of spe-1 to the M89 serotype. Recombinant SPE-L and rSPE-M were highly mitogenic for human peripheral blood lymphocytes with half maximum responses at 1 pg/ml and 10 pg/ml, respectively. The results from competitive binding experiments suggest that both proteins bind MHC class II at the β-chain, butnot at the α-chain. The most common target for both toxins were human Vβl.1 expressing T cells. Seroconversion against SPE-L and SPE-M was observed in healthy blood donors. The two novel OFRs identified in both, S.equi and S.pyogenes, code for proteins that show typical superantigen features. The seroconversion seen in some blood donors suggest that the proteins are indeed produced by the bacteria. Interestingly, the spe-l gene is highly associated with S. pyogenes M89, which is linked to acute rheumatic fever in New Zealand.

3 illus, 21 tables

004467   Prathap G (CSIR Centre for Mathematical Modelling and Computer Simulation, , Bangalore 560 037) : Indian science slows down-V: the slack in the university sector. Curr Sci 2004, 87(6), 732-4.

One of the reasons for the poor performance of Indian science since Independence has been the slack in the university sector. In this commentary, some hard evidence is collected to show how the university sector is performing relative to the other sectors in the S&T enterprise.

4 illus, 2 tables, 3 ref

004466   Prasad S;Chellam R;Krishnaswamy J;Goyal S P (Cen for Ecological Sci, Indian Inst of Sci, Bangalore 560 012) : Frugivory of Phyllanthus emblica at Rajaji National park, North West India. Curr Sci 2004, 87(9-10), 1188-90.

1 illus, 2 tables, 16 ref

004465   Paul D;Chauhan A;Pandey G;Jain R K (Inst of Microbial Technology, , Sector 39-A, Chandigarh 160 036) : Degradation of p-hydroxybenzoate via protocatechuate in Arthrobacter protophormiae RKJ100 and Burkholderia cepacia RKJ200. Curr Sci 2004, 87(9-10), 1263-8.

The use of microorganisms is expected to be an effective tool for remediation of environments contaminated with various aromatic compounds. Aromatic compounds are degraded via diverse ring-cleavage pathways by soil microorganisms. Two bacterial strains isolated from pesticide-contaminated agricultural fields and identified as Arthrobacter protophormiae RKJ100 and Burkholderia cepacia RKJ200 were found to be capable of utilizing p-hydroxybenzoate (PHB) as a sole source of carbon and energy. Thin layer chromatography, gas chromatography and gas chromatography-mass spectrometry of the ethyl acetate extracts from the culture medium showed the p[resence of protocatechuate. Both the strains follow the ortho-cleavage pathway for the cotabolism of PC, since it was found that the terminal aromatic intermediate of the degradation pathway is cleaved to β-ketoadipate. Plasmid cured derivatives of A. protophormiae and B. cepacia were also investigated to determine the location of the genes involved in PHB degradation.

5 illus, 1 table, 27 ref

004464   Parveen M;Santosh Kumar (Bioscience Dep Faculty of Life Sciences, Barkatullah University, Bhopal-462 026) : Effects of dichlorvos on acetylcholinesterase in brain of rats. Indian J Envir Toxicol 2001, 11(2), 76-7.

The LD₅₀ of dichlorvos in male rats (Rattus norvegicus) was found to be 21.4 mg/kg b.wt. Rats were given intraperitoneally the sublethal doses of dichlorvos @ 3, 5 and 7 mg/kg. body weight. The brain acetylcholinesterase activity decreased at all the dose levels with the maximum decrease (71.26%) at 7 mg./kg. b.wt. dose. Dichlorvos toxicity caused a significant increase in ACh content in all the animals. Behavioural changes and signs of toxicity characterised by salivation and tremors were also observed and these started within 24 hrs. post treatment.

1 illus, 1 table, 11 ref

004463   Opavski N;Dukic S;Mijac V;Ranin L (Department of Microbiology & Immunology, School of Medicine, University of Belgrade, Dr Subotica 1, 11000 Belgrade, Serbia) : Influence of decreased penicillin susceptibility on growth rate of beta haemolytic streptococci. Indian J med Res 2004, 119(May), 237-41.

