Drug-sensitive FGFR3 mutations in lung adenocarcinoma.

Background: Lung cancer is the leading cause of cancer-related deaths across the world. In this study, we present therapeutically relevant genetic alterations in lung adenocarcinoma of Indian origin. Materials and methods: Forty-five primary lung adenocarcinoma tumors were sequenced for 676 amplicons using RainDance cancer panel at an average coverage of 1500 × (reads per million mapped reads). To validate the findings, 49 mutations across 23 genes were genotyped in an additional set of 363 primary lung adenocarcinoma tumors using mass spectrometry. NIH/3T3 cells over expressing mutant and wild-type FGFR3 constructs were characterized for anchorage independent growth, constitutive activation, tumor formation and sensitivity to FGFR inhibitors using in vitro and xenograft mouse models. Results: We present the first spectrum of actionable alterations in lung adenocarcinoma tumors of Indian origin, and shows that mutations of FGFR3 are present in 20 of 363 (5.5%) patients. These FGFR3 mutations are constitutively active and oncogenic when ectopically expressed in NIH/3T3 cells and using a xenograft model in NOD/SCID mice. Inhibition of FGFR3 kinase activity inhibits transformation of NIH/3T3 overexpressing FGFR3 constructs and growth of tumors driven by FGFR3 in the xenograft models. The reduction in tumor size in the mouse is paralleled by a reduction in the amounts of phospho-ERK, validating the in vitro findings. Interestingly, the FGFR3 mutations are significantly higher in a proportion of younger patients and show a trend toward better overall survival, compared with patients lacking actionable alterations or those harboring KRAS mutations. Conclusion: We present the first actionable mutation spectrum in Indian lung cancer genome. These findings implicate FGFR3 as a novel therapeutic in lung adenocarcinoma.

Early screening to identify patients at risk of developing intra-abdominal hypertension and abdominal compartment syndrome.

BACKGROUND: To develop a screening tool to identify patients at risk of developing intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) within 24 h of a patient's admission to intensive care unit (ICU). METHODS: Prospective, observational study of 403 consecutively enrolled patients with an indwelling catheter, admitted to a mixed medical-surgical ICU in a tertiary referral, university hospital. Intra-abdominal pressure was measured at least twice daily and IAH and ACS defined as per consensus definitions. RESULTS: Thirty-nine per cent of patients developed IAH and 2% developed ACS. Abdominal distension, hemoperitoneum/pneumoperitoneum/intra-peritoneal fluid collection, obesity, intravenous fluid received > 2.3 l, abbreviated Sequential Organ Failure Assessment score > 4 points and lactate > 1.4 mmol/l were identified as independent predictors of IAH upon admission to ICU. The presence of three or more of these risk factors at admission identified patients that would develop IAH with a sensitivity of 75% and a specificity of 76%, the development of grades II, III and IV IAH with a sensitivity of 91% and a specificity of 62%. Patients that developed IAH required a significantly longer duration of mechanical ventilation and ICU care. Patients that developed grades II-IV IAH had a significantly higher rate of ICU mortality. CONCLUSION: IAH is a common clinical entity in the intensive care setting that is associated with morbidity and mortality. A screening tool, based on data readily available within a patient's first 24 h in ICU, was developed and effectively identified patients that required intra-abdominal pressure monitoring.

Intra-abdominal hypertension and abdominal compartment syndrome: pathophysiological and non-operative management.

Intra-abdominal hypertension (IAH) and abdominal compartment syndrome (ACS) are conditions that commonly manifest in critically ill patients. They are associated with a multiplicity of pathophysiological disturbances. This study retrospectively reviewed literature relating to IAH and ACS published in the last two decades to consolidate an understanding of the epidemiology, etiology, pathophysiology, diagnosis and non-operative management of these conditions. Additionally, the authors of this study have recently conducted a large study on intra-abdominal pressures of consecutive catheterised patients admitted to the Intensive Care Unit (N.=403). A preliminary analysis of this study has also been included.

A novel method of detection and classification of motor nerve late-wave activity.

In motor nerve conduction studies, important diagnostic information is provided by the late-wave responses, comprised of F-waves, A-waves, and repeaters. Late-waves in addition to contamination from power line interference and baseline disturbance, are of low amplitude and random in nature. This makes computer-based analysis of late-wave activity very challenging, especially the computer-based F-wave onset latency assignment. Currently available algorithms assign latency independently on a trace-by-trace basis without considering the information present in an entire ensemble of traces. A novel algorithm that takes into account the ensemble information for segmenting and classifying regions of late-wave data is described in this paper, which in turn can be used to improve the performance of computer-based F-wave onset latency assignment.

Depression in the elderly in Vellore, South India: the use of a two-question screen.

