Enhancing Metabolome Coverage in Data-Dependent LC-MS/MS Analysis through an Integrated Feature Extraction Strategy.
In untargeted metabolomics, conventional data preprocessing software (e.g., XCMS, MZmine 2, MS-DIAL) are used extensively due to their high efficiency in metabolic feature extraction. However, these programs present limitations in recognizing low-abundance metabolic features, thus hindering complete metabolome coverage from the analysis. In this work, we explored the possibility of enhancing the metabolome coverage of data-dependent liquid chromatography-tandem mass spectrometry (LC-MS/MS) results by rescuing metabolic features that are missed by conventional software. To achieve this goal, we first categorized the metabolic features into four confidence levels based on their chromatographic peak shapes and the presence of corresponding MS/MS spectra. We then assessed the false positives and quantitative accuracy of the metabolic features that contain MS/MS spectra but are not recognized by conventional software. Our results indicate that these missed features contain valid and important metabolic information and should be integrated into the conventional metabolomics results. Thus, we developed a data-preprocessing pipeline to extract low-abundance metabolic features and integrate them with the results from conventional programs. This integrated feature extraction strategy was tested on a set of fecal metabolomic data retrieved from mice who have undergone normal diet vs high-fat diet treatments. In our test data set, the integrated feature extraction approach increased the number of significant features being extracted by 24.4% and identified five additional metabolites bearing critical biological meanings. Our results show that this integrated feature extraction strategy remarkably improves the metabolome coverage beyond that of conventional data preprocessing, therefore facilitating the confirmation of metabolites of interest and accomplishment of a higher success rate in de novo metabolite identification.


[Effect of (-)-epigallocatechin-3-gallate and 5-AZA-CdR on expression of X-linked inhibitor of apoptosis protein-associated factor 1 in human leukemia HL-60 and K562 cells].
To study the expression of X-linked inhibitor of apoptosis protein (XIAP)-associated factor 1 (Xaf1) in human leukemia HL-60 and K562 cells treated with interferon alpha (INFalpha) and the demethylating agent 5-AZA-CdR, and observe the synergetic antitumor effect of Xaf1 inducer and (-)-epigallocatechin-3-gallate (EGCG). Human leukemia HL-60 and K562 cells were treated for 48 h with 1000 U/ml INFalpha and different doses of 5-AZA-CdR, and the mRNA expressions of both Xaf1 and XIAP were measured by RT-PCR. The leukemia cells were also treated with the optimal Xaf1 inducer in combination with EGCG, after which flow cytometry was employed to examine the changes in the members of the Bcl-2 family, mitochondrial transmembrane potential and apoptosis. As the dose increased, 5-AZA-CdR dose-dependently up-regulated the mRNA expression of Xaf1 in HL-60 and K562 cells; INFalpha treatment also resulted in increased Xaf1 expression, but 5 micromol/L 5-AZA-CdR showed the most potent effect. Neither INFalpha nor 5-AZA-CdR caused significant changes in XIAP expression. Combined treatment with 5-AZA-CdR and EGCG altered the expressions of Bcl-2 (Bcl-xl) and Bax in HL-60 and K562 cells, decreased the mitochondrial transmembrane potential and induced cell apoposis, and the two agents exhibited obvious synergistic effect. INFalpha and 5-AZA-CdR can induce Xaf1 mRNA expressions in HL-60 and K562 cells, and the effect of 5-AZA-CdR was dose-dependent. 5-AZA-CdR and EGCG induces apoptosis of leukemia cells in vitro, and they exhibits obvious synergetic effects.


Vasoactive intestinal polypeptide secreting islet cell tumors: a 15-year experience and review of the literature.
Vasoactive intestinal polypeptide (VIP)-secreting tumors of the pancreas represent a rare subtype of pancreatic islet cell tumors with an estimated incidence of 0.2 to 0.5 per million per year. We provide data on a relatively large series of patients with VIP-secreting tumors and review current literature regarding this specific entity. A retrospective review was performed of all patients with VIP-secreting tumors of the pancreas treated from 1977 to 1992 at our institution. Presenting signs, symptoms, mode of diagnosis, extent of disease, surgical resectability, tumor size, treatments, hormone levels, and survival were assessed. Eighteen patients were identified, 9 male and 9 female. Ages ranged from 23 to 74 years (mean 51 years). Secretory diarrhea was the most common symptom, occurring in 16 of 18 patients (89%). The most common tumor location was the tail of the pancreas (9 patients). Fourteen patients (78%) had liver metastasis at diagnosis. Curative resections were attempted in only 5 patients (28%). The mean survival was 3.6 years with the longest disease-free survival being 15 years and longest overall survival 15 years. VIP-secreting tumors are extremely rare entities and usually metastatic at the time of diagnosis. Despite advanced disease, these patients can have extended survival.


Stimulation of fibroblast growth factor receptor-1 occupancy and signaling by cell surface-associated syndecans and glypican.
The formation of distinctive basic FGF-heparan sulfate complexes is essential for the binding of bFGF to its cognate receptor. In previous experiments, cell-surface heparan sulfate proteoglycans extracted from human lung fibroblasts could not be shown to promote high affinity binding of bFGF when added to heparan sulfate-deficient cells that express FGF receptor-1 (FGFR1) (Aviezer, D., D. Hecht, M. Safran, M. Eisinger, G. David, and A. Yayon. 1994. Cell 79:1005-1013). In alternative tests to establish whether cell-surface proteoglycans can support the formation of the required complexes, K562 cells were first transfected with the IIIc splice variant of FGFR1 and then transfected with constructs coding for either syndecan-1, syndecan-2, syndecan-4 or glypican, or with an antisense syndecan-4 construct. Cells cotransfected with receptor and proteoglycan showed a two- to three- fold increase in neutral salt-resistant specific 125I-bFGF binding in comparison to cells transfected with only receptor or cells cotransfected with receptor and anti-syndecan-4. Exogenous heparin enhanced the specific binding and affinity cross-linking of 125I-bFGF to FGFR1 in receptor transfectants that were not cotransfected with proteoglycan, but had no effect on this binding and decreased the yield of bFGFR cross-links in cells that were cotransfected with proteoglycan. Receptor-transfectant cells showed a decrease in glycophorin A expression when exposed to bFGF. This suppression was dose-dependent and obtained at significantly lower concentrations of bFGF in proteoglycan-cotransfected cells. Finally, complementary cell-free binding assays indicated that the affinity of 125I-bFGF for an immobilized FGFR1 ectodomain was increased threefold when the syndecan-4 ectodomain was coimmobilized with receptor. Equimolar amounts of soluble syndecan-4 ectodomain, in contrast, had no effect on this binding. We conclude that, at least in K562 cells, syndecans and glypican can support bFGF-FGFR1 interactions and signaling, and that cell-surface association may augment their effectiveness.


Transforming healthcare through patient empowerment.
The United States faces tremendous challenges with its healthcare system. By any standard, it is expensive and performs poorly in most measures of health and thus, is in great need of reform. But how do we reform things without making the situation worse? Some of the more fundamental problems arise from the combination of a fee-for-service payment system for physicians with insurance-based financing care. This combination results in conflicts among the interests of patients, physicians and payers. This paper examines this issue from a decision analytic perspective, starting with a definition of the patient-centered view, and an assessment of the practicality of controlling costs by making healthcare more patient-centric. It then illustrates how fee-for-service models corrupt decision-making and other solutions designed to reign in the abuses of the fee-for-service model and also negatively impacts the quality of decision making for individual patients. Whatever the strategies for health reform, the degree of patient-centeredness of care is a benchmark that allows policy makers to understand how far they have had to deviate from optimal to achieve the desired ends of cost control.


Tryparedoxin peroxidases from Trypanosoma cruzi: high efficiency in the catalytic elimination of hydrogen peroxide and peroxynitrite.
During host cell infection, Trypanosoma cruzi parasites are exposed to reactive oxygen and nitrogen species. As part of their antioxidant defense systems, they express two tryparedoxin peroxidases (TXNPx), thiol-dependent peroxidases members of the peroxiredoxin family. In this work, we report a kinetic characterization of cytosolic (c-TXNPx) and mitochondrial (m-TXNPx) tryparedoxin peroxidases from T. cruzi. Both c-TXNPx and m-TXNPx rapidly reduced hydrogen peroxide (k=3.0 x 10⁷ and 6 x 10⁶ M⁻¹ s⁻¹ at pH 7.4 and 25 °C, respectively) and peroxynitrite (k=1.0 x 10⁶ and k=1.8 x 10⁷ M⁻¹ s⁻¹ at pH 7.4 and 25 °C, respectively). The reductive part of the catalytic cycle was also studied, and the rate constant for the reduction of c-TXNPx by tryparedoxin I was 1.3 x 10⁶ M⁻¹ s⁻¹. The catalytic role of two conserved cysteine residues in both TXNPxs was confirmed with the identification of Cys52 and Cys173 (in c-TXNPX) and Cys81 and Cys204 (in m-TXNPx) as the peroxidatic and resolving cysteines, respectively. Our results indicate that mitochondrial and cytosolic TXNPxs from T. cruzi are highly efficient peroxidases that reduce hydrogen peroxide and peroxynitrite, and contribute to the understanding of their role as virulence factors reported in vivo.


MMP-9 expression varies according to molecular subtypes of breast cancer.
In 2014, breast cancer remains a major cause of mortality worldwide mostly due to tumor relapse and metastasis. There is currently a great interest in identifying cancer biomarkers and signalling pathways mechanistically related to breast cancer progression. Matrix metalloproteinase-9 (MMP-9) is a member of matrix degrading enzymes involved in cancer development, invasion and metastasis. Our objective was to investigate MMP-9 expression in normal human breast tissue and to compare it to that of breast cancer of various histological grades and molecular subtypes. We also sought to correlate MMP-9 expression with the incidence of metastasis, survival rates and relapse in breast cancer patients. MMP-9 was first studied using in silico analysis on available DNA microarray and RNA sequencing data of human breast cancer tissues and human breast cancer cell lines. We next ascertained MMP-9 expression in both normal breast tissue and in human breast carcinoma tissue microarrays. Significant increase in MMP-9 expression was found in breast cancer cells where compared to normal breast tissue. A positive correlation could also be established between elevated levels of MMP-9 and breast cancer of high histological grade. Furthermore, our results indicate that not only MMP-9 is differentially expressed between each molecular subset but also, more importantly MMP-9 overexpression revealed itself as a startling feature of triple-negative and HER2-positive breast cancers. Lastly, the clinical relevance of MMP-9 overexpression is strongly supported by its significant association with a higher incidence of metastasis and relapse. Differential expression of MMP-9 reflects the extent of cellular differentiation in breast cancer cells and is closely related to the most aggressive subtypes of breast cancer. Hence, MMP-9 is a promising prognostic biomarker of high-grade breast cancer. In our opinion, MMP-9 expression could help segregate subsets of aggressive breast cancer into clinically meaningful subtypes.


