Synergistic effect of benzo triazole with polyethoxylated sorbitan monooleate in inhibiting corrosion of rebars and chloride diffusion through mortars.

It is found that mixture of 1,2,3 benzo triazole (BTAH) with polyethoxylated sorbitan monooleate, a non-ionic surface-active agent (NIS) effectively improves the properties of the cast concrete as well as significantly reduces the chloride induced corrosion of steel reinforced bars, when added in freshly prepared paste of mortar mixture. The addition of this mixture in the cast mortars is noted to reduce the water absorption in comparison to the control mortars cast using identical materials and under similar cast conditions. Electrochemical impedance spectroscopy and polarization studies of the rebars embedded in mortars and exposed in cement slurry have been performed to study the role of synergistic mixture on kinetics and mechanism of corrosion of rebars. The characterisation of corrosion products formed on the surface of rebars was carried out by X-ray diffraction, Scanning electron microscopy and EDX analysis. It is proposed that the synergistic boosting in protection is caused due to the shielding of NIS around anionic BTA-, thus minimizing their electrostatic repulsion. This facilitates the migration of additional ionic BTA towards the double layer which increases their concentration at the corroding interface leading to reduced susceptibility to corrosion.

Synergistic protection of borate and silicate salts composite for controlling the chloride-induced pitting and uniform corrosion of steel reinforcement bars embedded in mortars.

In this study, the efficacy of the combined effect of borate and silicate alkali metal salts added to mortars for controlling the chloride-induced uniform and localized corrosion of embedded steel rebars is examined. The individually added salts in mortars are found to have insignificant effects in terms of reducing the uniform corrosion rate and localized damage. However, their combination (0.50% sodium tetra borate + 0.10% sodium silicate added with respect to the weight of the binder) provides complete protection to reinforcements tested for long durations under wet/dry treatments with mortars in saline water and laboratory atmospheres. Electrochemical impedance spectroscopy, direct current cyclic polarization, polarization resistance, and visual observations are used for quantitative and qualitative evaluations of the protective effects of the tested additives. X-ray diffraction analysis, scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis of the corrosion products formed on the embedded steel surfaces help explain the possible mechanisms behind the considerable improvement in the inhibitive effects of a mixed composition of borate and silicate. This combination also improves the compressive strength and workability of the mixed concrete. The results reveal that the synergistic protection provided by a mixture of borate and silicate can be attributed to the co-deposition of an iron-boron + ferrosilicate + cortensitite (an iron-silicon phase) film on the rebar surface.

Role of phosphorus as micro alloying element and its effect on corrosion characteristics of steel rebars in concrete environment.

This communication reports the effect of phosphorus (P) added in micro concentration range in steel on kinetics, mechanism and growth of passive film in contact of chloride contaminated concrete. Electrochemical impedance spectroscopy, direct-current polarization, mass loss and Raman spectroscopic techniques were used to arrive at the findings. The results showed that an intentional addition of P in steel (0.064%) makes it more prone to uniform and localized corrosion (about 1.1 and 1.7 times) than the steel having low phosphorus (< 0.016%, present as tramp element) exposed under wet/dry conditions in simulated pore solution added with chloride and in the absence of this ion. A similar effect is also noted for the rebars embedded in mortars. Identification of corrosion products formed on steel rebars surface by Raman spectroscopy reveals thermodynamically stable maghemite and goethite phases on the surface of low P content steel. Unstable phase of lepidocrocite is recorded on the surface of higher phosphorus steel rebars. The findings are discussed with experimental evidence and taking clues from the published literature to arrive at plausible mechanism for this behaviour.

Chemotactic factor inducing Interleukin-8 (IL8) gene is transcriptionally elevated in experimental enterotoxaemia in goats caused by Clostridium perfringens type D.

