Interactions between the protein barnase and co-solutes studied by NMR.

Protein solubility and stability depend on the co-solutes present. There is little theoretical basis for selection of suitable co-solutes. Some guidance is provided by the Hofmeister series, an empirical ordering of anions according to their effect on solubility and stability; and by osmolytes, which are small organic molecules produced by cells to allow them to function in stressful environments. Here, NMR titrations of the protein barnase with Hofmeister anions and osmolytes are used to measure and locate binding, and thus to separate binding and bulk solvent effects. We describe a rationalisation of Hofmeister (and inverse Hofmeister) effects, which is similar to the traditional chaotrope/kosmotrope idea but based on solvent fluctuation rather than water withdrawal, and characterise how co-solutes affect protein stability and solubility, based on solvent fluctuations. This provides a coherent explanation for solute effects, and points towards a more rational basis for choice of excipients.

Predicting the Anatomical Location of Neck of Femur Fractures in Osteoporotic Geriatric Indian Population.

Introduction: Neck of femur fractures are quite common fractures in the elderly. Though a lot is spoken about the various modes of management of these fractures across different age groups, hardly any literary support mentioning their distribution, location and pattern can be found. In this study, we aim to find whether the Singh index, as a marker of osteoporosis on digital radiographs, can predict the location of neck of femur fractures in the elderly population. Materials and methods: We accessed 556 fractured hip radiographs in our institution over the past 5 years (20152020) and correlated with the Singh index, as a marker of degree of osteoporosis, on pre-operative pelvis digital radiographs. Mid coronal CT cuts were also corroborated with the radiographic findings. A control group was set up and 361 radiographs were evaluated in the study group. Results: A total of 124 transcervical fractures (73%) were in Singh index 4, while 76 subcapital fractures (70%) were in Singh index 3. A total of 166 fractures (66%) were found in transcervical region in the age group of 60 to 80 years, while 80 fractures (74%) were in the subcapital region in patients above 80 years. Conclusion: We concluded that transcervical fractures were more common in patients with Singh index 4 (p<0.001) and subcapital more common in patients with Singh index 3(p<0.001). There was also a shift in location of the fractures from the transcervical region to the subcapital region with age above 80 years (p<0.001).

Covid-19 vaccination coverage and break through infections in urban slums of Bengaluru, India: A cross sectional study.

BackgroundThe ongoing pandemic of Corona virus disease 2019(covid-19) is caused by severe acute respiratory syndrome Corona virus 2(SAR-COV-2). The world health organization declared it as public health emergency of international concern on January 2020, and later declared as pandemic on 11 March 2020.One of the high-risk groups for COVID-19 disease are people residing in urban overcrowded slums and as most of the population is migrant, they are less aware of the pandemic and have less access to health care facilities. Vaccinating these high-risk groups can decrease disease burden and control the ongoing pandemic. Objectives1] To estimate COVID 19 vaccination coverage 2] To assess the factors responsible for COVID - 19 vaccination coverage and vaccine hesitancy 3] To study AEFI pattern following COVID-19 vaccination 4] To determine the prevalence of breakthrough infections after COVID - 19 Vaccination in urban slums of Bengaluru, India. MethodologyA community based cross sectional study was conducted in Urban slums belonging to Urban Health and Training Centre, Department of community medicine, Akash Institute of Medical Sciences and Research Centre, Bengaluru Rural District, Karnataka, India. After obtaining Institutional ethical clearance and informed consent from study participants, data was collected from 1638 participants, fulfilling inclusion criteria using a predesigned, pretested, structured questionnaire. Data was entered in Microsoft excel and analyzed using SPSS version 24. Chi square test and Fischers exact test was applied and p <0.05 considered as statistically significant. ResultsIn the present study, 35.5% (583 out of 1638) of the study participants had taken COVID Vaccine, of which 533 (91.42%) were partially vaccinated and remaining 50 (8.5%) were fully Vaccinated. Majority i.e., 98.45% have taken vaccine at Govt health centers. 63.65% vaccinated with Covishield reported adverse events, whereas 18.6% vaccinated with Covaxin reported adverse events. Adverse events were more likely to be reported by women (74.7%) compared to men (58.6%), this observation was consistent across all age groups. Vaccination coverage was high among 18 - 45 years age group (37.75%), males (64.86%), Christians (47.05%) followed by Hindus (43.56%), graduates (95.67%), clerical and skilled workers (70.75%), Upper middle socioeconomic class (72.41%). This difference was statistically significant. Our study reported Break through infections in 7 out of total 583 vaccinated with a prevalence of 1.2%. The break through infections was very high among partially vaccinated (85.71%) as compared to fully vaccinated individuals (14.28%). This was observed among those vaccinated with Covaxin only. ConclusionThe COVID vaccine coverage was low in urban slums. The prevalence of Break through infections in our study was higher as compared to available data/reports in the country. Break through infections was very high among partially vaccinated as compared to fully vaccinated individuals. This study on break through infections on COVID vaccination is first study in South India on general population. The most important factor for vaccine hesitancy is the occurrence of mild or serious adverse effects following immunization, and this may be the biggest challenge in the global response against the pandemic.

