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Background: Although prediction of hospital readmissions has been studied in medical patients, it has received relatively little attention in surgical patient populations. Published predictors require information only available at the moment of discharge. The authors hypothesized that machine learning approaches can be leveraged to accurately predict readmissions in postoperative patients from the emergency department. Further, the authors hypothesize that these approaches can accurately predict the risk of readmission much sooner than hospital discharge., Methods: Using a cohort of surgical patients at a tertiary care academic medical center, surgical, demographic, lab, medication, care team, and current procedural terminology data were extracted from the electronic health record. The primary outcome was whether there existed a future hospital readmission originating from the emergency department within 30 days of surgery. Secondarily, the time interval from surgery to the prediction was analyzed at 0, 12, 24, 36, 48, and 60 h. Different machine learning models for predicting the primary outcome were evaluated with respect to the area under the receiver-operator characteristic curve metric using different permutations of the available features., Results: Surgical hospital admissions (N = 34,532) from April 2013 to December 2016 were included in the analysis. Surgical and demographic features led to moderate discrimination for prediction after discharge (area under the curve: 0.74 to 0.76), whereas medication, consulting team, and current procedural terminology features did not improve the discrimination. Lab features improved discrimination, with gradient-boosted trees attaining the best performance (area under the curve: 0.866, SD 0.006). This performance was sustained during temporal validation with 2017 to 2018 data (area under the curve: 0.85 to 0.88). Lastly, the discrimination of the predictions calculated 36 h after surgery (area under the curve: 0.88 to 0.89) nearly matched those from time of discharge., Conclusions: A machine learning approach to predicting postoperative readmission can produce hospital-specific models for accurately predicting 30-day readmissions via the emergency department. Moreover, these predictions can be confidently calculated at 36 h after surgery without consideration of discharge-level data.

Background and Aim: Brucellosis is an important zoonotic disease that affects fertility in farm animals. The risk factors of brucellosis have not been well studied. This study aimed to understand the seroprevalence and risk factors of brucellosis among livestock in Bangriposi block of Mayurbhanj district in Odisha, a region that borders Similipal wildlife reserve., Materials and Methods: Rose Bengal plate test (RBPT) was carried out to estimate the seroprevalence of the livestock in this region. Bivariate analysis was carried out to analyze the association between the variables and brucellosis. Binary logistic regression was performed to assess the risk factors associated with brucellosis in the livestock., Results: Based on RBPT, the seroprevalence of brucellosis among cattle and goats was estimated to be 1.1% and 11.2%, respectively. Binary logistic regression analysis indicates that study area, age, goats, animals with a history of abortion, and rearing practices were the major risk factors in this region., Conclusion: This is one of the first studies in India to shed light on risk factors of brucellosis, an important neglected disease that affects the health of animals and humans and nation's economy., (Copyright: © Behera, et al.)

Malaria parasites rely on a plastid organelle for survival during the blood stages of infection. However, the entire organelle is dispensable as long as the isoprenoid precursor, isopentenyl pyrophosphate (IPP), is supplemented in the culture medium. We engineered parasites to produce isoprenoid precursors from a mevalonate-dependent pathway, creating a parasite line that replicates normally after the loss of the apicoplast organelle. We show that carbon-labeled mevalonate is specifically incorporated into isoprenoid products, opening new avenues for researching this essential class of metabolites in malaria parasites. We also show that essential apicoplast proteins, such as the enzyme target of the drug fosmidomycin, can be deleted in this mevalonate bypass parasite line, providing a new method to determine the roles of other important apicoplast-resident proteins. Several antibacterial drugs kill malaria parasites by targeting basic processes, such as transcription, in the organelle. We used metabolomic and transcriptomic methods to characterize parasite metabolism after azithromycin treatment triggered loss of the apicoplast and found that parasite metabolism and the production of apicoplast proteins is largely unaltered. These results provide insight into the effects of apicoplast-disrupting drugs, several of which have been used to treat malaria infections in humans. Overall, the mevalonate bypass system provides a way to probe essential aspects of apicoplast biology and study the effects of drugs that target apicoplast processes., Competing Interests: The authors have declared that no competing interests exist.

Ribonucleic acid (RNA) viruses associated with chronic diseases in humans are major threats to public health causing high mortality globally. The high mutation rate of RNA viruses helps them to escape the immune response and also is responsible for the development of drug resistance. Chronic infections caused by human immunodeficiency virus (HIV) and hepatitis viruses (HBV and HCV) lead to acquired immunodeficiency syndrome (AIDS) and hepatocellular carcinoma respectively, which are one of the major causes of human deaths. Effective preventative measures to limit chronic and re-emerging viral infections are absolutely necessary. Each class of antiviral agents targets a specific stage in the viral life cycle and inhibits them from its development and proliferation. Most often, antiviral drugs target a specific viral protein, therefore only a few broad-spectrum drugs are available. This review will be focused on the selected viral target proteins of pathogenic viruses containing single-stranded (ss) RNA genome that causes chronic infections in humans (e.g. HIV, HCV, Flaviviruses). In the recent past, an exponential increase in the number of available three-dimensional protein structures (>150000 in Protein Data Bank), allowed us to better understand the molecular mechanism of action of protein targets and antivirals. Advancements in the in silico approaches paved the way to design and develop several novels, highly specific small-molecule inhibitors targeting the viral proteins., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

This is a case report of necrosis of more than two-third of the dorsal tongue in a 74-year-old male following prolonged oral intubation for vascular surgery. This necrosis progressed from the left tongue to involve much of the dorsal tongue bilaterally. A diagnosis of rhabdomyolysis was made evident by compartment syndrome of the legs with an elevated creatine kinase level of 89 789 u/l (units per litre). The literature also reveals that vasopressin has been linked with reported cases of tongue necrosis. Other possible aetiological factors were discussed in this finding., (Published by Oxford University Press and JSCR Publishing Ltd. All rights reserved. © The Author(s) 2019.)