The current study was designed to analyse the effects of experimental induction of enterotoxaemia through intra-duodenal inoculation of C. perfringens type D culture isolated from spontaneous outbreaks in goats. Twenty goats (6-9 month age) were divided into four groups and C. perfringens type D culture was inoculated intra-duodenally as per following: Group-I (whole cultures-WC), group-II (culture supernatant-CS), group-III (washed cells-WS), and group-IV (uninfected control-C). The treated animals were sacrificed after 72 h post infection (hpi), and necropsy showed gross changes including haemorrhages and congestion in the ileal and colon mucosa, pulmonary congestion and edema in lung. Kidney, brain and spleen exhibited severe to moderate congestion. Microscopic changes like haemorrhages, degenerative and necrotic changes in the mucosal epithelium of intestine and haemorrhages in kidney parenchyma were observed in the H&E stained sections. Lung alveolar sacs were filled with proteinaceous fluid. Immunohistochemistry revealed positive immunolabelling for etx (epsilon toxin) in the mucosa of intestine in WC and CS group. Control animals did not exhibit any significant gross or microscopic changes. PCR amplification of DNA extracted from intestinal tissues of WC and CS groups showed positive for etx gene demonstrating the production of epsilon toxin. Transcriptional responses in experimental groups were assessed by quantitative reverse transcription real time PCR (qRT-PCR). Genes including IL-1ß and IL2 showed up-regulation in all the experimental groups (WC, CS&WS). Specifically the toxin-based experimental groups (WC&CS) showed up-regulation of the gene responsible for chemotaxis viz. IL-8, while the washed cells group (WS) showed higher transcriptional response to Cathepsin-L (Cat-L) gene denoting the acute inflammatory response due to neutrophil elastase activity. These results take a cue on the evolving nature of the enterotoxaemia in goats due to various strains circulating in the field. The host response and its modulation due to the novel enterotoxaemia strains throws light on the current challenges in efficient control of the disease in goats.

Primary orbital reconstruction with selective laser melted core patient-specific implants: overview of 100 patients.

Contemporary advances in technology have enabled the transfer of industrial laser melting technology to surgery, and its use can improve the accuracy of orbital restoration. The aim of this study therefore was to evaluate the accuracy of primary orbital reconstruction with the use of selective laser melted, patient-specific implants and navigation. A total of 100 patients with complex orbital fractures were included. Planned orbital volumes were compared with those achieved, and angles were compared with the unaffected side. Analysis included the overlay of postoperative on planned images (iPlan® 3.0.5, Brainlab). The mean (SD) orbital volume of the unaffected side was 27.2 (2.8)ml in men and 25.0 (2.6)ml in women. Fractures that involved the posterior third of the orbital floor and comminuted fractures showed significant orbital enlargement (p=0.026). The mean (SD) reconstructed orbital volume was 26.9 (2.7)ml in men and 24.26 (2.5)ml in women. Three-dimensional analysis of the colour mapping showed minor deviations when compared with the unaffected side. The results suggest that a high degree of accuracy can be routinely achieved in these complex cases.

Prevalence and bacterial etiology of subclinical mastitis in goats reared in organized farms.

AIM: Assessment of the status of subclinical mastitis (SCM) in Jamunapari and Barbari goats in Indian organized farms, the involvement of bacterial pathogens and their sensitivity to antibiotics. MATERIALS AND METHODS: A total of 181 composite milk samples were aseptically collected from the apparently healthy Barbari (n=95) and Jamunapari (n=86) goats. The California mastitis test (CMT) and somatic cell count (SCC) were used to diagnose SCM. The milk samples with CMT scores of 0 and +1 were considered as negative, while the samples with the score of +2 or +3 were taken as positive, and further, the positive samples were used for the bacteriological examination. An antibiotic sensitivity test was performed by disk diffusion method using seven commercially available antibiotic discs. RESULTS: All the samples having CMT score of +2 or +3 demonstrated SCC more than 1 million. Overall, the prevalence of SCM in the goats was assessed as 19.89% (36/181). The prevalence of SCM in Barbari and Jamunapari goats was found as 24.21% (23/95) and 15.12% (13/86), respectively. Out of 11 isolates of Staphylococci, 9 isolates were identified as coagulase-negative Staphylococci (CNS), whereas 2 isolates were found as Staphylococcus aureus. The identified bacterial isolates (n=30) did not show antibiotic resistance. CONCLUSION: The current investigation showed the considerable prevalence of SCM among Jamunapari and Barbari goats which may have a negative impact on quantity and quality of the milk. CNS was found as the most prevalent cause of SCM in the goats. Negligible antibiotic resistance was found among the identified udder pathogens.

