Antimalarial Activity of Anacardium occidentale Leaf Extracts Against Plasmodium falciparum Transketolase (PfTK).

BACKGROUND: As per estimates by WHO in 2021 almost half of the world's population was at risk of malaria and > 0.6 million deaths were attributed to malaria. Therefore, the present study was aimed to explore the antimalarial activity of extracts derived from the leaves of the plant Anacardium occidentale L., which has been used traditionally for the treatment of malaria. Different extracts of A. occidentale leaves were prepared and tested for their inhibitory activity against recombinant P. falciparum transketolase (rPfTK) enzyme, in vitro. Further, growth inhibitory activity against cultivated blood stage P. falciparum parasites (3D7 strain), was studied using SYBR Green fluorescence-based in vitro assays. Acute toxicity of the hydro alcoholic extracts of leaves of A. occidentale (HELA) at different concentrations was evaluated on mice and Zebra fish embryos. HELA showed 75.45 ± 0.35% inhibitory activity against the recombinant PfTk and 99.31 ± 0.08% growth inhibition against intra-erythrocytic stages of P. falciparum at the maximum concentration (50 µg/ml) with IC50 of 4.17 ± 0.22 µg/ml. The toxicity test results showed that the heartbeat, somite formation, tail detachment and hatching of embryos were not affected when Zebra fish embryos were treated with 0.1 to 10 µg/ml of the extract. However, at higher concentrations of the extract, at 48 h (1000 µg/ml) and 96 h (100 µg/ml and 1000 µg/ml, respectively) there was no heartbeat in the fish embryos. In the acute oral toxicity tests performed on mice, the extract showed no toxicity up to 300 mg/kg body weight in mice. CONCLUSION: The hydro-alcoholic extract of leaves of A. occidentale L. showed potent antimalarial activity against blood stage P. falciparum. Based on the observed inhibitory activity on the transketolase enzyme of P. falciparum it is likely that this enzyme is the target for the development of bioactive molecules present in the plant extracts. The promising anti-malarial activity of purified compounds from leaves of A. occidentale needs to be further explored for development of new anti-malarial therapy.

A Cross-Sectional Study on Molecular Detection of Helicobacter pylori cytotoxin-associated gene A and 16SrRNA Gene from Gastric Biopsy Specimens.

INTRODUCTION: The aim of the study is relative proportion of cytotoxin-associated gene A (cagA) virulence marker in Helicobacter pylori isolates and gastric biopsy samples by polymerase chain reaction (PCR). METHODS: This cross-sectional study was conducted at a tertiary care hospital setting. Gastric biopsy tissues from 200 patients, suffering from upper gastrointestinal tract disorders, were examined for H. pylori infection using methods, such as hematoxylin and eosin (H and E) staining, 16S rRNA (Ribosomal ribonucleic acid), and cagA gene PCR. Chi-square and kappa statistics were used to find the association and agreement between the tests, respectively; P ≤ 0.05 was considered statistically significant. Screening tests' accuracy was calculated in terms of sensitivity and specificity along with positive and negative predictive values. RESULTS: Out of 200 patients, H. pylori was detected in 14.5%, 48.5%, and 31% patients by H and E staining, 16S rRNA, and cagA PCR, respectively. Sensitivity and specificity of cagA PCR as compared to H and E staining were 89.6% and 78.9%, respectively. CONCLUSIONS: CagA detection directly from biopsy specimen by PCR can potentially and rapidly determine the patient's status, especially when at a higher risk of peptic ulcer.

Triad of infective endocarditis, splenic abscess, and septicemia caused by Brucella melitensis.

A 40-year-old farmer from the district of North Karnataka who had received treatment for high fever of 8 days duration was admitted with fever, dyspnea, and poor general condition. Ultrasonography and echocardiogram revealed multiple splenic abscesses, vegetation on atrioventricular valve, aortic regurgitation (Grade I-II), and mitral valve regurgitation (Grade II-III), respectively. Brucella melitensis was detected in blood culture, and high titers of IgM and IgG anti-Brucella antibodies were observed in Brucella specific serological tests. The patient developed fulminant septicemia and succumbed due to multi-organ failure.

Utility of Serological Tests in the Era of Molecular Testing for Diagnosis of Human Brucellosis in Endemic Area with Limited Resources.