Educating leaders in hospital management: a new model in Sub-Saharan Africa.
QUALITY ISSUE: The vast majority of health system capacity-building efforts have focused on enhancing medical and public health skills; less attention has been directed at developing hospital managers despite their central role in improving the functioning and quality of health-care systems. Initial ASSESSMENT AND CHOICE OF INTERVENTION: Initial assessment of hospital management systems demonstrated weak functioning in several management areas. In response, we developed with the Ethiopian Ministry of Health (MoH) a novel Master of Hospital Administration (MHA) program, reflecting a collaborative effort of the MoH, the Clinton HIV/AIDS Initiative, Jimma University and Yale University. The MHA is a 2-year executive style educational program to develop a new cadre of hospital leaders, comprising 5% classroom learning and 85% executive practice. The MHA has been implemented with 55 hospital leaders in the position of chief executive officer within the MoH, with courses taught in collaboration by faculty of the North and the South universities. The program has enrolled two cohorts of hospital leaders and is working in more than half of the government hospitals in Ethiopia. Lessons learned include the need to: (i) balance education in applied, technical skills with more abstract thinking and problem solving, (ii) recognize the interplay between management education and policy reform, (iii) remain flexible as policy changes have direct impact on the project, (iv) be realistic about resource constraints in low-income settings, particularly information technology limitations, and (v) manage the transfer of knowledge for longer term sustainability.


[Tricuspid and pulmonary percutaneous valvulopathy in the cardiopathy of the carcinoid syndrome. A case report].
We present a 36-year-old woman with a malignant carcinoid syndrome, liver metastases and right heart involvement with severe tricuspid and pulmonic valve stenosis. She was admitted with severe heart failure and poor general status. A combined tricuspid and pulmonic valvuloplasty was performed and she exhibited great clinical improvement that was maintained during the follow-up. We point out the importance of two-dimensional echocardiography in the diagnosis of this pathology and the role of Doppler measurements for the evaluation of the valvuloplasty's results.


The information management and technology strategy for the National Health Service in England: an approach worth watching.
This paper is an analysis of England's National Health Service (NHS) Information and Management (IM&T) strategy as a result of a March-June 1994 on-site study, which included an extensive review of the literature as well as over 60 interviews with general practitioners, senior executives in acute care hospitals and community care units, representatives of District and Family Health Services Authorities (FHSA), and senior executives from the Information Management Group of the NHS Management Executive. The observations are those of an outsider who was initially skeptical but finished quite impressed with the level of national commitment being made to the use of information technology to support the radical health care reforms the country is experiencing.


The importance of skip lesions in temporal arteritis.
To determine the frequency of skip lesions in an unselected series of temporal artery biopsies and compare the results with other series. The study was a retrospective review of 102 consecutive temporal artery biopsies taken in a five year period (1992-1997) in one large hospital. 35 cases (34.3%) showed evidence of active cranial vasculitis with pathological evidence of inflammation of the intima or media, with or without giant cells. Three of these cases (8.5%) showed apparent skip lesions: normal intima, media, and adventitia in one segment while in other segments there was clear evidence of active vasculitis. Immunocytochemical stains for leucocyte common antigen (LCA) and CD15 were helpful in identifying the absence of intimal or medial inflammatory cell infiltrates within skip lesions. Skip lesions have been described in up to 28.3% of cases in some series, while others have not found evidence of skip lesions or have identified them in a much smaller percentage of cases. In this series skip lesions were relatively rare, accounting for 8.5% of cases of active vasculitis. The degree of inflammation in temporal arteritis is discontinuous. Immunostaining for inflammatory cells, for example LCA and CD15, may be helpful in identifying the presence of an inflammatory cell infiltrate in skip lesion segments of the temporal artery.


Characteristics of Serum Autoantibody Repertoire and Immune Subgroup Variation of Tuberculosis-Associated Obstructive Pulmonary Disease.
Studying the potential etiology and pathogenesis of tuberculosis-associated chronic obstructive pulmonary disease (TOPD) from an autoimmunity perspective may provide insights into peripheral blood autoantibodies and immune cells, as well as their interactions. This study examined the serum autoantibody repertoire in healthy individuals, patients with chronic obstructive pulmonary disease (COPD), patients with pulmonary tuberculosis (TB), and TOPD patients using the HuProtTM protein chip. Autoantigens in the peripheral blood of TOPD patients were verified using ELISA assay. Various epitopes and immune simulation were predicted using bioinformatic methods. Flow cytometry was employed to detect macrophages(Mφ), T cells, and innate lymphoid cells (ILCs) in the peripheral blood. COPD patients displayed distinct alterations in their IgG and IgM autoantibodies compared to the other groups. GeneOntology (GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses revealed that these autoantibodies were associated with regulating macrophages, T cells, and B cells. ELISA results confirmed the upregulation of expression of proliferating cell nuclear antigen (PCNA), Mitogen-Activated Protein Kinase 3 antigen (MAPK3), and threonine protein kinase 1 antigen (AKT1) proteins in the peripheral blood of TOPD patients. Bioinformatic analysis predicted multiple potential epitopes in Th, CTL, and B cells. Immune simulation results demonstrated that PCNA, MAPK3, and AKT1 can activate innate and adaptive immune responses and induce the expression of different cytokines, such as IFN-g and IL-2. Furthermore, data obtained from flow cytometry assay revealed an upregulation in the face of Th1 cells in the peripheral blood of TOPD patients. Tuberculosis infection can effectively induce autoimmune responses, contributing to increased expression of Th1 cells and associated cytokines, ultimately leading to immune dysregulation. Furthermore, the accumulation of pulmonary inflammatory response facilitates the progression of TOPD and is helpful for the clinical diagnosis and the development of targeted therapeutic drugs for this disease.


T cell expression of MyD88 is required for resistance to Toxoplasma gondii.
Resistance to Toxoplasma gondii depends on dendritic cells to recognize this pathogen and secrete IL-12, in turn promoting IFN-gamma production from responding T cells. The adaptor protein, myeloid differentiation primary-response gene 88 (MyD88), is important for most Toll-like receptor (TLR) signaling, as well as IL-1R/IL-18R signals. There is considerable evidence that MyD88 is required for the innate sensing of T. gondii and IL-12 responses. Although Myd88(-/-) mice challenged with T. gondii have defective IL-12 and Th1 effector responses and succumb to disease, administration of IL-12 to Myd88(-/-) mice partially restores the Th1 response and yet fails to prolong survival. This finding suggested that MyD88 may mediate signals within T cells important for resistance to this pathogen. To evaluate the role of MyD88 in T cells under noncompetitive conditions, bone marrow chimeras were generated, in which the T cells lacked MyD88, but MyD88-dependent innate immune responses were intact. Upon challenge with T. gondii, these chimeric mice were more susceptible to disease, developing severe toxoplasmic encephalitis and succumbing within 30 days. Splenocytes and brain mononuclear cells isolated from infected chimeric mice produced less IFN-gamma when cultured with a T. gondii-derived antigen. The increase in susceptibility observed was independent of signals via the IL-1R and IL-18R, suggesting a role for TLRs in MyD88-mediated T cell responses to T. gondii. These observations show that, in addition to a role for MyD88 in innate responses, T cell expression of MyD88 is necessary for prolonged resistance to a pathogen.


Mucins, mucus, and sputum.
Normal airway mucus lines the epithelial surface and provides an important innate immune function by detoxifying noxious molecules and by trapping and removing pathogens and particulates from the airway via mucociliary clearance. The major macromolecular constituents of normal mucus, the mucin glycoproteins, are large, heavily glycosylated proteins with a defining feature of tandemly repeating sequences of amino acids rich in serine and threonine, the linkage sites for large carbohydrate structures. The mucins are composed of two major families: secreted mucins and membrane-associated mucins. Membrane-associated mucins have been reported to function as cell surface receptors for pathogens and to activate intracellular signaling pathways. The biochemical and cellular functions for secreted mucin glycoproteins have not been definitively assigned. In contrast to normal mucus, sputum production is the hallmark of chronic inflammatory airway diseases such as asthma, chronic bronchitis, and cystic fibrosis (CF). Sputum has altered macromolecular composition and biophysical properties which vary with disease, but unifying features are failure of mucociliary clearance, resulting in airway obstruction, and failure of innate immune properties. Mucin glycoprotein overproduction and hypersecretion are common features of chronic inflammatory airway disease, and this has been the underlying rationale to investigate the mechanisms of mucin gene regulation and mucin secretion. However, in some pathologic conditions such as CF, airway sputum contains little intact mucin and has increased content of several macromolecules including DNA, filamentous actin, lipids, and proteoglycans. This review will highlight the most recent insights on mucus biology in health and disease.


Case finding for chronic obstructive pulmonary disease in people attending long-term condition clinics in primary care.
Despite increased interest and awareness of chronic obstructive pulmonary disease (COPD), nearly half of the people with COPD remain undiagnosed. Inviting people at risk for screening is unlikely to be effective as many will not attend. Co-morbidities are common in people with COPD but COPD is also a comorbidity of other long-term conditions and people with these conditions are under regular review in primary care clinics. This study aimed to develop a pilot programme to case find people with COPD among patients attending other long-term clinics in primary care. Twenty-three general practices were recruited to participate in South West England. All current or ex-smokers aged ≥35 attending a long-term condition clinic who were not known to have COPD were asked to complete a questionnaire designed to help identify people with COPD and to perform microspirometry. Practices were asked to collect data on up to 100 patients. One thousand three hundred and thirty-three patients were assessed. Four hundred and ten people (31%) were current smokers. Six hundred and thirteen (46%) had high questionnaire scores and 287 (22%) of these also had a forced expiratory volume in 1 second (FEV1) below the lower limit of normal (LLN). The mean FEV1 in these patients was 59.0% of predicted (range 22-79.0%). Two hundred and twenty-four had an FEV1 between 50% and 80% of predicted, 50 had an FEV1 between 30% and 50% of predicted. One hundred and sixteen (40%) of the people with an FEV1 below the LLN were still smoking and 55 accepted referral to cessation services. A total of 56% of the other smokers assessed but not thought to have COPD also accepted referral. Assessing symptoms and performing microspirometry in people attending long-term condition clinics in primary care is feasible and has a high yield of identifying people likely to have previously undiagnosed COPD.