Molecular etiopathology of naturally occurring reproductive diseases in female goats.

AIM: The aim of the present study was to investigate the molecular etiopathology of occurrence of reproductive diseases in female goats. Reproductive diseases in goats account for major economic losses to goat farmers in terms of valuable loss of offspring and animal productivity. MATERIALS AND METHODS: A total of 660 female genitalia were examined for pathological conditions (macroscopic and microscopic lesions). The etiopathological study was carried out for the presence of pathogenic organisms such as Brucella, Chlamydia, and Campylobacter in the uterus and ovary. Based on the microscopic lesions, suspected samples were subjected to diagnostic polymerase chain reaction (PCR) for various etiological agents employing 16srRNA genus specific primers for Campylobacter and Chlamydophila and OMP31 gene-based PCR for Brucella melitensis and nested PCR using ITS-1 gene primers for Toxoplasma gondii. For Brucella suspected samples, immunohistochemistry (IHC) was also performed. RESULTS: In studied female genitalia, 108 (16.30%) showed gross abnormalities with overall 23.32% occurrence of pathological conditions (macroscopic and microscopic lesions). Pathological involvement of the uterus was the highest 68 (62.96%), followed by the ovaries 27 (25%) and other organs. Major uterine condition observed was endometritis (5.60%). In uterine infections, 35 (5.30%) samples were found positive for Campylobacter spp., 12 (1.81%) samples for B. melitensis, and 3 (0.45%) samples were positive for Chlamydophila spp. Among the samples positive for B. melitensis by PCR, 3 were found positive by IHC also. Corynebacterium ovis was detected by PCR using specific primers in a case of hydrosalpinx. It was concluded that many pathological lesions in female genitalia of functional significance play a major role in infertility in goats. CONCLUSION: The present study concluded that many pathological lesions in female genitalia of functional significance play a major role in infertility in goats.

Comparative diagnostic evaluation of OMP31 gene based TaqMan® real-time PCR assay with visual LAMP assay and indirect ELISA for caprine brucellosis.

Brucellosis is one of the leading causes of abortion in domestic animals that imposes costs on both economy and society. The disease is highly zoonotic and poses risk to animal handlers due to its zoonotic nature. It causes stillbirth, loss of kids and abortion in last term of pregnancy. Reproductive damage includes infertility in does and orchitis and epididymitis in breeding bucks, which result in high financial losses to farmers and the agriculture industry as a whole. It requires highly sensitive and specific assays to diagnose the disease at field level. In the current study, a visual loop-mediated isothermal amplification (LAMP) assay and the TaqMan® real-time PCR were developed with high sensitivity and specificity. For the TaqMan® probe, real-time PCR primers were developed using Omp31 gene as target and primers were designed using discontiguous conserved sequences of Omp31 gene. The Omp31 probes were designed by attaching 6-FAM reporter dye at the 5' end and BHQ-1 quencher at the 3' end. Published primers were used for visual LAMP assay targeting the Omp25 gene. Sensitivity of the standardized visual LAMP assay and TaqMan® real-time PCR assay was determined by serial dilution of positive Brucella melitensis DNA (102 to 10-4 ng) obtained from standard culture. The TaqMan® probe real-time assay can detect as low as 100 fg of B. melitensis DNA, whereas culture from vaginal swab washings has a limit of detection (LOD) of only 1 cfu/ml. Similarly, the visual LAMP assay can detect as low as 10 fg of B. melitensis DNA as compared to an LOD of 30 cfu/ml from culture of vaginal swab washings. Both assays were compared with serological tests (serum tube agglutination test (STAT) and indirect enzyme-linked immunosorbent assay (iELISA)) for diagnostic sensitivity and specificity. Diagnostic sensitivities and specificities for TaqMan® real-time PCR vs. LAMP assays were 98 and 100% vs. 100 and 97.8%, respectively. Results of visual LAMP assay indicated that LAMP is a fast, specific, sensitive, inexpensive and suitable method for diagnosis of B. melitensis infection under field conditions. On the other hand, Omp31 TaqMan® probe real-time assay can be used in conjunction with the other field-based diagnostic tests due to its high specificity.