Transition of Children with Perinatally Acquired HIV-Infection into Adulthood: Social Outcome and Quality of Life.

OBJECTIVES: In India, access to free anti-retroviral therapy has improved the survival of perinatally human immunodeficiency virus (HIV) infected children resulting in the transition of many such children to adulthood. This study aims to understand the social-outcomes and quality of life (QOL) among these adults. METHODS: This cross-sectional study was conducted in two tertiary HIV care centers in south India. Perinatally HIV-infected adults aged >18 y were enrolled after obtaining consent. Data were collected by questionnaire based interviews for social outcomes and WHO Quality of Life-BREF (WHOQOL-BREF) for QOL. The social-outcome indicators monitored pertained to family support, educational qualification and occupational, economic, and marital status. RESULTS: The mean age of 107 participants was 18·9 ± 1·1y. The school drop-out rate was 58%. Sixty-two percent were double orphans. Forty-three-percent of the participants were employed with mean per-capita monthly income of Rs.4105 ± 2979 ($65·2 ± 47·3). Fourteen-percent of the participants were married, or in a relationship, and a majority of them, 93%, were females. For QOL, the mean raw score was highest for social relationship (15·79). Relative to studying subjects, a higher proportion of school dropouts scored poorly in social relationship (42% vs.14·8%; Chi-square = 5·28; p = 0·02) and environmental QOL (46% vs.19·6%;Chi-square = 8·09; p = 0·004). The proportion of subjects with a poor physical health QOL was higher among those with a per-capita monthly income above the national average than those with below the national average (69% vs.33·3%; Chi-square = 5·27; p = 0·02). CONCLUSIONS: Though clinico-immunological disease was stable in these perinatally HIV-infected young adults, their social-outcomes pertaining to education, occupation, income, and family support were poor. Factors like education, parental care, and income of the subjects were associated with poor QOL.

Olze et al. stages of radiographic visibility of root pulp and cameriere's third molar maturity index to estimate legal adult age in Hyderabad population.

CONTEXT: Predicting one's attainment of age of majority is a controversial issue and considered as important aspect in medicolegal cases. In India, individuals older than 18 years of age have full capacity regarding civil conduct and are tried as adults for criminal charges. AIMS AND OBJECTIVE: To compare the accuracy of Olze et al., stages of radiographic visibility of root pulp, and Cameriere's third molar maturity index (I 3M < 0.08) to estimate the age of majority. MATERIALS AND METHODS: A total of 615 digital orthopantomographs of children aged between 15 and 22 years. The lower left third molars were evaluated using ImageJ computer software. The effectiveness of both methods was evaluated using sensitivity, specificity, positive likelihood ratios (LR+), and LR negative (LR-). RESULTS: For I 3M < 0.08, the sensitivity, specificity, LR+, LR- were 0.76, 0.72, 2.79, 0.32 and 0.67, 0.76, 2.83, 0.43 in males and females respectively. For Stage 0, the sensitivity, specificity, LR+, LR- were 0.68, 0.86, 5.18, 0.36 and 0.72, 0.91, 8.63, 0.31, respectively. CONCLUSION: Stage 0 of Olze's radiographic root pulp visibility showed to be more accurate than cutoff value of I 3M< 0.08 in discriminating adults and minors of Hyderabad sample when a test of high sensitivity and specificity is required.