Low voltage, non-gassing electroosmotic pump (EOP) was assembled with poly(2-ethyl aniline) (EPANI)-Prussian blue nanocomposite electrode and commercially available hydrophilic PVDF membranes. The nanocomposite material combines excellent oxidation/reduction capacity of EPANI with exceptional stability by shuttling of proton between Prussian blue nanoparticles and EPANI redox matrix. The flow rate was highly dependent on the electrode composition but it was linear with applied voltage. The flow rate at 5 V for different nanocomposite, EPANI, EPANI-A, EPANI-B, and EPANI-C were 127.29, 187.41, 148.51, and 95.47 µL/min cm 2 , respectively, which increases substantially with increase in the Prussian blue content. The obtained best electro osmotic flux was 43 µL/min/V/cm 2 for EPANI-A. It was higher than most of the EOP assembled using polyquinone and polyanthraquinone redox polymers. The assembled EOP remained exceptionally stable until the electrode charge capacity was fully utilized. The best EOP produces a maximum stall pressure of 1.2 kPa at 2 V. These characteristics make it suitable for a variety of microfluidic/device applications., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Unlike the adult mammalian retina, Müller glia (MG) in the adult zebrafish retina are able to dedifferentiate into a "stem cell"-like state and give rise to multipotent progenitor cells upon retinal damage. We show that miR-216a is downregulated in MG after constant intense light lesioning and that miR-216a suppression is necessary and sufficient for MG dedifferentiation and proliferation during retina regeneration. miR-216a targets the H3K79 methyltransferase Dot1l, which is upregulated in proliferating MG after retinal damage. Loss-of-function experiments show that Dot1l is necessary for MG reprogramming and mediates MG proliferation downstream of miR-216a. We further demonstrate that miR-216a and Dot1l regulate MG-mediated retina regeneration through canonical Wnt signaling. This article reports a regulatory mechanism upstream of Wnt signaling during retina regeneration and provides potential targets for enhancing regeneration in the adult mammalian retina., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Interferon-regulated immune defenses protect mammals from pathogenically diverse obligate intracellular bacterial pathogens of the genus Chlamydia Interferon gamma (IFN-γ) is especially important in controlling the virulence of Chlamydia species and thus impacts the modeling of human chlamydial infection and disease in mice. How IFN-γ contributes to cell-autonomous defenses against Chlamydia species and how these pathogens evade IFN-γ-mediated immunity in their natural hosts are not well understood. We conducted a genetic screen which identified 31 IFN-γ - s ensitive (Igs) mutants of the mouse model pathogen Chlamydia muridarum Genetic suppressor analysis and lateral gene transfer were used to map the phenotype of one of these mutants, Igs4, to a missense mutation in a putative chlamydial inclusion membrane protein, TC0574. We observed the lytic destruction of Igs4-occupied inclusions and accompanying host cell death in response to IFN-γ priming or various proapoptotic stimuli. However, Igs4 was insensitive to IFN-γ-regulated cell-autonomous defenses previously implicated in anti- Chlamydia trachomatis host defense in mice. Igs4 inclusion integrity was restored by caspase inhibitors, indicating that the IFN-γ-mediated destruction of Igs4 inclusions is dependent upon the function of caspases or related prodeath cysteine proteases. We further demonstrated that the Igs4 mutant is immune restricted in an IFN-γ-dependent manner in a mouse infection model, thereby implicating IFN-γ-mediated inclusion destruction and host cell death as potent in vivo host defense mechanisms to which wild-type C. muridarum is resistant. Overall, our results suggest that C. muridarum evolved resistance mechanisms to counter IFN-γ-elicited programmed cell death and the associated destruction of intravacuolar pathogens. IMPORTANCE Multiple obligatory intracellular bacteria in the genus Chlamydia are important pathogens. In humans, strains of C. trachomatis cause trachoma, chlamydia, and lymphogranuloma venereum. These diseases are all associated with extended courses of infection and reinfection that likely reflect the ability of chlamydiae to evade various aspects of host immune responses. Interferon-stimulated genes, driven in part by the cytokine interferon gamma, restrict the host range of various Chlamydia species, but how these pathogens evade interferon-stimulated genes in their definitive host is poorly understood. Various Chlamydia species can inhibit death of their host cells and may have evolved this strategy to evade prodeath signals elicited by host immune responses. We present evidence that chlamydia-induced programmed cell death resistance evolved to counter interferon- and immune-mediated killing of Chlamydia -infected cells., (Copyright © 2019 Giebel et al.)