The study was undertaken to analyze the growth rates of BHS with decreased susceptibility to penicillin. Serial pasages in the medium with subinhibitory concentraction of penicillin were done to induce resistance to this antibiotic in 12 clinical isolates of BHS serogroups A, B, C and G. Both penicillin susceptible (parental) and variants with decreased susceptibility to penicillin (laboratoy strains) were grown in three different media and their growth rates were detemined by counting the number of bacterial colonies and by measuring optical density of bacterial culture. The lowest increase in minimal inhibitory concentratio (MIC) value for penicillin (8-16 times) was obtained in BHS group A isolates, while the increase in MIC values of BHS groups B, C and G strains was higher (64-128 times) and they reached the level of complete resistance. Laboratory variants differed significantly from parental in their morphological and cultural characteristics. There were no statistically significant differences between the growth rates of penicillin susceptible and variants with decreased susceptibility to penicillin, though a delay in multiplication of the laboratory strains during exponetial phase of growth was noted. Though significant differences in phenotypic characteristics of penicillin susceptible and laboratory variants were noted, the results of this study provides no support to the assumption that variants of BHS with decreased susceptibility to penicillin of BHS were incapable for normal growth. Further studies needs to be done to find out the association between the decreased susceptibility to penicillin in the BJS and decreased growth capability in these bacteria.

2 illus, 1 table, 8 ref

004462   Olive C;Batzloff M;Horvath A;Clair T;Yarwood P;Toth I;Good M F (Cooperative Research Centre for Vaccine Technology, The Queensland Institute of Medical ResearchPO Royal Brisbane Hospital, Brisbane, Queensland Australia) : Group a-streptococcal vaccine delivery by immunization with a self-adjuvanting M protein-based lipid core peptide construct. Indian J med Res 2004, 119(May), 88-94.

We investigated the lipid polylysine core peptide (LCP) system as a self-adjuvanting GAS vaccine delivery approach. An LCP-GAS construct was synthesised incorporating multiple copies of a protective peptide epitope (J8) from teh conserved carboxy terminal C-repeat region of the M protein. B10.BR mice were immunized parenterallywith the LCP-J8 construct, with or without conventional adjuvant, prior to the assessment of immunogenicity and the induction of serum opsonic antibodies. Our data demonstrated immunogenicity of LCP-J8 when coadministered in complete Freund's adjuvant (CFA), or administered in the absence of conventional adjuvant. In both cases, immunization led to the induction of high-titre J8 peptide-specific serum IgG antibody responses, and the induction of heterologous opsonic antibodies that did not cross-react with human heart tissue proteins. These data indicated the potential of a novel self-adjuvanting LCP vaccine delivery system incorporating a synthetic GAS M protein C-region peptide immunogen in the induction of broadly protective immune responses, and pointed to the potential application of this system in human vaccine development against infectious diseases.

3 illus, 28 ref

004461   Okhuni H (Clinical Lab Medca Japan Co Ltd, , Saitama, Tokyo, Japan) : Studies of recombinant streptococcal pyrogenic exotoxin B/cysteine protease (rSPE B/SCP) in the skin of guiena pigs & the release of histamine from cultured mast cells & basophilic leukocytes. Indian J med Res 2004, 119(May), 33-46.

Human mast cels and basophils were generated from CD34 positive cells isolated from cord blood and cultured in the presence of rIL-6,stem cell factor and/or rIL-3. The capacity of increasing capillary permeability of recombinant SPE B/SCP (rSPE B/SCP) was studied by using the skin of guinea pigs. Mitogenic activity to human T-cells of rSPE B/SCP was studied by incorporatio of ³H-thymidine. The levels of histamine in the plasma of patients with STSS and controls were measured by ELISA kit. rSPE B/SCP induced increased capillary permeability in the skin of guinea pigs, but both SPE A and SPE C did not exhibit such activity. Histamine was released from cultured human mast cells stimulated with rSPE B/SCP. Th rSPE B/SCP did not exhibit mitogenic activity to human T-cells. Three of the 7 patients with STSS showed higher levels of plasma histamine than those of normal subjects. The results suggest that increased capillary permeability and histamine release from mast cells induced by rSPE B/SCP might be involved in STSS and/or streptococcal infection of skin and mucous membrane.

