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Initial COVID-19 surges in the United States created a need for technology to supplement human resources to increase efficiency and efficacy.Resolve to Save Lives worked with jurisdictions to co-design four technology solutions-Epi Viaduct data pipeline, Epi Contacts contact elicitation webform, Epi Locator contact information search plugin and Epi Viewpoint case management system-to expand the capacity of case investigation and contact tracing (CI/CT) teams. We assessed impact on reducing CI/CT time intervals for COVID-19 using product data and user feedback.Epi Viaduct accelerated the transfer of approximately 7,400,000 records from an electronic lab reporting (ELR) system in a single jurisdiction to the respective case management system (CMS) from more than 2.5 hours to less than one minute and reduced time to remove duplicate lab results from multiple days to less than six hours. Epi Contacts focused on increasing efficacy of contact elicitation and, during a single period, 10% of index cases (9,440/96,319) completed Epi Contacts for a total of approximately 18,700 contacts elicited. User interviews indicated the tool increased speed of CI/CT workflows. In total, 134,410 searches were run in Epi Locator by 7,320 distinct users-75% of which returned one or more person matches. A simple CMS, Epi Viewpoint, was developed and completed, but not deployed.Systems to mount large-scale population-based contact tracing programs were developed and implemented during the COVID-19 pandemic and can be adapted for CI/CT programs aiming to control the spread of other communicable diseases such as STDs.

Ebola virus (EBOV) disease is characterized by lymphopenia, breach in vascular integrity, cytokine storm, and multiorgan failure. The pathophysiology of organ involvement, however, is incompletely understood. Using [18F]-DPA-714 positron emission tomography (PET) imaging targeting the translocator protein (TSPO), an immune cell marker, we sought to characterize the progression of EBOV-associated organ-level pathophysiology in the EBOV Rhesus macaque model. Dynamic [18F]-DPA-714 PET/computed tomography imaging was performed longitudinally at baseline and at multiple time points after EBOV inoculation, and distribution volumes (Vt) were calculated as a measure of peripheral TSPO binding. Using a mixed-effect linear regression model, spleen and lung Vt decreased, while the bone marrow Vt increased over time after infection. No clear trend was found for liver Vt. Multiple plasma cytokines correlated negatively with lung/spleen Vt and positively with bone marrow Vt. Multiplex immunofluorescence staining in spleen and lung sections confirmed organ-level lymphoid and monocytic loss/apoptosis, thus validating the imaging results. Our findings are consistent with EBOV-induced progressive monocytic and lymphocytic depletion in the spleen, rather than immune activation, as well as depletion of alveolar macrophages in the lungs, with inefficient reactive neutrophilic activation. Increased bone marrow Vt, on the other hand, suggests hematopoietic activation in response to systemic immune cell depletion and leukocytosis and could have prognostic relevance. In vivo PET imaging provided better understanding of organ-level pathophysiology during EBOV infection. A similar approach can be used to delineate the pathophysiology of other systemic infections and to evaluate the effectiveness of newly developed treatment and vaccine strategies.

PURPOSE The current diagnostic testing algorithm for Lynch syndrome (LS) is complex and often involves multiple follow-up germline and somatic tests. We aimed to describe the results of paired tumor/germline testing performed on a large cohort of patients with colorectal cancer (CRC) and endometrial cancer (EC) to better determine the utility of this novel testing methodology. MATERIALS AND METHODS We retrospectively reviewed a consecutive series of patients with CRC and EC undergoing paired tumor/germline analysis of the LS genes at a clinical diagnostic laboratory (N = 702). Microsatellite instability, MLH1 promoter hypermethylation, and germline testing of additional genes were performed if ordered. Patients were assigned to one of five groups on the basis of prior tumor screening and germline testing outcomes. Results for each group are described. RESULTS Overall results were informative regarding an LS diagnosis for 76.1% and 60.8% of patients with mismatch-repair–deficient (MMRd) CRC and EC without and with prior germline testing, respectively. LS germline mutations were identified in 24.8% of patients in the group without prior germline testing, and interestingly, in 9.5% of patients with previous germline testing; four of these were discordant with prior tumor screening. Upon excluding patients with MLH1 promoter hypermethylation and germline mutations, biallelic somatic inactivation was seen in approximately 50% of patients with MMRd tumors across groups. CONCLUSION Paired testing identified a cause for MMRd tumors in 76% and 61% of patients without and with prior LS germline testing, respectively. Findings support inclusion of tumor sequencing as well as comprehensive LS germline testing in the LS testing algorithm. Paired testing offers a complete, convenient evaluation for LS with high diagnostic resolution.

Ebola virus (EBOV) causes neurological symptoms yet its effects on the central nervous system (CNS) are not well-described. Here, we longitudinally assess the acute effects of EBOV on the brain, using quantitative MR-relaxometry, 18F-Fluorodeoxyglucose PET and immunohistochemistry in a monkey model. We report blood–brain barrier disruption, likely related to high cytokine levels and endothelial viral infection, with extravasation of fluid, Gadolinium-based contrast material and albumin into the extracellular space. Increased glucose metabolism is also present compared to the baseline, especially in the deep gray matter and brainstem. This regional hypermetabolism corresponds with mild neuroinflammation, sporadic neuronal infection and apoptosis, as well as increased GLUT3 expression, consistent with increased neuronal metabolic demands. Neuroimaging changes are associated with markers of disease progression including viral load and cytokine/chemokine levels. Our results provide insight into the pathophysiology of CNS involvement with EBOV and may help assess vaccine/treatment efficacy in real time., The neurological effects of Ebola disease in the acute stage are not well-described. Here, the authors use longitudinal in vivo neuroimaging and immunohistochemistry to assess pathological changes in the central nervous system in a non-human primate model of Ebola virus infection.

