MEASUREMENT OF CORTISOL AND CORTISONE IN HUMAN SALIVA BY UPLC-MS/MS

Objective: To develop and validate a simple and rapid assay for simultaneous measurement of cortisol and cortisone in human saliva by ultra-performance liquid chromatography-tandem mass spectrometry. Methods: Chromatographic analysis was performed on an Atlantis dC18 column (2.1 x 100 mm, 3 µm) using a mobile phase consisting of acetonitrile and 2 mmol ammonium-acetate (50:50, v; v) that was delivered at a flow rate of 0.3 ml/min. The eluents were monitored using electrospray ionization in the positive ion mode set at transition set of mass-to-charge (m/z): 363.11 → 121.00, 361.18 → 163.11, and 367.19 → 121.24 for cortisol, cortisone and internal standard (IS), respectively the method was validated for linearity, accuracy, precision, and recovery, according to international guidelines. Results: The retention times of cortisol, cortisone and internal were about 1.38, 1.43 and 1.38 min, respectively. Cortisol level and cortisone level relationship to the ratio of their respective peak-area to IS’s peak-area was linear (range of 0.5-100 ng/ml). Coefficients of variation and inaccuracy were, ≤9.9% and-0.3 to 6.9 for cortisol and ≤8.4 and-1.5 to 4.8 for cortisone, respectively. Extraction recoveries for cortisol, cortisone, and the IS were 90%, 94%, and 98%, respectively. Cortisol and cortisone stability was evaluated in processed saliva samples (stored at room temperature for 24 h) and unprocessed saliva samples (stored at room temperature for 24 h or at-20 °C for 10 w) and after 3 freeze-thaw cycles was ≥ 86%. Conclusion: The proposed method is simple, precise, and accurate for the rapid simultaneous measurement of cortisol and cortisone levels in saliva. The assay was successfully applied to determine levels of cortisol and cortisone in human saliva samples obtained from healthy volunteers

Delayed surgical therapy reduces mortality in patients with acute necrotizing pancreatitis

Objective: To review the trends in management and analyze the factors influencing outcomes of acute necrotizing pancreatitis.Methods: It wa a retrospective analytical study. All adult patients with computed tomography with proven necrotizing pancreatitis managed at the department of surgery, Aga Khan University Hospital, Karachi were included in this study extending from January 1998 to January 2008. Outcome variables were hospital stay, complication rate and in-hospital mortality. Data analysis was carried out using SPSS version 16. For comparison, Pearson chi-square test, Fisher\u27s exact test, t-test and ANOVA were used, where applicable. A p-value less than 0.05 was considered statistically significant. Results: Of 1479 patients, 47 patients were included. Median age was 48 (range: 38-56) years with 31(66%) males and 16(34%) females. Overall out of 18(38%) that underwent necrosectomy, 16 had infected acute necrotizing pancreatitis while the rest were negative. Computed Tomography and/or FNAC identified 18 infective acute necrotizing pancreatitis patients, 16 underwent necrosectomy, one patient expired without surgery and the other was managed conservatively. Enteral nutrition was widely used with rising trend of oral feeding from 2006 onwards. Complication rate was 63%. Overall mortality remained 9/47(19.7%), where in infected ANP as well as in postoperative patients, mortality was 7/18 (38.9%). The patients with early pancreatic necrosectomy had prolonged hospital stay, more complications and statistically significant increase in in-hospital mortality. Conclusion: Better outcomes were achieved in infected acute necrotizing pancreatitis with delayed pancreatic necrosectomy and the other contributing factor could be early enteral nutritional therapy

CALLOVIAN TO OXFORDIAN BENTHIC FORAMINIFERA FROM LER DOME, KUTCH BASIN (GUJARAT, INDIA): SYSTEMATIC, ECOSTRATIGRAPHY AND PALAEOENVIRONMENTAL RECONSTRUCTION