Defective oxidative metabolism of heart mitochondria from genetically diabetic mice.
Long chain saturated beta-hydroxy fatty acid content and oxidative metabolism were studied in hearts of diabetic mice (C57BL/KsJ db/db) with a progressive cardiomyopathy at intervals of 7, 10, 16, and 26 wk of age. Total beta-hydroxy fatty acid (BHFA) content increases progressively with age in diabetic hearts with a mean value of 143.5 nmol/g dry wt as compared with a mean of 59.6 nmol/g dry wt in control hearts. There was also a redistribution of BHFA in myocardium of diabetic mice when compared with controls, with a relative decrease in beta-hydroxymyristate and an increase of beta-hydroxypalmitate. Oxidative phosphorylation studies using isolated mitochondria from diabetic mice demonstrated depressed state 3 oxidation rates with both palmityl carnitine and pyruvate as substrates. While mitochondrial NADH-oxidase activity was not statistically different from that of controls, there was a significant decrease in mitochondrial total NAD + NADH content in diabetic hearts. In addition, treatment of myocardial tissue with lanthanum demonstrated an abnormal permeability of sarcolemmal, intercalated disc as well as mitochondrial membranes in myocytes of diabetic mice. The data indicate that deficiencies in total NAD + NADH content can account for the depressed state 3 oxidation of palmitylcarnitine and pyruvate in diabetic mice that in turn may explain the abnormal accumulation of BFHA. The latter could play a role in altering the permeability of cardiac cell membranes.


Worklife expectancies of fixed-term Finnish employees in 1997-2006.
Fixed-term employment is prevalent in the Finnish labor force. This form of employment contract is marked by fragmentary work periods, demands for flexibility in workhours, and concern for multiple insecurities. A nonpermanent employee may also incur adverse health consequences. Yet there exist no exact statistics on the duration of fixed-term employment. This paper estimated the future duration of the time that a Finn is expected to be engaged in irregular work. Multistate regression modeling and stochastic analysis were applied to aggregated data from surveys conducted among the labor force by Statistics Finland in 1997-2006. In 2006, a Finnish male was expected to work a total of 3.8 years in fixed-term employment, combined over consecutive or separate time spans; this time amounts to 8% of his remaining work career from entry into the work force until final retirement. For a woman the expectancy was greater, 6.5 years or 13%. For the age interval 20-29 years, the total was 16% for men and 23% for women. The type and duration of employment is influenced by security factors and economic cycles, both of which affect men and women differently. Over the past decade, fixed-term employment increased consistently in the female labor contingent, and it was more pronounced during economic slowdowns. This labor market development calls for standards for flexibility and guarantees for security in the fragmented future worklives of fixed-term employees.


Microtubule contribution to the reflectance of the retinal nerve fiber layer.
The reflectance of the retinal nerve fiber layer (RNFL) arises from cylindrical light scattering elements distributed throughout its thickness. To determine whether these elements include axonal microtubules, eyecup preparations of the toad Bufo marinus were treated with the microtubule depolymerizing agent colchicine. Some eyecups were incubated for various times in different colchicine concentrations and then fixed and prepared for electron microscopy (EM). In other eyecups the reflectance of the RNFL at 440 nm was measured by means of imaging microreflectometry. After a period of baseline measurements, the bathing solution was changed either to one of identical composition (control experiments) or to one containing 10 mM colchicine. Measurements then continued for 2 to 3 hours. Quantitative EM showed that colchicine caused the density of axonal microtubules to decrease by approximately one third in 2 hours. Reflectometry showed, in control experiments, that the RNFL reflectance remained near baseline for the duration of an experiment. In contrast, after changing to colchicine solution, the RNFL reflectance declined to a level 37% to 55% below baseline. Microtubules made a major contribution to the 440-nm reflectance of the unmyelinated axons of the toad RNFL and could make a similar contribution to the reflectance of human RNFL.


Cubital Tunnel Syndrome: Current Concepts.
Cubital tunnel syndrome is the second most common upper extremity compressive neuropathy. In recent years, rates of surgical treatment have increased, and the popularity of in situ decompression has grown. Nonsurgical treatment, aiming to decrease both compression and traction on the ulnar nerve about the elbow, is successful in most patients with mild nerve dysfunction. Recent randomized controlled trials assessing rates of symptom resolution and ultimate success have failed to identify a preferred surgical procedure. Revision cubital tunnel surgery, most often consisting of submuscular transposition, may improve symptoms. However, ulnar nerve recovery after revision cubital tunnel surgery is less consistent than that after primary cubital tunnel surgery.


The dynamics of syntax acquisition: facilitation between syntactic structures.
This paper sets out to show how facilitation between different clause structures operates over time in syntax acquisition. The phenomenon of facilitation within given structures has been widely documented, yet inter-structure facilitation has rarely been reported so far. Our findings are based on the naturalistic production corpora of six toddlers learning Hebrew as their first language. We use regression analysis, a method that has not been used to study this phenomenon. We find that the proportion of errors among the earliest produced clauses in a structure is related to the degree of acceleration of that structure's learning curve; that with the accretion of structures the proportion of errors among the first clauses of new structures declines, as does the acceleration of their learning curves. We interpret our findings as showing that learning new syntactic structures is made easier, or facilitated, by previously acquired ones.


[Evaluation and quantification of biologic risk in the hospital setting with ATP determination on work surfaces].
The purpose of this study was to establish whether a method of determining ATP levels using the HY-Lite system could be reliably applied in the various hospital workplaces. We estimated the correlation between ATP levels and the number of C.F.U. (Colony Forming Units), observed correlation r = 0.96. We established hygienically acceptable values for each environment, and drew up reference tables. The study reveals that conditions of high bacterial and fungine contamination can occur, albeit the agents are not normally pathogenic is so far as they consist of common environmental pollutants. Consequently, these results emphasize the need to apply strict sanitization and hygienic measures, that health workers must be suitably trained to deal with follow. The results of the sampling appear to suggest that condition of potential biological risk for workers are largely due to incorrect work procedure in workplaces where pathologic agents are present.


Pancreas allograft rejection: analysis of concurrent renal allograft biopsies and posttherapy follow-up biopsies.
Pancreas and kidney allograft function is routinely monitored with serum studies (amylase, lipase, and creatinine). Increased levels commonly prompt tissue biopsy, to diagnose cause of graft dysfunction. Historically, pancreas allografts were infrequently biopsied, although serum enzymes and renal rejection may be poor surrogates for pancreas status. Pancreas allograft biopsies at our center were reviewed and reclassified according to University of Maryland (UMD) and Banff criteria; C4d immunostaining was performed. Findings were correlated with clinical data and renal allograft biopsies. Fifty-six pancreas allograft biopsies from 27 patients were evaluated. UMD and Banff grading were similar, although two UMD "indeterminate" biopsies were Banff grade 1 rejection. There were 21 concurrent pancreas and renal biopsies, all from simultaneous pancreas-kidney allograft recipients. Thirteen pairs were concordant for rejection; eight pairs were discordant for rejection (38%); six pairs showed pancreas rejection without kidney rejection, and two pairs showed the converse. Fourteen patients had a total of 21 follow-up pancreas allograft biopsies. Seven biopsies showed a lower grade of rejection on follow-up biopsy, 4 biopsies showed more severe rejection, and 10 had unchanged grade. In only 9 of these 21 (43%) cases, did the interval serum amylase or lipase trend parallel the subsequent biopsy diagnosis. With a high biopsy discordance rate, our data suggest that renal allograft rejection is a poor surrogate for pancreas allograft status. Likewise, serum amylase and lipase levels do not predict response to rejection therapy. Surveillance or posttherapy pancreas allograft biopsies may be a useful means to monitor pancreas allograft status.


Anal sex, vaginal sex and HIV risk among female sex workers in Papua New Guinea.
Female sex workers (FSW) are considered one of the key affected populations in Papua New Guinea at risk of acquiring HIV. An integrated bio-behavioral survey of sex workers in Port Moresby was conducted to determine the nature and extent of this risk. About half (51.1 %) of the 411 FSW who reported having any sexual intercourse with clients had engaged in both anal and vaginal intercourse with clients in the last 6 months. In spite of having poorer HIV knowledge (OR95 % CI = 0.14-0.34), FSW who had anal intercourse with clients were significantly more likely to have used a condom at the last vaginal intercourse with a client (OR95 % CI = 1.04-2.87). Similarly, FSW who had anal intercourse with regular and casual partners were significantly more likely to have used a condom at the last vaginal intercourse. Those who engaged in both anal and vaginal intercourse with clients had similar condom use for both vaginal and anal intercourse, with the majority (78.1 %) using a condom at the last occasion for both vaginal and anal intercourse. These FSW may have different risk and protective factors that affect their use of condom during sexual intercourse. Further research is needed to investigate this difference between those who practice anal intercourse and those who do not in order to provide evidence for better programming.


Large Anomalous Nernst Effects at Room Temperature in Fe3 Pt Thin Films.
Heat current in ferromagnets can generate a transverse electric voltage perpendicular to magnetization, known as anomalous Nernst effect (ANE). ANE originates intrinsically from the combination of large Berry curvature and density of states near the Fermi energy. It shows technical advantages over the conventional longitudinal Seebeck effect in converting waste heat to electricity due to its unique transverse geometry. However, materials showing giant ANE remain to be explored. Herein, a large ANE thermopower of Syx ≈ 2 µV K-1 at room temperature in ferromagnetic Fe3 Pt epitaxial films is reported, which also show a giant transverse thermoelectric conductivity of αyx ≈ 4 A K-1 m-1 and a remarkable coercive field of 1300 Oe. The theoretical analysis reveals that the strong spin-orbit interaction in addition to the hybridization between Pt 5d and Fe 3d electrons leads to a series of distinct energy gaps and large Berry curvature in the Brillouin zone, which is the key for the large ANE. These results highlight the important roles of both Berry curvature and spin-orbit coupling in achieving large ANE at zero magnetic field, providing pathways to explore materials with giant transverse thermoelectric effect without an external magnetic field.