Current surrogacy research primarily focuses on commercial surrogacy with a particular emphasis on experiences of surrogate mothers, whereas intended parents' voices are dominated by western perspectives. Indigenous voices are only a whisper. This study presents another side of the surrogacy story by including the voices of intended parents residing in India, elicited through eight in-depth interviews. We assert there is need to understand Indian intended parents' socio-spatial experiences in the presence of a changing surrogacy law in India and the socio-cultural importance of childbearing and parenthood to move towards relational reproductive justice. By recognising the relational nature of surrogacy reproduction and drawing on the concepts of ethics of care in light of power and stigma discussions, this paper demonstrates how stigma is experienced by intended parents, its effect on their reproductive journey and wellbeing, as well as how stigma hinders achieving the vision of relational reproductive justice. We suggest that, in order to make surrogacy a positive experience for the people involved in surrogacy, there is a need to address stigma and view surrogacy as a relationship.

Background The germline cancer predisposition genes associated with increased risk of each clinical subtype of breast cancer, defined by estrogen receptor (ER), progesterone receptor (PR), and HER2, are not well defined. Methods A total of 54 555 invasive breast cancer patients with 56 480 breast tumors were subjected to clinical hereditary cancer multigene panel testing. Heterogeneity for predisposition genes across clinical breast cancer subtypes was assessed by comparing mutation frequencies by gene among tumor subtypes and by association studies between each tumor subtype and reference controls. Results Mutations in 15 cancer predisposition genes were detected in 8.6% of patients with ER+/HER2-; 8.9% with ER+/HER2+; 7.7% with ER-/HER2+; and 14.4% of ER-/PR-/HER2- tumors. BRCA1, BRCA2, BARD1, and PALB2 mutations were enriched in ER- and HER2- tumors; RAD51C and RAD51D mutations were enriched in ER- tumors only; TP53 mutations were enriched in HER2+ tumors, and ATM and CHEK2 mutations were enriched in both ER+ and/or HER2+ tumors. All genes were associated with moderate (odds ratio > 2.00) or strong (odds ratio > 5.00) risks of at least one subtype of breast cancer in case-control analyses. Mutations in ATM, BARD1, BRCA1, BRCA2, CHEK2, PALB2, RAD51C, RAD51D, and TP53 had predicted lifetime absolute risks of at least 20.0% for breast cancer. Conclusions Germline mutations in hereditary cancer panel genes confer subtype-specific risks of breast cancer. Combined tumor subtype, age at breast cancer diagnosis, and family history of breast and/or ovarian cancer information provides refined categorical estimates of mutation prevalence for women considering genetic testing.

Introduction Optimal therapeutic cancer discovery pipelines rely on high throughput, reproducibility and minimal bias for candidate discovery and clinical trials. Acute Myeloid Leukemia (AML) cell-based assays, combining flow cytometry (FCM) immunophenotyping with genomic sequencing, provide comprehensive characterization. However, sequential manual gating by expert flow analysts has been shown to have interlaboratory variation, weaknesses with higher dimensional feature spaces due to increased fluorescent channels, and limited scalability for throughput due to the necessary expert interpretation. Automated analysis of Flow Cytometry Standard (FCS) data using a combination of unsupervised and supervised algorithms that combine quality control, gating and clustering strategies offers a more scalable and sustainable option. Given these established breakthroughs, we investigated the use of these informatic tools, combined with sequencing data, to create a therapeutic discovery platform. Methods Peripheral blood and bone marrow samples with abnormal myeloid population and normal controls were exposed to multiple chemotherapeutic small molecules at varying concentrations. Samples were flow cytometrically evaluated with an 8-color tube of antibodies targeting various antigens allowing for apoptosis, viability and blast characterization using standardized lab protocols with a flow cytometer, followed by hematopathologist review. Furthermore, the samples underwent molecular sequencing and annotation with the MyAML® assay to interpret results in the context of AML molecular classification. The computational pipeline utilized Bioconductor version 3.11 open-source packages for quality control, Logicle transformation, apoptosis and viability analyses utilizing a supervised gating strategy. Once viability was established, a Self-Organizing Map (unsupervised dimensionality reduction) algorithm was used to cluster cells into different cell populations. Moreover, a neural network utilizing TensorFlow combined with the API, Keras, was used to detect cell populations from these clusters. The neural network classifier was trained, validated and tested on these clusters compared to expert interpretation. Results Six bone marrow and three peripheral blood samples with the appropriate controls were evaluated via the discovery pipeline. Each sample was exposed to various chemotherapeutic molecules for a total of 203 samples for the pipeline with 50,000 cells per sample. Sequencing demonstrated these samples reflected various European LeukemiaNet (ELN) molecular risk categories. The correlation between the total viable cells in 59 samples as determined by manual and automated gating processes was 98% (r 2=0.98) (Figure 1a). Heatmaps were generated for data visualization of therapeutic efficacy using this viable cell data as demonstrated in one example, Figure 1b, which was consistent with biological hypotheses. Beyond total cell viability detection, cell populations were determined. The validation and test accuracy of our lymphocyte classification model were both 99%, and F1 statistic for model's testing performance was 0.98. Figure 2a displays the ROC curve. The classification of lymphocyte cell population using our combined unsupervised learning/neural network approach for 59 samples had a correlation of 97% (r 2=0.97) (Figure 2b). Conclusions Based on a preliminary comparison with manual gating by an expert flow analyst, we show an automated scalable protocol gives comparable results; therefore, it would be an appropriate alternative to use to screen multiple cells for downstream therapeutic analysis with increased throughput for both cell viability and cell population identification. Figure 1 Figure 1. Disclosures Patay: Invivoscribe: Current Employment, Current holder of stock options in a privately-held company. Zlotnicki: Invivoscribe: Current Employment. Shah: Invivoscribe: Current Employment, Current holder of stock options in a privately-held company. Apilado: Invivoscribe: Current Employment. Andrews: Invivoscribe: Current Employment, Current holder of stock options in a privately-held company.