Comparative evaluation of three noninvasive gingival displacement systems: An in vivo study.

AIM: An attempt is made to investigate clinical efficacy of cord, paste system, and a strip gingival retractile materials. This study aims to evaluate and compare the gingival retraction efficacy of retraction strip along with conventional retraction cord and paste system. MATERIAL AND METHODS: This in vivo experimental study was carried out on 30 patients. Three different gingival retraction systems were used to evaluate the amount of vertical and lateral displacement.Based on selection criteria, 30 individuals requiring fixed dental prosthesis with respect to mandibular first molar were selected. Tooth preparation for metal ceramic restoration with subgingival finish line was performed. Gingival displacement was accomplished with ultrapak cord, merocel strip, and magic foam cord immediately, 7 and 14 days after the tooth preparation, respectively. The amount of gingival displacement in vertical and lateral directions was measured at mesiobuccal, midbuccal, and distobuccal regions of the prepared tooth. The vertical retraction was measured intraorally by using digital vernier caliper, and postgingival displacement impression was used to measure lateral gingival retraction. Stereomicroscopic images of impression under ×10 resolution were transferred to image analyzer to measure the lateral displacement. The obtained data analyzed in one-way ANOVA and Bonferroni test were used to determine the significant difference at P < 0.05 level. RESULTS: ANOVA test showed the significant difference between the materials tested with respect to the mean vertical and lateral gingival retraction (P = 0.001). Multiple comparisons by Bonferroni test revealed a significant difference in vertical and lateral displacement among the materials tested. CONCLUSIONS: Merocel strip provided the maximum amount of vertical and lateral tissue displacement, followed by ultrapak cord and least with magic foam cord which was statistically significant.

Depression and suicidal ideation among adolescents.

No abstract provided.

A Molecular Docking and Dynamics Approach to Screen Potent Inhibitors Against Fosfomycin Resistant Enzyme in Clinical Klebsiella pneumoniae.

Klebsiella pneumoniae, BA6753 was cultured from a patient in the Clinical Microbiology Laboratory of Christian Medical College. K. pneumoniae, BA6753 has a multidrug resistance plasmid encoding novel FosA variant-7, fosfomycin resistance enzyme. Minimal side effects and a wide range of bactericidal activity of fosfomycin have resulted in its expanded clinical use that prompts the rise of fosfomycin-resistant strains. At present, there are no effective inhibitors available to conflict the FosA-medicated fosfomycin resistance. To develop effective FosA inhibitors, it is crucial to understand the structural and dynamic properties of resistance enzymes. Hence, the present study focuses on the identification of potent inhibitors that can effectively bind to the fosfomycin resistance enzyme, thus predispose the target to inactivate by the second antibiotic. Initially, a series of active compounds were screened against the resistant enzyme, and the binding affinities were confirmed using docking simulation analysis. For efficient activity, the binding affinity of the resistance enzyme ought to be high with the inhibitor than the fosfomycin drug. Consequently, the enzyme-ligand complex which showed higher binding affinity than the fosfomycin was employed for subsequent analysis. The stability of the top scoring enzyme-ligand complex was further validated using molecular dynamics simulation studies. On the whole, we presume that the compound 19583672 demonstrates a higher binding affinity for the resistance enzyme comparing to other compounds and fosfomycin. We believe that further enhancement of the lead compound can serve as a potential inhibitor against resistance enzyme in drug discovery process. J. Cell. Biochem. 118: 4088-4094, 2017. © 2017 Wiley Periodicals, Inc.

A cephalometric study to establish the relationship of the occlusal plane to the three different ala-tragal lines and the Frankfort horizontal plane in different head forms.