[Chlamydia infection in patients with and without cervical intra-epithelial lesions].
To determine the frequency of Chlamydia trachomatis infection in patients with and without intraepithelial cervical lesions attended at specialized ambulatory in Recife (2007) and its association with biologic and demographic variables, habits, reproductive, clinical and gynecologic characteristics. A cross-sectional study was conducted including 70 women (35 with cytological cervical lesions and 35 with normal smears). Colposcopy and biopsy when necessary were performed and Chlamydia infection was determined by direct immunofluorescence. Analysis variables were age, race, precedence, schooling, marital status, menarche, age at first sexual intercourse, parity, number of sexual partners, vaginal discharge, previous Pap smear, STD episodes, cervical cauterization, contraceptive methods, familial antecedents of cervical cancer, alcohol intake, use of illicit and immunosuppressive drugs, Papanicolaou result and cervical Chlamydia l infection. To determine the strength of association prevalence ratio (PR) and its 95% confidence interval were calculated. Multivariate analysis was performed for controlling potentially confounding variables. the frequency of Chlamydia l infection was significantly greater in patients with intraepithelial cervical lesions (80.0% vs. 14.3%; PR=5.60; 95% CI=2.44-12.82). When the factors associated with Chlamydia l infection were analyzed, the only variable that remained significantly associated after multivariate analysis was previous episode of STD (OR=63.47; 95% CI=13.93-289.09). Chlamydia trachomatis infection is associated with intraepithelial cervical lesions and history of STD should be considered for treatment and follow-up of these patients.


The Development, Optimization and Future of Prime Editing.
Prime editing is a rapidly developing method of CRISPR/Cas-based genome editing. The increasing number of novel PE applications and improved versions demands constant analysis and evaluation. The present review covers the mechanism of prime editing, the optimization of the method and the possible next step in the evolution of CRISPR/Cas9-associated genome editing. The basic components of a prime editing system are a prime editor fusion protein, consisting of nickase and reverse transcriptase, and prime editing guide RNA, consisting of a protospacer, scaffold, primer binding site and reverse transcription template. Some prime editing systems include other parts, such as additional RNA molecules. All of these components were optimized to achieve better efficiency for different target organisms and/or compactization for viral delivery. Insights into prime editing mechanisms allowed us to increase the efficiency by recruiting mismatch repair inhibitors. However, the next step in prime editing evolution requires the incorporation of new mechanisms. Prime editors combined with integrases allow us to combine the precision of prime editing with the target insertion of large, several-kilobase-long DNA fragments.


Acute lymphoblastic leukemia following myelodysplastic syndrome.
A 58-year-old man presented with anemia and neutropenia. The bone marrow examinations showed changes consistent with myelodysplastic syndrome. Twenty months later, acute lymphoblastic leukemia developed. This case appears to represent myelodysplastic (preleukemic) syndrome terminating in acute lymphoblastic leukemia.


Benzoyl peroxide: percutaneous penetration and metabolic disposition.
The transepidermal penetration and metabolic disposition of 14C-benzoyl peroxide were assessed in vitro (excised human skin) and in vivo (rhesus monkey). In vitro, the benzoyl peroxide penetrated into the skin, through the stratum corneum or the follicular openings, or both, and was recovered on the dermal side as benzoic acid. In vivo, benzoic acid was recovered from urine in amounts equivalent to 45% and 98% of the radiolabel following, respectively, topical and intramuscular administration of small amounts of 14C-benzoyl peroxide. We conclude that benzoyl peroxide penetrates as such into the skin layers and is converted therein to benzoic acid, which, in turn is absorbed into the systemic circulation. Renal clearance of the metabolite is sufficiently rapid as to preclude its hepatic conjugation with glycine, since following topical administration to rhesus monkeys, no hippuric acid was found in the urine, as could have been expected had a significant amount of benzoic acid passed through the liver.


Household Behavior with Respect to Meat Consumption: Differences between Households with and without Children.
Meat consumers around the world are increasingly paying attention to product quality and safety, and are starting to reduce their meat consumption, especially with regard to red meat. This trend is prevalent in households with children who prefer health-certified meat products. Our study compares meat consumption habits in households with and without children or adolescences (0-18 years). A structured questionnaire was distributed to 401 retail purchasers at 12 different points of sales of meat in the Piedmont region in northwest Italy. Socio-demographic variables and quantitative-qualitative meat consumption habits of retail purchasers were investigated. One part of the questionnaire analyzed the relative importance of 12 meat choice purchasing attributes by employing the Best-Worst scaling methodology, a type of choice experiment. Our research found that households without children (subset B) have higher weekly meat consumption habits than those with children (subset A). Alternatively, the households with children (subset A) have a diet characterized by a greater variety of protein sources, such as legumes and fish. Both of the considered subsets preferred trusted butchers for meat buying, with supermarkets as a second choice. However, only consumers of subset A bought meat from farm butchers. Our team performed a consumer analysis to identify meat consumption patterns in the two considered subsets. Simultaneously, a Best-Worst analysis evidenced several choice attributes with different relevance for the two investigated samples segmentation in three clusters of purchase.


[Microcirculatory changes in the cheek pouch mucosa of hamsters in the dynamics of experimental traumatic shock].
Acute experiments conducted on hamsters demonstrated significant disturbances of the microcirculation in the mucosa of the cheek pouch during a severe traumatic shock after a standardized mechanical injury of the hip. All the animals died in the course of the first 24 hours after this trauma. If the animals died not earlier than in one hour after the trauma the microcirculation changes were distinctly phasic in character; particularly there was seen a phase of temporary relative adaptation and stabilization of the peripheral circulation, invariably followed by the phase of decompensation, the terminal phase and death. In cases with a rapid lethal issue in the course of one hour no distinct phasic character of the microcirculation changes was observed, but there was a more or less rapid aggravation of all he indices. In difference from the majority of other investigators, no marked intravascular erythrocyte aggregation was seen by the authors in the experiments described.


Comparison of visual and automated Deki Reader interpretation of malaria rapid diagnostic tests in rural Tanzanian military health facilities.
Although microscopy is a standard diagnostic tool for malaria and the gold standard, it is infrequently used because of unavailability of laboratory facilities and the absence of skilled readers in poor resource settings. Malaria rapid diagnostic tests (RDT) are currently used instead of or as an adjunct to microscopy. However, at very low parasitaemia (usually < 100 asexual parasites/µl), the test line on malaria rapid diagnostic tests can be faint and consequently hard to visualize and this may potentially affect the interpretation of the test results. Fio Corporation (Canada), developed an automated RDT reader named Deki Reader™ for automatic analysis and interpretation of rapid diagnostic tests. This study aimed to compare visual assessment and automated Deki Reader evaluations to interpret malaria rapid diagnostic tests against microscopy. Unlike in the previous studies where expert laboratory technicians interpreted the test results visually and operated the device, in this study low cadre health care workers who have not attended any formal professional training in laboratory sciences were employed. Finger prick blood from 1293 outpatients with fever was tested for malaria using RDT and Giemsa-stained microscopy for thick and thin blood smears. Blood samples for RDTs were processed according to manufacturers' instructions automated in the Deki Reader. Results of malaria diagnoses were compared between visual and the automated devise reading of RDT and microscopy. The sensitivity of malaria rapid diagnostic test results interpreted by the Deki Reader was 94.1% and that of visual interpretation was 93.9%. The specificity of malaria rapid diagnostic test results was 71.8% and that of human interpretation was 72.0%. The positive predictive value of malaria RDT results by the Deki Reader and visual interpretation was 75.8 and 75.4%, respectively, while the negative predictive values were 92.8 and 92.4%, respectively. The accuracy of RDT as interpreted by DR and visually was 82.6 and 82.1%, respectively. There was no significant difference in performance of RDTs interpreted by either automated DR or visually by unskilled health workers. However, despite the similarities in performance parameters, the device has proven useful because it provides stepwise guidance on processing RDT, data transfer and reporting.


Effect of synthetic metalloproteinase inhibitor or citrate on neutrophil chemotaxis and the respiratory burst.
A topical synthetic metalloproteinase inhibitor (SIMP) has been reported to reduce the incidence of corneal ulcerations in the alkali-injured eye by the putative mechanism of collagenase inhibition. The current study was performed to determine whether SIMP has a direct inhibitory effect on the activation of neutrophils and to compare any such effect with that of citrate, a known inhibitor of polymorphonuclear leukocyte (PMNL) activity. The effect of SIMP or citrate was tested on chemotaxis of PMNL and the respiratory burst in vitro. Synthetic metalloproteinase inhibitor (1 mM) or citrate (12 mM) produced significant inhibition of chemotaxis of PMNL and the accompanying behavioral characteristics of motility. Synthetic metalloproteinase inhibitor, but not citrate, generated a dramatic respiratory burst when incubated with resting PMNL. Both SIMP and citrate inhibited the respiratory burst of PMNL in the presence of opsonized zymosan. For a brief, initial period each substance enhanced the respiratory burst generated by the metabolic stimulant from alkali-degraded corneas, followed by a rapid decline in activity that was associated with cell death. Although SIMP has been shown to exhibit powerful collagenase inhibition, its inhibitory effect on chemotaxis of PMNL might be the primary mechanism in reducing the incidence of ulceration in the alkali-injured cornea. Very significant reduction in the accumulation of PMNL in SIMP-treated corneas supports this conviction. Activation by SIMP of the respiratory burst in resting PMNL is of concern, but overall its beneficial effect is favorable, as demonstrated in prior alkali-injured animal models. The dual effect of inhibition of chemotaxis of PMNL and activity of collagenase makes SIMP a potential drug for combating ulceration in the alkali-injured cornea.


[Ischemic stoke in the elderly].
Stroke can affect individuals of any age but mostly occurs in elderly people. For each decade after the age of 55 years, the stroke rates doubles in both men and women. Worldwide, the aging of the population increases the importance of primary and secondary prevention interventions for stroke among the elderly group.


Phosphoinositide binding by par-3 involved in par-3 localization.
Electrostatic interactions between lipids and proteins control many cellular events. We found that phospholipids, including phosphatidylinositol 3-phosphate, phosphatidylinositol 4,5-bisphosphate, and phosphatidylinositol 3,4,5-triphosphate, bound to the C-terminal coiled-coil region of par-3 at conserved, basic residues. We identified K1013 and K1014 as the phosphoinositide binding site, because the K1013E/K1014E mutation of rat par-3 abolished its lipid binding. Importantly, the K1013E/K1014E par-3 mutant exhibited significantly weaker localization at the cell-cell junctions than the wild-type par-3. Fluorescence recovery after photo-bleaching analyses confirmed the faster turnover of mutant par-3 at cell-cell junctions. The treatment of cells with an inhibitor of phosphatidylinositol 3-kinases partially increased the turnover of par-3. These data suggested that the putative phospholipid binding by par-3 is important for its localization at cell-cell junctions.