Background: The malarial parasite Plasmodium falciparum is an auxotroph for purines, which are required for nucleic acid synthesis during the intra-erythrocytic developmental cycle (IDC) of the parasite. The capabilities of the parasite and extent to which it can use compensatory mechanisms to adapt to purine deprivation were studied by examining changes in its metabolism under sub-optimal concentrations of hypoxanthine, the primary precursor utilized by the parasite for purine-based nucleic acid synthesis., Methods: The concentration of hypoxanthine that caused a moderate growth defect over the course of one IDC was determined. At this concentration of hypoxanthine (0.5 μM), transcriptomic and metabolomic data were collected during one IDC at multiple time points. These data were integrated with a metabolic network model of the parasite embedded in a red blood cell (RBC) to interpret the metabolic adaptation of P. falciparum to hypoxanthine deprivation., Results: At a hypoxanthine concentration of 0.5 μM, vacuole-like structures in the cytosol of many P. falciparum parasites were observed after the 24-h midpoint of the IDC. Parasites grown under these conditions experienced a slowdown in the progression of the IDC. After 72 h of deprivation, the parasite growth could not be recovered despite supplementation with 90 µM hypoxanthine. Simulations of P. falciparum metabolism suggested that alterations in ubiquinone, isoprenoid, shikimate, and mitochondrial metabolism occurred before the appearance of these vacuole-like structures. Alterations were found in metabolic reactions associated with fatty acid synthesis, the pentose phosphate pathway, methionine metabolism, and coenzyme A synthesis in the latter half of the IDC. Furthermore, gene set enrichment analysis revealed that P. falciparum activated genes associated with rosette formation, Maurer's cleft and protein export under two different nutrient-deprivation conditions (hypoxanthine and isoleucine)., Conclusions: The metabolic network analysis presented here suggests that P. falciparum invokes specific purine-recycling pathways to compensate for hypoxanthine deprivation and maintains a hypoxanthine pool for purine-based nucleic acid synthesis. However, this compensatory mechanism is not sufficient to maintain long-term viability of the parasite. Although P. falciparum can complete a full IDC in low hypoxanthine conditions, subsequent cycles are disrupted.

Acetyl-CoA carboxylase (ACC) is a biotin-dependent enzyme that is the target of several classes of herbicides. Malaria parasites contain a plant-like ACC, and this is the only protein predicted to be biotinylated in the parasite. We found that ACC is expressed in the apicoplast organelle in liver- and blood-stage malaria parasites; however, it is activated through biotinylation only in the liver stages. Consistent with this observation, deletion of the biotin ligase responsible for ACC biotinylation does not impede blood-stage growth, but results in late liver-stage developmental defects. Biotin depletion increases the severity of the developmental defects, demonstrating that parasite and host biotin metabolism are required for normal liver-stage progression. This finding may link the development of liver-stage malaria parasites to the nutritional status of the host, as neither the parasite nor the human host can synthesize biotin., Competing Interests: The authors declare no conflict of interest., (Copyright © 2018 the Author(s). Published by PNAS.)

The evolution of drug resistance is a recurrent problem that has plagued efforts to treat and control malaria. Recent emergence of artemisinin resistance in Southeast Asia underscores the need to develop novel antimalarials and identify new targetable pathways in Plasmodium parasites. Transmission-blocking approaches, which typically target gametocytes in the host bloodstream or parasite stages in the mosquito gut, are recognized collectively as a strategy that when used in combination with antimalarials that target erythrocytic stages will not only cure malaria but will also prevent subsequent transmission. We tested four derivatives of (+)-usnic acid, a metabolite isolated from lichens, for transmission-blocking activity against Plasmodium falciparum using the standard membrane feeding assay. For two of the derivatives, BT37 and BT122, we observed a consistent dose-response relationship between concentration in the blood meal and oocyst intensity in the midgut. To explore their mechanism of action, we used the murine model Plasmodium berghei and found that both derivatives prevent ookinete maturation. Using fluorescence microscopy, we demonstrated that in the presence of each compound zygote vitality was severely affected, and those that did survive failed to elongate and mature into ookinetes. The observed phenotypes were similar to those described for mutants of specific kinases (NEK2/NEK4) and of inner membrane complex 1 (IMC1) proteins, which are all vital to the zygote-to-ookinete transition. We discuss the implications of our findings and our high-throughput screening approach to identifying next generation, transmission-blocking antimalarials based on the scaffolds of these (+)-usnic acid derivatives.

Objectives: The aim of this double blinded randomized controlled clinical trial was to evaluate the efficacy of two local anaesthetic solutions, 'Plain lignocaine' and 'Lignocaine with vasoconstrictor', on pain during administration and post-extraction wound healing in patients undergoing therapeutic extractions., Materials and Methods: Fifty patients indicated for therapeutic extraction of upper and lower premolars for orthodontic purpose were recruited for the study. Using a split-mouth study design, anaesthesia was achieved using lignocaine with adrenaline on the control side and plain lignocaine on the study side. Pain perception was measured by modified visual analogue scale and wound healing was assessed by Landry's Wound Healing Index. Sample allocation was done by simple randomization. The outcome parameters compared were (1) pain during administration of LA and (2) post-operative healing after extraction. Data analysis involved Chi-square test to compare proportions between treatment groups and independent sample t-test to compare mean values between treatment groups. SPSS version 22.0 was used to analyse the data., Results: The study group demonstrated a statistically significant wound healing on day1 and day 3 between the study and control group with p < 0.001. In the control group 68% had severe pain, whereas only 2% had severe pain in the study group. The proportions between the two groups were statistically significant (p < 0.001)., Conclusion: The patients who received Plain Lignocaine perceived less pain during injection of local anaesthetic solution when compared to patients who received lignocaine with vasoconstrictor. The early post-operative wound healing was better in patients anaesthetized by Plain Lignocaine.

Over the last three decades, phage display technology has been used for the display of target-specific biomarkers, peptides, antibodies, etc. Phage display-based assays are mostly limited to the phage ELISA, which is notorious for its high background signal and laborious methodology. These problems have been recently overcome by designing a dual-display phage with two different end functionalities, namely, streptavidin (STV)-binding protein at one end and a rheumatoid arthritis-specific autoantigenic target at the other end. Using this dual-display phage, a much higher sensitivity in screening specificities of autoantibodies in complex serum sample has been detected compared to single-display phage system on phage ELISA. Herein, we aimed to develop a novel, rapid, and sensitive dual-display phage to detect autoantibodies presence in serum samples using quartz crystal microbalance with dissipation monitoring as a sensing platform. The vertical functionalization of the phage over the STV-modified surfaces resulted in clear frequency and dissipation shifts revealing a well-defined viscoelastic signature. Screening for autoantibodies using antihuman IgG-modified surfaces and the dual-display phage with STV magnetic bead complexes allowed to isolate the target entities from complex mixtures and to achieve a large response as compared to negative control samples. This novel dual-display strategy can be a potential alternative to the time consuming phage ELISA protocols for the qualitative analysis of serum autoantibodies and can be taken as a departure point to ultimately achieve a point of care diagnostic system.