BACKGROUND: The aim of the study is to compare the relationship of the occlusal plane to 3 different ala-tragal lines, namely the superior, middle and inferior lines, in individuals having different head forms and its relation to the Frankfort horizontal plane. METHODOLOGY: A total of 75 lateral cephalometric radiographs of subjects with natural dentition, having full complement of teeth, between the age group of 18-25 were screened and selected. Lateral cephalogram were made for each subjects in an open mouth position. Prior to making the lateral cephalogram, radiopaque markers were placed on the superior, middle and inferior tragus points and on the inferior border of the ala of the nose. Cephalometric tracing was done over each cephalogram. RESULTS: In mesiocephalic head form the middle ala-tragal line was most parallel to the occlusal plane having a mean angle of (1.96°). In dolichocephalic headform, the superior ala-tragal line was most parallel to the occlusal plane having a mean angle of (0.48°). In brachycephalic head form, the middle ala-tragal line was most parallel to the occlusal plane having a mean angle of (2.08°). The mean angulations of occlusal plane to FH plane is 11.04°, 10.16° and 10.60° in mesiocephalic, dolichocephalic and brachycephalic head forms, respectively. RESULTS AND CONCLUSION: The study concludes that the middle ala-tragal line can be used as a reference for the mesiocephalic head form and the superior ala-tragal line for the dolichocephalic and brachycephalic head form as a reference to establish the occlusal plane.

Parental concerns on disclosure of HIV status to children living with HIV: children's perspective.

Disclosure is an important component of comprehensive management of children living with HIV infection (CLHIV). Many parental concerns are barriers for disclosure in children and only few studies addresses children's perspective on these concerns. Our study aims to understand children's perspective on parental concerns for disclosure and assess the knowledge of HIV. A questionnaire-based cross-sectional study involving CLHIV between 10 and 18 years attending HIV clinic in southern India, was conducted. Data were collected by directly interviewing only the children after obtaining consent from parents/caregivers. Initial open-ended questions were asked to assess the disclosure status and only fully disclosed children were included. Out of 362 enrolled eligible children, the prevalence of full and partial disclosure was 36.7% and 24%, respectively. The mean age of disclosure was 10.4 years (SD ± 2.6) and non-parental family members in an informal setting were the most common source of disclosure (38.3%). Forty-six percentages of parents were unaware of their child's disclosure status. Only 2% had disclosed their status to others who were not part of their care. Among disclosed children, 33.8% became upset or sad upon knowing their status, 12% faced discrimination and 41.4% had complete knowledge about their illness. Though the prevalence of disclosure among CLHIV was high, a majority of them had incomplete knowledge about HIV infection. The parental concerns as reported in literature like the child is too young to disclose, concerns about coping, fear of stigma and discrimination and child disclosing to others were not expressed by children.

Predictors of Mortality and Mortality Rate in a Cohort of Children Living with HIV from India.

OBJECTIVES: To study the predictors of mortality and mortality rate in a clinical cohort of Children Living with Human Immunodeficiency Virus infection (CLHIV) from India. METHODS: This retrospective cohort analysis of CLHIV aged between 2 mo and 18 y registered during January 2004 through December 2014 at Pediatric Centre of Excellence (PCOE), Indira Gandhi Institute of Child Health (IGICH), was conducted using standard data collection sheet. Demographic and clinical characteristics of all eligible children were analyzed. The primary outcome measured was mortality. The authors also analyzed the cause of death and baseline parameters associated with death to study the predictors of mortality. RESULTS: Out of 1289 CLHIV registered in the PCOE during the study period, 834 (64.7 %) CLHIV, with or without antiretroviral therapy (ART) care, were included. The total time contributed by the study participants was 2872.8 child-years. The mortality rate in these children was 4.9/100 child-years. A significantly higher mortality rate of 28.2 % was found in children < 5 y, 38.6 % in children with advanced WHO clinical staging, 35.2 % among severely immunosuppressed children and 22.3 % in severely malnourished children. Tuberculosis accounted for 28 % of deaths. Univariate Cox regression analysis showed treatment status, age <5 y, baseline WHO clinical stage 3 and 4, severe immune suppression and severe malnutrition were strongly associated with mortality. CONCLUSIONS: The mortality rate in the index study cohort was 4.9/100 child-years and tuberculosis was the major cause of death. Younger age, baseline advanced clinical and immunological staging were predictors of mortality. Even though mortality was significantly higher in Pre-ART children, treatment status was not found to be an independent predictor of mortality.