Analysis of the foraminiferal assemblages of the Chari Formation (Middle-Upper Jurassic transition) exposed at Ler Dome, Kutch Basin (India) allows one to interpret the incidence of different environmental parameters, especially the effect of sea-level changes in this group of microorganisms. The overall deposition of the Chari Formation took place in an open marine environment in the middle to outer shelf, having normal salinity and well-oxygenated bottom waters according to the lithofacies and the composition of the foraminiferal and macroinvertebrate assemblages. Changes in the diversity, abundance of foraminifera, and proportion of specialist forms were associated mainly with the availability of labile organic matter on the sea floor. The changes in trophic resources were associated with fluctuations in the type of sedimentation, which ranges from carbonates to siltstones and sandstones. During the regressive phase, a relatively high input of food resources, probably phytodetritus, was associated with siliciclastic sedimentation and commonly related with increased abundance and diversity of foraminifera, including specialist forms. During the transgressive phase, the influx of food resources from emerged areas and shallow environments decreased; sedimentation was more calcareous, with an accumulation of ammonoid shells that indicates hemipelagic conditions. The decrease in food resources for benthic foraminifera is reflected by a lesser diversity and abundance, and lower proportions of specialist foraminifera

Digital breast tomosynthesis versus additional diagnostic mammographic views for the evaluation of asymmetric mammographic densities

Introduction: Many young females present with an advanced stage of breast cancer, which has a negative effect on the prognosis. Digital breast tomosynthesis is a new emerging imaging technique that aids in improving the specificity of mammography with subsequent early detection of breast cancer, especially in women with radiographically dense breasts. Tomosynthesis is subjectively preferred to conventional mammography and may offer superior diagnostic accuracy for the evaluation of breast lesions.Method: Two breast radiologists retrospectively reviewed asymmetric densities using protocols that were institutional review board-approved in 185 patients aged 18 - 70 years (mean: 48 years) who underwent diagnostic mammography and tomosynthesis. Each asymmetric density was interpreted once with tomosynthesis and once with supplemental mammographic views; both modes included the mediolateral oblique and craniocaudal views in a fully crossed and balanced design by using a five-category Breast Imaging Reporting and Data System (BI-RADS) assessment and a probability-of-malignancy score. If the abnormality persisted and appeared benign or completely disappeared on both modalities, the agreement between additional views and tomosynthesis was determined by calculating Kappa value. If there was a discrepancy between additional views and tomosynthesis, the abnormality was subjected to ultrasound. In our study, 89 asymmetric densities were subjected to ultrasound.Results: In a total of 182 cases, 84 (46.15%) were categorized as BIRADS-0; 97 (53.30 %) as BIRADS-I, and one (0.55 %) as BIRADS-II on an additional view. Among the asymmetric densities categorized as BIRADS-0 on additional mammography views, digital breast tomosynthesis categorized 72, six, five, and one patient as BIRADS-0, BIRADS-I, BIRADS-II, and BIRADS-IV, respectively. For densities categorized as BIRADS-I (97) on additional view, digital breast tomosynthesis categorized 10 and 87 densities as BIRADS-0 and BIRADS-I, respectively. No change in the BIRADS category was observed among BIRADS-II and BIRADS-IV. A significant difference was observed with the chi-square test among BIRADS categories assigned by an additional view and digital breast tomosynthesis with a p-value of \u3c 0.001. There was, however, a substantial agreement among additional views and tomosynthesis with a kappa value of 0.767.Conclusion: Our study results suggest that tomosynthesis may be equivalent to, if not more equivalent to, additional imaging in the assessment of mammographically-detected asymmetric densities, thus improving BI-RADS classification and patient management