BACKGROUND: The culture has always been the gold standard test for diagnosis of human brucellosis but the conventional Brucella diagnostic tests viz. serology and culture are often beset with poor specificity & sensitivity respectively. The culture positivity rates for Brucella vary from 92% for bone marrow to 10% for non-blood samples and also dependent on the type of sample. The primary immune-determinant for Brucella species is the cell wall surface lipopolysaccharide, which is antigenically similar to other gram-negative rods. Hence, Brucella serological tests cross react with Escherichia coli 0116 and 0157, Salmonella urbana, Yersinia enterocolitica 0:9, Vibrio cholerae, Xanthomonas maltophilia and Afipia clevellandensis infections, which are common in developing countries also having higher incidence of brucellosis. AIM: The aim of the study was evaluation of conventional serological techniques and PCR for diagnosis of human brucellosis in and around north Karnataka which is endemic for brucellosis and patients often present with elevated base line antibody titers and confounding clinical manifestations. MATERIALS AND METHODS: Blood/serum samples of 400 patients suffering from acute undifferentiated fever (AUF) were subjected to culture, Brucella slide agglutination test (SAT), standard tube agglutination test (STAT coupled with 2 ME) and PCR. RESULTS: Of the 400 AUF patients, anti-Brucella antibodies were detected by SAT and STAT in serum of 35 and 34 patients respectively. IS711 gene for Brucella was identified in 32 patients by PCR. Twenty samples yielded Brucella in culture on biphasic medium with average incubation period of 9 days. All patients having titer of ≥ 160IU / ml in STAT were found positive by PCR also. CONCLUSION: Brucella STAT corroborated well with PCR results in all those cases where antibodies were present at least one dilution above cut-off value of 80 IU/ml. We probably need to raise cut-off titers to ≥160 IU/ml because of endemic region. The SAT was upheld as very good quick, easy to perform and economical screening test for human brucellosis. SAT as rapid screening test and STAT as more definitive test can be very well adopted by laboratories working in resource scarce settings for diagnosis of human brucellosis in absence of PCR even for population with normally elevated antibodies levels due to residing in Brucella endemic areas.

Microbial flora in chronic periodontitis: study at a tertiary health care center from north karnataka.

BACKGROUND AND OBJECTIVE: Periodontitis is a major public health problem in India with a prevalence of 60-80%. If untreated it acts as a risk factor for systemic diseases. Data on anaerobic periodontal microflora in the Indian population is very scarce. Hence, this study was undertaken to know the nature of oral microbiota in chronic periodontitis in this region of India and also the semiquantitative study in pre- and post-treatment group and to determine antibiotic susceptibility pattern for aerobic isolates. MATERIALS AND METHODS: The present study was conducted on 60 cases. Material was collected from the subgingival pockets in patients with chronic periodontitis attending the Periodontology, Outpatient Department. Clinical samples were transported to the laboratory in fluid thioglycollate medium. Initially Gram's stain and Fontana stains were done. Aerobic, anaerobic, and microaerophilic culture were put up. Antibiotic sensitivity test was done for aerobic isolates. RESULTS: Sixty samples yielded 121 isolates of which 78.34% were polymicrobial, 11.66% were monomicrobial and oral commensals were grown in 10% cases. Out of 121 isolates 91.74% were anaerobic, 7.43% were aerobic and 0.83% were microaerophilic. Fusobacterium species was the most common isolate among anaerobes. Using "paired t-test" "P" value was significant indicating significant reduction in colony count after phase-I periodontal therapy. CONCLUSION: This study has shown that anaerobic bacteria are important cause of chronic periodontitis, along with aerobes and microaerophilic organisms. Fusobacterium spp, Bacteroides fragilis, Porphyromonas spp and Prevotella intermedia are the most common anaerobic pathogens. Bacterial culture methods are still economical and gold standard.

Diagnosis of Trichomonas Vaginalis from Vaginal Specimens by Wet Mount Microscopy, In Pouch TV Culture System, and PCR.

BACKGROUND: In recent years, trichomoniasis has emerged as the most common sexually transmitted disease and limited data are available on the effective screening technique for the diagnosis of Trichomonas vaginalis. AIM: The aim was to compare and evaluate different diagnostic methods like wet mount microscopy, In Pouch TV culture, and Polymerase chain reaction (PCR) to establish which method or combination of methods was most effective for detection of Trichomonas vaginalis in vaginal swab specimens. SETTINGS AND DESIGN: This is a cross-sectional study. MATERIALS AND METHODS: A total of 200 patients complaining of vaginal discharge were included in the study. Three vaginal swabs were screened for trichomoniasis by wet mount microscopy, In Pouch TV culture system and PCR, using TVK3 and TVK7 specific primers. RESULTS: Of the 200 cases studied, 36 (18%) were positive by wet mount microscopy, 44 (22%) by In Pouch TV culture system and 60(30%) by PCR. Sensitivity and specificity of wet mount were 60% and 100%, respectively, whereas sensitivity and specificity of the In Pouch TV culture system were 73.33% and 100%, respectively when compared to PCR. CONCLUSION: Comparison of different methods showed that at least two techniques, such as wet mount microscopy and culture have a better chance of detection of T. vaginalis infection. Diagnosis of trichomoniasis by PCR was found to be highly specific and sensitive, but its availability and cost effectiveness limit its use in routine diagnostic laboratories.

PCR confirmation of cutaneous manifestation due to Brucella melitensis.

Human brucellosis, a zoonotic disease, is endemic in the Belgaum district, Karnataka, India. A male patient presented with a generalized itchy rash. Blood was sent for venereal disease research laboratory testing. Screening was carried out for Brucella antibodies following hospital policy and diagnosis was confirmed by PCR.