Luteolin induces caspase-14-mediated terminal differentiation in human epidermal keratinocytes.

Recent studies have demonstrated the role of caspase-14 in terminally differentiated keratinocytes, and its expression may decrease the magnitude of tumors in the epidermis. In the present study, we assessed the potential of luteolin (LUT) to elicit the expression of caspase-14 in terminal differentiation of human keratinocytes. The semi-qualitative RT-PCR data revealed a significant level of caspase-14 expression in LUT-treated human immortalized keratinocytes (HaCaT) with respect to untreated cells. The quantitative data (ELISA) further supported the potency of LUT to induce caspase-14 expression at 3.19 ng/ml when compared to 1.29 ng/ml of vitamin D3 (positive control). Further, the enhanced expression of human involucrin gene in LUT-treated HaCaT cells confirmed its ability to drive terminal differentiation in these cells. These preliminary results provide first-hand information about the in vitro potential of LUT to elicit the expression of caspase-14, thereby inducing terminal differentiation in human keratinocytes.

Influence of natural and synthetic antioxidants on the degradation of Soybean oil at frying temperature.

The effect of several natural and synthetic antioxidants to retard the formation of polar compounds and thermo-oxidation at prolonged frying temperatures was studied. Antioxidants, including butyl hydroxyl toluene (BHT), ter-butyl hydroxyquinone (TBHQ), α- and γ-tocopherols, tocopherol mixture (α, ß, γ and δ), sesamol, ß-sitosterol, ß-sitostanol, γ-oryzanol, curcumin, rosemary extract and sucrose acetate isobutyrate (SAIB) were tested in refined soybean oil without added any additives recovered from refinery. Rosemary extract and SAIB were showed a considerable effect on both polar compound formation and secondary oxidation. These compounds increased the oxidative stability of oil for more than 30 % compared to conventional synthetic antioxidants. Oils treated with SAIB showed higher color retention after 6 h heating compared to the oils added with BHT, TBHQ and tocopherols. Curcumin, sesamol and γ-oryzanol showed higher antioxidant potential compared to other antioxidants. Preliminary results obtained from this study have clearly demonstrated that SAIB and rosemary extracts are more commercially viable antioxidants to increase the stability of frying oils.

Antioxidant, DNA protective efficacy and HPLC analysis of Annona muricata (soursop) extracts.

Annona muricata is a naturally occurring edible plant with wide array of therapeutic potentials. In India, it has a long history of traditional use in treating various ailments. The present investigation was carried out to characterize the phytochemicals present in the methanolic and aqueous leaf extracts of A. muricata, followed by validation of its radical scavenging and DNA protection activities. The extracts were also analyzed for its total phenolic contents and subjected to HPLC analysis to determine its active metabolites. The radical scavenging activities were premeditated by various complementary assays (DRSA, FRAP and HRSA). Further, its DNA protection efficacy against H2O2 induced toxicity was evaluated using pBR322 plasmid DNA. The results revealed that the extracts were highly rich in various phytochemicals including luteolin, homoorientin, tangeretin, quercetin, daidzein, epicatechin gallate, emodin and coumaric acid. Both the extracts showed significant (p < 0.05) radical scavenging activities, while methanolic extract demonstrated improved protection against H2O2-induced DNA damage when compared to aqueous extract. A strong positive correlation was observed for the estimated total phenolic contents and radical scavenging potentials of the extracts. Further HPLC analysis of the phyto-constituents of the extracts provides a sound scientific basis for compound isolation.

Bullous Lichen Planus treated with Oral Minipulse Therapy: A Rare Case Report.

