An endoscopic and histopathological assessment and correlation of endoscopic score with clinical activity index (CIBDAI) in the diagnosis of canine idiopathic inflammatory bowel disease.

Background: Inflammatory bowel disease (IBD) in dogs is often characterized by a relapsing and remitting clinical course. Determination of inflammatory activity is important for assessing the disease extent, severity, and tailoring appropriate treatment. Aims: The study was conducted to record the macroscopic and microscopic changes associated with IBD to assess the usefulness of endoscopy in the diagnosis of the disease and to correlate the clinical activity index (CIBDAI) with endoscopic score. Methods: Thirty-three dogs with idiopathic IBD were selected after thorough examination and exclusion. Gastroduodenoscopy and colonoscopy were performed to document the gross macroscopic intestinal lesions. Histopathology of endoscopic aided biopsy samples was used to confirm the disease. Results: Mucosal erythema and increased friability were the most predominant endoscopic findings in the stomach, duodenum, and colon of IBD dogs. Lymphoplasmacytic infiltration was predominant in the mucosal samples on histopathology and diffuse form of IBD is more common in canines. Gastroduodenoscopy and colonoscopy in combination with endoscopically guided biopsy and histopathology are of value in the assessment and diagnosis of IBD. There was no correlation between the clinical inflammatory bowel disease activity index (CIBDAI) with the endoscopic score. Conclusion: A diffuse form of IBD and colitis is more common in dogs in comparison to human IBD where the disease manifests in two distinct forms. Colonoscopy with ileal biopsy could act as a gold standard in the confirmation of diffuse IBD in dogs. CIBDAI can be used as a reliable measure of clinical signs of inflammation and histopathology can be used as a definitive diagnosis of intestinal inflammation.

Quantitative trait loci mapping for stomatal traits in interspecific hybrids of Eucalyptus.

Eucalyptus is an important industrial species with tolerance to drought and salt stress. Genetic improvement activities including quantitative trait loci (QTL) mapping for pulping and adventitious rooting traits are in progress, but no information is available on the genomic regions on adaptive traits such as stomatal characteristics. In this study, an interspecific cross between Eucalyptus tereticornis and E. grandis was generated for the development of genetic map and QTL identification for stomatal traits. Simple sequence repeats (SSRs), inter-simple sequence repeats (ISSRs) and sequence related amplified polymorphism (SRAP)markers were used for genotyping the F1 individuals. Parent-specific geneticmaps (female, 1023.56 cM;male, 1049.64cM) and consensus map (1049.4 cM) were developed. QTL analysis was carried out to identify the chromosomal regions affecting stomatal density, area and pore length in adaxial and abaxial leaf surface. Seven QTLs were identified with phenotypic variation of 11.36 to 27.30% for stomatal density, area and pore length. Correlation of stomatal traits when combined with growth and wood properties would have greater implications for generation of stress tolerant eucalypt hybrids with higher productivity and adaptability.

Cat fur mite Lynxacarus radovskyi in India.

A Persian cat with the complaint of lustreless dry coat, salt and pepper appearance was brought to the dermatology unit of Madras Veterinary College, Chennai. Microscopical examination of the tape impression and hair coat brushings revealed Lynxacarus radovskyi mites with eggs adhering to the hair shaft. The cat was treated with Ivermectin @ 300 µg/kg body weight subcutaneously once a week for 5 weeks after getting owner's consent. The cat was free from the parasites 5 weeks post treatment. Fomite and close contact might aid the spread of infestation. Isolation coupled with full course of treatment favours complete recovery from the infestation.

Detection and sequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates.