QUANTITATION OF AMLODIPINE IN HUMAN PLASMA BY LCMS/MS ASSAY

­­­­­­­­­­­­­­­­­Objective: To develop and validate a simple, precise, and rapid liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for quantification of amlodipine in human plasma.Methods: Chromatographic analysis was performed on Atlantis dC18 column (2.1 x 100 mm, 3 µm) with a mobile phase consisting of acetonitrile and 10 mM formic acid (80:20, v: v) that was delivered at a flow rate of 0.3 ml/min. The eluents were monitored using electrospray ionization in the positive ion mode set at transition 409 → 238.4 and 254.3 → 43.9 for amlodipine and tizanidine hydrochloride (IS), respectively. The method was validated for linearity, accuracy, precision, and recovery as per US-FDA guidelines.Results: The retention times of amlodipine and tizanidine (IS) were 1.26 and 1.22 respectively. The relationship between amlodipine concentration and peak height ratio of amlodipine to the IS was linear (R2³ 0.9868) in the range of 0.2–20 ng/ml, and the intra-and inter-day coefficient of variations and bias were ≤14.4% and ≤13.6% and ≤13.7% and ≤11.2%, respectively.Conclusion: The proposed method is simple, precise, and accurate for rapid measurement of amlodipine level using 0.5 ml human plasma. Further, the assay was successfully applied to determine amlodipine level in human plasma samples obtained from a healthy volunteer

Genomes of <i>Helicobacter pylori</i> from native Peruvians suggest admixture of ancestral and modern lineages and reveal a western type cag-pathogenicity island

Background: Helicobacter pylori is presumed to be co-evolved with its human host and is a highly diverse gastric pathogen at genetic levels. Ancient origins of H. pylori in the New World are still debatable. It is not clear how different waves of human migrations in South America contributed to the evolution of strain diversity of H. pylori. The objective of our 'phylogeographic' study was to gain fresh insights into these issues through mapping genetic origins of H. pylori of native Peruvians (of Amerindian ancestry) and their genomic comparison with isolates from Spain, and Japan. Results: For this purpose, we attempted to dissect genetic identity of strains by fluorescent amplified fragment length polymorphism (FAFLP) analysis, multilocus sequence typing (MLST) of the 7 housekeeping genes (atpA, efp, ureI, ppa, mutY, trpC, yphC) and the sequence analyses of the babB adhesin and oipA genes. The whole cag pathogenicity-island (cagPAI) from these strains was analyzed using PCR and the geographic type of cagA phosphorylation motif EPIYA was determined by gene sequencing. We observed that while European genotype (hp-Europe) predominates in native Peruvian strains, approximately 20% of these strains represent a sub-population with an Amerindian ancestry (hsp-Amerind). All of these strains however, irrespective of their ancestral affiliation harbored a complete, 'western' type cagPAI and the motifs surrounding it. This indicates a possible acquisition of cagPAI by the hsp-Amerind strains from the European strains, during decades of co-colonization. Conclusion: Our observations suggest presence of ancestral H. pylori (hsp-Amerind) in Peruvian Amerindians which possibly managed to survive and compete against the Spanish strains that arrived to the New World about 500 years ago. We suggest that this might have happened after native Peruvian H. pylori strains acquired cagPAI sequences, either by new acquisition in cagnegative strains or by recombination in cag positive Amerindian strains

NIR-dye based mucoadhesive nanosystem for photothermal therapy in breast cancer cells

Breast cancer is one of the leading causes of mortality in women, worldwide. The average survival rate of patients suffering from advanced breast cancer is about 27% for five years. Photothermal therapy employing biodegradable nanoparticle are extensively researched for enhanced anticancer therapy in breast cancer treatment. In the current study, we report a chitosan based mucoadherant and biodegradable niosome nanoparticle entrapping near infrared (NIR) dye (IR 806) for the treatment of breast cancer. Niosome entrapping IR 806 (NioIR) showed encapsulation efficacy of about 56 ± 2%. The prepared nanoparticles (NioIR) were further coated with chitosan (NioIR-C) to impart mucoadhesive property to the nanosystem. NioIR-C showed minimal degradation following NIR laser irradiation, thus enhancing its photothermal stability. They also exhibited efficient photothermal transduction, when compared with IR 806 dye. NioIR-C were biocompatible when treated with normal cell lines (NIH 3T3 and L929) and showed cytotoxicity towards breast cancer cell lines (MCF-7 and MDA-MB 231). When triggered with NIR laser, NioIR-C showed photothermal cell death (approximately 93%). The presence of chitosan coating on NioIR led to mucoadherence potential that further enhances the therapeutic effect on breast cancer cells when compared with IR 806 dye and NioIR. Thus NioIR-C can be a promising nanosystem for effective treatment of breast cancer using photothermal therapy. © 2020 Elsevier B.V