Advances in antiretroviral therapy have resulted in significantly decreased HIV-related mortality. HIV-associated neurocognitive disorders, however, continue to be a major problem in infected patients. The neuropathology underlying HIV-associated neurocognitive disorders has not been well characterized, and evidence suggests different contributing mechanisms. One potential mechanism is the induction of oxidative stress. Using the HIV-1 transgenic (Tg) rat model of HIV, we found increased striatal NADPH oxidase-4 and neuronal nitric oxide synthase expression in the adult (7- to 9-month-old) Tg rat compared with control rats but not in the young (1-month-old) Tg rats. This was accompanied by increased 3-nitrotyrosine (3-NT) immunostaining in the adult Tg rats, which worsened significantly in the old Tg rats (18 to 20 months old). There was, however, no concurrent induction of the antioxidant systems because there was no change in the expression of the nuclear factor-erythroid 2-related factor 2 and its downstream targets (thioredoxin and glutathione antioxidant systems). Colocalization of 3-NT staining with neurofilament proteins and evidence of decreased tyrosine hydroxylase and dopamine transporter expression in the old rats support dopaminergic involvement. We conclude that the HIV-1 Tg rat brain shows evidence of nitrosative stress without appropriate oxidation-reduction adaptation, whereas 3-NT modification of striatal neurofilament proteins likely points to the ensuing dopaminergic neuronal loss and dysfunction in the aging HIV-1 Tg rat.

Purpose: Increased incidence of depression in HIV+ patients is associated with lower adherence to treatment and increased morbidity/mortality. One possible underlying pathophysiology is serotonergic dysfunction. In this study, we used an animal model of HIV, the SIV-infected macaque, to longitudinally image serotonin transporter (SERT) expression before and after inoculation, using 11C-DASB (SERT ligand) PET imaging.Methods: We infected seven rhesus macaques with a neurovirulent SIV strain and imaged them at baseline and multiple time points after inoculation (group A). Pyrosequencing methylation analysis of the SERT promoter region was performed. We also measured SERT mRNA/protein in brain single-cell suspensions from another group (group B) of SIV-infected animals (n = 13).Results: Despite some animals showing early fluctuations, 86% of our group A animals eventually showed a net increase in midbrain/thalamus binding potential (BPND) over the course of their disease (mean increased binding between last time point and baseline = 30.2% and 32.2%, respectively). Repeated-measures mixed-model analysis showed infection duration to be predictive of midbrain BPND (p = 0.039). Thalamic BPND was statistically significantly associated with multiple CSF cytokines (P < 0.05). There was higher SERT protein levels in the second group (group B) of SIV-infected animals with SIV encephalitis (SIVE) compared to those without SIVE (p = 0.014). There were no longitudinal changes in SERT gene promoter region percentage methylation between baselines and last time points in group A animals.Conclusion: Upregulated SERT leading to lower synaptic levels of serotonin is a possible mechanism of depression in HIV+ patients, and extrapolating our conclusions from SIV to HIV should be sought using translational human studies.

Neurological and cognitive problems are a common complication of HIV infection and are prevalent even in treated individuals. Although the molecular processes underlying brain involvement with HIV are not completely understood, inflammation is suspected to play a significant role. Our work presents an in vivo assessment of neuroinflammation in an animal model of HIV, the simian immunodeficiency virus (SIV)-infected rhesus macaque. Using positron emission tomography (PET) imaging, we identified changes in brain inflammation after inoculation with SIV over time. Interestingly, we found decreased binding of the PET ligand in the presence of very high cerebrospinal fluid (CSF) viral loads. These findings were supported by immunostaining which showed marked glial loss instead of inflammation. This study provides insight into glial and neuronal changes associated with very high CSF viral load and could reflect similar changes occurring in HIV-infected patients., The exact cause of neurocognitive dysfunction in HIV-positive patients despite successful control of the infection in the periphery is not completely understood. One suggested mechanism is a vicious cycle of microglial activation and release of proinflammatory chemokines/cytokines that eventually leads to neuronal loss and dysfunction. However, the exact role of microglial activation in the earliest stages of the infection with high cerebrospinal fluid (CSF) viral loads (VL) is unclear. In this study, we imaged the translocator protein (TSPO), a mitochondrial membrane receptor known to be upregulated in activated microglia and macrophages, in rhesus macaques before and multiple times after inoculation with a neurotropic simian immunodeficiency virus (SIV) strain (SIVsm804E), using 18F-DPA714 positron emission tomography (PET). The whole-brain standardized uptake values of TSPO at equilibrium reflecting total binding (SUVT) and binding potentials (BPND) were calculated and correlated with CSF and serum markers of disease, and a corresponding postmortem immunostaining analysis was also performed. SUVT was found to be inversely correlated with both CSF VL and monocyte chemoattractant protein 1 (MCP-1) levels. In SIV-infected macaques with very high CSF VL at necropsy (>106 copies/ml), we found decreased TSPO binding by PET, and this was supported by immunostaining which showed glial and neuronal apoptosis rather than microglial activation. On the other hand, with only moderately elevated CSF VL (∼104 copies/ml), we found increased TSPO binding as well as focal and diffuse microglial activation on immunostaining. Our results in the SIV-infected macaque model provide insights into the relationship between HIV neuropathology and CSF VL at various stages of the disease.