BACKGROUND: Staphylococcus pseudintermedius (SP) is the major pathogenic species of dogs involved in a wide variety of skin and soft tissue infections. The accessory gene regulator (agr) locus of Staphylococcus aureus has been extensively studied, and it influences the expression of many virulence genes. It encodes a two-component signal transduction system that leads to down-regulation of surface proteins and up-regulation of secreted proteins during in vitro growth of S. aureus. The objective of this study was to detect and sequence analyzing the AgrA, B, and D of SP isolated from canine skin infections. MATERIALS AND METHODS: In this study, we have isolated and identified SP from canine pyoderma and otitis cases by polymerase chain reaction (PCR) and confirmed by PCR-restriction fragment length polymorphism. Primers for SP agrA and agrBD genes were designed using online primer designing software and BLAST searched for its specificity. Amplification of the agr genes was carried out for 53 isolates of SP by PCR and sequencing of agrA, B, and D were carried out for five isolates and analyzed using DNAstar and Mega5.2 software. RESULTS: A total of 53 (59%) SP isolates were obtained from 90 samples. 15 isolates (28%) were confirmed to be methicillin-resistant SP (MRSP) with the detection of the mecA gene. Accessory gene regulator A, B, and D genes were detected in all the SP isolates. Complete nucleotide sequences of the above three genes for five isolates were submitted to GenBank, and their accession numbers are from KJ133557 to KJ133571. AgrA amino acid sequence analysis showed that it is mainly made of alpha-helices and is hydrophilic in nature. AgrB is a transmembrane protein, and AgrD encodes the precursor of the autoinducing peptide (AIP). Sequencing of the agrD gene revealed that the 5 canine SP strains tested could be divided into three Agr specificity groups (RIPTSTGFF, KIPTSTGFF, and RIPISTGFF) based on the putative AIP produced by each strain. The AIP of SP contains serine and produce lactone ring structured AIP. CONCLUSION: Presence of AgrA, B, and D in all SP isolates implies the importance of this regulatory system in the virulence genes expression of the SP bacteria. SP isolates can be typed based on the AgrD auto-inducible protein sequences as it is being carried out for typing of S. aureus isolates. However, further studies are required to elucidate the mechanism of controlling of virulence genes by agr gene locus in the pathogenesis of soft tissue infection by SP.

Automatic diagnosis of mammographic abnormalities based on hybrid features with learning classifier.

Breast cancer screening is currently performed by mammography, which is limited by overlying anatomy and dense breast tissue. Computer-aided detection (CAD) systems can serve as a double reader to improve radiologist performance. In this paper, we have applied a novel approach to segmentation of suspicious region by mammogram and classification based on hybrid features with learning classifier. We formulated differentiation of lesion from normal tissue as a supervised learning problem, and applied this learning method to develop the classification algorithm. The algorithm has been verified with 164 mammograms in the mini Mammographic Image Analysis Society database. The experimental results show that the detection method has a sensitivity of 94.5% at 0.26 false positives per image. The efficiency of algorithm is measured using free receiver operating characteristics curve and the results are highlighted. We conclude that CAD technology with learning classifier has the potential to help radiologists with the task of discriminating between lesion and normal tissues.

Antioxidant activities of bark extract from mangroves, Bruguiera cylindrica (L.) Blume and Ceriops decandra Perr.

OBJECTIVES: The antioxidant activities of two Indian mangrove plants, Bruguiera cylindrica and Ceriops decandra, were investigated. MATERIALS AND METHODS: Total phenolics and total flavonoid contents of the mangroves were determined using folin-ciocalteu reagent method and aluminium chloride method, respectively. Antioxidant capacity was assessed by the following methods: 1,1-diphenyl-2-picryl hydroxyl (DPPH.) quenching assay; 2,2'- azinobis-3-ethylbenzothiozoline-6-sulfonic acid (ABTS.(+)) cation decolorization test; scavenging capacity towards hydroxyl ion radicals (.OH); reductive capacity; and antihemolytic activity. RESULTS: The mangroves yielded 233.3 ± 0.062 and 283.31 ± 0.04 mg gallic acid equivalent/g phenolic contents and 11.6 ± 0.12 and 15.1 ± 0.02 mg quercetin equivalent/g flavonoid contents. The methanol extracts of both mangroves exhibited high antiradical activity against DPPH., ABTS.(+), and .OH radicals. The reductive capacity of the extracts increased with increasing concentration of samples. The extracts also inhibited H(2)O(2) induced hemolysis in cow blood erythrocytes. The antioxidant activities were found stronger than that of the reference standard, butylated hydroxy toluene (BHT). The antioxidant activity of mangrove plants was correlated with total phenolics and flavonoid contents. CONCLUSION: Both plants can be considered as good sources of natural antioxidants for medicinal uses. Further studies are necessary to isolate active principles responsible for the overall antioxidant activity of the extracts.