3 illus, 4 ref

004460   Norton R;Smith H V;Wood N;Siegbrecht E;Rose A;Ketheesan N (Microbiology, Queensland Health Pathology Services, Townsville Hospital, Townsville Australia) : Invasive group A sterptococcal disease in North Queensland (1996-2001). Indian J med Res 2004, 119(May), 148-51.

Characterizes clinical and microbiological features of isolates obtained from invasive GAS infections in North Queensland, Australia between 1996 and 2001. Clinical and demographic data were collected prospectively. Isolates were biotyped, emm sequenced, M typed and tested for antibiotic sensitivity using E-test. Detection of the presence of the streptococcal pyrogenic exotoxin (spe) and fibronectin binding protein (prtF1) genes was also carried out. There were 109 isolates from blod and sterile sites. All isolates were sensitive to penicillin. Tetracycline and erythromycin resistance was seen in 11 and 2.7 per cent of isolates respectively. The isolates were evenly distributed by age and sex. The overall mortality was 7 per cent and there were 18 cases of streptococcal toxic shock syndrome (STSS) in which the mortality was 22 per cent. Indigenous patients had a crude incidence rate of 82.5 per 100,000 per year compared with 10.3 per 100,000 per year in the non-indigenous patients. There was no predominance of emm/ M type of association of spe type with STSS. There was also no relationship between the presence of the prtF1 gene and invasive disease. Invasive group A streptococci from North Queensland are similar to those from the Northern Territory of Australia in that no single strain is predominant. The indigenous population is overrepresented. Invasivences and the development of streptococal toxic shock is not related to the presence of the prtF1 gene or spe a or c.

2 tables, 16 ref

004459   Nagesha G K;Manohar B;Udaya Sankar K (Food Engineering Dep, Central Food Technological Research Institute, Mysore-57- -13) : Enzymatic esterification of free fatty acids of hydrolyzed soy deodorizer distillate in supercritical carbon dioxide. J Supercritical fluid 2004, 32(1-3), 137-145.

004458   Moyo S R;Maeland J A (Department of Medical Microbiology, Faculty of Medicine, University of Zimbabwe Medical School, P O Box A178, Avondale, Harare, Zimbabwe) : Recognition of different epitopes on the Cα & R4 proteins of group B streptococci by normal human & antibodies induced in animals. Indian J med Res 2004, 119(May), 224-7.

Levels of protective antibodies against Cα and R4 proteins of the group B streptococci (GBS) have been measured in humans. The findings indicated that the human anti Cα-and anti-R4 antibodies may recognize targets which are different from those recognized by antibodies raised in animals. In the present study normal human serum antibodies which target the GBS proteins Cα and R4 and immune anti-Cα and anti-R4 antibodies raised in animals were compared. The antigens were prepared by extractionof whole cells of GBS with trypsin and purified, and the testing was done by ELISA and Western blotting. The immune antibodies showed specificity for the corresponding protein and targeted proteins which had been denatured by hot sodium dodecyl sulphate (SDS) or by heating in a nearly neutral buffer. The human antibodies targeted asite(s) common to Cα and R4 and failed to bind to the denatured proteins. The bulk of antibodies in sera from healthy pregnant women was directed against the SDS and heat labile determinant(s). The present results indicated that the immune antibodies were directed against sequential epitopes and the normal human serum antibodies against epitopes determined by molecular conformation.

2 illus, 2 tables, 8 ref

004457   Motlova J;Strakova L;Urbaskova P;Sak P;Sever T (National Reference Laboratory for Streptococci & Enterococci, , Prague) : Vaginal & rectal carriage of Streptococcus agalactiae in the Czech republic: incidence, serotypes distribution & susceptibility to antibiotics. Indian J med Res 2004, 119(May), 84-7.