BACKGROUND: Depression in the elderly is a common and disabling condition. The aim of the study was to evaluate the sensitivity and specificity of a two-question screen to identify depression and common mental disorders in the elderly. METHOD: Residents of a ward in the town of Vellore were identified by a door-to-door survey from which 204 subjects aged over 60 years were selected for the study by systematic random sampling. They were screened using the two-question screen. The Revised Clinical Interview Schedule (CIS-R) was employed to confirm the diagnosis. RESULTS: The prevalence of depression and common mental disorder, using the CIS-R standard, was found to be 31.5%. The two-question screen has a sensitivity of 93.8% and specificity of 48.2%. CONCLUSIONS: The high sensitivity of the two-question screen makes it a useful screening method which can be employed by health workers in the field.

Detection and classification of motor nerve late-wave activity.

The health of peripheral nerves is commonly assessed using nerve conduction studies, in which the nerve is stimulated electrically and the evoked responses are analyzed. In motor nerve studies, important diagnostic information is provided by the late-wave responses, comprised of F-waves, A-waves and repeaters. Computer-based analysis of late-wave activity is challenging due to the random nature and low amplitude of these signals. This paper describes an algorithm for segmenting portions of late-wave activity and classifying the type of evoked response. The algorithm may be used to improve the robustness of computer-based late-wave assessment.

Estimation of individual evoked potential components using iterative independent component analysis.

Independent component analysis (ICA) has been successfully employed in the study of single-trial evoked potentials (EPs). In this paper, we present an iterative temporal ICA methodology that processes multielectrode single-trial EPs, one channel at a time, in contrast to most existing methodologies which are spatial and analyze EPs from all recording channels simultaneously. The proposed algorithm aims at enhancing individual components in an EP waveform in each single trial, and relies on a dynamic template to guide EP estimation. To quantify the performance of this method, we carried out extensive analyses with artificial EPs, using different models for EP generation, including the phase-resetting and the classical additive-signal models, and several signal-to-noise ratios and EP component latency jitters. Furthermore, to validate the technique, we employed actual recordings of the auditory N100 component obtained from normal subjects. Our results with artificial data show that the proposed procedure can provide significantly better estimates of the embedded EP signals compared to plain averaging, while with actual EP recordings, the procedure can consistently enhance individual components in single trials, in all subjects, which in turn results in enhanced average EPs. This procedure is well suited for fast analysis of very large multielectrode recordings in parallel architectures, as individual channels can be processed simultaneously on different processors. We conclude that this method can be used to study the spatiotemporal evolution of specific EP components and may have a significant impact as a clinical tool in the analysis of single-trial EPs.

Comparison between ICA and wavelet-based denoising of single-trial evoked potentials.

Analysis of evoked potentials (EPs) on a single-trial basis allows the study of the dynamical characteristics of brain activity. However, single-trial responses are buried into the more prominent ongoing electroencephalographic (EEG) activity, and thus advanced procedures are needed to obtain the activity only of the cortical generators that are activated by the experimental task under study. We compare the effectiveness of two methods at removing extraneous activity from single-trial EPs, namely, a recently-proposed iterative procedure based on independent component analysis (ICA) and wavelet denoising, using simulated data and actual EP recordings from normal subjects, more specifically the auditory N100-P200 complex.

Developmental changes in expression of myotonic dystrophy protein kinase in the rat central nervous system.

Myotonic dystrophy protein kinase (DMPK) is the protein product of the genetic locus associated with myotonic dystrophy, in which alterations of muscle excitability, cardiac conduction defects, mental retardation, and cognitive deficiencies are inherited as an autosomal dominant trait. DMPK belongs to a novel protein serine/threonine kinase family, but its regulation and physiological functions have not been specified. In a first step toward understanding the functions of DMPK in the central nervous system, we have characterized its localization and developmental pattern of expression in rat brain and spinal cord by using a monospecific rabbit antiserum produced against bacterially expressed DMPK. Expression of DMPK begins after birth and increases gradually to peak at postnatal day 21 with antibody labeling of neuronal cell types in many regions. After postnatal day 21 and proceeding to the adult, the pattern of expression becomes more restricted, with localization to certain regions or cell groups in the central nervous system. Electron microscopy reveals localization within adult spinal motor neurons to the endoplasmic reticulum and dendritic microtubules. The adult localizations suggest that DMPK may function in membrane trafficking and secretion within neurons associated with cognition, memory, and motor control.

Effects of Mycobacterium tuberculosis on the bioelectric properties of the alveolar epithelium.

