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Introduction: The maternal and fetal outcomes are poorer in pregnancies associated with obstetric complications. A newly recognized marker of hypoxia, serum ischemia-modified albumin (IMA), which serves as an early predictive marker of myocardial infarction in the emergency department, is now being studied in obstetric complications of pregnancy. Aims and Objectives: (1) To study the levels of maternal serum IMA in the early 2nd trimester (12–20 weeks) to predict the occurrence of uncomplicated and complicated pregnancy post 20 weeks of gestation. (2) To evaluate the IMR ratio (IMAR) (serum IMA-to-human serum albumin ratio) and its significance in the obstetric complications of pregnancy. Methodology: This prospective cohort study was conducted on 150 antenatal women with gestational age from 12 to 20 weeks. A complete detailed history of all participants was taken and they were followed up to the time of delivery for the specific complications (i.e., pregnancy-induced hypertension [PIH], preeclampsia, eclampsia, and gestational diabetes in mother and stillbirth, preterm birth, and fetal growth restriction [FGR] in the fetus) that developed during pregnancy. IMA levels and IMA ratio were obtained and correlated with the development of the defined complications of pregnancy. The data were recorded, tabulated, and subjected to statistical analysis. Results: A significant relationship between IMA value and the incidence of PIH, preeclampsia, and eclampsia and between IMAR value and the incidence of preeclampsia and eclampsia was seen among the study population (P < 0.05). We observed that IMA and IMAR values were higher in women who did not have any fetal complications than in women who had fetal complications, namely, stillbirth, preterm birth, and FGR. However, this difference was not significant (P > 0.05). Conclusion: Serum IMA and IMAR levels were higher in women with PIH, preeclampsia, and eclampsia. The most common pregnancy outcome was small for gestation age. No significant link was found between serum IMA levels or IMAR and the severity of eclampsia, gestational diabetes, stillbirth, preterm, and FGR.

Background: The Global Typhoid Genomics Consortium was established to bring together the typhoid research community to aggregate and analyse Salmonella enterica serovar Typhi (Typhi) genomic data to inform public health action. This analysis, which marks 22 years since the publication of the first Typhi genome, represents the largest Typhi genome sequence collection to date (n=13,000). Methods: This is a meta-analysis of global genotype and antimicrobial resistance (AMR) determinants extracted from previously sequenced genome data and analysed using consistent methods implemented in open analysis platforms GenoTyphi and Pathogenwatch. Results: Compared with previous global snapshots, the data highlight that genotype 4.3.1 (H58) has not spread beyond Asia and Eastern/Southern Africa; in other regions, distinct genotypes dominate and have independently evolved AMR. Data gaps remain in many parts of the world, and we show the potential of travel-associated sequences to provide informal ‘sentinel’ surveillance for such locations. The data indicate that ciprofloxacin non-susceptibility (>1 resistance determinant) is widespread across geographies and genotypes, with high-level ciprofloxacin resistance (≥3 determinants) reaching 20% prevalence in South Asia. Extensively drug-resistant (XDR) typhoid has become dominant in Pakistan (70% in 2020) but has not yet become established elsewhere. Ceftriaxone resistance has emerged in eight non-XDR genotypes, including a ciprofloxacin-resistant lineage (4.3.1.2.1) in India. Azithromycin resistance mutations were detected at low prevalence in South Asia, including in two common ciprofloxacin-resistant genotypes. Conclusions: The consortium’s aim is to encourage continued data sharing and collaboration to monitor the emergence and global spread of AMR Typhi, and to inform decision-making around the introduction of typhoid conjugate vaccines (TCVs) and other prevention and control strategies. Funding: No specific funding was awarded for this meta-analysis. Coordinators were supported by fellowships from the European Union (ZAD received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 845681), the Wellcome Trust (SB, Wellcome Trust Senior Fellowship), and the National Health and Medical Research Council (DJI is supported by an NHMRC Investigator Grant [GNT1195210]).