Prevalence of hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus and hepatitis E virus as causes of acute viral hepatitis in North India: a hospital based study.

CONTEXT: Acute viral hepatitis (AVH) is a major public health problem and is an important cause of morbidity and mortality. AIM: The aim of the present study is to determine the prevalence of hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV) and hepatitis E virus (HEV) as causes of AVH in a tertiary care hospital of North India. SETTINGS AND DESIGN: Blood samples and clinical information was collected from cases of AVH referred to the Grade I viral diagnostic laboratory over a 1-year period. SUBJECTS AND METHODS: Samples were tested for hepatitis B surface antigen, anti-HCV total antibodies, anti-HAV immunoglobulin M (IgM) and anti-HEV IgM by the enzyme-linked immunosorbent assay. PCR for nucleic acid detection of HBV and HCV was also carried out. Those positive for HBV infection were tested for anti-HDV antibodies. STATISTICAL ANALYSIS USED: Fisher's exact test was used and a P < 0.05 was considered to be statistically significant. RESULTS: Of the 267 viral hepatitis cases, 62 (23.22%) patients presented as acute hepatic failure. HAV (26.96%) was identified as the most common cause of acute hepatitis followed by HEV (17.97%), HBV (16.10%) and HCV (11.98%). Co-infections with more than one virus were present in 34 cases; HAV-HEV co-infection being the most common. HEV was the most important cause of acute hepatic failure followed by co-infection with HAV and HEV. An indication towards epidemiological shift of HAV infection from children to adults with a rise in HAV prevalence was seen. CONCLUSIONS: To the best of our knowledge, this is the first report indicating epidemiological shift of HAV in Uttar Pradesh.

TB or not TB? Difficulties in the diagnosis of tuberculosis in HIV-negative immigrants to Germany.

BACKGROUND: Immigrants to Germany and their children are at particular risk for tuberculosis (TB). - METHODS: 35 Patients (10 male / 25 female aged 2 - 59 years (median 33 years) originating mostly from high incidence countries in Asia (19 (54.3%)) in Africa (14 (40.0%) and East Europe (2 (5.7%)), attended at the Tropical Medicine Unit were analysed. - RESULTS: Primary clinical presentation was most frequently lymphadenitis (13 (37.1%)). Other organs involved included bones (7 (20.0%)), central nervous system (5 (14.3%)), urogenital organs (3 (8.6%)), lung (3 (8.6%)), mediastinum, (2 (5.7%)) and abdomen (2 (5.7%)). ESR was abnormal in 21/28 (75.0%), CRP in 20/35 (57.1%), and protein electrophoresis in 22/26 (84.6%) cases. The tuberculin skin test was strongly positive in all 15 cases where the test had been performed. Tuberculosis interferon gamma release assay (TB-IGRA) was positive in all 35 cases (100%). PCR for nucleic acids of Mycobacterium (M.) tuberculosis complex was positive in only 7/20 (35.0%) cases. M. tuberculosis was identified in 32/35 (91.4%), M. bovis in 2 (5.7%) cases. 1 case was diagnosed clinically. All patients were negative for HIV. Typical histopathology was seen in the 29 cases, where biopsies had been taken. Chest-X-ray did not reveal specific pulmonary lesions in the majority of cases (22/35 (62.9%)). Diagnosis of TB was mostly delayed (4 to 299 weeks, (median 8)). The most frequent primary suspicion was a malignancy (17/35 (48.6%)) while TB was initially suspected in 5 cases only. Diagnosis of TB is impeded by its multifaceted presentation especially in immigrants.