Linagliptin improves glycemic control after 1 year as add-on therapy to basal insulin in Asian patients with type 2 diabetes mellitus.
To evaluate the efficacy and long-term safety of linagliptin added to basal insulin in Asian patients with type 2 diabetes mellitus (T2DM) inadequately controlled by basal insulin with/without oral agents. This was a post hoc analysis of Asian patients from a global ≥52 week study in which patients on basal insulin were randomized (1:1) to double-blind treatment with linagliptin 5 mg once daily or placebo (NCT00954447). Basal insulin dose remained stable for 24 weeks, after which adjustments could be made according to the investigator's discretion to improve glycemic control. The primary endpoint was the mean change in glycated hemoglobin (HbA1c) from baseline to 24 weeks. Data were available for 154 Asian patients (80 linagliptin, 74 placebo). Baseline HbA1c (standard deviation [SD]) was 8.6 (0.9)% (70 [10] mmol/mol). The placebo-corrected mean change (standard error [SE]) in HbA1c from baseline was -0.9 (0.1)% (-10 [1] mmol/mol) (95% confidence interval [CI]: -1.2, -0.7; p<0.0001) at Week 24 and -0.9 (0.1)% (-10 [1] mmol/mol) (95% CI: -1.1, -0.6; p<0.0001) at Week 52. The frequency of adverse events (linagliptin 81.3%, placebo 91.9%) and hypoglycemia (Week 24: linagliptin 25.0%, placebo 25.7%; treatment end: linagliptin 28.8%, placebo 35.1%) was similar between groups. By Week 52, changes (SE) in mean body weight were similar in both groups (linagliptin -0.67 [0.26] kg, placebo -0.38 [0.25] kg). This study was limited by the post hoc nature of the analysis and the small number of patients in the subgroup. However, the results suggest that linagliptin significantly improves glycemic control in Asian patients with T2DM inadequately controlled by basal insulin, without increasing the risk for hypoglycemia or weight gain. ClinicalTrials identifier: NCT00954447.


Circulating macrophage migration inhibitory factor levels and its polymorphisms in systemic lupus erythematosus: A meta-analysis.
The present study aimed to systemically review the evidence regarding the relationship between circulating macrophage migration inhibitory factor (MIF) levels and systemic lupus erythematosus (SLE), as well as the associations between several polymorphisms in the MIF gene and SLE susceptibility. We performed a meta-analysis of serum/plasma levels of MIF in SLE patients and controls and evaluated evidence of associations between the MIF -173 C/G allele and -794CATT5-8 polymorphisms and the associated risk for SLE. Nine studies were included in this meta-analysis. Meta-analysis indicated that MIF levels were significantly higher in the SLE group than in the control group (SMD = 1.154, 95% CI = 0.369-1.938, P = 0.004). Stratification by ethnicity showed significantly higher MIF levels in the SLE group representing Asian populations (SMD = 1.911, 95% CI = 0.871-2.951, P < 0.001). MIF levels were significantly higher in the SLE group than in the control group in the age-and/or sex matched population, but not in the unmatched population (SMD = 1.236, 95% CI = 0.579-1.893, P < 0.001; SMD = 1.118, 95% CI = -0.027-2.263, P = 0.056). However, results of the meta-analysis showed no association between SLE and the MIF -173 C allele, the -794CATT7 allele, and the -794CATT7-MIF-173C haplotype with high heterogeneity. Our meta-analysis demonstrated significantly higher circulating MIF levels in patients with SLE, but no evidence of associations between MIF -173 C/G and -794CATT5-8 polymorphisms and SLE susceptibility.


[The diagnostic possibilities of saliva].
Saliva is a clinically informative biological fluid which contains multitude of bio-markers. This characteristic makes it possible to carry out numerous analyzes for developing mode to test patient in situ, express-tests included. The diagnostic by saliva is a new area of more simple application both markers and analyzers that can be useful in diagnostic of diseases of oral cavity, oncological diseases included. The using of saliva expands perspectives for making clinical diagnosis and establishment of dynamics and monitoring of disease.


Aggressive hemangioma starting from l5 body, treated with mass embolization and radiation therapy with protons: a case report.
A hemangioma is a benign tumor of endothelial cells, multiplying into the medullary spaces of the cancellous bone. This tumor is in most cases not symptomatic, and in this case, they do not require any specific treatment. Pain and neurological symptoms derived from hemangiomas as shown in 2-11% of the cases. The 55% of these cases presents low back pain, while the 45% presents neurological deficit from compression of the spinal cord, peripheral nerves or both. We present a clinical case report of a young woman, affected by aggressive L5 hemangioma causing a spinal canal stenosis with associated sciatalgic symptoms. We performed a review of the current literature on the treatment options, giving the rationale of our treatment choice (mass embolization and radiation therapy with protons).


An intronic PICALM polymorphism, rs588076, is associated with allelic expression of a PICALM isoform.
Although genome wide studies have associated single nucleotide polymorphisms (SNP)s near PICALM with Alzheimer's disease (AD), the mechanism underlying this association is unclear. PICALM is involved in clathrin-mediated endocytosis and modulates Aß clearance in vitro. Comparing allelic expression provides the means to detect cis-acting regulatory polymorphisms. Thus, we evaluated whether PICALM showed allele expression imbalance (AEI) and whether this imbalance was associated with the AD-associated polymorphism, rs3851179. We measured PICALM allelic expression in 42 human brain samples by using next-generation sequencing. Overall, PICALM demonstrated equal allelic expression with no detectable influence by rs3851179. A single sample demonstrated robust global PICALM allelic expression imbalance (AEI), i.e., each of the measured isoforms showed AEI. Moreover, the PICALM isoform lacking exons 18 and 19 (D18-19 PICALM) showed significant AEI in a subset of individuals. Sequencing these individuals and subsequent genotyping revealed that rs588076, located in PICALM intron 17, was robustly associated with this imbalance in D18-19 PICALM allelic expression (p = 9.54 x 10-5). This polymorphism has been associated previously with systolic blood pressure response to calcium channel blocking agents. To evaluate whether this polymorphism was associated with AD, we genotyped 3269 individuals and found that rs588076 was modestly associated with AD. However, when both the primary AD SNP rs3851179 was added to the logistic regression model, only rs3851179 was significantly associated with AD. PICALM expression shows no evidence of AEI associated with rs3851179. Robust global AEI was detected in one sample, suggesting the existence of a rare SNP that strongly modulates PICALM expression. AEI was detected for the D18-19 PICALM isoform, and rs588076 was associated with this AEI pattern. Conditional on rs3851179, rs588076 was not associated with AD risk, suggesting that D18-19 PICALM is not critical in AD. In summary, this analysis of PICALM allelic expression provides novel insights into the genetics of PICALM expression and AD risk.


Acute psychostimulant challenge in primary obsessive-compulsive disorder.
The effects of acute oral administration of methylphenidate 40 mg versus dextroamphetamine 30 mg versus matched placebo were compared in 11 patients with primary obsessive-compulsive disorder. Dextroamphetamine but not methylphenidate had a significantly greater antiobsessive-compulsive effect as measured by the Comprehensive Psychiatric Rating Scale--Obsessive-Compulsive Subscale, as compared with placebo. This effect appeared unrelated to their effect on depression although a differential effect of the two psychostimulants on anxiety was observed. Although both these stimulants affect serotonin, the differences noted between dextroamphetamine and methylphenidate suggest that catecholamines may be implicated in the pathophysiology of obsessive-compulsive disorder.


[Characteristics of the structural organization of the phylogenetically new fields in the human neocortex].
The myeloarchitectonics of the specific human fields 40, 39, 37c, 37b of neocortex and also fields 17 and 19 which are the cortex end of visual analyzer has been studied with the help of neurohistological and quantitative methods. According to the character of the fibrous bundles, their diameter, the quantity of the fibrous in them and the diameter of the fibrous gradual changes from field 17 to field 19 and further to field 39 have been shown. It is an evidence of the succession between phylogenetically older and younger fields of the cortex. The peculiarities of the field quantitative myeloarchitectonics differing in the function and genesis are marked by heterochronous growth of these fields square in the process of evolution.


Minimal hepatic changes in rats fed alcohol and a high casein diet.
This study was designed to determine whether prolonged ingestion of alcohol in combination with a nutritious diet can produce hepatic cirrhosis in the rat. Male Wistar-strain rats were fed a high protein liquid diet in which alcohol comprised 35.5% of total calories. Rats maintained on this diet for 19 months had normal growth. There was no evidence of cirrhosis after 19 months. Histological examination showed no hepatocellular necrosis, inflammation, or increased connective tissue. Ultrastructural study showed no substantial changes, except for occasional mitochondrial enlargement in cells containing increased lipid. Biochemical analyses at 19 months showed moderate increase in hepatic triglyceride levels and slight increase in hepatic collagen content in the experimental groups.


Therapeutic strategies and outcomes of MDR and pre-XDR-TB in Italy: a nationwide study.
BACKGROUND: Treatment outcomes in multidrug-resistant TB (MDR-TB) patients are suboptimal in several low-incidence countries.METHODS: The primary outcome measure was the proportion of successfully treated patients in Italy during an 18-year period. Secondary outcomes were treatment outcomes in certain drug-containing regimens and the possibility for the WHO shorter MDR-TB regimen.RESULTS: In the 191 patients included (median age at admission: 33 years; 67.5% male, following drug-resistance patterns were found: MDR-TB in 68.6%, pre-extensively drug-resistant TB (pre-XDR-TB) in 30.4% and XDR-TB in 1.1% patients. The most frequently prescribed drugs were fluoroquinolones in 84.6% cases, amikacin in 48.7%, linezolid in 34.6% and meropenem/clavulanic acid in 29.5%. The median duration of treatment was 18 months. Treatment success was achieved in 71.2% patients, of whom, 44% were cured and 27.2% completed treatment. Treatment success rates did not statistically differ between the MDR- (68.8%) and pre-XDR-TB (77.6%) groups (P = 0.26). Treatment success rates had large variability between North and South of Italy (81.3% vs. 53.3%). Only 22.5% of the cases would have been eligible for shorter MDR-TB regimensCONCLUSION: Our study highlights variability in treatment outcomes in MDR- and pre-XDR-TB patients. Study findings confirmed the potential utility of linezolid and, for patients with limited oral options, meropenem/clavulanic acid and amikacin.