Background: During October 2020, Delta variant was detected for the first time in India and rampantly spread across the globe. It also led to second wave of pandemic in India which affected millions of people. However, there is limited information pertaining to the SARS-CoV-2 strain infecting the children in India. Methods: Here, we assessed the SARS-CoV-2 lineages circulating in the pediatric population of India during the second wave of the pandemic. Clinical and demographic details linked with the nasopharyngeal/oropharyngeal swabs (NPS/OPS) collected from SARS-CoV-2 cases (n = 583) aged 0–18 year and tested positive by real-time RT-PCR were retrieved from March to June 2021. Results: Symptoms were reported among 37.2% of patients and 14.8% reported to be hospitalized. The E gene CT value had significant statistical difference at the point of sample collection when compared to that observed in the sequencing laboratory. Out of these 512 sequences 372 were VOCs, 51 were VOIs. Most common lineages observed were Delta, followed by Kappa, Alpha and B.1.36, seen in 65.82%, 9.96%, 6.83% and 4.68%, respectively in the study population. Conclusion: Overall, it was observed that Delta strain was the leading cause of SARS-CoV-2 infection in Indian children during the second wave of the pandemic. We emphasize on the need of continuous genomic surveillance in SARS-CoV-2 infection even amongst children.

Introduction: The introduction of curriculum based medical education by National Medical Commission (NMC) aims to produce graduates of global standard through curricular reforms. Early Clinical Exposure (ECE) is one of the proposed reforms in the new curriculum, which when designed and integrated into the teaching programmes will enhance the attitude and communication skills of the students. Aim: To develop, implement and evaluate the ECE module in biochemistry and also evaluate the faculty and student feedback for ECE as teaching module and ways to improve them. Materials and Methods: The cross-sectional study, was conducted from June 2020 to August 2020, in the Department of Biochemistry, CMC and hospital Ludhiana, Punjab, India. Study was conducted on 70 phase 1 Bachelor of Medicine and Bachelor of Surgery (MBBS) students admitted in year 2020. The faculty along with the students were sensitised to the ECE teaching module. The ECE module pertaining to the two selected topics (Type 2 Diabetes Mellitus (T2DM) and Jaundice) was developed and discussed under case-based sessions and clinical implementation sessions. After these sessions the feedback was taken from the faculty and the students. The results were collected, analysed and expressed in percentages and charts. Results: In present study, out of total 75 students (a batch of 75 out of which five students were absent) 29 (38.67%) were boys and 46 (61.33%) were girls. Majority of the students 35 (50%) strongly agreed that ECE increased their attention in class, and helped them to understand the topic in a better manner- 36 (51.4%) agreed, with a better correlation of the topic- 31 (44.3%) agreed. whereas the majority 3 (75%) of the faculty agreed that ECE is important for understanding the clinical relevance of the topic and 3 (75%) agreed that its more time consuming. All the faculty agreed 4 (100%) that it is better teaching method as compared to the traditional method. Conclusion: ECE is better and acceptable way of teaching and learning then traditional. It motivates students to read and helps them understand the clinical relevance in a better way. It increases their attention in the class and helps to retain the topic for long

Abstract Background In previous studies, five vasoactive drugs were investigated for their effect on the recovery process after extended liver resection without observing relevant improvements. We hypothesized that an analysis of gene expression could help to identify potentially druggable pathways and could support the selection of promising drug candidates. Methods Liver samples obtained from rats after combined 70% partial hepatectomy and right median hepatic vein ligation (n = 6/group) sacrificed at 0 h, 24 h, 48 h, and 7days were selected for this study. Liver samples were collected from differentially perfused regions of the median lobe (obstruction-zone, border-zone, normal-zone). Gene expression profiling of marker genes regulating hepatic hemodynamics, vascular remodeling, and liver regeneration was performed with microfluidic chips. We used 3 technical replicates from each sample. Raw data were normalized using LEMming and differentially expressed genes were identified using LIMMA. Results The strongest differences were found in obstruction-zone at 24 h and 48 h postoperatively compared to all other groups. mRNA expression of marker genes from hepatic hemodynamics pathways (iNOS,Ptgs2,Edn1) was most upregulated. Conclusion These upregulated genes suggest a strong vasoconstrictive effect promoting arterial hypoperfusion in the obstruction-zone. Reducing iNOS expression using selective iNOS inhibitors seems to be a promising approach to promote vasodilation and liver regeneration.