Cracked tooth syndrome (CTS), the term was coined by Cameron in 1964, which refers to an incomplete fracture of a vital posterior tooth extending to the dentin and occasionally into the pulp. CTS has always been a nightmare to the patient because of its unpredictable symptoms and a diagnostic dilemma for the dental practitioner due to its variable, bizarre clinical presentation. The treatment planning and management of CTS has also given problems and challenges the dentist as there is no specific treatment option. The management of CTS varies from one case to another or from one tooth to another in the same individual based on the severity of the symptoms and depth of tooth structure involved. After all, the prognosis of such tooth is still questionable and requires continuous evaluation. This article aims at presenting a series three cases of CTS with an overview on the clinical presentation, diagnosis and the different treatment options that varies from one case to another.

The ability of certain species of Chlamydia to inhibit the biogenesis of phagolysosomes permits their survival and replication within macrophages. The survival of macrophage-adapted chlamydiae correlates with the multiplicity of infection (MOI), and optimal chlamydial growth occurs in macrophages infected at an MOI of ≤1. In this study, we examined the replicative capacity of Chlamydia muridarum in the RAW 264.7 murine macrophage cell line at different MOIs. C. muridarum productively infected these macrophages at low MOIs but yielded few viable elementary bodies (EBs) when macrophages were infected at a moderate (10) or high (100) MOI. While high MOIs caused cytotoxicity and irreversible host cell death, macrophages infected at a moderate MOI did not show signs of cytotoxicity until late in the infectious cycle. Inhibition of host protein synthesis rescued C. muridarum in macrophages infected at a moderate MOI, implying that chlamydial growth was blocked by activated defense mechanisms. Conditioned medium from these macrophages was antichlamydial and contained elevated levels of interleukin 1β (IL-1β), IL-6, IL-10, and beta interferon (IFN-β). Macrophage activation depended on Toll-like receptor 2 (TLR2) signaling, and cytokine production required live, transcriptionally active chlamydiae. A hydroxyl radical scavenger and inhibitors of inducible nitric oxide synthase (iNOS) and cathepsin B also reversed chlamydial killing. High levels of reactive oxygen species (ROS) led to an increase in cathepsin B activity, and pharmacological inhibition of ROS and cathepsin B reduced iNOS expression. Our data demonstrate that MOI-dependent TLR2 activation of macrophages results in iNOS induction via a novel ROS- and cathepsin-dependent mechanism to facilitate C. muridarum clearance., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Pathogenically diverse Chlamydia spp. can have surprisingly similar genomes. Chlamydia trachomatis isolates that cause trachoma, sexually transmitted genital tract infections (chlamydia), and invasive lymphogranuloma venereum (LGV) and the murine strain Chlamydia muridarum share 99% of their gene content. A region of high genomic diversity between Chlamydia spp. termed the plasticity zone (PZ) may encode niche-specific virulence determinants that dictate pathogenic diversity. We hypothesized that PZ genes might mediate the greater virulence and gamma interferon (IFN-γ) resistance of C. muridarum compared to C. trachomatis in the murine genital tract. To test this hypothesis, we isolated and characterized a series of C. muridarum PZ nonsense mutants. Strains with nonsense mutations in chlamydial cytotoxins, guaBA-add, and a phospholipase D homolog developed normally in cell culture. Two of the cytotoxin mutants were less cytotoxic than the wild type, suggesting that the cytotoxins may be functional. However, none of the PZ nonsense mutants exhibited increased IFN-γ sensitivity in cell culture or were profoundly attenuated in a murine genital tract infection model. Our results suggest that C. muridarum PZ genes are transcribed--and some may produce functional proteins--but are dispensable for infection of the murine genital tract., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The present investigation elucidates the isolation and characterization of bioactive compound from the ethanolic leaf extract of Andrographis lineata (EtALL) which suppress the differentiation of 3T3-L1 adipocytes. The ethanolic leaf extract was subjected to bioassay guided fractionation in 3T3-L1 cell lines. Five fractions were isolated from the EtALL extract by column chromatography. All the Fractions (I-V) along with EtALL were screened for adipogenesis activity (Oil-Red-O staining).The fraction which showed maximum adipogenesis activity was purified by thin layer chromatography. The bioactive Fraction IV was found to have maximum adipogenic (96.83%) activity and the activity was comparable to Rosiglitazone. The spectroscopic data analysis reveals that, the isolated bioactive compound was Dimethyl 3, 3', 4, 4'-tetrahydroxy-δ-truxinate (DTδT), a combination of truxillic and truxinic acid derivative. DTδT showed insulin mimicking (131.2%), sensitizing (810.02%) and adipogenic activity (80.23%). Hence our present study concluded that, Dimethyl 3, 3', 4, 4'-tetrahydroxy-δ-truxinate isolated from the ethanolic leaf extract of Andrographis lineata stimulates glucose uptake, potentiates insulin-stimulated glucose in 3T3-L1 adipocytes without increasing adiposity.