The type I IFNs (IFN-α and -β) are important for host defense against viral infections. In contrast, their role in defense against nonviral pathogens is more ambiguous. In this article, we report that IFN-β signaling in murine bone marrow–derived macrophages has a cell-intrinsic protective capacity against Mycobacterium tuberculosis via the increased production of NO. The antimycobacterial effects of type I IFNs were mediated by direct signaling through the IFN-α/β–receptor (IFNAR), as Ab-mediated blocking of IFNAR1 prevented the production of NO. Furthermore, M. tuberculosis is able to inhibit IFNAR-mediated cell signaling and the subsequent transcription of 309 IFN-β–stimulated genes in a dose-dependent way. The molecular mechanism of inhibition by M. tuberculosis involves reduced phosphorylation of the IFNAR-associated protein kinases JAK1 and TYK2, leading to reduced phosphorylation of the downstream targets STAT1 and STAT2. Transwell experiments demonstrated that the M. tuberculosis–mediated inhibition of type I IFN signaling was restricted to infected cells. Overall, our study supports the novel concept that M. tuberculosis evolved to inhibit autocrine type I IFN signaling to evade host defense mechanisms.

The development of targeted therapies for both germline and somatic DNA mutations has increased the need for molecular profiling assays to determine the mutational status of specific genes. Moreover, the potential of off-label prescription of targeted therapies favors classifying tumors based on DNA alterations rather than traditional tissue pathology. Here we describe the analytical validation of a custom probe-based NGS tumor panel, TumorNext, which can detect single nucleotide variants, small insertions and deletions in 142 genes that are frequently mutated in somatic and/or germline cancers. TumorNext also detects gene fusions and structural variants, such as tandem duplications and inversions, in 15 frequently disrupted oncogenes and tumor suppressors. The assay uses a matched control and custom bioinformatics pipeline to differentiate between somatic and germline mutations, allowing precise variant classification. We tested 170 previously characterized samples, of which > 95% were formalin-fixed paraffin embedded tissue from 8 different cancer types, and highlight examples where lack of germline status may have led to the inappropriate prescription of therapy. We also describe the validation of the Affymetrix OncoScan platform, an array technology for high resolution copy number variant detection for use in parallel with the NGS panel that can detect single copy amplifications and hemizygous deletions. We analyzed 80 previously characterized formalin-fixed paraffin-embedded specimens and provide examples of hemizygous deletion detection in samples with known pathogenic germline mutations. Thus, the TumorNext combined approach of NGS and OncoScan potentially allows for the identification of the "second hit" in hereditary cancer patients.

Cash payments and other arrangements between patent holders and their competitors in exchange for a delayed market entry interfere with free-market principles and keep product prices at a premium, which affects payers, governments, healthcare providers and especially patients.

Background Although rates of severe HIV-associated neurocognitive disorders have declined in the post-antiretroviral treatment (ART) era, subtle deficits persist, possibly exacerbated by treatment non-adherence. The actual effects of ART interruption/initiation on brain glucose metabolism as a reflection of viral replication and neuroinflammation remain unclear. Our study investigates how treatment initiation and interruption alter brain glucose metabolism in SIV-infected macaques, using 18F-FDG PET in correlation with plasma and CSF viral loads (VL) and cytokine levels. Methods SIV-infected macaques (n = 7) underwent ART initiation only, ART interruption only, or both. Five uninfected animals served as controls. 18F-FDG PET imaging was performed at baseline and 1, 3, and 6 months after treatment modification. Mean and maximum standardized uptake values (SUV) for the whole-brain and subregions were calculated. Plasma and CSF VL and cytokine levels were measured. Paired t tests evaluated acute changes in whole-brain SUV from baseline to 1 month, while mixed-effect linear regression models evaluated changes over multiple timepoints and correlated SUV values with disease markers. Results ART interruption was associated with increased SUVmean and SUVmax acutely, after 1 month (SUVmean 95% CI [0.044–0.786 g/ml], p = 0.037; SUVmax 95% CI [0.122–3.167 g/ml], p = 0.041). The correlation between SUV and time, however, was not significant when evaluated across all timepoints. Increased SUVmean and SUVmax correlated with decreased CD4+ and CD8+ T-cell counts and increased plasma VL. SUVmax was positively associated with increases in CSF VL, and there were borderline positive associations between SUVmax and IL-2, and between SUVmean and IL-15. The treatment initiation group showed no associations between imaging and disease biomarkers despite viral suppression, reduced cytokine levels, and increased CD4+ and CD8+ T-cell counts. Conclusions ART interruption is associated with increased brain glucose metabolism within 1 month of treatment cessation, which, in concert with increased levels of pro-inflammatory cytokines in the CSF, may reflect neuroinflammation in the setting of viral rebound. Although we cannot assert neurologic damage in association with cerebral hypermetabolism, it is a concerning outcome of ART non-adherence. Treatment initiation, meanwhile, did not result in significant changes in brain metabolism. HIV-induced neuroinflammation may require a longer period to abate than our follow-up period allowed. Electronic supplementary material The online version of this article (10.1186/s12974-018-1244-z) contains supplementary material, which is available to authorized users.