A total of 586 women at childbirth were screened for GBS carriage in vaginal and anorectal regions using the non-enrichment and selective culture media. The isolates were serotyped by precipitation with antisera raised agaist varios serotypes and antigenic extracts prepared according to Lancefield's modification. Mueller Hinton agar with 5 per cent defibrinated sheep blood was used for antimicrobial susceptibility testing. MIC values were evaluated according to the NCCLS criteria. Using selective media, GBS was detected in 172 (29.3%) of 586 women screened, vaginal and anorectal colonization was found in 21.7 and 24.4 per cent of them, respectively, concomitant vaginal and anorectal colonization was recorded in 16.5 per cent of the women studied. Serotypes III (33.2%), Ia (22.0%) and V (13.9%) prevailed among 172 isolates tested. All isolates were susceptible to penicillin, ampicillin and cefotaxime. The rates of GBS resistance to tetracycline, erythromycin and clindamycin were 83.9, 3.8 and 3.2 per cent, respectively. GBS carriage in pregnant women in the Czech Republic is rather high as compared with that reported in literature. The most frequent serotypes III, Ia and V, identified in GBS-colonized pregnant women in the Czech Republic, were among those predominant in the USA and Western Europe. Our findings confirm uniform susceptibility of GBS isolates from pregnant women to penicillin and other beta-lactam antibiotics tested. Resistance to erythromycin remains low in Czech Republic.

1 illus, 1 table, 15 ref

004456   Mihaila -Amrouche L;Schlegel L;Bouvet A (Centre National de Reference des Streptocoques-, Service de Microbiologie, Hotel Dieu, Assistance Publique Hopitaux de Paris) : Impact of susceptibility to antibiotics of streptococci & enterococci isolated from patients with infective endocarditis on antibiotic treatment. Indian J med Res 2004, 119(May), 80-3.

Streptococci and enterococci are the most frequent pathogens causing infective endocarditis. In order to update the recommendations for both curative and prophylaxis treatment, the susceptibility to antibiotics of the most prevalent species of Streptococcaceae isolated from the patients with infective endocarditis was determined. Streptococcal and enterococcal isolates (n=133) isolated from confirmed cases of infective endocarditis during a one-year prospective survey conducted in 1999 in France were studied. The identification of 106 streptococci and 27 enterococci to the species level was carried out by conventional methods. Their susceptibility to ten antibiotics used in curative or prophylactic treatment was measured. Minimal inhibitory concentrations were determined by agar dilution method. All the streptococcal and enterococcal isolates were susceptible to 4 mg/l or less of penicillin or amoxicillin. High levels of resistance to aminoglycosides were observed in two species, Streptococcus gallolyticus subsp. gallolyticus and Enterococcus faecallis. All isolates were susceptible to glycopeptides. Resistance to erythromycin, clindamycin,and pristinamycin was restricted to some species. Curative treatments recommended for streptococcal or enterococcal endocarditis, including penicillin, amoxicillin or vancomycin in association with gentamicin were found to be appropriate for 98.5 per cent of cases. The emergence of erythromycin resistance in oral streptococci led to the use of pristinamycin in oral prophylactic treatment in patients allergic to β-lactams.

2 tables, 15 ref

004455   Menon T;Shanmugasundaram S;Palani KUmar M; Girish KUmar C P (Department of Microbiology, Dr ALM PGIBMS, University of Madras, Department of Cardiology, Stanley Medical College, Chennai, ) : Group a streptococcal infections of the pharynx in a rural population in South India. Indian J med Res 2004, 119(May), 171-3.