Paratyphoid fever caused by S. Paratyphi A is endemic in parts of South Asia and Southeast Asia. The proportion of enteric fever cases caused by S. Paratyphi A has substantially increased, yet only limited data is available on the population structure and genetic diversity of this serovar. We examined the phylogenetic distribution and evolutionary trajectory of S. Paratyphi A isolates collected as part of the Indian enteric fever surveillance study "Surveillance of Enteric Fever in India (SEFI)." In the study period (2017-2020), S. Paratyphi A comprised 17.6% (441/2503) of total enteric fever cases in India, with the isolates highly susceptible to all the major antibiotics used for treatment except fluoroquinolones. Phylogenetic analysis clustered the global S. Paratyphi A collection into seven lineages (A-G), and the present study isolates were distributed in lineages A, C and F. Our analysis highlights that the genome degradation events and gene acquisitions or losses are key molecular events in the evolution of new S. Paratyphi A lineages/sub-lineages. A total of 10 hypothetically disrupted coding sequences (HDCS) or pseudogenes-forming mutations possibly associated with the emergence of lineages were identified. The pan-genome analysis identified the insertion of P2/PSP3 phage and acquisition of IncX1 plasmid during the selection in 2.3.2/2.3.3 and 1.2.2 genotypes, respectively. We have identified six characteristic missense mutations associated with lipopolysaccharide (LPS) biosynthesis genes of S. Paratyphi A, however, these mutations confer only a low structural impact and possibly have minimal impact on vaccine effectiveness. Since S. Paratyphi A is human-restricted, high levels of genetic drift are not expected unless these bacteria transmit to naive hosts. However, public-health investigation and monitoring by means of genomic surveillance would be constantly needed to avoid S. Paratyphi A serovar becoming a public health threat similar to the S. Typhi of today.

Purpose: Magnetic resonance image guided radiation therapy (MR-IGRT) has been used at our institution since 2014. We report on more than 2 years of clinical experience in treating patients with the world's first MR-IGRT system. Methods and materials: A clinical service was opened for MR-IGRT in January 2014 with an MR-IGRT system consisting of a split 0.35T magnetic resonance scanner that straddles a ring gantry with 3 multileaf collimator-equipped 60Co heads. The service was expanded to include online adaptive radiation therapy (ART) MR-IGRT and cine gating after 6 and 9 months, respectively. Patients selected for MR-IGRT were enrolled in a prospective registry between January 2014 and June 2016. Patients were treated with a variety of radiation therapy techniques including intensity modulated radiation therapy and stereotactic body radiation therapy (SBRT). When applicable, online ART was performed and gating on sagittal 2-dimensional cine MR was used. The charts of patients treated with MR-IGRT were reviewed to report on the clinical and treatment characteristics of the initial patients who were treated with this novel technique. Results: A total of 316 patients have been treated with the MR-IGRT system, which has been integrated into a high-volume clinic. The cases were most commonly selected for improved soft tissue visualization, ART, and cine gating. Seventy-six patients were treated with 3-dimensional conformal radiation therapy, 146 patients with intensity modulated radiation therapy, and 94 patients with SBRT. The most commonly treated disease sites were the abdomen (28%), breast (26%), pelvis (22%), thorax (19%), and head and neck (5%). Sixty-seven patients were treated with online ART over a total of 244 adapted fractions. Cine treatment gating was used for a total of 81 patients. Conclusions: MR-IGRT has been successfully implemented in a high-volume radiation clinic and provides unique advantages in the treatment of a variety of malignancies. Additional clinical trials are in development to formally evaluate MR-IGRT in the treatment of multiple disease sites with techniques such as SBRT and ART.