Objective: To find out the knowledge, attitude, practice, and barriers of cervical cancer screening in mid-western rural, Nepal. Methods: A hospital-based cross-sectional study was conducted. Women aged 20 or more were interviewed using a structured questionnaire regarding the socio-demographic information, knowledge, attitude, practice, and barriers to the cervical cancer screening. Results: Total of 360 participants were recruited for this study, mean age was 30.13±10.4 years. More than 87% of participants had inadequate knowledge, but around 72% had a favorable attitude towards cervical cancer screening. There was a significant portion of women (86.4%) had never done any cervical cancer screening test. Despite being higher literacy rate of Brahmin and Chhetri ethnic group, they were less likely to attend the cervical cancer screening than Dalit and Janajati (p

The current algorithm for Lynch syndrome diagnosis is highly complex with multiple steps which can result in an extended time to diagnosis while depleting precious tumor specimens. Here we describe the analytical validation of a custom probe-based NGS tumor panel, TumorNext-Lynch-MMR, which generates a comprehensive genetic profile of both germline and somatic mutations that can accelerate and streamline the time to diagnosis and preserve specimen. TumorNext-Lynch-MMR can detect single nucleotide variants, small insertions and deletions in 39 genes that are frequently mutated in Lynch syndrome and colorectal cancer. Moreover, the panel provides microsatellite instability status and detects loss of heterozygosity in the five Lynch genes; MSH2, MSH6, MLH1, PMS2 and EPCAM. Clinical cases are described that highlight the assays ability to differentiate between somatic and germline mutations, precisely classify variants and resolve discordant cases.

Background: Diabetes mellitus (DM) is one of the most common chronic diseases globally. Diabetic neuropathy is the most common & troublesome complication. But exact pathogenesis is not yet known. Comparatively there are few studies showing relation between glycemic status & diabetic neuropathy. Hence present study was conducted, which was aimed to assess the same in lower limbs in type 2 DM. Methods: 60 type 2 diabetes mellitus male patients were selected from diabetic OPD. 30 were having glycated hemoglobin (HbA1c) 6%-9% (group B), 30 were having HbA1c > 9% (group C). They were compared with age and sex matched 30 normal healthy controls (group A). Conduction velocity and amplitude of bilateral sural sensory nerve action potential (SNAP) and peroneal compound muscle action potential (CMAP) were recorded. Glycated hemoglobin was measured using ion exchange resin method. Results: Group B and group C had significantly lesser means of conduction velocity and amplitude of sural SNAP (p

Introduction In vivo imaging biomarkers of various HIV neuropathologies, including dopaminergic dysfunction, are still lacking. Towards developing dopaminergic biomarkers of brain involvement in HIV, we assessed the pre and postsynaptic components of the dopaminergic system in the HIV-1 transgenic rat (Tg), a well-characterized model of treated HIV + patients, using small-animal PET imaging. Methods Fifteen to 18 month-old Tg and wild type (WT) rats were imaged with both [18F]-FP-CMT, a dopamine transporter (DAT) ligand (n = 16), and [18F]-Fallypride, a D2/D3 dopamine receptor (D2/D3DR) ligand (n = 16). Five to 8 month-old Tg and WT rats (n = 18) were also imaged with [18F]-FP-CMT. A subset of animals was imaged longitudinally at 7 and 17 months of age. Multiplex immunohistochemistry staining for DAT, tyrosine hydroxylase, D2DR, D3DR, GFAP, Iba1 and NeuN was performed on a subgroup of the scanned animals. Results [18F]-FP-CMT and [18F]-Fallypride binding potential (BPND) values were significantly lower in 15–18 month-old Tg compared to age-matched WT rats (p Conclusions We found presynaptic and postsynaptic dopaminergic dysfunction/loss in older Tg compared to WT rats. We believe this to be related to neurotoxicity of viral proteins present in the Tg rats’ serum and brain. Advances in knowledge Our findings confirm prior reports of neurobehavioral abnormalities suggestive of dopaminergic dysfunction in this model. They also suggest similarities between the Tg rat and HIV + patients as far as dopaminergic dysfunction. Implications for patient care The Tg rat, along with the above-described quantitative PET imaging biomarkers, can have a role in the evaluation of HIV neuroprotective therapies prior to human translation.