Comment on "The great Sumatra-Andaman earthquake of 26 December 2004".

Lay et al. (Research Articles, 20 May 2005, p. 1127) estimated a 600-km length for the tsunami source region. Adding tide-gauge data from Paradip, the northernmost of the Indian east-coast stations and therefore the most critical constraint on the northern extent of the source, we estimate that its length was greater by approximately 30%.

A prognostic insight on in vivo expression of interleukin-6 in uterine cervical cancer.

Interleukin-6 (IL-6) is a multifunctional cytokine with a growth promoting role in cancers. In an attempt to evaluate the prognostic significance of IL-6, we have studied the localization of this cytokine by immunohistochemical methods in normal, premalignant, and invasive cervical cancers. We stained frozen sections with a monoclonal antibody specific for human IL-6 and demonstrated that tumor cells show positivity with high intensity and stromal cells with moderate intensity. T cells and macrophages, components of the stroma that secrete IL-6, were also looked at. While T cells showed a weak positive correlation, CD68-positive macrophages showed significant positive correlation (P < 0.05) with IL-6 positivity in stroma. The patients were followed up for 40 months after treatment with standard radiotherapy regimen and showed residual tumor where pretreatment IL-6 levels were high. Our results suggest its possible candidature as a prognostic indicator of cervical cancer.

Identification and characterisation of a group of cervical carcinoma patients with profound downregulation of intratumoral Type 1 (IFNgamma) and Type 2 (IL-4) cytokine mRNA expression.

Type 1 cytokines, such as interferon gamma (IFNgamma) and interleukin-2 (IL-2), increase T cell-mediated immune responses and are considered to be beneficial for antitumour immunity. Type 2 cytokines, such as IL-4, IL-5, and IL-10, inhibit Type 1 responses and promote humoral responses. We have previously reported an association between low intratumoral IFNgamma mRNA levels and poor clinical outcome in patients with invasive cervical carcinoma. In this study, by using quantitative polymerase chain reaction (PCR), we identified a group of cervical carcinoma patients with undetectable intratumoral T cell-derived cytokine mRNAs, as IFNgamma, IL-4 and IL-17 expression could not be detected in 5, 25 and 8 of the 52 biopsies analysed, respectively. Global downregulation of Type 1 and Type 2 cytokines was observed in a subgroup of patients who more frequently presented advanced stage tumours. Biopsies of patients with no IFNgamma gene expression did not appear to be less infiltrated by T cells than control biopsies with measurable IFNgamma gene expression. These results clearly demonstrate that, in some clinical situations, the decrease in intratumoral Type 1 cytokines is not associated with a Type 2 polarisation, but rather reflects global deactivation of T cells at the tumour site. These data provide support for immunotherapy protocols designed to reverse the anergic state of T cells in cancer.

Characterization of multidrug resistance and monitoring of tumor response by combined 31P and 1H nuclear magnetic resonance spectroscopic analysis.

A combined 31P and 1H nuclear magnetic resonance (NMR) spectroscopic study was carried out in drug-sensitive and adriamycin- and mitoxantrone-resistant P388 murine leukemic cells. Typical spectral changes characteristic of multidrug resistance were observed in resistant cells compared to drug-sensitive cells. Quantitative comparison of phosphate metabolites ATP, phosphocreatine, phosphomonoesters and phosphodiesters revealed a significant alteration in the metabolism of resistant cells. The elevated levels of energy metabolites supported the energy-dependent process of drug efflux in resistant cells. An increased rate of glycolysis in resistant cells as indicated by the elevated lactate level further supported this. The near total loss of energy metabolites and marked decrease in phospholipid metabolites in sensitive cells upon treatment with drug compared to unaltered metabolite levels in resistant cells suggested that the spectral changes can reveal the subtle differences in tumor response between drug-sensitive and -resistant cells. The results substantiate the potential of this non-invasive method to characterize the multidrug resistance phenotype and monitor tumor response.