Geometric control of capillary architecture via cell-matrix mechanical interactions.
Capillary morphogenesis is a multistage, multicellular activity that plays a pivotal role in various developmental and pathological situations. In-depth understanding of the regulatory mechanism along with the capability of controlling the morphogenic process will have direct implications on tissue engineering and therapeutic angiogenesis. Extensive research has been devoted to elucidate the biochemical factors that regulate capillary morphogenesis. The roles of geometric confinement and cell-matrix mechanical interactions on the capillary architecture, nevertheless, remain largely unknown. Here, we show geometric control of endothelial network topology by creating physical confinements with microfabricated fences and wells. Decreasing the thickness of the matrix also results in comparable modulation of the network architecture, supporting the boundary effect is mediated mechanically. The regulatory role of cell-matrix mechanical interaction on the network topology is further supported by alternating the matrix stiffness by a cell-inert PEG-dextran hydrogel. Furthermore, reducing the cell traction force with a Rho-associated protein kinase inhibitor diminishes the boundary effect. Computational biomechanical analysis delineates the relationship between geometric confinement and cell-matrix mechanical interaction. Collectively, these results reveal a mechanoregulation scheme of endothelial cells to regulate the capillary network architecture via cell-matrix mechanical interactions.


A role for the metal binding domain in determining the DNA sequence selectivity of Fe-bleomycin.
Previous studies of Fe-bleomycin-mediated DNA cleavage have established that the bithiazole moiety + C-terminal substituent of bleomycin are required for DNA binding, while the metal binding domain is responsible for O2 activation. Although recent studies have indicated that the metal binding domain also participates in DNA unwinding, and in determining the sequence and strand selectivity of DNA cleavage, no study has defined the structural domain that bears primary responsibility for the observed pattern of bleomycin-mediated DNA degradation. Presently, by the use of four synthetic analogs of bleomycin demethyl A2 having the functional domains connected by rigid spacers of varying lengths, the source of DNA cleavage specificity has been determined. When the four analogs cleaved 242- and 127-base pair 5'-32P-end-labeled DNA restriction fragments containing isolated Fe-bleomycin cleavage sites, all four produced cleavage at the same preferred sites. Because the (oligo)glycine spacers altered the distance between the domains by as much as 14 A, the identical cleavage patterns argue that the primary determinant of sequence specificity for these analogs is the metal binding domain.


Endocrine profile and neuroendocrine challenge tests in transgenic mice expressing antisense RNA against the glucocorticoid receptor.
A transgene expressing antisense RNA complementary to a fragment of the glucocorticoid receptor cDNA was incorporated into the mouse genome and resulted in a transgenic animal that has decreased glucocorticoid receptor function. The transgenic mice showed basal plasma ACTH and corticosterone levels similar to those of the normal control animals. We have further investigated changes in HPA axis regulation by use of different neuroendocrine challenge tests including a dexamethasone suppression test (DST). In comparison to normal mice, a tenfold higher dose of dexamethasone (i.e. 20 micrograms/100 g body weight) was required to suppress the basal corticosterone levels of transgenic mice. Dexamethasone (2 micrograms/100 g body weight) produced a long-lasting suppression of plasma ACTH and corticosterone levels in control mice, whereas in transgenic animals only a short-lasting decrease in ACTH levels was apparent. Corticotropin-releasing hormone (CRH) administration resulted in an enhanced response in plasma ACTH levels in transgenic mice, whereas the corticosterone response was markedly reduced. The discrepancy between ACTH and corresponding corticosterone secretions in transgenic mice could be attributed, in part, to a reduced sensitivity of the adrenal gland to stimulation by ACTH. Pituitaries of transgenic mice contained about 50% less proopiomelanocortin (POMC) mRNA than those of control animals. No significant differences were noted in the ACTH or protein contents of normal and transgenic mice pituitary glands although a slight increase in protein content of the transgenic mouse adrenal gland was apparent. In conclusion, transgenic mice with impaired GR function show major disturbances in HPA axis regulation which seem to be caused by the primary defect in conjunction with secondary modifications in, amongst others, pituitary CRH receptor system(s), sympathetic output and adrenal development. This mouse is therefore a useful model to study the consequences of life-long defective GR function and HPA axis regulation in general.


Bedside Clinician's Guide to Pulmonary Artery Catheters.
Pulmonary artery catheters provide important information about cardiac function, mixed venous oxygenation, and right-sided pressures and potentially provide temporary pacing ability. To provide bedside clinicians with guidance for techniques to insert right heart monitors and devices, describe risk factors for difficult insertion and contraindications to placement, and provide updates on new technologies that may be encountered in the intensive care unit. An extensive literature review was performed. Experienced clinicians were asked to identify topics not addressed in the literature. Advanced imaging techniques such as transesophageal echocardiography or fluoroscopy can supplement traditional pressure waveform-guided insertion when needed, and several other techniques can be used to facilitate passage into the pulmonary artery. Caution is warranted when attempting insertion in patients with right-sided masses or preexisting conduction abnormalities. New technologies include a pacing catheter that anchors to the right ventricle and a remote monitoring device that is implanted in the pulmonary artery. Bedside clinicians should be aware of risk factors such as atrial fibrillation with dilated atria, decreased ventricular function, pulmonary hypertension, and right-sided structural abnormalities that can make pulmonary artery catheter insertion challenging. Clinicians should be familiar with advanced techniques and imaging options to facilitate placement. The overall risk of serious complications with right heart catheter placement and manipulation is low and often outweighed by its benefits, specifically pressure monitoring and pacing.


Zonisamide attenuates MPP+-induced oxidative toxicity through modulation of Ca2+ signaling and caspase-3 activity in neuronal PC12 cells.
Parkinson's disease is an incurable progressive neurological condition caused by a degeneration of dopamine-producing cells characterized by motor and non-motor symptoms. The major mechanisms of the antiepileptic actions of ZNS are inhibition of voltage-gated Na(+) channel, T-type voltage-sensitive Ca(2+) channel, Ca(2+)-induced Ca(2+) releasing system, and neuronal depolarization-induced glutamate release; and enhancement of release of inhibitory neurotransmitters; however, the detailed mechanism of antiparkinsonian effects of ZNS remains to be clarified. We aimed to investigate to the effect of ZNS on the oxidative stress, cell viability, Ca(2+) signaling, and caspase activity that induced by the MPP(+) model of Parkinson's in neuronal PC12 cells. Neuronal PC12 cells were divided into four groups namely, control, ZNS, MPP(+), and ZNS+MPP(+) groups. The dose and duration of ZNS and MPP(+) were determined according to cell viability (MTT) analysis which used to assess the cell viability. The cells in ZNS, MPP(+), and ZNS+MPP(+) groups were incubated for 5 h with 100 μM ZNS, 10 h with 100 μM MPP(+), and 10 h with ZNS and MPP(+), respectively. Lipid peroxidation and cytosolic free Ca(2+) concentrations were higher in the MPP(+) group than in control although their levels were lower in ZNS and the ZNS+MPP(+) groups than in control. Reduced glutathione and glutathione peroxidase values were lower in the MPP(+) group although they were higher in the ZNS and the ZNS+MPP(+) groups than in control. Caspase-3 activity was lower in the ZNS group than in the MPP(+) group. In conclusion, ZNS induced modulator effects on the oxidative stress, intracellular Ca(2+), and the caspase-3 values in an experimental model of Parkinson disease.


Inhibitory cerebello-olivary projections and blocking effect in classical conditioning.
The behavioral phenomenon of blocking indicates that the informational relationship between the conditioned stimulus and the unconditioned stimulus is essential in classical conditioning. The eyeblink conditioning paradigm is used to describe a neural mechanism that mediates blocking. Disrupting inhibition of the inferior olive, a structure that conveys unconditioned stimulus information (airpuff) to the cerebellum prevented blocking in rabbits. Recordings of cerebellar neuronal activity show that the inferior olive input to the cerebellum becomes suppressed as learning occurs. These results suggest that the inferior olive becomes functionally inhibited by the cerebellum during conditioning, and that this negative feedback process might be the neural mechanism mediating blocking.


Aspects of electrolyte transport across isolated dog retinal pigment epithelium.
Transport of Na and Cl across the isolated dog retinal pigment epithelium (RPE) choroid was investigated. Under the short-circuit condition, a net Na flux was observed from choroid to retina and a net Cl flux was determined in the opposite direction. The current created by the net flux of these two ions was larger than the short-circuit current (SCC). Addition of 10(-5) M ouabain to the apical side inhibited net fluxes of both Na and Cl, whereas it reduced the SCC 84%. Addition of 10(-4) M furosemide to the apical side inhibited net Cl flux but had no effect on the net Na transport. The 10(-4) M furosemide reduced the SCC 38%. These drugs had no effect when applied to the basal side. Thus the transport of both Na and Cl depends on the Na-K-ATPase in the apical membrane of the dog RPE. A furosemide-sensitive neutral carrier at the apical membrane is suggested for the transport of Cl. Replacement of HCO3 with SO4 in the bathing solution caused an increase in the SCC, indicating the choroid-to-retina movement of HCO3 across the short-circuited dog RPE choroid.


APP accumulates with presynaptic proteins around amyloid plaques: A role for presynaptic mechanisms in Alzheimer's disease?
In Alzheimer's disease (AD), the distribution of the amyloid precursor protein (APP) and its fragments other than amyloid beta, has not been fully characterized. Here, we investigate the distribution of APP and its fragments in human AD brain samples and in mouse models of AD in reference to its proteases, synaptic proteins, and histopathological features characteristic of the AD brain, by combining an extensive set of histological and analytical tools. We report that the prominent somatic distribution of APP observed in control patients remarkably vanishes in human AD patients to the benefit of dense accumulations of extra-somatic APP, which surround dense-core amyloid plaques enriched in APP-Nter. These features are accentuated in patients with familial forms of the disease. Importantly, APP accumulations are enriched in phosphorylated tau and presynaptic proteins whereas they are depleted of post-synaptic proteins suggesting that the extra-somatic accumulations of APP are of presynaptic origin. Ultrastructural analyses unveil that APP concentrates in autophagosomes and in multivesicular bodies together with presynaptic vesicle proteins. Altogether, alteration of APP distribution and its accumulation together with presynaptic proteins around dense-core amyloid plaques is a key histopathological feature in AD, lending support to the notion that presynaptic failure is a strong physiopathological component of AD.