Macrophage-derived cytokines largely influence the behavior of hepatocytes during an inflammatory response. We previously reported that both TNFα and IL-1β, which are released by macrophages upon LPS stimulation, affect Fas ligand (FasL)-induced apoptotic signaling. Whereas TNFα preincubation leads to elevated levels of caspase-3 activity and cell death, pretreatment with IL-1β induces increased caspase-3 activity but keeps cells alive. We now report that IL-1β and TNFα differentially influence NF-κB activity resulting in a differential upregulation of target genes, which may contribute to the distinct effects on cell viability. A reduced NF-κB activation model was established to further investigate the molecular mechanisms which determine the distinct cell fate decisions after IL-1β and TNFα stimulation. To study this aspect in a more physiological setting, we used supernatants from LPS-stimulated bone marrow-derived macrophages (BMDMs). The treatment of hepatocytes with the BMDM supernatant, which contains both IL-1β and TNFα, sensitized to FasL-induced caspase-3 activation and cell death. However, when TNFα action was blocked by neutralizing antibodies, cell viability after stimulation with the BMDM supernatant and FasL increased as compared to single FasL stimulation. This indicates the important role of TNFα in the sensitization of apoptosis in hepatocytes. These results give first insights into the complex interplay between macrophages and hepatocytes which may influence life/death decisions of hepatocytes during an inflammatory reaction of the liver in response to a bacterial infection.

Background: The photopatch test is used to detect photoallergic reactions to various antigens such as sunscreens and drugs. Photosensitive dermatitis can be caused due to antigens like parthenium, fragrances, rubbers and metals. The photopatch test does not contain these antigens. Therefore, the Indian Standard Series (ISS) along with the Standard photopatch series from Chemotechnique Diagnostics, Sweden was used to detect light induced antigens. Aim: To detect light induced antigens in patients with photosensitive dermatitis. Methods: This study was done in a descriptive, observer blinded manner. Photopatch test and ISS were applied in duplicate on the patient's back by the standard method. After 24 hours, readings were recorded according to ICDRG criteria. One side was closed and other side irradiated with 14 J/cm2 of UVA and a second set of readings were recorded after 48 hrs. Result: The highest positivity was obtained with parthenium, with 18 out of 35 (51%) patients showing a positive patch test reaction with both photoallergic contact dermatitis and photoaggravation. Four patients (11%) showed positive patch test reaction suggestive of contact dermatitis to potassium dichromate and fragrance mix. Six patients had contact dermatitis to numerous antigens such as nickel, cobalt, chinoform and para-phenylenediamine. None of these patients showed photoaggravation on patch testing. Conclusion: Parthenium was found to cause photoallergy, contact dermatitis with photoaggravation and contact allergy. Hence, photopatch test and UV irradiated patch test can be an important tool to detect light induced antigens in patients with photosensitive dermatitis.

Introduction: Fungi have emerged as major opportunistic pathogens. Candida species account for nearly 96% of all opportunistic mycoses and is an important cause of bloodstream infections. There has been a progressive shift from the predominance of Candida albicans to nonalbicans Candida species as the major cause of candidemia all over the world. Resistance to antifungal drugs is more in nonalbicans Candida species. Hence, speciation and antifungal susceptibility testing is the need of the hour. Materials and Methods: This retrospective study was conducted in a multispecialty center in North India from January 1, 2014 to March 31, 2015. The blood culture samples that were positive for Candida species were processed further. Species identification was done by standard microbiological techniques. Antifungal drug susceptibility was done by disk diffusion method (Clinical and Laboratory Standards Institute M44-A2). Results: A total of 10893 samples were processed, 1440 (13.2%) blood cultures were positive. Candida species was isolated from 105 (7.3%) samples, of which 15 (14.3%) were C. albicans and 90 (85.7%) were nonalbicans Candida. Nonalbicans Candida included Candida tropicalis (50.5%), Candida glabrata (19.0%), Candida parapsilosis (14.3%) and one isolate each of Candida guillermondi and Candida krusei. Majority of the Candida spp. isolates were resistant to clotrimazole (55.5%) fluconazole (42%) and itraconazole (69%) and and ketoconazole (38%). All isolates were uniformly sensitive to amphotericin B. Conclusion: There is predominance of nonalbicans Candida species in hospital setting. A high index of suspicion, early diagnosis, and a prompt and appropriate therapy remains the cornerstone of treatment.

