Experimental Evolution Studies in Φ6 Cystovirus.

Experimental evolution studies, in which biological populations are evolved in a specific environment over time, can address questions about the nature of spontaneous mutations, responses to selection, and the origins and maintenance of novel traits. Here, we review more than 30 years of experimental evolution studies using the bacteriophage (phage) Φ6 cystovirus. Similar to many lab-studied bacteriophages, Φ6 has a high mutation rate, large population size, fast generation time, and can be genetically engineered or cryogenically frozen, which facilitates its rapid evolution in the laboratory and the subsequent characterization of the effects of its mutations. Moreover, its segmented RNA genome, outer membrane, and capacity for multiple phages to coinfect a single host cell make Φ6 a good non-pathogenic model for investigating the evolution of RNA viruses that infect humans. We describe experiments that used Φ6 to address the fitness effects of spontaneous mutations, the consequences of evolution in the presence of coinfection, the evolution of host ranges, and mechanisms and consequences of the evolution of thermostability. We highlight open areas of inquiry where further experimentation on Φ6 could inform predictions for pathogenic viruses.

Development and diagnostic evaluation of indirect enzyme linked immunosorbent assay for Epidermophyton floccosum infection in humans.

BACKGROUND: Epidermophyton floccosum is an anthrophophilic dermatophyte widely distributed in the tropics and subtropics. It can invade keratinized tissues of humans and cause superficial mycoses called dermatophytosis (ringworm). OBJECTIVE: The main objective of this study was to develop an in-house indirect enzyme-linked immunosorbent assay (ELISA) and to evaluate its performance for the immunological diagnosis of E. floccosum infection in humans. METHODS: An in-house indirect ELISA was developed using partially purified E. floccosum antigens, pre immunized rabbit serum as negative control, immunized rabbit polyclonal antibodies as positive control, enzyme labeled goat anti rabbit antibodies and goat anti human antibodies. A total of 50 serum samples from E. floccosum infected patients as confirmed by direct microscopy and culture and 30 samples from humans without history of dermatophyte infection that served as controls were used to evaluate the performance of an in-house indirect ELISA developed in this study. Analytical and diagnostic performance characteristics were determined to evaluate its diagnostic value. RESULTS: The diagnostic sensitivity, specificity, positive and negative predictive values of E. floccosum indirect ELISA were 90.00 %, 83.33 %, 90.83 %, 83.83 % respectively. The performance of indirect ELISA assay was compared with gold standard diagnostic tests such as KOH hydrolysis test and fungal culture. The correlation coefficients of E. floccosum indirect ELISA with KOH hydrolysis and fungal culture method were 0.612** and 0.826** (P < 0.01) respectively indicating significant correlation between these tests. CONCLUSION: This revealed the great potentiality of E. floccosum indirect ELISA in early, specific and precise detection of E. floccosum infection in humans.

Elicitor mediated enhancement of shoot biomass and lupeol production in Hemidesmus indicus (L.) R. Br. ex. Schult. and Tylophora indica (Burm. F.) Merrill using yeast extract and salicylic acid.

Hemidesmus indicus (L.) R. Br. ex Schult. and Tylophora indica (Burm. F.) Merrill shoot cultures were treated with different concentrations of yeast extract (YE; 25-200 mg/L) and salicylic acid (SA; 50-200 µM), and their effect on lupeol production was assessed. The maximum dry weight (DW) biomass was recorded when H. indicus shoots were treated with SA (50 µM) and T. indica shoots with YE (200 mg/L). Highest lupeol yield (335.40 ± 0.04 µg/g DW) was obtained in H. indicus shoots after treatment with 50 µM of SA for 3 weeks. Whereas in T. indica, maximum lupeol content (584.26 ± 8.14 µg/g DW) was recorded by giving treatment with 25 µM of SA for 6 weeks.

Development of indirect ELISA and its evaluation in comparison with KOH hydrolysis and fungal culture for the immuno diagnosis of Trichophyton rubrum and Trichophyton mentagrophytes infection in humans.