[Data bank on post-stroke patients: experience in organization and use].
The bank of clinical data concerning restorative therapy of neurological patients was characterized. These data were included in computer program "System of management of data bank on poststroke patients" which was elaborated in the department of V.M.Bekhterev's St-Petersburg Psychoneurological Institute. Information obtained in process of answering special questionnaire formed this data bank which included social-, demographic, anamnestic, clinical-anatomical data, information about both initial functional state of the patient and its dynamics as well as therapeutic methods applied and follow-up data about the patients which entered the hospital repeatedly, 450 case reports were accumulated in the bank. Practice proved informative value of the bank's materials and its suitability for statistic analysis. Data bank may be used for scientific and statistic purposes as well for control and prognosis of efficiency of treatment of the patients with consequences of cerebral strokes.


Dengue Virus Infection of Human Retinal Müller Glial Cells.
Retinopathy is a recently recognized complication of dengue, affecting up to 10% of hospitalized patients. Research on the pathogenesis has focused largely on effects of dengue virus (DENV) at the blood-retinal barrier. Involvement of retinal Müller glial cells has received little attention, although this cell population contributes to the pathology of other intraocular infections. The goal of our work was to establish the susceptibility of Müller cells to infection with DENV and to identify characteristics of the cellular antiviral, inflammatory, and immunomodulatory responses to DENV infection in vitro. Primary human Müller cell isolates and the MIO-M1 human Müller cell line were infected with the laboratory-adapted Mon601 strain and DENV serotype 1 and 2 field isolates, and cell-DENV interactions were investigated by immunolabelling and quantitative real-time polymerase chain reaction. Müller cells were susceptible to DENV infection, but experiments involving primary cell isolates indicated inter-individual variation. Viral infection induced an inflammatory response (including tumour necrosis factor-α, interleukin [IL]-1β, and IL-6) and an immunomodulatory response (including programmed death-ligand [PD-L]1 and PD-L2). The type I interferon response was muted in the Müller cell line compared to primary cell isolates. The highest infectivity and cell responses were observed in the laboratory-adapted strain, and overall, infectivity and cell responses were stronger in DENV2 strains. This work demonstrates that Müller cells mount an antiviral and immune response to DENV infection, and that this response varies across cell isolates and DENV strain. The research provides a direction for future efforts to understand the role of human retinal Müller glial cells in dengue retinopathy.


Regulation of Hippo signaling by Jun kinase signaling during compensatory cell proliferation and regeneration, and in neoplastic tumors.
When cells undergo apoptosis, they can stimulate the proliferation of nearby cells, a process referred to as compensatory cell proliferation. The stimulation of proliferation in response to tissue damage or removal is also central to epimorphic regeneration. The Hippo signaling pathway has emerged as an important regulator of growth during normal development and oncogenesis from Drosophila to humans. Here we show that induction of apoptosis in the Drosophila wing imaginal disc stimulates activation of the Hippo pathway transcription factor Yorkie in surviving and nearby cells, and that Yorkie is required for the ability of the wing to regenerate after genetic ablation of the wing primordia. Induction of apoptosis activates Yorkie through the Jun kinase pathway, and direct activation of Jun kinase signaling also promotes Yorkie activation in the wing disc. We also show that depletion of neoplastic tumor suppressor genes, including lethal giant larvae and discs large, or activation of aPKC, activates Yorkie through Jun kinase signaling, and that Jun kinase activation is necessary, but not sufficient, for the disruption of apical-basal polarity associated with loss of lethal giant larvae. Our observations identify Jnk signaling as a modulator of Hippo pathway activity in wing imaginal discs, and implicate Yorkie activation in compensatory cell proliferation and disc regeneration.


Effects of host sociality on ectoparasite population biology.
Theory predicts a positive relationship between parasite infection intensity and host density. However, this generalization is complicated in natural systems by differences in life history among parasite taxa, e.g., transmissibility. Accordingly, predictions relating host density to parasite load should be specific to each parasite taxon. To illustrate this, we studied parasites that differed greatly in life history in the context of the Galapagos hawk's (Buteo galapagoensis) variably cooperative mating system. Two louse (Phthiraptera) species were collected: Colpocephalum turbinatum (Amblycera), with 53 host species, and Degeeriella regalis (Ischnocera), with 10 host species, although B. galapagoensis was the only known Galapagos host. Sixty territorial adult male hawks from 26 groups of 1-6 males were quantitatively sampled for lice. Average abundance and intensity of C. turbinatum but not D. regalis were significantly larger in large groups of hawks than small groups. Males from the same polyandrous group harbored significantly correlated abundances of C. turbinatum but not D. regalis. Prevalence, average abundance, and intensity of C. turbinatum were significantly higher than D. regalis. These are the first results to demonstrate significant differences in a suite of population responses between these louse suborders in the context of host sociality.


Chirped-Pulse millimeter-Wave spectroscopy for dynamics and kinetics studies of pyrolysis reactions.
A Chirped-Pulse millimeter-Wave (CPmmW) spectrometer is applied to the study of chemical reaction products that result from pyrolysis in a Chen nozzle heated to 1000-1800 K. Millimeter-wave rotational spectroscopy unambiguously determines, for each polar reaction product, the species, the conformers, relative concentrations, conversion percentage from precursor to each product, and, in some cases, vibrational state population distributions. A chirped-pulse spectrometer can, within the frequency range of a single chirp, sample spectral regions of up to ∼10 GHz and simultaneously detect many reaction products. Here we introduce a modification to the CPmmW technique in which multiple chirps of different spectral content are applied to a molecular beam pulse that contains the pyrolysis reaction products. This technique allows for controlled allocation of its sensitivity to specific molecular transitions and effectively doubles the bandwidth of the spectrometer. As an example, the pyrolysis reaction of ethyl nitrite, CH3CH2ONO, is studied, and CH3CHO, H2CO, and HNO products are simultaneously observed and quantified, exploiting the multi-chirp CPmmW technique. Rotational and vibrational temperatures of some product molecules are determined. Subsequent to supersonic expansion from the heated nozzle, acetaldehyde molecules display a rotational temperature of 4 ± 1 K. Vibrational temperatures are found to be controlled by the collisional cooling in the expansion, and to be both species- and vibrational mode-dependent. Rotational transitions of vibrationally excited formaldehyde in levels ν4, 2ν4, 3ν4, ν2, ν3, and ν6 are observed and effective vibrational temperatures for modes 2, 3, 4, and 6 are determined and discussed.


Association between use of antidepressants or benzodiazepines and the risk of subsequent fracture among those aged 65+ in the Netherlands.
This is the first study to examine the association between antidepressant and benzodiazepine use following a MOF and risk of subsequent fracture in those 65+. Using national data, drug use following MOF showed that the 1-year fully adjusted risk of subsequent MOF in those on antidepressants was more than doubled. We evaluated the association between the use of antidepressants or benzodiazepines and the risk of a subsequent major osteoporotic fracture. A cohort study was performed using the Dutch PHARMO Database Network. Between 2002 and 2011, a total of 4854 patients sustained a first major osteoporotic fracture after the age of 65 years, of which 1766 sustained a hip fracture. Incidence rates and adjusted hazard ratios were calculated using Cox proportional hazards models. Within 1 year following a major osteoporotic fracture, 15% (95% CI 13.7-15.7) and 31% (95% CI 30.1-32.8) of patients were dispensed an antidepressant or benzodiazepine, respectively. Current use of antidepressants in the first year following a major osteoporotic fracture was associated with subsequent fracture (adjusted HR 2.17 (95% CI 1.37-3.43)). Recent and past use of antidepressants were also associated with an increased risk of subsequent fracture. When the complete follow-up period was included, only the current use of antidepressants was associated with subsequent fracture following a major osteoporotic fracture (adjusted HR 1.48; 95% CI 1.06-2.06). Current benzodiazepine use was not associated with an increased risk of fracture within 1 year following a major osteoporotic fracture (adjusted HR 1.18; 95% CI 0.76-1.81) or during the complete follow-up period (adjusted HR 1.18; 95% CI 0.90-1.55). This study provides evidence that antidepressants should be used with caution following a major osteoporotic fracture. It provides needed insights that can be used to inform clinicians when assessing subsequent fracture risk in patients.


Ascending projections from the area around the spinal cord central canal: A Phaseolus vulgaris leucoagglutinin study in rats.
A single small iontophoretic injection of Phaseolus vulgaris leucoagglutinin labels projections from the area surrounding the spinal cord central canal at midthoracic (T6-T9) or lumbosacral (L6-S1) segments of the spinal cord. The projections from the midthoracic or lumbosacral level of the medial spinal cord are found: 1) ascending ipsilaterally in the dorsal column near the dorsal intermediate septum or the midline of the gracile fasciculus, respectively; 2) terminating primarily in the dorsal, lateral rim of the gracile nucleus and the medial rim of the cuneate nucleus or the dorsomedial rim of the gracile nucleus, respectively; and 3) ascending bilaterally with slight contralateral predominance in the ventrolateral quadrant of the spinal cord and terminating in the ventral and medial medullary reticular formation. Other less dense projections are to the pons, midbrain, thalamus, hypothalamus, and other forebrain structures. Projections arising from the lumbosacral level are also found in Barrington's nucleus. The results of the present study support previous retrograde tract tracing and physiological studies from our group demonstrating that the neurons in the area adjacent to the central canal of the midthoracic or lumbosacral level of the spinal cord send long ascending projections to the dorsal column nucleus that are important in the transmission of second-order afferent information for visceral nociception. Thus, the axonal projections through both the dorsal and the ventrolateral white matter from the CC region terminate in many regions of the brain providing spinal input for sensory integration, autonomic regulation, motor and emotional responses, and limbic activation.


Relations between Structure and Zn(II) Binding Affinity Shed Light on the Mechanisms of Rad50 Hook Domain Functioning and Its Phosphorylation.
The metal binding at protein-protein interfaces is still uncharted territory in intermolecular interactions. To date, only a few protein complexes binding Zn(II) in an intermolecular manner have been deeply investigated. The most notable example of such interfaces is located in the highly conserved Rad50 protein, part of the Mre11-Rad50-Nbs1 (MRN) complex, where Zn(II) is required for homodimerization (Zn(Rad50)2). The high stability of Zn(Rad50)2 is conserved not only for the protein derived from the thermophilic archaeon Pyrococcus furiosus (logK12 = 20.95 for 130-amino-acid-long fragment), which was the first one studied, but also for the human paralog studied here (logK12 = 19.52 for a 183-amino-acid-long fragment). As we reported previously, the extremely high stability results from the metal-coupled folding process where particular Rad50 protein fragments play a critical role. The sequence-structure-stability analysis based on human Rad50 presented here separates the individual structural components that increase the stability of the complex, pointing to amino acid residues far away from the Zn(II) binding site as being largely responsible for the complex stabilization. The influence of the individual components is very well reflected by the previously published crystal structure of the human Rad50 zinc hook (PDB: 5GOX). In addition, we hereby report the effect of phosphorylation of the zinc hook domain, which exerts a destabilizing effect on the domain. This study identifies factors governing the stability of metal-mediated protein-protein interactions and illuminates their molecular basis.