Background & aimsKnowledge about innate antimicrobial defense of the liver is limited. We investigated hepatic expression and regulation of antimicrobial peptides with focus on the human beta defensin-1 (hBD-1).MethodsRadial diffusion assay was used to analyze antimicrobial activity of liver tissue. Different defensins including hBD-1 and its activator thioredoxin-1 (TXN) were analyzed in healthy and cholestatic liver samples by qPCR and immunostaining. Regulation of hBD-1 expression was studied in vitro and in vivo using bile duct-ligated mice. Regulation of hBD-1 via bilirubin and bile acids (BAs) was studied using siRNA.ResultsWe found strong antimicrobial activity of liver tissue against Escherichia coli. As a potential mediator of this antimicrobial activity we detected high expression of hBD-1 and TXN in hepatocytes, whereas other defensins were minimally expressed. Using a specific antibody for the reduced, antimicrobially active form of hBD-1 we found hBD-1 in co-localization with TXN within hepatocytes. hBD-1 was upregulated in cholestasis in a graded fashion. In cholestatic mice hepatic AMP expression (Defb-1 and Hamp) was enhanced. Bilirubin and BAs were able to induce hBD-1 in hepatic cell cultures in vitro. Treatment with siRNA and/or agonists demonstrated that the farnesoid X receptor (FXR) mediates basal expression of hBD-1, whereas both constitutive androstane receptor (CAR) and FXR seem to be responsible for the induction of hBD-1 by bilirubin.ConclusionhBD-1 is prominently expressed in hepatocytes. It is induced during cholestasis through bilirubin and BAs, mediated by CAR and especially FXR. Reduction by TXN activates hBD-1 to a potential key player in innate antimicrobial defense of the liver.

This article presents a multi-theoretical model to address processes of authorship development in the English as a foreign language teaching profession. Working within a sociocultural perspective of second-language teacher education, the authors examined six experienced nonnative English-speaking teacher-researchers. Perceptions of their key moments in the profession were analyzed using a mixed-method approach that combined document analysis with personal narratives. The findings indicated that both authoring (products and activities in their profession) and self-authorship (transformational processes throughout their careers) contributed to constructing these English as a foreign language professionals as scholars.

Background: RNA interference (RNAi) has tremendous potential for investigating gene function and for developing new therapies. Primary human hepatocytes (PHH) are the “gold standard” for studying the regulation of hepatic metabolism in vitro. However, application of RNAi in PHH has some technical hurdles. The objective of this study was to develop effective and robust protocol for transduction of PHH with lentiviral vectors. Methods: We used lentiviral vectors to transduce PHH for introduction of short hairpin RNAs (shRNAs) targeting constitutive androstane receptor (CAR), peroxisome proliferator activated receptor alpha (PPARα), and microRNA, miR-143. Infection efficiency was quantitatively analyzed by flow cytometry and microscopy. Target gene expression was assessed using quantitative real-time (qRT-PCR) method. Results: Lentiviral vector transduction resulted in ≥95% of infected cells at low multiplicity of infection (MOI) of 3, which did not impair cellular viability. We demonstrated the feasibility of this technique in studies on targeting nuclear receptors, PPARα and CAR, with shRNAs as well as in lentivirus-mediated overexpression and knock-down of miRNA-143 experiments. Conclusions: We developed an efficient and robust protocol with standardized procedures for virus production, method of titer determination, and infection procedure for RNAi in primary human hepatocytes based on delivery of shRNAs, microRNAs or anti-microRNAs in different laboratory settings. This approach should be useful to study not only the regulation via nuclear receptors but also other biological, pharmacological, and toxicological aspects of drug metabolism.