Genomes of Helicobacter pylori from native Peruvians suggest admixture of ancestral and modern lineages and reveal a western type cag-pathogenicity island

BACKGROUND: Helicobacter pylori is presumed to be co-evolved with its human host and is a highly diverse gastric pathogen at genetic levels. Ancient origins of H. pylori in the New World are still debatable. It is not clear how different waves of human migrations in South America contributed to the evolution of strain diversity of H. pylori. The objective of our 'phylogeographic' study was to gain fresh insights into these issues through mapping genetic origins of H. pylori of native Peruvians (of Amerindian ancestry) and their genomic comparison with isolates from Spain, and Japan. RESULTS: For this purpose, we attempted to dissect genetic identity of strains by fluorescent amplified fragment length polymorphism (FAFLP) analysis, multilocus sequence typing (MLST) of the 7 housekeeping genes (atpA, efp, ureI, ppa, mutY, trpC, yphC) and the sequence analyses of the babB adhesin and oipA genes. The whole cag pathogenicity-island (cagPAI) from these strains was analyzed using PCR and the geographic type of cagA phosphorylation motif EPIYA was determined by gene sequencing. We observed that while European genotype (hp-Europe) predominates in native Peruvian strains, approximately 20% of these strains represent a sub-population with an Amerindian ancestry (hsp-Amerind). All of these strains however, irrespective of their ancestral affiliation harbored a complete, 'western' type cagPAI and the motifs surrounding it. This indicates a possible acquisition of cagPAI by the hsp-Amerind strains from the European strains, during decades of co-colonization. CONCLUSION: Our observations suggest presence of ancestral H. pylori (hsp-Amerind) in Peruvian Amerindians which possibly managed to survive and compete against the Spanish strains that arrived to the New World about 500 years ago. We suggest that this might have happened after native Peruvian H. pylori strains acquired cagPAI sequences, either by new acquisition in cag-negative strains or by recombination in cag positive Amerindian strains

A rural community-based investigation to assess mHealth for family planning advice in district Multan

Background: Globally, Pakistan is the 5th populous country. To increase the use of family planning services by public private business model, Punjab Population Innovation Fund selected “Connect4fp” project implemented by International Rescue Committee organization. The purpose of this research was to determine the impact of mobile health(mHealth) advice on family planning in project area. This project was implemented in 9 Union councils of district Multan, Pakistan. The innovative idea of this project was the use of mobile phones to spread family planning knowledge among community in the form of Interactive Voice Response (IVR) as well as Short Message Service (SMS) (i.e., text messages). Methods: A cross-sectional study targeted 197 men as well as women (mostly married women of reproductive age) receiving mobile messages or recorded robocalls from the project area. The purpose was to assess their family planning knowledge and awareness by using structured Likert scale questionnaire. Secondly the viewpoint about mHealth and its effect on family planning was determined from 40 private providers and 27 Family planning Champions (who were part of the project) by telephonic interviews. Their response was quantified by using Likert scale. Descriptive statistics were calculated of the study’s main variables. The association of family planning knowledge of community members was determined to their contraceptive use by using Chi square. Results:  The results specified that 60% community members included in this study told that their family planning knowledge is increased due to this mHealth program. 36% Private providers while 75% FP Champions were agreed that family planning patients are increased due to this mHealth program. Conclusion: Mobile messages and robocalls about family planning sent in local language positively contributed to increase FP knowledge and awareness in community. This may trigger communication about family planning within couples, as a result increase use of family planning services.