Trauma of the palate in children is a common presentation in emergency departments. Children have a tendency to place objects in their mouths, which leaves them at risk of traumatising the oral cavity. Most traumatic injuries, including those that penetrate the palate, can be treated conservatively, but some require surgical intervention. Careful assessment, appropriate intervention, and follow up by experienced clinicians are essential to ensure correct management and prevent long-term complications. This article presents two case studies involving children who had experienced traumatic injuries to the palate that required surgical intervention.

Background: Adult zebrafish spontaneously regenerate their retinas after damage. Although a number of genes and signaling pathways involved in regeneration have been identified, the exact mechanisms regulating various aspects of regeneration are unclear. microRNAs (miRNAs) were examined for their potential roles in regulating zebrafish retinal regeneration., Results: To investigate the requirement of miRNAs during zebrafish retinal regeneration, we knocked down the expression of Dicer in retinas prior to light-induced damage. Reduced Dicer expression significantly decreased the number of proliferating Müller glia-derived neuronal progenitor cells during regeneration. To identify individual miRNAs with roles in neuronal progenitor cell proliferation, we collected retinas at different stages of light damage and performed small RNA high-throughput sequencing. We identified subsets of miRNAs that were differentially expressed during active regeneration but returned to basal levels once regeneration was completed. We then knocked down five different miRNAs that increased in expression and assessed the effects on retinal regeneration. Reduction of miR-142b and miR-146a expression significantly reduced INL proliferation at 51 h of light treatment, while knockdown of miR-7a, miR-27c, and miR-31 expression significantly reduced INL proliferation at 72 h of constant light., Conclusions: miRNAs exhibit dynamic expression profiles during retinal regeneration and are necessary for neuronal progenitor cell proliferation., (© 2014 The Authors. Developmental Dynamics published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.)

Background & Objectives: The human gut microbiota play a significant role in nutritional processes. The concept of probiotics has led to widespread consumption of food preparations containing probiotic microbes such as curd and yogurt. Curd prepared at home is consumed every day in most homes in southern India. In this study the home-made curd was evaluated for lactic acid bacteria (LAB) with probiotic potential., Methods: Fifteen LAB (12 lactobacilli, 1 l0 actococcus , 2 Leuconostoc) and one yeast isolated from home-made curd were evaluated for resistance to acid, pepsin, pancreatin and bile salts; antimicrobial resistance; intrinsic antimicrobial activity; adherence to Caco-2 epithelial cells; ability to block pathogen adherence to Caco-2 cells; ability to inhibit interleukin (IL)-8 secretion from HT-29 epithelial cells in response to Vibrio cholerae; and ability to induce anti-inflammatory cytokine expression in THP-1 monocyte cells., Results: Lactobacillus abundance in fermenting curd peaked sharply at 12 h. Nine of the strains survived exposure to acid (pH 3.0) for at least one hour, and all strains survived in the presence of pancreatin or bile salts for 3 h. None showed haemolytic activity. All were resistant to most antimicrobials tested, but were sensitive to imipenem. Most strains inhibited the growth of Salmonella Typhimurium while five inhibited growth of V. cholerae O139. Seven strains showed adherence to Caco-2 cells ranging from 20-104 per cent of adherence of an adherent strain of Escherichia coli, but all inhibited V. cholerae adherence to Caco-2 cells by 20-100 per cent. They inhibited interleukin-8 secretion from HT-29 cells, in response to V. cholerae, by 50-80 per cent. Two strains induced IL-10 and IL-12 messenger ribonucleic acid (mRNA) expression in THP-1 cells., Interpretation & Conclusions: LAB in curd had properties consistent with probiotic potential, but these were not consistent across species. LAB abundance in curd increased rapidly at 12 h of fermentation at room temperature and declined thereafter.

Damage of the zebrafish retina triggers a spontaneous regeneration response that is initiated by Müller Glia (MG) dedifferentiation and asymmetric cell division to produce multipotent progenitor cells. Subsequent expansion of the progenitor pool by proliferation is critical for retina regeneration. Pax6b expression in the progenitor cells is necessary for their proliferation, but exact regulation of its expression is unclear. Here, we show that miR-203 is downregulated during regeneration in proliferating progenitor cells. Elevated miR-203 levels inhibit progenitor cell expansion without affecting MG dedifferentiation or progenitor cell generation. Using GFP-reporter assays and gain and loss of function experiments in the retina, we show that miR-203 expression must be suppressed to allow pax6b expression and subsequent progenitor cell proliferation., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)

Zebrafish are capable of robust and spontaneous regeneration of injured retina. Constant intense light exposure to adult albino zebrafish specifically causes apoptosis of rod and cone photoreceptor cells and is an excellent model to study the molecular mechanisms underlying photoreceptor regeneration. However, this paradigm has only been applied to lesion zebrafish of the nonpigmented albino genetic background, which precludes the use of numerous transgenic reporter lines that are widely used to study regeneration. Here, we explored the effectiveness of constant intense light exposure in causing photoreceptor apoptosis and stimulating regeneration in normally pigmented zebrafish retinas. We show that constant intense light exposure causes widespread photoreceptor damage in the dorsal-central retinas of pigmented zebrafish. Photoreceptor loss triggers dedifferentiation and proliferation of Müller glia as well as progenitor cell proliferation. We also demonstrate that the timeline of regeneration response is comparable between the albino and the pigmented retinas.

M13 filamentous bacteriophage has been used in displaying disease-specific antibodies, biomarkers, and peptides. One of the major drawbacks of using phage in diagnostic assays is the aspecific adsorption of proteins leading to a high background signal and decreasing sensitivity. To deal with this, we developed a genetically pure, exchangeable dual-display phage system in which biomarkers and streptavidin-binding protein (SBP) are displayed at opposite ends of the phage. This approach allows for sample purification, using streptavidin-coated magnetic beads resulting in a higher sensitivity of signal detection assays. Our dual-display cassette system approach also allows for easy exchange of both the anchor protein (SBP) and the displayed biomarker. The presented principle is applied for the detection of antibody reactivity against UH-RA.21 which is a good candidate biomarker for rheumatoid arthritis (RA). The applicability of dual-display phage preparation using a helper plasmid system is demonstrated, and its increased sensitivity in phage ELISA assays using patient serum samples is shown.