Context: Medical students undergo tremendous stress during various stages of the Bachelor of Medicine, Bachelor of Surgery (MBBS) course. Academic examinations have been considered as one of the most acute stresses experienced by the students. Nitric oxide (NO) is an important physiological messenger and effector molecule in many biological systems. There is evidence that a sustained overproduction of NO via inducible NO synthase (iNOS) is responsible, at least in part, for some of the neurodegenerative changes caused by stress . Aims: To investigate the relationship if any between plasma NO and psychological stress caused by academic pressure in first year MBBS students. Settings and Design: A 2-year prospective longitudinal study. Materials and Methods: A total of 94 first year medical students after informed consent were enrolled in the study. They were evaluated twice during their first year academic program. First evaluation was done 2 months after their joining the MBBS course and second on the day of their first professional university practical exam. On each evaluation, a history was taken, general physical examination done, and a blood sample was drawn for plasma NO, which was measured using Griess reaction. Statistical Analysis Used: For comparison of means of plasma NO values between the two evaluations, the paired Student′s ′t′- test was used. A ′P′ < 0.05 was considered as statistically significant. Results: The mean NO values increased from 14.76 ± 10.30 during first evaluation to 22.07 ± 13.02 during second evaluation. This increase was statistically significant (P = 0.000). Conclusions: Plasma NO showed a statistically significant increase in levels during the time of examination stress. As plasma NO had a positive correlation with stress, this can be considered as a suitable biomarker for academic stress assessment.

Given the important role of angiogenesis in liver pathology, the current study investigated the role of Runt-related transcription factor 1 (RUNX1), a regulator of developmental angiogenesis, in the pathogenesis of non-alcoholic steatohepatitis (NASH). Quantitative RT-PCRs and a transcription factor analysis of angiogenesis-associated differentially expressed genes in liver tissues of healthy controls, patients with steatosis and NASH, indicated a potential role of RUNX1 in NASH. The gene expression of RUNX1 was correlated with histopathological attributes of patients. The protein expression of RUNX1 in liver was studied by immunohistochemistry. To explore the underlying mechanisms, in vitro studies using RUNX1 siRNA and overexpression plasmids were performed in endothelial cells (ECs). RUNX1 expression was significantly correlated with inflammation, fibrosis and NASH activity score in NASH patients. Its expression was conspicuous in liver non-parenchymal cells. In vitro, factors from steatotic hepatocytes and/or VEGF or TGF-β significantly induced the expression of RUNX1 in ECs. RUNX1 regulated the expression of angiogenic and adhesion molecules in ECs, including CCL2, PECAM1 and VCAM1, which was shown by silencing or over-expression of RUNX1. Furthermore, RUNX1 increased the angiogenic activity of ECs. This study reports that steatosis-induced RUNX1 augmented the expression of adhesion and angiogenic molecules and properties in ECs and may be involved in enhancing inflammation and disease severity in NASH.

Macrophages are cells with remarkable plasticity. They integrate signals from their microenvironment leading to context-dependent polarization into classically (M1) or alternatively (M2) activated macrophages, representing two extremes of a broad spectrum of divergent phenotypes. Thereby, macrophages deliver protective and pro-regenerative signals towards injured tissue but, depending on the eliciting damage, may also be responsible for the generation and aggravation of tissue injury. Although incompletely understood, there is emerging evidence that macrophage polarization is critical for these antagonistic roles. To identify activation-specific expression patterns of chemokines and cytokines that may confer these distinct effects a systems biology approach was applied. A comprehensive literature-based Boolean model was developed to describe the M1 (LPS-activated) and M2 (IL-4/13-activated) polarization types. The model was validated using high-throughput transcript expression data from murine bone marrow derived macrophages. By dynamic modeling of gene expression, the chronology of pathway activation and autocrine signaling was estimated. Our results provide a deepened understanding of the physiological balance leading to M1/M2 activation, indicating the relevance of co-regulatory signals at the level of Akt1 or Akt2 that may be important for directing macrophage polarization.