Throat swabs were collected from children (5-14 yr) in the village of Orathur, Tamil Nadu and cultured on tryptose blood agar plates. Beta haemolytic streptococci were serogrouped using Streptex kit and biotyped based on their ability to ferment carbohydrates and production of β-glucuronidase enzyme. Blood samples were also collected and antibodies to streptolysin O demonstrated by latex agglutination tests. All the children were examined by a paediatrician; ECG and echocardiography were performed to assess cardiac function. Eighty of the 310 children included in the study had symptoms of acute respiratory infections; 16 of them grew beta haemolytic streptococci of which 8 belonged to group A (10%). Biotype 4 was most common. Antistreptolysin O (ASO) test did not correlate with culture results. Two of 310 children had rheumatic heart disease but both were culture negative. Pharyngeal cariage of group A streptococci was common in this population. The prevalence of rheumatic heart disease was 0.6 per cent. The study emphasized the need for active surveillance and characterization of GAS isolates.

2 tables, 7 ref

004454   Melin P;Verscharaegen;Mahieu L;Claeys G;De Mol P (Belgian Reference Laboratory for Group B Streptococci, Medical MIcrobiology, University Hospital of Liege, Belgium) : Towards a belgian consensus for prevention of perinatal group B streptococcal disease. Indian J med Res 2004, 119(May), 197-200.

Reports here the experience and findings of the meeting. In November 2001, ostetricians, neonatologists, microbiologists and infectious diseases specialists were invited to participate in a GBS symposium. International and Belgian speakers presented epidemiological aspects, argued comparative cost-effectivences of different approaches for prevention and debated technical and practical problems. Management of neonates with risk factors for GBS disease and progress in GBS vaccines were also included in the programme. Further results about Belgian obstetricians' practice and compliance to a policy for prevention of neonatal GBS diseases, as answered in tow mail surveys, were commented and discussed. In an interactive session at the end, each participant was asked to vote on the key points related to the different steps of the ideal prevention strategy to recommend. For the main questions, 94 per cent of participants choose a screening-based approach and 94 per cent shifted from the current use of ampicillin to penicillin as first choice for antimicrobial prophylaxis. Further, 79 per cent voted for an approach with integrated neonatal prophylaxis for selected neonates at high risk for GBS disease and 47 per cent voted for a strategy based on an intrapartum rapid screening-based approach. The state of the question by different speakers, the data from Belgian epidemiology, and the debate about cost-effectiveness of different approaches led to massive vote in favour of the universal screening-based approach. Based on these results, a working group has been appointed by the Ministry of Health to draft and edit Belgian recommendations for the prevention of perinatal GBS disease.

15 ref

004453   McMillan D J;Davies M R;Browning C L;Good M F;Sriprakash K S (The Cooperative Research Centre for Vaccine Technology, The Queensland Institute of Medical Research, PO Royal Brisbane Hospital, QLD 4029, Australia) : Prospecting for new group a streptococcal vaccine candidates. Indian J med Res 2004, 119(May), 121-5.

Most group A streptococcal (GAS) vaccine strategies focused on the surface M protein of the GAS. However, vaccine based on M protein have some drawbacks. Two approaches to identify new proteins and peptides that may have utility as vaccine candidates. A whole gel elution procedure was used to separate GAS surface antigens into 9 size fractionated pools. Mice were vaccinated with each pool and antibody titre, opsonic ability and protective capacity measured. In an alternative approach BioInformatics was used to identify putative GAS surface proteins. Peptides fromwithin these proteins were then selected on the basis of predicted antigenicity or location. These peptides were conjugated to keyhole lymphocyanin (KLH) and immunogenicity measured in a mouse model. One pool of GAS surfaceproteins (approximately 29kDa) induced antibodies that were both opsonic and potentially protective. Immunoflourescent microscopy demonstrated that these antibodies bound to the surface of M1 GAS. AMino acid sequencing subsequently identified superoxide dismutase as the major antigen in this pool. A BioInformatic search of the M1 GAS genome and subsequent analysis identified several peptides that fulfilled criteria as potential vaccine candidates. Each peptide when conjugated to KLH was able to induce a strong antibody response. Several new antigens were identified that may have potential as vaccine targets. A future GAS vaccine may have multiple peptide epitopes, providing protection against multiple GAS strains.