Single-stranded DNA (ssDNA) is a prerequisite for electrochemical sensor-based detection of parasite DNA and other diagnostic applications. To achieve this detection, an asymmetric polymerase chain reaction method was optimised. This method facilitates amplification of ssDNA from the human lymphatic filarial parasite Wuchereria bancrofti. This procedure produced ssDNA fragments of 188 bp in a single step when primer pairs (forward and reverse) were used at a 100:1 molar ratio in the presence of double-stranded template DNA. The ssDNA thus produced was suitable for immobilisation as probe onto the surface of an Indium tin oxide electrode and hybridisation in a system for sequence-specific electrochemical detection of W. bancrofti. The hybridisation of the ssDNA probe and target ssDNA led to considerable decreases in both the anodic and the cathodic currents of the system's redox couple compared with the unhybridised DNA and could be detected via cyclic voltammetry. This method is reproducible and avoids many of the difficulties encountered by conventional methods of filarial parasite DNA detection; thus, it has potential in xenomonitoring.

There has been an increasing emphasis on health care efficiency and costs and on improving quality in health care settings such as hospitals or clinics. However, there has not been sufficient work on methods of improving access and customer service times in health care settings. The study develops a framework for improving access and customer service time for health care settings. In the framework, the operational concept of the bottleneck is synthesized with queuing theory to improve access and reduce customer service times without reduction in clinical quality. The framework is applied at the Ronald Reagan UCLA Medical Center to determine the drivers for access and customer service times and then provides guidelines on how to improve these drivers. Validation using simulation techniques shows significant potential for reducing customer service times and increasing access at this institution. Finally, the study provides several practice implications that could be used to improve access and customer service times without reduction in clinical quality across a range of health care settings from large hospitals to small community clinics.

Cherubism was first described by Jones in 1933 as "familial multilocular cystic disease of jaws." Renamed as cherubism in 1938 because of classical characteristics of full round cheeks and upward cast of the eyes to the angelic look of the cherubs immortalized by renaissance art. It is characterized by progressive painless bilateral swelling of jaws involving either maxilla or mandible producing chubby face. It is uncommon fibro-osseous disorder of bone. Mutation in the gene encoding SH3-binding protein 2 (SH3BP2) plays a role in the disease. There are indications that the gene SH3BP2 plays a role in regulating the increased osteoblast and osteoclast activities that are seen in normal tooth eruption and point mutations in the gene could cause pathologic activation of osteoclasts. The purpose of this paper is to present the uncommon form of cherubism and to review the clinicoradiographic, histopathologic features and treatment so as to facilitate diagnosis of disease.

Background: Under the Revised National Tuberculosis Control Programme of India, patients with new smear-positive pulmonary tuberculosis are treated with a thrice-weekly regimen of antitubercular drugs (2H(3)R(3)Z(3)E(3)/4H(3)R(3) [H isoniazid, R rifampicin, Z pyrazinamide and E ethambutol]) for 6 months. We conducted a retrospective analysis of the efficacy andtolerability of this regimen under clinical trial conditions in HIV-negative patients with newly diagnosed smear-positive pulmonary tuberculosis., Methods: We retrospectively analysed the data on patients assigned to the control regimen (2H (3)R(3)Z(3)E(3)/4H(3)R(3)) in two clinical trials during 2001-06 at the National Institute for Research in Tuberculosis, Chennai, India., Results: Of the 268 patients treated with this regimen, data for efficacy analysis were available for 249. At the end of treatment, of 249 patients, 238 (96%) had a favourable status. Treatment failure occurred in the remaining 11: 7 in whom the organisms were initially drug-susceptible and 4 with initial drug resistance. Of the 238 patients who had a favourable status at the end of treatment, 14 (6%) had recurrence of tuberculosis during the following 24 months. In the intention-to-treat analysis, 245 (94%) of 262 patients had a favourable status at the end of treatment. Of the 28 patients with initial drug resistance, 24 (86%) had a favourable outcome. Only 4 of these 24 patients were found to have recurrence of tuberculosis in 2 years of follow-up. Among the 221 patients initially infected with drug-susceptible organisms, drug resistance did not develop in any of the 7 patients in whom the treatment failed or the 10 who had recurrence of tuberculosis. Further, 5 of the 7 patients in whom the treatment failed continued to excrete drug-susceptible bacilli at 6 months. Adverse drug reactions were observed in 38 (14%) of the 262 patients. Only 3 (1.1%) needed a modification in the treatment., Conclusion: This thrice-weekly 6-month regimen of antitubercular drugs, when administered under full supervision, is associated with a high rate of favourable treatment outcomes in HIV-negative patients with newly diagnosed sputum smearpositive pulmonary tuberculosis. There are few adverse drug reactions in these patients., (Copyright 2012, NMJI.)