Trichophyton is a keratinophilic fungus that can invade keratinized tissues of humans and cause superficial mycoses called dermatophytosis. The objective of this study was to develop and evaluate indirect ELISA in comparison with gold standard methods such as direct microscopic examination of KOH mounts and fungal culture for the diagnosis of Trichophyton infection in humans. The present investigation reports the production and partial purification of T. rubrum mycelial antigens and production of specific polyclonal antibodies. It also reports the development and optimization of indirect ELISA and evaluation of its potential in comparison with gold standard methods for the diagnosis of Trichophyton infection in humans. The diagnostic sensitivity and specificity of Trichophyton indirect ELISA was 93.75% and 93.33% respectively. The positive and negative predictive values were high as well, found to be 93.75% and 90.00% respectively indicating usefulness of the assay. In all comparisons, the correlation coefficient (r) value was >0.5 indicating strong correlation between KOH hydrolysis test, fungal culture method and indirect ELISA. A significant correlation coefficient of 0.856 (P < 0.0001) was obtained between indirect ELISA and fungal culture method. This shows a good agreement between fungal culture method and indirect ELISA. The present study clearly shows diagnostic performance of Trichophyton indirect ELISA developed in this study is efficient as fungal culture method for the diagnosis of Trichophyton infection in humans.

The prevalence of depression, anxiety, and stress among high school adolescent's children in public and private schools in Rangareddy district Telangana state: A cross-sectional study.

BACKGROUND: The depression, anxiety, and stress is a major educational problem among students all around the world. The purposes of this study were to determine the depression, anxiety, and stress among the high school adolescent children in public and private schools in Rangareddy district, Telangana state. MATERIALS AND METHODS: The present study was a cross-sectional study. The study sample considered 120 children from private schools and 259 children from public schools were studied. The study tool used was a questionnaire containing DASS (Depression, Anxiety, and Stress scale and sociodemographic characteristics. The data were expressed as proportion and mean values (Analysis of variance, independed t-test, and Pearson correlation) spatially tests were used at a significance level of P < 0.05). RESULTS: The overall prevalence of depression in children from the public school was found to be only 5%, that of anxiety was 20.8%, and that of stress was 15.8%; the overall prevalence of depression in children from private schools was found to be 1.9%, that of anxiety was 20.5%, and that of stress was 20.1%. CONCLUSION: In both schools, anxiety and atress were found to be more among study participants. Depression was observed to be less. But if they have more anxiety and stress and if not taken proper care at this point of time, it might get converted into a state of Depression.

Development and evaluation of indirect enzyme linked immunosorbent assay for the serological diagnosis of Microsporum canis infection in humans.

PURPOSE: The aim of this study was to develop and evaluate the diagnostic potential of indirect enzyme-linked immunosorbent assay (I-ELISA) for the rapid and precise diagnosis of Microsporum canis infection in humans. BASIC PROCEDURES: The present study reports the production, partial purification and SDS-PAGE analysis of M. canis mycelial antigens and production of specific polyclonal antibodies. It also reports the development and optimization of indirect ELISA and evaluation of its potential for the diagnosis of M. canis infection in humans. MAIN FINDINGS: An I-ELISA showed the sensitivity of 94.55% and specificity of 93.33%. Positive and negative predictive values were 96.30% and 90.32% respectively. Receiver operating characteristic (ROC) curve analysis of the data showed higher diagnostic accuracy. The area under the curve (AUC) was 0.925. A significant correlation coefficient of 0.8771 (P<0.0001) was obtained between I-ELISA and fungal culture method. PRINCIPAL CONCLUSIONS: In conclusion, the present study clearly shows the detection of specific antibodies by indirect ELISA using M. canis antigens. The assay is sensitive, specific and easy to perform, could enable rapid and more convenient diagnosis of dermatophytosis in humans.

Perception, knowledge, and attitude toward mental disorders and psychiatry among medical undergraduates in Karnataka: A cross-sectional study.