Cloning, expression, purification, crystallization and preliminary X-ray studies of a secreted lectin (Rv1419) from Mycobacterium tuberculosis.

A secreted lectin, Rv1419, from Mycobacterium tuberculosis has been cloned, expressed, purified and crystallized and the crystals have been characterized. This represents the first X-ray investigation of a lectin or lectin-like molecule from the pathogen. The cubic crystals contain one molecule in the asymmetric unit. Sequence comparisons indicate that the lectin has a ß-trefoil fold and belongs to a well characterized family of carbohydrate-binding modules. Structural analysis of the crystals is in progress.

Investigations on anti-Aspergillus properties of bacterial products.

AIMS: To investigate the anti-Aspergillus properties of bacterial products. METHODS AND RESULTS: In the present study, 12 bacterial strains were screened for antifungal activity against Aspergilli. The culture supernatant and lysates of Pseudomonas aeruginosa, Bacillus cereus, Escherichia coli (BL21, DH5alpha, HB101, XL Blue), Klebsiella pneumoniae, Streptomyces thermonitrificans, Streptococcus pneumoniae, Enterobacter aerogenes, Staphylococcus aureus and Salmonella typhi were examined for antifungal activity in protein concentration ranging from 1000.0 to 7.8 microg ml-1 using microbroth dilution assay. The lysate of Salm. typhi and E. coli BL21 exhibited the maximum activity against Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. Their in vitro minimum inhibitory concentrations (MICs) were found to be 15.6-31.2 microg ml-1 by microbroth dilution and spore germination inhibition assays. In disc diffusion assay, a concentration of 3.1 microg disc-1 of Salm. typhi lysate showed significant activity against Aspergilli. Escherichia coli BL21 exhibited similar activity at 6.2 microg disc-1. The work on identification of molecule endowed with antimycotic properties is in progress. CONCLUSION: The products of Salm. typhi and E. coli demonstrated significant activity against Aspergillus species. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that E. coli has been reported for anti-Aspergillus activity. It could be an important source of biologically active compounds useful for developing better new antifungal drugs/or probiotics.

Unusual sugar specificity of banana lectin from Musa paradisiaca and its probable evolutionary origin. Crystallographic and modelling studies.

The crystal structure of a complex of methyl-alpha-D-mannoside with banana lectin from Musa paradisiaca reveals two primary binding sites in the lectin, unlike in other lectins with beta-prism I fold which essentially consists of three Greek key motifs. It has been suggested that the fold evolved through successive gene duplication and fusion of an ancestral Greek key motif. In other lectins, all from dicots, the primary binding site exists on one of the three motifs in the three-fold symmetric molecule. Banana is a monocot, and the three motifs have not diverged enough to obliterate sequence similarity among them. Two Greek key motifs in it carry one primary binding site each. A common secondary binding site exists on the third Greek key. Modelling shows that both the primary sites can support 1-2, 1-3, and 1-6 linked mannosides with the second residue interacting in each case primarily with the secondary binding site. Modelling also readily leads to a bound branched mannopentose with the nonreducing ends of the two branches anchored at the two primary binding sites, providing a structural explanation for the lectin's specificity for branched alpha-mannans. A comparison of the dimeric banana lectin with other beta-prism I fold lectins, provides interesting insights into the variability in their quaternary structure.

Purification, crystallization and preliminary X-ray structure analysis of the banana lectin from Musa paradisiaca.

The banana lectin from Musa paradisiaca, MW 29.4 kDa, has been isolated, purified and crystallized. The trigonal crystals contain one dimeric molecule in the asymmetric unit. The structure has been solved using molecular replacement to a resolution of 3 A. The structure of the subunit is similar to that of jacalin-like lectins.