Aortic arch surgery requires meticulous teamwork in the true perioperative sense. Planning and communication at all phases from preoperative evaluation through intraoperative management to postoperative care should be well coordinated between surgical anesthesia perfusion and intensive care unit teams. This case report discusses intraoperative management of a case of Marfanrsquos syndrome with anomalous right subclavian artery from the anesthesiologistrsquos perspective. Bentall with Total Arch Replacement using Long Elephant Trunk for Marfanrsquos syndrome with anomalous right subclavian artery was performed. Stanford type A dissection of the aorta extending down till the bifurcation can affect multi-organs system besides brain. An intraoperative strategy should aim multi-organ protection to avoid morbidity and mortality. Cerebral protection using retrograde and antegrade cerebral perfusion and systemic hypothermic circulatory arrest to protect multi organs made surgery and post-operative recovery uneventful.

ObjectivesTo describe the technique of robotic-assisted video endoscopic inguinal lymphadenectomy (R-VEIL) in patients with carcinoma vulva and discuss the advantages of the technique and oncological outcome.MethodsTwelve patients of squamous cell cancer of vulva underwent 22 R-VEIL procedures from February 2011 to February 2015. Their preoperative, intraoperative, and postoperative data were retrospectively analysed.ResultsThe mean age of patients was 61 years (range, 32–78 years). The mean operative time was 69.3 minutes (range, 45–95 minutes). The mean blood loss was 30 mL (range, 15–50 mL). No intraoperative complication was observed. The mean drain output was 119 mL (range, 50–250 mL), and the drains were removed at a mean of 13.9 days (range, 8–38 days). The average number of superficial and deep inguinofemoral lymph nodes retrieved was 11 (range, 4–26). Two patients had positive lymph nodes on histopathology (16.67%). Postoperative complications were lymphocele (6 groins), chronic lower limb lymphedema (6 cases), prolonged lymphorrhea (1 groin), and cellulitis (2 groins). Over a follow-up period ranging from 7 to 67 months, 1 patient developed recurrence in the inguinal nodes and died 7 months after the recurrence.ConclusionsThe R-VEIL allows the removal of inguinal lymph nodes within the same limits as the open procedure for inguinal lymph node dissection and has a potential to reduce the surgical morbidity associated with the open procedure. Long-term oncological results are not available though our initial results appear promising. Prospective multi-institutional studies are required to prove its efficacy over open inguinal lymph node dissection.

Ovarian carcinoma usually presents at an advanced stage with diffuse intra abdominal manifestations. Inguinal lymph node metastasis is rare event in ovarian cancer. We report 7 cases who presented with inguinal lymphadenopathy as the initial manifestation between January 2014 to January 2016. All patients underwent tru-cut biopsy from inguinal area. Morphology and IHC were suggestive of ovarian origin or female genital Tract origin. Two patients underwent primary debulking surgery while four patients were managed by neo-adjuvant chemotherapy followed by interval cytoreductive surgery owing to relatively poor performance status at presentation. One patient underwent secondary debulking in which inguinal Lymph node was positive for metastatic deposits.

Ovarian cancers tend to recur in 50–70 % cases. Metastases from ovarian cancer primarily involve the peritoneum rather than parenchymal sites; thus, the presence of small-volume recurrence or metastatic deposits on the visceral surfaces poses a challenge for interpretation of CT and MR images. PET/CT utilizes its property of higher accumulation in malignant cells to provide both anatomic and functional information for diagnosing malignant tumors. The objectives of the study were to find the correlation between PET/CT findings and final histopathological diagnosis in suspected ovarian cancer recurrences. PET/CT was done in cases with rising or above normal CA-125. These patients with abnormal PET/CT findings were taken up for a secondary cytoreductive surgery, and histopathological reports were taken as the standard against which PET/CT positive findings were compared. Of the 52 patients with suspected recurrence, 40 patients with a PET-CT scan with findings suggestive of an avid uptake underwent surgery. Of the 40 patients who underwent a secondary cytoreductive surgery, 38 had histopathologically confirmed recurrence. PET-CT detected a total of 86 lesions, of which 38 were in the lymph nodes, 32 were peritoneal lesions and 14 were pelvic, 2 were metastatic in the parenchyma of liver. Mean diameter of the lesions detected was 2.2 cm (range 0.3–6.2 cm). PET-CT accurately identified 72 of the 80 histopathologically proven lesions. The overall lesion-based sensitivity of PET-CT is 90 %, specificity 66.7 %, positive predictive value being 83.7 % and negative predictive value 77.7 % and the overall accuracy being 81.9 %. Correlation between PET/CT and histopathological disease: κ (Cohen value) = 0.88 suggests excellent correlation. Selected patients with ovarian cancer recurrence may benefit from PET-CT at the time of even no. or minimal CA-125 elevation by early detection and successful cytoreduction.

Background: Chronic obstructive pulmonary disease (COPD) is the fourth most common cause of death in the world, for which smoking is a common cause. It is preferable to diagnose COPD at an earlier stage and to assess its progression so that mortality and morbidity of the disease could be reduced. Hence, we conducted this study to assess parameters of body plethysmography in Indian population where the data are lacking and to assess whether the use of body plethysmography can detect COPD earlier. Subjects and Methods: The study was approved by the Ethics Committee at B. J Government Medical College, Pune. In this comparative randomized cross-sectional study, healthy control subjects (CN), smokers without COPD diagnosis (SM) who were smoking for more than 5 pack-years and smokers with COPD who were further classified depending upon GOLD criteria as mild COPD (C1), moderate COPD (C2), and severe COPD (C3) (n = 30 each group) were considered. All the participants were males who gave written informed consent. Subject underwent routine spirometry (FEV1, FVC, FEV1/FVC, PEFR, and FEF25-75%) along with body plethysmography where sGaweff, sGawtot, residual volume (RV), total lung capacity (TLC), and inspiratory capacity (IC) were recorded. Statistical Analysis: The differences in lung function were compared between healthy controls and smokers and also between the three groups of COPD severity (GOLD guidelines) employing univariate analysis of variance and Bonferroni's post hoc test. Results: Spirometry could not differentiate between smokers without COPD and healthy controls. However, three parameters on body plethysmography (IC, sGawtot, and sGaweff) were sensitive enough to detect differences between smokers without COPD and healthy controls. Conclusion: Using body plethysmography, the vexed question troubling the clinician, which of the smokers progress to COPD and who do not before they develop irreversible changes can perhaps be answered if further large-scale multicenter studies and long-term follow-up studies confirm the findings in this study.