CONTEXT: Globally, psychiatry as a subject, psychiatrists as professionals, and patients with psychiatric disorders are subjected to cultural stereotypes and negative attitude by the general population. What is of alarming concern is that these prejudices exist within the medical community as well. AIMS: This study aims at evaluating the perception, knowledge, and attitude toward psychiatric disorders, therapeutic modalities used in psychiatry, psychiatry as a subject and psychiatrists as professionals among undergraduate medical students in Karnataka. SETTINGS AND DESIGN: This is a descriptive, cross-sectional type of study conducted in three medical colleges located in Karnataka. MATERIALS AND METHODS: A sample of 500 students from all three professional phases of MBBS was selected using purposive sampling. A semistructured prevalidated questionnaire was used to assess the perception, knowledge, and attitude of undergraduate medical students toward psychiatric disorders and psychiatry. STATISTICAL ANALYSIS: Data were analyzed using Statistical Package for Social Sciences, version 16.0. RESULTS: The undergraduate medical student population had significant shortcomings in knowledge and attitude pertaining to psychiatric disorders, more glaring in the initial years of education. A comparatively positive opinion was obtained regarding psychiatry as a subject and psychiatrists as professionals, which may reflect the changing trends and concepts, both in society and medical community. CONCLUSION: This study highlights the need for better educational measures at undergraduate level in order to shape a positive attitude of the health care providers towards psychiatry, which is essential for ensuring better care for patients as well as reduction of stigma surrounding psychiatric disorders.

Estimation of N-terminal telopeptides of type I collagen in periodontal health, disease and after nonsurgical periodontal therapy in gingival crevicular fluid: A clinico-biochemical study.

AIM: This study explored gingival crevicular fluid (GCF) N-terminal telopeptides of type I collagen (NTx) levels in periodontal health, disease and after nonsurgical periodontal therapy along with its association with the clinical parameters. MATERIALS AND METHODS: Study comprised of three groups of 10 subjects each: Healthy (Group I), gingivitis (Group II), and periodontitis (Group III), while Group III patients after scaling and root planning (SRP) constituted Group IV. Gingival index (GI), probing pocket depth (PPD), clinical attachment loss (CAL), and radiological parameters were recorded. GCF samples were analyzed by competitive-enzyme-linked immunosorbent assay. RESULTS: Samples in Group III and Group IV tested positive for NTx whereas in Group I and Group II, NTx was not detected. Mean NTx levels were higher in Group III (6.79 ± 0.94 nanomole bone collagen equivalents per liter [nm BCE/L]) compared to Group IV (5.73 ± 0.95 nm BCE/L) which was statistically significant. Positive correlation was seen between the clinical parameters and the NTx levels in Group III and IV. CONCLUSION: As NTx is specific bone turnover marker, it is detected only in periodontitis Group and the values decline after SRP. Failure to detect NTx in Group I and II, relates to the minimum or no resorption at the sample sites.

Solid lipid nanoparticles and nanosuspension formulation of Saquinavir: preparation, characterization, pharmacokinetics and biodistribution studies.

Solid lipid nanoparticles (SLNs) and nanosuspensions (NSs) have shown great promise for improving bioavailability of poorly water-soluble drugs. This study was aimed to develop SLNs and NS of Saquinavir (SQ) for improvement in bioavailability. These formulations were characterized and their pharmacokinetics and biodistribution in mice were evaluated. Saquinavir-loaded SLNs (SQSLNs) showed particle size 215 ± 9 nm and entrapment efficiency 79.24 ± 1.53%, while solid-state studies (differential scanning calorimetry and X-ray diffraction) indicated entrapment of the drug in SLNs. Saquinavir NS (SNS) showed particle size 344 ± 16 nm with fourfold increase in saturation solubility and its solid-state studies showed reduction in crystallinity. Pharmacokinetics and biodistribution studies of orally administered SQSLN and SNS in mice exhibited higher plasma level concentration compared to saquinavir microsuspension (SMS). The relative bioavailabilities for SNS and SQSLN were 37.39% and 66.53%, respectively, compared to 18.87% bioavailability obtained after administration of SMS, indicating suitability of nanoparticulate formulations for improving bioavailability.

Stereoselective synthesis of an iodinated resveratrol analog: preliminary bioevaluation studies of the radioiodinated species.

Stereoselective synthesis of an E-hydroxystilbene has been carried out using the McMurry reaction. Synthesis of a monoiodinated hydroxystilbene has been carried out by a McMurry cross-coupling reaction. For the purpose of biological evaluation, the facile electrophilic substitution route has been attempted to radioiodinate it with (125)I. The HPLC pattern of the radioiodinated hydroxystilbene, which could be obtained in >90% radiochemical purity, was found to be identical to that of its non-radioactive analog that has been independently prepared using the McMurry cross-coupling route. In vitro cell uptake studies were carried out in breast cancer cells MCF7, overexpressing estrogen receptors. In vivo biodistribution studies in female Swiss mice show a uterine uptake of 0.85±0.4% ID/g at 3h.p.i. with a uterus to muscle ratio of 2.83. Uptake in the thyroid was insignificant indicating good in vivo stability of the radioiodinated hydroxystilbene.