Oral Lichen planus (OLP) is a common mucocutaneous disorder with a multifactorial aetiology, affecting the women more commonly than men. Most OLP are asymptomatic, except the atrophic and erosive forms.Till date many treatment modalities are implicated to treat this disorder, but no therapy is considered as the single most effective, without side-effects and remission of the lesion. As the treatment of OLP is challenging to the oral practitioners, here we report a case of successful management of extensive, symptomatic bullous and erosive oral lichen planus with a novel treatment protocol- oral minipulse therapy with betamethasone.

Acceleration of pro-caspase-3 maturation and cell migration inhibition in human breast cancer cells by phytoconstituents of Rheum emodi rhizome extracts.

The aggressive nature of estrogen receptor (ER)-negative breast cancer subtype obligates for innovative targeted therapies. The present study aimed to investigate the phytoconstituents and specific anticancer activities of Rheum emodi rhizome, a known food source used locally to treat various ailments. Petroleum ether extracts (hot [PHR] and cold [PCR]) of R. emodi, exhibited significant free radical scavenging potentials through DPPH and reducing power assays, rendering them as good sources of antioxidants. The extracts, PHR and PCR had shown significant (P < 0.05) cancer-cell-specific cytotoxicity in the assayed cells (MDA-MB-231 [breast carcinoma] and WRL-68 [non-tumoral]) at 100 µg/ml, and 50 and 100 µg/ml concentrations respectively. Extracts also induced fervent apoptosis in ER-negative cells (MDA-MB-231) compared to ER-positive subtype (MCF-7), and found to involve CPP32/caspase-3 in its apoptosis induction mechanism. Moreover, extracts had an inevitable potential to inhibit the migration of metastatic breast cancer cells (MDA-MB-231) in vitro. Further, the active principles of extracts were identified through HPLC and GC-MS analysis to reveal major polyphenolics, 4,7-Dimethyl-(octahydro)indolo[4,3-fg]quinolin-10-one, 5-Oxo-isolongifolene, Valencene-2, and other quinone, quinoline and anthraquinone derivatives. The extracts are thus good candidates to target malignant ER-negative breast cancer, and the identified phytoconstituents could be eluted in further exploratory studies for use in dietary-based anti-breast cancer therapies.

Isolation of proline-based cyclic dipeptides from Bacillus sp. N strain associated with rhabditid [corrected] entomopathogenic nematode and its antimicrobial properties.

Entomopathogenic nematodes (EPN) are well-known as biological control agents and are found to have associated bacteria which can produce a wide range of bioactive secondary metabolites. We report herewith isolation of six proline containing cyclic dipeptides cyclo(D-Pro-L-Leu), cyclo(L-Pro-L-Met), cyclo(D-Pro-L-Phe), cyclo(L-Pro-L-Phe), cyclo(L-Pro-L-Tyr) and cyclo(L-Pro-D-Tyr) from ethyl acetate extract of the Luria Broth (LB) cell free culture filtrate of Bacillus sp. strain N associated with a new EPN Rhabditis sp. from sweet potato weevil grubs collected from Central Tuber Crops Research Institute farm. Antimicrobial studies of these 2,5-diketopiperazines (DKPs) against both medicinally and agriculturally important bacterium and fungi showed potent inhibitory values in the range of µg/mL. Cyclic dipeptides showed significantly higher activity than the commercial fungicide bavistin against agriculturally important fungi, viz., Fusarium oxysporum, Rhizoctonia solani, and Pencillium expansum. The highest activity of 2 µg/mL by cyclo(L-Pro-L-Phe) was recorded against P. expansum, a plant pathogen responsible for causing post harvest decay of stored apples and oranges. To our knowledge, this is the first report on the isolation of these DKPs from Rhabditis EPN bacterial strain Bacillus sp.

Cytotoxicity, apoptosis induction and anti-metastatic potential of Oroxylum indicum in human breast cancer cells.