During various inflammatory processes circulating cytokines including IL-6, IL-1β, and TNFα elicit a broad and clinically relevant impairment of hepatic detoxification that is based on the simultaneous downregulation of many drug metabolizing enzymes and transporter genes. To address the question whether a common mechanism is involved we treated human primary hepatocytes with IL-6, the major mediator of the acute phase response in liver, and characterized acute phase and detoxification responses in quantitative gene expression and (phospho-)proteomics data sets. Selective inhibitors were used to disentangle the roles of JAK/STAT, MAPK, and PI3K signaling pathways. A prior knowledge-based fuzzy logic model comprising signal transduction and gene regulation was established and trained with perturbation-derived gene expression data from five hepatocyte donors. Our model suggests a greater role of MAPK/PI3K compared to JAK/STAT with the orphan nuclear receptor RXRα playing a central role in mediating transcriptional downregulation. Validation experiments revealed a striking similarity of RXRα gene silencing versus IL-6 induced negative gene regulation (rs = 0.79; P

In the months after July 1936, acts of anticlerical violence and iconoclasm underscored attempts to reconfigure radically social relations and the physical landscape in many parts of Republican Spain. Taking the destruction of the Sacred Heart of Jesus as its analytical focus, this text examines the connection between the grassroots battle waged by anticlerical workers against the Catholic Church’s public presence during the peacetime Republican years, and the severity and forms of the war’s anticlerical destruction.

Gene expression analysis is an essential part of biological and medical investigations. Quantitative real-time PCR (qPCR) is characterized with excellent sensitivity, dynamic range, reproducibility and is still regarded to be the gold standard for quantifying transcripts abundance. Parallelization of qPCR such as by microfluidic Taqman Fluidigm Biomark Platform enables evaluation of multiple transcripts in samples treated under various conditions. Despite advanced technologies, correct evaluation of the measurements remains challenging. Most widely used methods for evaluating or calculating gene expression data include geNorm and ΔΔCt, respectively. They rely on one or several stable reference genes (RGs) for normalization, thus potentially causing biased results. We therefore applied multivariable regression with a tailored error model to overcome the necessity of stable RGs.We developed a RG independent data normalization approach based on a tailored linear error model for parallel qPCR data, called LEMming. It uses the assumption that the mean Ct values within samples of similarly treated groups are equal. Performance of LEMming was evaluated in three data sets with different stability patterns of RGs and compared to the results of geNorm normalization. Data set 1 showed that both methods gave similar results if stable RGs are available. Data set 2 included RGs which are stable according to geNorm criteria, but became differentially expressed in normalized data evaluated by a t-test. geNorm-normalized data showed an effect of a shifted mean per gene per condition whereas LEMming-normalized data did not. Comparing the decrease of standard deviation from raw data to geNorm and to LEMming, the latter was superior. In data set 3 according to geNorm calculated average expression stability and pairwise variation, stable RGs were available, but t-tests of raw data contradicted this. Normalization with RGs resulted in distorted data contradicting literature, while LEMming normalized data did not.If RGs are coexpressed but are not independent of the experimental conditions the stability criteria based on inter- and intragroup variation fail. The linear error model developed, LEMming, overcomes the dependency of using RGs for parallel qPCR measurements, besides resolving biases of both technical and biological nature in qPCR. However, to distinguish systematic errors per treated group from a global treatment effect an additional measurement is needed. Quantification of total cDNA content per sample helps to identify systematic errors.

Sustained inflammation may increase the susceptibility of hepatocytes to apoptotic cell death and therefore exacerbate liver damage. Here we report that the pro-inflammatory cytokine IL-1β sensitizes primary murine hepatocytes to Fas ligand (FasL)-induced caspase-3/-7 activity. This process was dependent on JNK1/2 and the BH3-only proteins Bim and Bid. Mathematical modeling revealed that incubation of hepatocytes with IL-1β depleted the anti-apoptotic Bcl-2 protein pool and thus shifted hepatocytes to mitochondrial type II apoptosis following Fas activation. As a consequence, IL-1β and FasL treatment enhanced cytochrome c release. Surprisingly, despite increased caspase-3/-7 activation, FasL-induced cell death was reduced by IL-1β pre-treatment. This protective effect was independent of JNK1/2, Bim or Bid. Furthermore, elevated caspase-3/-7 activity upon IL-1β and FasL treatment did not result in enhanced PARP cleavage. The protective effect of IL-1β was seen after 3 h of pre-incubation, indicating an anti-apoptotic transcriptional response. Indeed, NF-κB DNA binding was increased in response to IL-1β plus FasL and gene-expression profiling of NF-κB regulated genes revealed a transcriptional and translational upregulation of the caspase-8 inhibitor A20. A mathematical model was developed to explain the contradictious occurrence of both increased caspase-3/-7 activity and elevated cell viability by including a heterogeneous distribution of Bcl-2 proteins and variations in Fas signaling resulting in different subpopulations of hepatocytes.