Accurate clinical staging of oral squamous cell cancer can be quite difficult to achieve especially if nodal involvement is identified. Radiologically-assisted clinical staging is more accurate and informs the clinician of loco-regional and distant metastasis.In this study, we compared clinical TNM (cTNM) staging (not including ultrasonography) to pathological TNM (pTNM) staging in 245 patients presenting with carcinoma of the oral cavity and the oro-pharyngeal region. Tumour size differences and nodal involvement were highlighted. US reports of the neck were then added to the clinical staging and results compared.Tumour size was clinically underestimated in 4 T1, 2 T2 and 2 T3 oral diseases. Also 20 patients that were reported as nodal disease free had histological proven N1 or N2 nodal involvement; while 3 patients with cTNM showing N1 disease had histologically proven N2 disease.Overall the agreement between the 2 systems per 1 site was 86.6% (Kappa agreement = 0.80), per 2 sites 90.0% (Kappa agreement = 0.68) and per 3 sites 90.5% (Kappa agreement 0.62).An accurate clinical staging is of an utmost importance. It is the corner stone in which the surgical team build the surgical treatment plan and decide whether an adjuvant therapy is required to deal with any possible problem that might arise. The failure to achieve an accurate staging may lead to incomplete surgical planning and hence unforeseen problems that may adversely affect the patient's survival.

Background: The use of tobacco is known to increase the incidence of developing oral cancer by 6 times, while the additive effect of drinking alcohol further increases the risk leading to higher rate of morbidity and mortality. In this short communication, we prospectively assessed the effect of tobacco smoking and alcohol drinking in oral cancer patients on the overall mortality from the disease, as well as the effect of smoking and drinking reduction/cessation at time of diagnosis on mortality in the same group., Materials and Methods: A cohort, involved 67 male patients who were diagnosed with oral squamous cell carcinoma, was included in this study. The smoking and drinking habits of this group were recorded, in addition to reduction/cessation after diagnosis with the disease. Comparisons were made to disease mortality at 3 and 5 years., Results: Follow-up resulted in a 3-year survival of 46.8% and a 5-year survival of 40.4%. Reduction of tobacco smoking and smoking cessation led to a significant reduction in mortality at 3 (P < 0.001) and 5 (P < 0.001) years. Reduction in drinking alcohol and drinking cessation led to a significant reduction in mortality at 3 (P < 0.001) and 5 (P < 0.001) years., Conclusion: Chronic smoking and drinking does have an adverse effect on patients with oral cancer leading to increased mortality from cancer-related causes. Reduction/cessation of these habits tends to significantly reduce mortality in this group of patients. Smoking and drinking cessation counseling should be provided to all newly diagnosed oral cancer patients.

Chlamydia trachomatis is an obligate intracellular bacterial pathogen that infects hundreds of millions of individuals globally, causing blinding trachoma and sexually transmitted disease. More effective chlamydial control measures are needed, but progress toward this end has been severely hampered by the lack of a tenable chlamydial genetic system. Here, we describe a reverse-genetic approach to create isogenic C. trachomatis mutants. C. trachomatis was subjected to low-level ethyl methanesulfonate mutagenesis to generate chlamydiae that contained less then one mutation per genome. Mutagenized organisms were expanded in small subpopulations that were screened for mutations by digesting denatured and reannealed PCR amplicons of the target gene with the mismatch specific endonuclease CEL I. Subpopulations with mutations were then sequenced for the target region and plaque-cloned if the desired mutation was detected. We demonstrate the utility of this approach by isolating a tryptophan synthase gene (trpB) null mutant that was otherwise isogenic to its parental clone as shown by de novo genome sequencing. The mutant was incapable of avoiding the anti-microbial effect of IFN-γ-induced tryptophan starvation. The ability to genetically manipulate chlamydiae is a major advancement that will enhance our understanding of chlamydial pathogenesis and accelerate the development of new anti-chlamydial therapeutic control measures. Additionally, this strategy could be applied to other medically important bacterial pathogens with no or difficult genetic systems.

Background: The treatment of tuberculosis (TB) with category I regimen of the Revised National Tuberculosis Control Programme (RNTCP) for patients with diabetes mellitus (DM) needs evaluation., Objective: To assess the cure and relapse rates in 3 years, among the new smear-positive TB patients with Type-2 DM (DMTB) treated with CAT-I regimen (2E3H3R3Z3/4R3H3) of RNTCP., Methodology: TB suspects attending the diabetology units and the TB research centre (TRC) Chennai, were investigated. Eligible DMTB cases were enrolled. Baseline estimation of cardiac, renal, liver function tests and glycosylated-HBA1c were undertaken. All patients received 2E3H3R3Z3/4R3H3 under supervision at TRC. Clinical and sputum (smear and culture) examinations and monitoring of diabetic status were undertaken every month up to 24 months, then once in 3 months up to 36 months., Results: Of 100 patients admitted, 7 were excluded for various reasons from analysis. Of 93 patients, 87 (94%) had a favourable response at the end of treatment. Pre and post treatment mean glycosylated-HBA1c were 9.7% and 8.4% (>7% poor control). During follow-up period, 6 died and one lost to follow-up. Of the remaining, four relapsed., Conclusion: Category-I regimen, recommended for all the new smear-positive patients in the Indian TB programme, is effective in the treatment of DMTB patients, despite poor control of diabetes.

Background: No study has surveyed the factors that influence morbidly obese patients' preference for a particular bariatric operation., Method: 469 consecutive patients in 2 major bariatric surgery centers in the United States (US, 124) and Australia (AU, 345) were prospectively studied to determine referral pattern and reason for their choice of operation., Results: The predominant operation was laparoscopic adjustable gastric banding (LAGB) in both US (75%) and AU (83%) centers. Gender (70% female), BMI (45 kg/m2) and age (42.5 years) were similar in both cohorts. In Australia, 53% had referral initiated by primary doctors and 25% by another patient, while in the US, 43% by another patient and 27% by the Internet. Safety of the operation (43%) was the highest-rated factor in choosing LAGB. LAGB being "least invasive" was most significant in the US (46%), and "surgical safety" in Australia (45%). In the US, Rouxen-Y gastric bypass was preferred due to "lack of a foreign body" (31%) and "inability to cheat" (28%), while in Australia, "dumping" was the most significant reason (50%). Duodenal switch (BPD/DS) was selected in 11% of patients, primarily because of "durability of the weight loss" (51%). Surprisingly, only 1 patient in the US group selected BPD/DS because the pylorus remains intact., Conclusion: Safety and invasiveness had the greatest impact on patient choice for bariatric operation in two different countries. This information may help clinicians better understand their patients' concerns, and their treatment choices.