Despite clinical advances in anticancer therapy, there is still a need for novel anticancer metabolites, with higher efficacy and lesser side effects. Oroxylum indicum (L.) Vent. is a small tree of the Bignoniaceae family which is well known for its food and medicinal properties. In present study, the chemopreventive properties of O. indicum hot and cold non-polar extracts (petroleum ether and chloroform) were investigated with MDA- MB-231 (cancer cells) and WRL-68 (non-tumor cells) by XTT assay. All the extracts, and particularly the petroleum ether hot extract (PHO), exhibited significantly (P<0.05) higher cytotoxicity in MDA-MB-231 when compared to WRL-68 cells. PHO was then tested for apoptosis induction in estrogen receptor (ER)-negative (MDA-MB-231) and ER-positive (MCF-7) breast cancer cells by cellular DNA fragmentation ELISA, where it proved more efficient in the MDA-MB-231 cells. Further, when PHO was tested for anti-metastatic potential in a cell migration inhibition assay, it exhibited beneficial effects. Thus non-polar extracts of O. indicum (especially PHO) can effectively target ER-negative breast cancer cells to induce apoptosis, without harming normal cells by cancer-specific cytotoxicity. Hence, it could be considered as an extract with candidate precursors to possibly harness or alleviate ER-negative breast cancer progression even in advanced stages of malignancy.

Apoptosis-induced cell death due to oleanolic acid in HaCaT keratinocyte cells--a proof-of-principle approach for chemopreventive drug development.

Oleanolic acid (OA) is a naturally occurring triterpenoid in food materials and is a component of the leaves and roots of Olea europaea, Viscum album L., Aralia chinensis L. and more than 120 other plant species. There are several reports validating its antitumor activity against different cancer cells apart from its hepatoprotective activity. However, antitumor activity against skin cancer has not been studied well thus far. Hence the present study of effects of OA against HaCaT (immortalized keratinocyte) cells--a cell-based epithelial model system for toxicity/ethnopharmacology-based studies--was conducted. Radical scavenging activity (DPPH·) and FRAP were determined spectrophotometrically. Proliferation was assessed by XTT assay at 24, 48 and 72 hrs with exposure to various concentrations (12.5-200 µM) of OA. Apoptotic induction potential of OA was demonstrated using a cellular DNA fragmentation ELISA method. Morphological studies were also carried out to elucidate its antitumor potential. The results revealed that OA induces apoptosis by altering cellular morphology as well as DNA integrity in HaCaT cells in a dose-dependent manner, with comparatively low cytotoxicity. The moderate toxicity observed in HaCaT cells, with induction of apoptosis, possibly suggests greater involvement of programmed-cell death-mediated mechanisms. We conclude that OA has relatively low toxicity and has the potential to induce apoptosis in HaCaT cells and hence provides a substantial and sound scientific basis for further validation studies.

Quantitative assessment of the relative antineoplastic potential of the n-butanolic leaf extract of Annona muricata Linn. in normal and immortalized human cell lines.

Natural products have been the target for cancer therapy for several years but there is still a dearth of information on potent compounds that may protect normal cells and selectively destroy cancerous cells. The present study was aimed to evaluate the cytotoxic potential of n-butanolic leaf extract of Annona muricata L. on WRL-68 (normal human hepatic cells), MDA-MB-435S (human breast carcinoma cells) and HaCaT (human immortalized keratinocyte cells) lines by XTT assay. Prior to cytotoxicity testing, the extract was subjected to phytochemical screening for detecting the presence of compounds with therapeutic potential. Their relative antioxidant properties were evaluated using the reducing power and DPPH* radical scavenging assay. Since most of the observed chemo-preventive potential invariably correlated with the amount of total phenolics present in the extract, their levels were quantified and identified by HPLC analysis. Correlation studies indicated a strong and significant (P<0.05) positive correlation of phenolic compounds with free radical scavenging potential. The results revealed that the extract was moderately cytotoxic to normal cells with a mean IC50 value of 52.4 µg when compared with those obtained for cancerous cells (IC50 values of 29.2 µg for MDA-MB-435S and 30.1 µg for HaCaT respectively). The study confirms the presence of therapeutically active antineoplastic compounds in the n-butanolic leaf extract of Annona muricata. Isolation of the active metabolites from the extract is in prospect.