BACKGROUND: Influenza A virus is one of world's major uncontrolled pathogen, causing seasonal epidemic as well as global pandemic. This was evidenced by recent emergence and continued prevalent 2009 swine origin pandemic H1N1 Influenza A virus, provoking first true pandemic in the past 40 years. In the course of its evolution, the virus acquired many mutations and multiple unidentified molecular determinants are likely responsible for the ability of the 2009 H1N1 virus to cause increased disease severity in humans. Availability of limited data on complete genome hampers the continuous monitoring of this type of events. Outbreaks with considerable morbidity and mortality have been reported from all parts of the country. METHODS/RESULTS: Considering a large number of clinical cases of infection complete genome based sequence characterization of Indian H1N1pdm virus and their phylogenetic analysis with respect to circulating global viruses was undertaken, to reveal the phylodynamic pattern of H1N1pdm virus in India from 2009-2011. The Clade VII was observed as a major circulating clade in phylogenetic analysis. Selection pressure analysis revealed 18 positively selected sites in major surface proteins of H1N1pdm virus. CONCLUSIONS: This study clearly revealed that clade VII has been identified as recent circulating clade in India as well globally. Few clade VII specific well identified markers undergone positive selection during virus evolution. Continuous monitoring of the H1N1pdm virus is warranted to track of the virus evolution and further transmission. This study will serve as a baseline data for future surveillance and also for development of suitable therapeutics.

Ninety-eight newly diagnosed cases of PML-RARαpositive APL were treated with a regimen of single agent ATO. FLT3 activating mutations were seen in 33% and an additional cytogenetic finding was noted in 23.2%. FLT3 activating mutations were significantly associated with a bcr3 PML-RARαisoform (p=0.012) and a delay in achieving a molecular remission (p=0.022). Neither FLT3 activating mutations nor secondary cytogenetic changes had an impact on clinical outcome.

A 47-year-old Indian woman with dark hair developed dramatic alopecia of the occipito-vertical area two weeks after a fluoroscopically-guided endovascular procedure for treatment of cerebral aneurysm. There was spontaneous repopulation of hair in 14 weeks. Neuro-radiological intervention procedures are becoming commoner by the day in India, and the rare but documented possibility of such reactions occurring in patients should be kept in mind by the treating surgeon as well as dermatologist. Necessary counseling regarding this uncommon side-effect is of essence, especially when the radiation dose exceeds 3 Gy. We believe this is the first case ever reported in Indian dermatology literature.

This article examines a series of dialogues by Pedro de Ribadeneyra (1527-1611) that recount stories of men who left the Society of Jesus only to endure a wide variety of misfortunes thereafter. These dialogues reveal a certain anxiety within the Society of Jesus concerning men who abandoned their vocation. When compared with Jesuit hagiographies, the stories of these men who left the Society show that the Jesuits were concerned that proximity to family could present temptations too poweful to overcome for many Jesuits. Ultimately, the rhetorical and propagandistic nature of the text presents the defectors as foils to showcase the holiness of the Society and its saints. Keywords: familial obligation, Pedro de Ribadeneyra, hagiography, divine punishment, rhetoric, vice and virtue, Introduction For centuries the historians of the Society of Jesus fell into two polemical parties. On the one hand, the apologists, composed primarily of Jesuits themselves, praised the Society's holiness, [...]