Enhanced neurogenesis during trimethyltin-induced neurodegeneration in the hippocampus of the adult rat.
The occurrence of neurogenesis in the hippocampus of the adult rat during trimethyltin (TMT)-induced neurodegeneration was investigated using bromodeoxyuridine (BrdU). Fifteen days after TMT intoxication, BrdU-labeled cells were significantly more numerous in the hippocampus of treated animals, gradually decreasing towards the control value 21 days after intoxication in the dentate gyrus (DG), while in the CA3/hilus region BrdU-labeled cells were still more numerous in TMT-treated rats. In order to investigate the fate of newly-generated cells double labeling experiments using neuronal or glial markers were performed. Colocalization of the neuronal marker NeuN was detected in many BrdU-positive cells in the DG, while in the CA3/hilus region no colocalization of NeuN and BrdU could be observed. No colocalization of BrdU and the astroglial marker GFAP or the microglial marker OX-42 was detected either in the DG and or in the CA3/hilus region. The results indicate an enhancement of endogenous neurogenesis in the hippocampus during TMT-induced neurodegeneration, with the development of a subpopulation of regenerated cells into neurons in the DG, while in the CA3/hilus region the population of newly-generated cells should be regarded as undifferentiated.


Distribution of Pasteurella haemolytica and Pasteurella multocida in the bovine lung following vaccination and challenge exposure as an indicator of lung resistance.
Experimental calves were vaccinated with virulent strains of Pasteurella haemolytica or Pasteurella multocida or with phosphate-buffered saline solution either by an aerosol method or by subcutaneous injection. Calves were subsequently challenge exposed by intrapulmonic inoculation of the homologous virulent Pasteurella species. Sections obtained from the resulting pulmonic lesion were stained, using a fluorescent antibody technique, to determine relative number, location, and integrity of the challenge organism. The resistance of the calf to challenge exposure, as determined by other factors, was compared with the capacity of the components of the lung to engulf or destroy pasteurellae. Calves vaccinated with an aerosol of the bacterium were most resistant to challenge exposure; most bacteria were engulfed or degraded by the phagocytic cells. Vaccination by subcutaneous injection was less effective in inducing resistance. Tissue sections from these calves contained many more extracellular intact bacteria and fewer intracellular intact or degraded bacteria than were seen in the sections of calves vaccinated by the aerosol method. The control calves were the least resistant; bacteria seen in tissue sections from these calves were numerous, predominantly extracellular, and intact. A group of nonvaccinated calves experimentally inoculated with infectious bovine rhinotracheitis virus 5 days before intrapulmonic challenge exposure with P haemolytica developed severe pulmonic lesions. The lesions were larger and more invasive and contained many more extracellular bacteria than did the lungs of calves in control groups. As in other nonvaccinated calves, there were few intracellular bacteria; however, unlike in other calves, the extracellular bacteria were seen in large numbers, particularly in alveolar lumens.


IFNL4 rs368234815 and rs117648444 variants predict off-treatment HBsAg seroclearance in IFN-treated HBeAg-negative chronic hepatitis B patients.
Robust baseline predictors of interferon (IFN) response in HBeAg-negative chronic hepatitis B (CHB) patients are not currently available. The recently described rs368234815 TT/ΔG dinucleotide and rs117648444 nonsynonymous P70S polymorphisms in IFN lambda 4 (IFNL4) gene, which are strongly associated with response to IFN in hepatitis C virus (HCV) infection, could be also useful in IFN-treated CHB patients. Here we assessed whether IFNL4 rs368234815 and rs117648444 polymorphisms predict IFN-induced HBsAg clearance in CHB patients. We sequenced the IFNL4 gene on genomic DNA collected from 126 HBeAg-negative CHB patients treated with IFN and followed up for a median of 11 (1-23) years. The 15-year cumulative probability of HBsAg loss in the 62 carriers of the rs368234815 TT/TT genotype, which abolishes the IFNλ4 protein production, was comparable to that of 19 patients carrying the rs117648444 T allele predicted to produce an impaired IFNλ4-S70 protein (39% vs 42%, P = .827). In contrast, these 81 patients, either not producing IFNλ4 or producing an impaired IFNλ4-S70 protein, had a significantly higher 15-year probability of HBsAg loss compared to the 45 subjects predicted to encode only the fully functional IFNλ4-P70 (42% vs 11% P = .003). At multivariate analysis, combination of the rs368234815 and rs117648444 genotypes strongly predicted HBsAg clearance (HR 5.90, 95% CI 1.70-20.9, P = .006) together with pretreatment serum HBV DNA levels (HR 0.57, 95% CI 0.39-0.83, P = .003). IFNL4 rs368234815 and rs117648444 functional variants are worth to be investigated as pretreatment combined predictors of IFN response in HBeAg-negative CHB patients.


New and improved proteomics technologies for understanding complex biological systems: addressing a grand challenge in the life sciences.
This White Paper sets out a Life Sciences Grand Challenge for Proteomics Technologies to enhance our understanding of complex biological systems, link genomes with phenotypes, and bring broad benefits to the biosciences and the US economy. The paper is based on a workshop hosted by the National Institute of Standards and Technology (NIST) in Gaithersburg, MD, 14-15 February 2011, with participants from many federal R&D agencies and research communities, under the aegis of the US National Science and Technology Council (NSTC). Opportunities are identified for a coordinated R&D effort to achieve major technology-based goals and address societal challenges in health, agriculture, nutrition, energy, environment, national security, and economic development.


Characterization of plant sulfiredoxin and role of sulphinic form of 2-Cys peroxiredoxin.
The antioxidant function of 2-Cys peroxiredoxin (Prx) involves the oxidation of its conserved peroxidatic cysteine to sulphenic acid that is recycled by a reductor agent. In conditions of oxidative stress, the peroxidatic cysteine can be overoxidized to sulphinic acid inactivating the Prx. An enzyme recently discovered, named sulfiredoxin (Srx), reduces the sulphinic 2-Cys Prx (Prx-SO(2)H). To explore the physiological functions of Srx in plants we have cloned, expressed and purified to homogeneity a Srx from Arabidopsis thaliana (AtSrx), as well as five variants by site-directed mutagenesis on amino acids involved in its activity. The activity of sulfiredoxin, determined by a new method, is dependent on the concentration of the sulphinic form of Prx and the conserved Srx is capable of regenerating the functionality of both pea and Arabidopsis Prx-SO(2)H. Molecular modelling of AtSrx and the facts that the R28Q variant shows a partial inactivation, that the activity of the E76A variant is equivalent to that of the native enzyme and that the double mutation R28Q/E76A abolishes the enzymatic activity suggests that the pair His100-Glu76 may be involved in the activation of C72 in the absence of R28. The knock-out mutant plants without Srx or 2-Cys Prx exhibited phenotypical differences under growth conditions of 16 h light, probably due to the signalling role of the sulphinic form of Prx. These mutants showed more susceptibility to oxidative stress than wild-type plants. This work presents the first systematic biochemical characterization of the Srx/Prx system from plants and contributes to a better understanding of its physiological function.


Nutrient cross-feeding in the microbial world.
The stability and function of a microbial community depends on nutritional interactions among community members such as the cross-feeding of essential small molecules synthesized by a subset of the population. In this review, we describe examples of microbe-microbe and microbe-host cofactor cross-feeding, a type of interaction that influences the forms of metabolism carried out within a community. Cofactor cross-feeding can contribute to both the health and nutrition of a host organism, the virulence and persistence of pathogens, and the composition and function of environmental communities. By examining the impact of shared cofactors on microbes from pure culture to natural communities, we stand to gain a better understanding of the interactions that link microbes together, which may ultimately be a key to developing strategies for manipulating microbial communities with human health, agricultural, and environmental implications.


Wisconsin card sorting test in children with temporal lobe epilepsy.
To search for the origin of frontal lobe dysfunction identified by the Wisconsin card sorting test (WCST) in temporal lobe epilepsy (TLE) patients, we investigated the WCST performance among 19 children with TLE (with hippocampal atrophy (HA group N=12), without structural lesions (NSL group N=7)), 15 patients with frontal lobe epilepsy (FLE group), and age-matched normal controls (N group). The paired verbal association learning test (PVALT) and Benton visual retention test (BVRT) were also performed. HA group and FLE groups achieved significantly fewer categories and demonstrated more perseverative errors on the WCST than NSL and N groups. In addition, category achievement in WCST showed significant inverse correlation to age at the initial status convulsivus in the HA group (P<0.05). The achievement on PVALT and BVRT did not show any significant differences between HA and FLE groups (P>0.05). Thus, the frontal lobe dysfunction in the HA group is found to exist as early as 7 years old, when they seem to have only a short seizure history or to receive a little electrical interference from the temporal lobe focus to the frontal region. These facts would underscore the importance of prefrontal dysfunction persisting from the early insults and only becoming apparent after maturation of the prefrontal region in patients with mesial TLE.


The Fatty Acid Methyl Ester (FAME) profile of Phytophthora agathidicida and its potential use as diagnostic tool.
Phytophthora diseases cause devastation to crops and native ecosystems worldwide. In New Zealand, Phytophthora agathidicida is threatening the survival of kauri, an endemic, culturally and ecologically important tree species. The current method for detecting P. agathidicida is a soil bating assay that is time-consuming and requires high levels of expertise to assess, thus limiting the analytical sample throughput. Here, we characterized the fatty acid methyl ester (FAME) profile of P. agathidicida. We also compared it with the FAME profile of P. cinnamomi and assessed the efficacy of FAME analysis as a diagnostic tool for detecting the pathogen in soil samples. In FAME analysis, the total fatty acid content is isolated from a sample and converted to FAMEs for analysis, a process that takes less than a day. Unique fatty acid acyl chains can serve as biomarkers for specific organisms. We detected 12 fatty acids in P. agathidicida, two of which (20:4ω6 and 20:5ω3) show promise as potential Phytophthora specific biomarkers. Collectively, these findings advance our fundamental understanding of P. agathidicida biology and provide a promising technique to increase the rate of sample processing and the speed of pathogen detection for P. agathidicida in soil.