Background: Postoperative follow-up after bariatric surgery is important. Because of the need for adjustments, follow-up after gastric banding may have a greater impact on weight loss than after Roux-en-Y gastric bypass. We reviewed all patients at 1 year after these two operations., Methods: During the first year after surgery, laparoscopic adjustable gastric banding (LAGB) patients were followed every 4 weeks and Roux-en-Y gastric bypass (RYGBP) patients were followed at 3 weeks postoperatively and then every 3 months. The number of follow-up visits for each patient was calculated, and 50% compliance for follow-up and weight loss was compared., Results: Between October 2000 and September 2002, 216 LAGB and 139 RYGBP operations were performed. Of these patients, 186 LAGB patients and 115 RYGBP patients were available for 1-year follow-up. Age and BMI were similar for each group. Overall excess weight loss (EWL) after LAGB was 44.5%. 130 (70%) returned 6 or less times in the first year and achieved 42% EWL. 56 patients (30%) returned more than 6 times and had 50% EWL (P=0.005). Overall %EWL after RYGBP was 66.1%. 53 patients (46%) returned 3 or less times in the first year, achieving 66.1% EWL. 62 patients (54%) returned more than 3 times after surgery and achieved 67.6% EWL (P=NS)., Conclusion: Patient follow-up plays a significant role in the amount of weight lost after LAGB, but not after RYGBP. Patient motivation and surgeon commitment for long-term follow-up is critical for successful weight loss after LAGB surgery.

Despite the endemic distribution of visceral leishmaniasis in certain parts of our country, there are only a few reports of this infection in renal transplant recipients. We report one renal transplant recipient from non-endemic area with visceral leishmaniasis and graft dysfunction that responded to treatment with stibogluconate. The infection should be considered in the differential diagnosis of a febrile transplant recipient with pancytopenia and allograft dysfunction.

A total of 446 lymph node biopsy specimens showing histological evidence of tuberculosis were classified into four groups based on the organization of the granuloma, the type and numbers of participating cells and the nature of necrosis. These were, hyperplastic (22.4%)--a well-formed epithelioid cell granuloma with very little necrosis, reactive (54.3%)--a well-formed granuloma consisting of epithelioid cells, macrophages, lymphocytes and plasma cells with fine, eosinophilic caseation necrosis, hyporeactive (17.7%)--a poorly organized granuloma with macrophages, immature epithelioid cells, lymphocytes and plasma cells and coarse, predominantly basophilic caseation necrosis and nonreactive (3.6%)--unorganized granuloma with macrophages, lymphocytes, plasma cells and polymorphs with non caseating necrosis. Though the number of bacilli in the sections differed in each group, there were no differences in culture positivity, Mantoux reaction or the clinical features. It is likely that the spectrum of histological responses seen in tuberculous lymphadenitis is the end result of different pathogenic mechanisms underlying the disease.

This paper reports the feasibility of involving unpaid National Service Scheme (NSS) male student volunteers in a city-based tuberculosis (TB) programme in supplying drugs and retrieving non-complaint TB patients. Twenty-five students were selected after assessing their attitude and were trained on TB drug delivery, home visits and motivation of non-compliant patients. Twenty-three sputum positive patients identified in a medical camp were started on an 8-month short-course chemotherapy regimen. Students supplied the drugs on a weekly basis and defaulters were visited. The treatment completion rate was 83% and defaulter retrieval was 57%. All patients had sputum smear conversion by 2 months and one relapsed during the 24-month follow-up.

Lymph node biopsy specimens, obtained from 297 paediatric and adult patients with tuberculous lymphadenitis at Madurai, were transported in selective Kirchner's liquid medium (KL-T) to the Tuberculosis Research Centre, Madras and processed for culture. Mycobacterium tuberculosis was isolated from 201 (68%) specimens. Of the 192 specimens received within 4 days of resection, 134 (69.8%) yielded M. tuberculosis on culture and of the 105 specimens received after 5 days, 67 (63.8%) were culture positive; the difference was not statistically significant. By incubating KL-T alone further, after removing the gland for processing, it was found that mere contact with the excised node during transportation was enough to retrieve 77 (38.3%) of the total of 201 positive isolates obtained, the delay did not affect the culture positivity rate. Thus, lymph node specimens for culture of tubercle bacilli can be stored in the refrigerator for up to 15 days and transported in KL-T at ambient temperature for 18-20 h without any loss in culture positivity.

A direct sensitivity test for rifampicin has been standardised for early detection of resistance for the mycobacterium tuberculosis smear positive sputum samples. Indirect sensitivity tests set up from primary cultures of the same samples served as controls. The direct test showed 95 per cent agreement with the standard indirect test and as such 74 per cent and 90 per cent of the resistant strains were detected by the fourth week and fifth week, respectively, with an overall gain of 4-5 wks time. Resistance could be detected earlier for multibacillary specimens. This direct sensitivity test on Lowenstein Jensen (LJ) medium offers a feasible alternative for laboratories which lack facilities to perform drug susceptibility tests by the rapid but sophisticated and costly BACTEC method. The method is simple to perform, economic, reliable and amenable to confirmation by the indirect test, if needed.