1 illus, 2 tables, 5 ref

004452   McDaniel L S;Thornton J A;Olga McDaniel D (Department of Microbiology, The University of Mississippi Medical Center, Jackson, MS 39216, USA) : Use of cDNA microarrays to analyze responses to pneumococcal virulence factors. Indian J med Res 2004, 119(May), 99-103.

To elucidate the role of microbial virulence factors in the host response to the pneumococcus, cDNA microarray analysis was used to identify genes in THP-1 cells, a human monocytic cell line, that are responsive to pneumococcal virulence factors. S. pneumoniae D39, a serotype 2 pneumococcus, and PLN an isogenic mutant of D39 that does not expres pneumolysin were used. Gene expression profiles elicited by both wild-type and mutant were compared with that of THP-1 cells not exposed to pneumococci. Results obtained from microarray analysis were confirmed and further characterized using reverse transcriptase (RT)-PCR, real-time RT-PCR, and ELISA. Genes in THP-1 cells that were responsive to the pneumococcus independent of the presence of the specific virulence factor, pneumolysin, were identified. THP-1 cell genes that were differentially expressed independent of pneumolysin included the ones involved in cell-to-cell signaling and antipathogen responses. Those that were responsive to pneumolysin included genes encoding adhesion molecules, chemokines, cytokine receptos, and cel cycle and apoptosis proteins. The global transcriptional response of naive monocytes to contact with the pneuococus was characterized and the utility of cDNA microarray analysis in elucidating the role of specific factors in host-pathogen interactions were demonstrated.

1 illus, 3 tables, 11 ref

004451   McArthur J;Schulze K;Chin J;Currie B J; Sriprakash K S;Talay S R;Chhatwal G S;Guzman C A;Walker M J (Department of Biological Sciences, University of Wollongong, Australia) : Immune responses of a liposome/ISCOM vaccine adjuvant against streptococcal fibronectin binding protein 1 (Sfb 1) in mice. Indian J med Res 2004, 119(May), 115-20.

The fibronectin binding protein Sfb 1 of streptococcus pyogenes is a well characterised antigen which induces protection against lethal challenge with group A streptococcus (GAS) when adjuvanted with cholera toxin B-submit (CTB). As an alternative to CTB adjuvanted intranasal immunisations investigated the immune responses generated in mice using Sfb 1 incorporated in to the skin and mucosal adjuvant SAMA4. Mice (BALB/c) were vaccinated intradermally with 100 μl of either SAMA4 (adjuvant only group) or SAMA4/Sfb 1 and were boosted 7 days later. Mice vaccinated with CTB based vaccines were immunised by intranasal inoculation with a mixture containing 30 μg Sfb 1 and 10 μg CTB on days 1, 3,5 and 15. At 14 days after the last booster immunisation the immune response was characterised and mice were challenged with 10⁸ CFU of S. pyogenes. Mice vaccinated with SAMA4/Sfb 1 elicited a Sfb1-specific IgG response in the sera that was significantly higher than that seen in control mice and mice immunised with the adjuvant only (P<60>0.05). No significant differences were seen for specific IgA antibodies in the sera in all groups examined. Compared with non-immunised and adjuvant only immunised controls, mice immunised with the Sfb1/SAMA4 vaccine exhibited a significant increase (P<60>0.05) in the number of Sfb1 reactive spleen cells in lymphoproliferation assays which were three fold higher than those seen for mice vaccinated with the Sfb1/CTB vaccine. Mice vaccinated with CTB/Sfb1 had the highest level of protection (80%) as where mice vaccinated with SAMA4 and SAMA4/Sfb1 displayed no protection (20% and 40%). These data suggest that the SAMA4 adjuvant used in this study fails to elicit protective immunity in BALB/c mice when used to adjuvant the known protective antigen Sfb1.

3 illus, 23 ref

004450   Mathew G;Gundu Rao T K;Sohoni P S;Karanth R V (Dep of Earth Sci, Indian Inst of Technol, Bombay Powai Mumbai 400 076) : ESR dating of interfault gypsum from Katrol hill range, Kachchh, Gujarat: implications for neotectonism. Curr Sci 2004, 87(9-10), 1269-74.