To investigate the hypothesis that Mycobacterium tuberculosis penetrates the alveolar epithelium by downregulating its barrier properties, we evaluated the interactions between M. tuberculosis and rat alveolar epithelial cell monolayers that are believed to share electrophysiologic properties of the human alveolar epithelium. Nonproteinaceous components of M. tuberculosis caused marked declines in electrical resistance and equivalent short-circuit current of the alveolar epithelial cell monolayers, indicating a reduction in the capacity to maintain tight intercellular junctions and to actively reabsorb sodium. M. tuberculosis elicited production of TNF-alpha mRNA and protein by alveolar epithelial cells, and the effects of recombinant TNF-alpha on the bioelectric properties of the alveolar epithelial paralleled those of M. tuberculosis. Furthermore, the effects of M. tuberculosis on alveolar epithelial resistance were abrogated by neutralizing anti-TNF-alpha antibodies. These results indicate that M. tuberculosis elicits production of TNF-alpha, which in turn reduces the bioelectric barrier properties of the alveolar epithelium. These findings provide insight into potential mechanisms by which M. tuberculosis establishes infection and disease in the lung.

Immune response to recombinant mycobacterial proteins in patients with tuberculosis infection and disease.

The capacity of four Mycobacterium tuberculosis recombinant antigens to elicit proliferation and cytokine production by human T cells was evaluated. Proliferative responses of peripheral blood mononuclear cells (PBMC) to all antigens were greater in healthy tuberculin reactors than in pulmonary tuberculosis patients, and proliferative responses of pleural fluid cells were greater than those of PBMC from patients with tuberculous pleuritis. The proliferative responses to the four recombinant antigens were similar in all patient groups, and there was no selective unresponsiveness to any antigen in pulmonary tuberculosis patients. The 38-kDa antigen induced less interferon-gamma than did the 10-, 30-, and 65-kDa antigens, and all four antigens induced similar amounts of interleukin-10. These results suggest that none of the four recombinant antigens are immunodominant, and that the 10-, 30-, and 65-kDa antigens are similar in their capacity to induce a potentially protective Th1-like response.

Interleukin-10 downregulates Mycobacterium tuberculosis-induced Th1 responses and CTLA-4 expression.

To characterize the mechanism by which interleukin 10 (IL-10) inhibits Th1 responses to intracellular pathogens, we evaluated the interaction between IL-10 and Mycobacterium tuberculosis-induced gamma interferon (IFN-gamma) production by peripheral blood mononuclear cells from persons across the spectrum of tuberculous infection. M. tuberculosis-induced IFN-gamma production was highest in healthy tuberculin reactors, intermediate in human immunodeficiency virus (HIV)-negative tuberculosis patients, and lowest in HIV-infected tuberculosis patients. Neutralizing antibodies to IL-10 increased IFN-gamma production in HIV-infected and HIV-negative tuberculosis patients by enhancing monocyte IL-12 production. Expression of the T-cell-costimulatory molecule CTLA-4 was depressed in M. tuberculosis-stimulated peripheral blood mononuclear cells from tuberculosis patients, and anti-IL-10 and Il-12 upregulated expression of CTLA-4. These findings provide evidence that intracellular pathogens can inhibit Th1 responses and downregulate expression of specific costimulatory molecules.

T-cell cytokine responses in human infection with Mycobacterium tuberculosis.

Compared with healthy tuberculin reactors, Mycobacterium tuberculosis-stimulated peripheral blood mononuclear cells from tuberculosis patients had diminished production and mRNA expression of the Th1 cytokines gamma interferon and interleukin 2 (IL-2), with no change in production and mRNA expression for the Th2 cytokines IL-4, IL-10, and IL-13. These results were confirmed by evaluation of T cells and CD4+ cells. At the level of systemic T cells, development of tuberculosis is associated with diminished Th1 but not enhanced Th2 responses.

T cell cytokine responses in persons with tuberculosis and human immunodeficiency virus infection.

Tuberculosis causes more extensive and life-threatening disease in patients with HIV infection than in immunocompetent persons. To investigate the hypothesis that these severe manifestations of tuberculosis may be due to alterations in cytokine production, we evaluated cytokine patterns in HIV-infected tuberculosis patients. Upon stimulation with Mycobacterium tuberculosis in vitro, PBMC from HIV-infected tuberculosis patients had reduced proliferative and type 1 responses, compared with HIV-seronegative tuberculosis patients. The reduction in proliferative responses was independent of the CD4 cell count, but the reduced type 1 response was a direct result of CD4 cell depletion. There was no enhancement of type 2 cytokine production in HIV-infected patients, although production of IL-10 was prominent in all tuberculosis patients. In HIV-infected tuberculosis patients, M. tuberculosis-induced proliferative responses were significantly enhanced by neutralizing antibodies to IL-10 but not by antibodies to IL-4 or by recombinant IL-12. The M. tuberculosis-induced type 1 response was augmented both by antibodies to IL-10 and by recombinant IL-12. Tuberculosis in the context of HIV infection is characterized by diminished type 1 responses, probably induced by immunosuppressive cytokines produced by macrophages/monocytes, rather than by type 2 cells.