The ESX-5 secretion system of Mycobacterium tuberculosis is important for bacterial virulence and for the secretion of the large PE/PPE protein family, whose genes constitute 10% of the M. tuberculosis genome. A four-gene region of the ESX-5 system is duplicated three times in the M. tuberculosis genome, but the functions of these duplicates are unknown. Here we investigated one of these duplicates: the region carrying the esxI , esxJ , ppe15 , and pe8 genes (ESX-5a). An ESX-5a deletion mutant in the model system M. marinum background was deficient in the secretion of some members of the PE/PPE family of proteins. Surprisingly, we also identified other proteins that are not members of this family, thus expanding the range of ESX-5 secretion substrates. In addition, we demonstrated that ESX-5a is important for the virulence of M. marinum in the zebrafish model. Furthermore, we showed the role of the M. tuberculosis ESX-5a region in inflammasome activation but not host cell death induction, which is different from the case for the M. tuberculosis ESX-5 system. In conclusion, the ESX-5a region is nonredundant with its ESX-5 paralog and is necessary for secretion of a specific subset of proteins in M. tuberculosis and M. marinum that are important for bacterial virulence of M. marinum . Our findings point to a role for the three ESX-5 duplicate regions in the selection of substrates for secretion via ESX-5, and hence, they provide the basis for a refined model of the molecular mechanism of this type VII secretion system.

AIMS AND OBJECTIVES: study was carried out to focus on effective and meaningful learning that will help the students’ overall development and to improve the continuity of students’ learning. MATERIAL AND METHOD: 150 students of 1st M.B.B.S, year 2012-2013 were selected and integrated teaching in subjects including anatomy, physiology, biochemistry, and pharmacology was conducted after selecting few important topics. Pre and post test evaluation was carried out. Questionnaire was provided for the feedback of the students. RESULTS: 75.2% students liked the session of integrated teaching and found it very useful. 80.6% of students appreciated content, method and presentation of integrated teaching module. 83.67% students wanted integrated teaching program for regular teaching every month. Out of 144 students, only 7 students were not in favor of integrated teaching. 85% students felt that integrated teaching would be beneficial to them for their summative examinations. CONCLUSION: The students recognized that integrated teaching in different medical subjects was useful and interesting and that should be continued as frequently as possible. Integrated teaching should be implemented in medical curriculum for effective teaching.

The need to enhance the relevance and quality of pre- and in-service teacher education in India has long been recognised in official commentaries. Despite the structural innovation of District Institutes of Education and Training to enhance systemic responsiveness to local contexts, training messages mediated through DIETs are largely not having the expected impact on classroom processes. Using qualitative data generated from six case study sites, this paper examines in-service training in DIETs and how they have responded to teacher development challenges. It finds that formal teacher education continues to have low ‘ecological validity’, and highlights tensions in the recruitment and technical expertise of DIET staff, and in their attitudes towards elementary teachers, that constrain engagement with local contexts. The paper then considers several aspects of elementary teachers’ local knowledge which underline the need for a review of the dominant ‘skills and knowledge’ approach to teacher development pursued by DIETs. It argues that training must shift its focus from skills to an engagement with the pressing question of teachers’ will to adopt training messages in current contexts. The paper closes with discussions of how teachers’ professional agency can be promoted, and further considerations of how teacher education can be strengthened in pursuit of quality in a context of decentralisation.

Due to the success of combined anti-retroviral therapy, HIV-1 infected patients are living longer with a better quality of life. Nonetheless, the prevalence of human immunodeficiency virus associated neurocognitive (HAND) disorder continues to be a widespread problem, and a more rapid progression of neurocognitive impairment caused by ageing in individuals with HIV has been reported. HAND is characterized by an increase in reactive oxygen, and reactive nitrogen species at the cellular level. Motor and behavioral dysfunction are among the common symptoms and it is believed that oxidative and nitrosative stress contribute to mechanistic changes leading to neurocognitive impairment. Our previous imaging, histological, motor and behavioral assessments support a dysregulation in dopaminergic function as the HIV-1 Transgenic Rat (HIV-Tg) ages. The goal of this study is to demonstrate the presence of oxidative and nitrosative stress within the neuroanatomic areas associated with dopaminergic dysfunction in the HIV-1 Tg rat brain. Herein, we report pathology suggestive of pre-synaptic dopaminergic neuronal loss (decreased striatal tyrosine hydroxylase and DAT staining with a decreased NeuN staining in the substantia nigra), increases in striatal Nox4 and neuronal nitric oxide synthase expression with an associated increase in 3-nitrotyrosine (3-NT) modification of striatal axonal neurofilament proteins of aged HIV-1 Tg rats compared to age-matched non-Tg controls. Finally, this increase in free radical mediated neuronal pathology occurred without significant induction of the transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) and redox adaptation of the Nrf2 responsive Thioredoxin and Glutathione antioxidant systems. Our findings suggest that in the aging HIV-1 Tg rat, possibly due to chronic exposure to HIV-1 proteins and lack of appropriate redox adaptation, increased nitrosative stress and 3-NT modification of striatal axonal neurofilament proteins is hastened possibly contributing to dopaminergic neuronal structural changes and secondary dopaminergic dysfunction.