Chronological investigations on interfault fibrous gypsum using the technique of Electron Spin Resonance (ESR) dating, essentially to demonstrate the robustness of the technique as a dating tool on fibrous gypsum precipitates in a semi-arid region have been presented. Samples of fibrous gypsum collected from Katrol hill range, Kachchh consist of those from fractures and bedding planes, and those along the region resulted in erosion,unloding and dissolution of near-surface evaporite deposists and formation of extensional joints parallel to bedding and across. These openings became favourable sites for the fibrous gypsum to develop perpendicular to the wall of factures as well as fault plane. ESR chronometry studies deduce that gypsum growth along the bedding plane and fractures took place around 300 ka, while ESR dating studies on fault plane gypsum indicate an age around 70 ka. The young age of the normal fault plane indicates that it developed on account of the ongoing compressive stresses which resulted in normal faults perpendicular to the principal stress and later got immediately filled by sulphate-rich solutions. The ongoing compressive stresses active on the region have finally resulted in further uplift of the footwall back limb to its present topography

6 illus, 1 table, 22 ref

004449   Malke H;Steiner K (Institute for Molecular Biology, Friedrich Schiller University Jena, D-07745 Jena, Germany) : Control of streptokinase gene expression in group A & C streptococci by two-component regulators. Indian J med Res 2004, 119(May), 48-56.

Group A streptococci (GAS) and human isolates of group C streptococci (GCS) have the stable capacity to produce the plasminogen activator streptokinase, albeit with varying efficiency. This property is subject to control by two-two-component regulatory systems, FasCAX and CovRS, which act as activator and repressor, respectively. The work aims at balancing these oposing activities in GAS and GCS, and at clarifying the phylogenetic position of the FasAresponse regulator, the less understood regulator of the two systems. The GCS strain H46A and GAS strain NZ131 were used. Escherichia coli JM 109 was used as host for plasmid construction. Streptokinase activity of various wild type and mutant strains was measured. Phylogenetic tress of streptococcal FasA homologues were established. The streptokinase activities of the GAS strain NZ131 and the GCS strain H46A were attributable to more efficient CovR repressor action in NZ131 than in H46A. The FasA activator, on the other hand, functioned about equally efficient in the two strains. Phylogenetically, FasA homologues clustered distinctly in the proposed FasA-BlpR-ComE family of streptococcal response regulators and used the LytTR domain for DNAbinding. Assessing the apparent streptokinase activity of streptoccal strains require the dissection of the activities of the cov and fas systems. Although experimental evidence is still missing, FasA is closely related to a widely distributed family of streptococcal response regulators that is involved in behavioral processes, such as quorum sensing.

3 illus, 23 ref

004448   Madhavan H N;Malathi J;Rinku Joseph P;Yuichi M;Abraham S JK;Yoshioka H (Vision Res Foundation, , Sankara Nethralaya, 18 College Road, Chennai 600 006) : A study on the growth of continuous culture cell lines embedded in mebiol gel. Curr Sci 2004, 87(9-10), 1275-7.

Mebiol gel is an aqueous solution of newly developed thermo-reversible gelation polymer that turns into hydrogel upon warming and liquefies upon cooling. A preliminary experimental study of the growth pattern of Vero, DEp-2, HeLa, BHK-21, McCoy and CHO cell lines embedded in Mebiol gel was performed in 24-well tissue-culture plates with controls without the gel. The migration rate of cells through the gel, the rate of monolayer formation outside the gel and change in pH of themedium were determined. Rate of migration of cells to the margin of the gel varied from 4 to 8 h and the rate of monolayer formation outside the gel varied from 24 to 48 h between the cell lines. Cells grown out-side the gel were healthy, without showing any signs of degeneration or cytotoxicity in the presence of Mebiol gel and the pH was maintained during the seven-day study period compared to the controls. The results of the experiments designed in this preliminary study may be useful to develop methods to harvest single cells or groups of cells outside the gel for specifically planned procedures and develop techniques to preserve viable cells within the gel at room temperature.

3 illus, 7 ref