Gingival crevicular fluid and plasma levels of neuropeptide Substance-P in periodontal health, disease and after nonsurgical therapy.

BACKGROUND AND OBJECTIVE: The level of Substance-P in gingival crevicular fluid has been found to correlate with clinical measures of periodontal disease. The present study was designed to assess the relationship between clinical parameters and levels of Substance-P in the gingival crevicular fluid from inflamed gingiva, periodontitis sites and after treatment of periodontitis sites, and to correlate them to the Substance-P levels of plasma. MATERIAL AND METHODS: Thirty, age- and gender-matched subjects were divided into three groups (healthy, gingivitis and chronic periodontitis) based on modified gingival index scores and clinical attachment loss. A fourth group consisted of 10 subjects from the periodontitis group, 6-8 wk after initial therapy. Plasma and gingival crevicular fluid samples were collected and quantified for Substance-P using an enzyme immunoassay. RESULTS: The mean concentration of Substance-P, both in gingival crevicular fluid and plasma, was observed to be highest in the periodontitis group (45.13 pg/mL in gingival crevicular fluid and 67.8 pg/mL in plasma) and lowest in the healthy group (6.07 pg/mL in gingival crevicular fluid and below the detection level in plasma). The mean Substance-P concentration in the gingivitis group (11.42 pg/mL in gingival crevicular fluid and 38.8 pg/mL in plasma) and in the after-treatment group (7.58 pg/mL in gingival crevicular fluid and 39.7 pg/mL in plasma) lay between the highest and lowest values. In all groups the gingival crevicular fluid levels showed a statistically significant positive correlation with that of plasma and clinical attachment loss. CONCLUSION: Substance-P levels were highest in the gingival crevicular fluid from sites with periodontal destruction; however, periodontal treatment resulted in the reduction of Substance-P levels. Gingival crevicular fluid and plasma Substance-P levels showed a positive correlation in all of the groups.

Immunotoxins: a review of their use in cancer treatment.

ABSTRACT: Antibody therapies have become an important component in the management of malignant disease. Antibodies can block tumour growth factors or their receptors, activate immunological attack on the tumour, and are used to deliver payloads such as radioisotopes, cytotoxic drugs or toxins. Immunotoxins are a new class of antitumour agents consisting of tumour- selective ligands (generally monoclonal antibodies [MoAbs]) linked to highly toxic protein molecules and take the advantage of the exquisite specificity of antibodies to selectively target drug delivery and the potency of toxins to kill the target cells. Toxins are modified to remove their normal tissue-binding domains by genetic engineering. Analysis of the aminoacid sequence of the region specific for immunogenecity and the signal transduction mechanisms involved in the interaction of immunotoxins with tumour cells will give the clue for the development of most efficient immunotoxins.

Seroprevalence study of HIV-I, HIV-II and HTLV-I among patients at the Dermato-Venereology Clinic of the Lagos University Teaching Hospital.

BACKGROUND: Seroprevalence studies of HIV-I and HIV-II that have been reported in Nigeria were among commercial sex workers and blood donors. There are no data from STD patients and dermatologic patients. METHODS: A seroprevalence study of HIV-I, HIV-II and HTLV-I was prospectively conducted among STD clinic attendees and among patients with dermatoses which have been linked with HIV disease. The studies were done in 1992 and 1994. RESULTS: Some patients had more than one seropositive type. In 1992, the percentages of seropositive results to HIV-I, HIV-II and HTLV-I were 31, 19 and 50, respectively, and in 1994 the percentages of HIV-I and HTLV-I were 65 and 35, respectively. CONCLUSIONS: Patients should be routinely screened for HTLV-I, in addition to HIV-I and HIV-II, among blood donors and also neurology clinic attendees in Nigeria. They should also be screened for retroviral infections when they present with dermatoses clinically suggestive of papular urticaria, onchodermatitis, or papulonecrotic tuberculids.