We morphometrically evaluated 5-micron HE-stained sections from 28 surgically resected high-grade pulmonary neuroendocrine neoplasms, including 16 small cell lung carcinomas (SCLCs) and 12 large cell neuroendocrine carcinomas (LCNECs). For each case, 200 tumor nuclei and 20 to 100 normal lymphocytes were measured. The frequency distributions of tumor cell/lymphocyte (TC/L) size ratios were plotted in bins ranging from 1 to 6, classified into 6 histogram types with TC/L size ratio peaks ranging from 2 to 6 (A-E) and a histogram with a wide distribution (F). SCLCs fit histograms A through E; LCNECs, A through F. Morphometry demonstrated considerable nuclear size overlap in high-grade neoplasms. Approximately one third of SCLCs exhibited considerable numbers of neoplastic cells that were larger than 3 normal lymphocytes, while 4 of 12 LCNECs had a predominant number of small cells. Ten tumors exhibited a B histogram with a "borderline" peak TC/L of 3. The rule that a TC/L size ratio larger than 3 helps distinguish "large" from "small" neoplastic cells was confirmed in only 9 of 28 cases. The use of more generic terminology such as "high-grade neuroendocrine carcinoma" or "grade III neuroendocrine carcinoma" for SCLC and LCNEC is discussed.

In India, due to smoking, environmental pollution, use of bio mass as a fuel in rural areas etc, there is increase in incidence of pulmonary disorders. Hence, a review of pulmonary function tests is needed. This study has therefore been carried out to reassess the pulmonary function values in normal, Indian population and to check their variations in various obstructive and restrictive pulmonary disorders. Forced spirometry is one of the best test for assessing pulmonary disorders. This simple test provides written record of forced vital capacity (FVC), % forced expiratory volume in 1st second (FEV1%) and peak expiratory flow rate (PEFR). Method: Ours is a cross sectional, comparative study in which we measured FVC, FEV1% and PEFR in 30 normal subjects with the help of computerized spirometer. We also measured the same values in patients of pulmonary disorders and collected data of 30 patients having restrictive and 30 patients having obstructive lung disease. Thereafter, we compared the pulmonary function tests, FVC, FEV1% and PEFR in three groups. Result: Our observations show that as compared to normal values, in obstructive lung diseases, FVC remained same but there is decrease in FEV1% and PEFR. And in restrictive group of lung diseases, FVC is less than normal but FEV1% and PEFR are same as that of normal subjects. Conclusion: By our study we can conclude that pulmonary function tests like FVC, FEV1% and PEFR can be used to distinguish between obstructive and restrictive group of diseases.

Mycobacterium tuberculosis extracellular DNA gains access to the host cell cytosol via the ESX-1 secretion system. It is puzzling that this extracellular DNA of M. tuberculosis does not induce activation of the AIM2 inflammasome because AIM2 recognizes cytosolic DNA. In this study, we show that nonvirulent mycobacteria such as Mycobacterium smegmatis induce AIM2 inflammasome activation, which is dependent on their strong induction of IFN-β production. In contrast, M. tuberculosis, but not an ESX-1–deficient mutant, inhibits the AIM2 inflammasome activation induced by either M. smegmatis or transfected dsDNA. The inhibition does not involve changes in host cell AIM2 mRNA or protein levels but led to decreased activation of caspase-1. We furthermore demonstrate that M. tuberculosis inhibits IFN-β production and signaling, which was partially responsible for the inhibition of AIM2 activation. In conclusion, we report a novel immune evasion mechanism of M. tuberculosis that involves the ESX-1–dependent, direct or indirect, suppression of the host cell AIM2 inflammasome activation during infection.

The production of IL-1β during the infection with Mycobacterium tuberculosis (Mtb) is important for successful host immune defense. In macrophages and dendritic cells the host cell inflammasome is crucial for generation of secreted IL-1β in response to Mtb infections. In these cell types Mtb infection only activates the NLRP3-inflammasome. New reports demonstrate that nitric oxide has an important function in the negative regulation of the NLRP3-inflammasome to reduce tissue damage during Mtb infections. The type I interferon, IFN-β, is induced after Mtb infections and can also suppress NLRP3-inflammasome activation. In contrast, IFN-β increases activity of the AIM2-inflammasome after infection with intracellular pathogens such as Francisella tularensis and Listeria monocytogenes. Recent results demonstrate that non-tuberculous mycobacteria but not virulent Mtb induce the AIM2-inflammasome in an IFN-β dependent matter. Indeed, Mtb inhibits AIM2-inflammasome activation via its ESX-1 secretion system. This novel immune evasion mechanism may help Mtb to allow the induction of low levels of IFN-β to suppress the NLRP3-inflammasome without activating the AIM2-inflammasome.