Inhibition by quercetin and luteolin of chromosomal alterations induced by salted, deep-fried fish and mutton in rats.

Previous studies from our laboratory have shown the clastogenic effects of long-term feeding on deep-fried fish and mutton in rat bone marrow cells. We report the chemopreventive action of two flavanoids, quercetin (Qn) and luteolin (Ln) against the induced mutagenicity by fish and mutton extracts. Groups of rats were treated with flavanoids through pre-, simultaneous- and post-treatment regimens and killed at the end of treatment. The bone marrow was removed and analysed for the presence of micronuclei (MN) and chromosome aberrations (CA). Pre-treatment showed most effectively a good inhibition of mutagenicity at every dose tested. Luteolin was a better protective agent than quercetin. It protected the cells against genetic damage to 93% in the micronucleus assay and to 95% in the chromosome aberrations induced by fish extract (p < 0.001 in both the groups). Mutton extract-induced micronuclei and chromosome aberrations were protected 85% and 90%, respectively, by luteolin and 79% and 76%, respectively, by quercetin. Our results tend to suggest that quercetin and luteolin are potential chemopreventive compounds.

Studies of the activities of lysosomal enzymes in serum and buccal pouch tissue of hamsters during 7,12-dimethylbenz[a]anthracene-induced carcinogenesis.

Alterations in the activities of certain lysosomal enzymes such as beta-D-galactosidase, beta-D-glucosidase, beta-D-glucuronidase, alpha-L-fucosidase, N-acetyl-beta-D-glucosaminidase, cathepsins B and D were studied in serum and tissue homogenates of buccal mucosa of hamsters treated with 0.5%, 7,12-dimethylbenz[a]anthracene (DMBA) in liquid paraffin. Among the enzymes studied, the activities of beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase showed significant elevation both in serum and tissue homogenates fro papilloma onwards and the elevations were progressive with the development of carcinomas. The elevations in the activities of alpha-D-fucosidase and cathepsin D were found to be significant from papillomatous tissue onwards whereas in serum they showed higher activities only in carcinoma stages. The activities of beta-D-glucosidase, beta-D-glucuronidase and cathepsin B in both serum and in tissue homogenate were elevated markedly only in carcinoma stages. It is suggested that beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase may be used as diagnostic markers for premalignant and malignant lesions of oral mucosa.

Induction of genotoxicity by salted deep-fried fish and mutton.

Salted, sun-dried and deep-fried fish and mutton were screened for their mutagenicity by the Ames test, single cell gel electrophoresis assay (SCG), chromosomal aberrations (CA), and micronucleus test. Fish and mutton given at 20% in the diet to the rats daily for 2 months resulted in cytogenetic damage which could not be repaired on withdrawal. However, the temporary damage was reversed at a 10% dose upon withdrawal after the same period. The maximum chromatid damage, found as breaks and gaps, was in agreement with the increased number of strand breaks. Treated rats also showed DNA strand breaks in hepatocytes and lymphocytes, more so in hepatocytes. Lime and onion extracts inhibited the nitrosation of fish and mutton, and were antitoxic.

Mutagenic potential of acute exposure to organophosphorus and organochlorine compounds.

The cytogenetic and cytotoxic effects of the pesticides methyl parathion, Bayleton and Hinosan were evaluated in mammalian test systems. The frequency of chromosome aberrations and micronuclei in bone marrow cells and the arginase enzyme profile in the liver tend to show the genotoxicity of organophosphorus and organochlorine pesticides in a single-exposure response study. Methyl parathion was the most hazardous among the three, showing definite pathology in the livers of treated rats.

Intervention of adriamycin induced free radical damage.

The cumulative cardiotoxicity of Adriamycin (ADR) is a constraint on its pharmacological use. The generation of drug induced oxygen radicals in heart cells lead to cardiac lipid membrane peroxidation. We studied the free radical scavenging potential of two compounds Oleanolic acid (OA) isolated from Eugenia jumbolana and Ursolic acid (UA) isolated from Ocimum sanctum against ADR induced lipid peroxidation both in liver and heart microsomes in vitro. In our attempt in the management of cardiotoxicity, we have identified OA as a strong protector against ADR induced lipid peroxidation and UA as a mild protector. Protection with OA was 49% and 21% in liver and heart microsomes respectively. On combined treatment, it increased to 69%. UA showed only 13% and 17% protection in liver and heart microsomes. Two methods for the microsome preparation, Calcium aggregation (CA) and Differential centrifugation (DC) were also compared. CA seems to give a better microsomal preparation though the protection was about the same.

Evaluation of sialic acid in lipoprotein fractions in fibrosarcoma bearing rats by 14C-glucose chase.

The changes in sialoglycoconjugates synthesis after administration of 14C-glucose to fibrosarcoma bearing rats were studied. After 1 hr treatment 14C-glucose tumor bearing rat synthesized hexosamine which contained more radioactivity when compared to sialic acid. On fractionation of lipoproteins, low density lipoproteins (LDL) contained more sialic acid. This was further confirmed by separating the lipoproteins, as more radioactivity incorporation was observed in the LDL fraction. It is noteworthy that inspite of the tumor, high density lipoproteins (HDL) did not show any change.

Protective effect of oleanolic acid and ursolic acid against lipid peroxidation.

In a search for plant products against cancer, the protective effect of two plant products, ursolic acid isolated from Ocimum sanctum and oleanolic acid from Eugenia jumbolana against free radical induced damage was studied. Three different standard systems viz., ascorbic acid, carbon tetrachloride, ADP/Iron were used to induce lipid peroxidation in isolated rat liver microsomes in vitro. Both oleanolic acid and ursolic acid offered remarkable protection of 90% and 60% respectively. Both the compounds did not induce lipid peroxidation by themselves that improved the therapeutic application.

Solid tumour chemotherapy using implantable collagen-poly (HEMA) hydrogel containing 5-fluorouracil.

Implantable collagen-poly(HEMA) hydrogels containing the anticancer drug 5-fluorouracil (5-FU) have been prepared and evaluated for their efficacy towards a solid tumour fibrosarcoma in Wistar rats. The tumour was developed by inoculation of a 10% tumour-cell suspension in the anterior aspect of the hind limb. Four groups were studied--untreated control, intratumoural injection of free 5-FU, subcutaneous implantation of placebo and implantation of 5-FU-bearing hydrogel pellets (10 mm diameter x 1 mm thick) containing the drug in close proximity to the tumour. The hydrogel showed an improved antitumour activity over free 5-FU as evidenced by the gross tumour weight assessments and by [3H]thymidine incorporation in-vitro. This was attributed to the controlled and slow release of 5-FU compared with free 5-FU over the same period of treatment. The implantation of hydrogel could thus be a potential alternative to free 5-FU therapy in the treatment of solid tumours such as fibrosarcoma.

Computerised algorithm of tumour-associated markers to monitor haematopoietic malignancy.

Hexosaminidase, polyamines and lipid-bound sialic acid are a few tumour-associated markers which are significantly elevated both in urine and serum of patients with haematopoietic malignancy, in direct correlation to dedifferentiation. Our computer database incorporates these parameters and predicts the clinical response based on the baseline values of these tumour-associated markers. This completely menu-driven user-friendly database can be routinely applied in tumour management and can access a billion records. The results of our evaluation confirm that most clinical assessments by a physician working blind are in agreement with the computer prediction, thus encouraging the use of microcomputers in patient management.