Sustainable Laboratory Capacity Building After the 2014 Ebola Outbreak in the Republic of Guinea.

Background: The 2014-2016 West Africa Ebola virus disease outbreak heavily impacted the Republics of Guinea, Sierra Leone, and Liberia. The outbreak uncovered the weaknesses of the public health systems, including inadequately trained and insufficient health personnel as well as limited and poorly equipped health infrastructures. These weaknesses represent significant threats to global health security. In the wake of the outbreak, affected countries made urgent requests for international engagement to help strengthening the public health systems. Methods: This work describes the successful multi-year implementation of a laboratory capacity building program in the Republic of Guinea. The program integrated biorisk and quality management systems training, infectious diseases diagnostic training, facility engineering and maintenance training, and mentorship to strengthen Guinea's bio-surveillance capacity. Results: The major outcome of these efforts was an established and local staff-operated public health laboratory that performs disease surveillance and reporting and diagnostic of priority diseases and pathogens of security concerns. Conclusions: This work has improved the Guinea country's capabilities to address country public health issues and preparedness to respond to future infectious disease threats.

Prediction of Violence, Suicide Behaviors and Suicide Ideation in a Sample of Institutionalized Offenders With Schizophrenia and Other Psychosis.

This study examined the predictive validity of the Spanish version of the Suicide Risk Assessment Manual (S-RAMM) and the Historical-Clinical-Risk Management-20 (HCR-20) in a sample of violent offenders with schizophrenia and other psychosis, who had committed violent crimes and had been sentenced to compulsory psychiatric treatment by the criminal justice system. Patients were prospectively monitored within the institution for 18 months. During the follow-up period, 25% of offenders were involved in any suicidal behavior including acts of self-harm, suicidal ideation and suicide attempts and 34% were physically or verbally violent. The S-RAMM and HCR-20 risk assessment tools were strongly correlated and were able to predict suicidal behavior and violence with a moderate-large effect size (AUCs = 0.81-0.85; AUCs = 0.78-0.80 respectively). Patients scoring above the mean on the S-RAMM (>20-point cut-off) had a five times increased risk of suicide related events (OR = 5.05, 95% CI = 2.6-9.7) and sevenfold risk of violence in the HCR-20 (>21-point cut-off) (OR = 7.13, 95% CI = 2.0-21.2) than those scoring below the mean. Offenders at high risk for suicide and violence had significantly more suicide attempts (p < 0.001) and more prior sentences for violent crimes (p < 0.001). These results support the use of the S-RAMM and HCR-20 for clinical practice by providing evidence of the utility of these measures for predicting risk for suicidal and violent behavior in mentally disordered offenders.

Children's First Experience of Taking Anabolic-Androgenic Steroids can Occur before Their 10th Birthday: A Systematic Review Identifying 9 Factors That Predicted Doping among Young People.

Taking performance-enhancing drugs (PEDs) can cause serious and irreversible health consequences, which can ultimately lead to premature death. Some young people may take PEDs without fully understanding the ramifications of their actions or based on the advice from others. The purpose of this systematic review was to identify the main factors that predicted doping among young people. The literature was systematically reviewed using search engines, manually searching specialist journals, and pearl growing. Fifty-two studies, which included 187,288 young people aged between 10 and 21 years of age, 883 parents of adolescent athletes, and 11 adult coaches, who were interviewed regarding young athletes, were included in this review. Nine factors predicted doping among young people: gender; age; sports participation; sport type; psychological variables; entourage; ethnicity; nutritional supplements; and health harming behaviors. In regards to psychological variables, 22 different constructs were associated with doping among young people. Some psychological constructs were negatively associated with doping (e.g., self-esteem, resisting social pressure, and perfectionist strivings), whereas other were positively associated with doping (e.g., suicide risk, anticipated regret, and aggression). Policy makers and National Anti-Doping Organizations could use these findings to help identify athletes who are more at risk of doping and then expose these individuals to anti-doping education. Based on the current findings, it also appears that education programs should commence at the onset of adolescence or even late childhood, due to the young age in which some individuals start doping.

Representation of information about family relatives as structured data in electronic health records.

BACKGROUND: The ability to manage and leverage family history information in the electronic health record (EHR) is crucial to delivering high-quality clinical care. OBJECTIVES: We aimed to evaluate existing standards in representing relative information, examine this information documented in EHRs, and develop a natural language processing (NLP) application to extract relative information from free-text clinical documents. METHODS: We reviewed a random sample of 100 admission notes and 100 discharge summaries of 198 patients, and also reviewed the structured entries for these patients in an EHR system's family history module. We investigated the two standards used by Stage 2 of Meaningful Use (SNOMED CT and HL7 Family History Standard) and identified coverage gaps of each standard in coding relative information. Finally, we evaluated the performance of the MTERMS NLP system in identifying relative information from free-text documents. RESULTS: The structure and content of SNOMED CT and HL7 for representing relative information are different in several ways. Both terminologies have high coverage to represent local relative concepts built in an ambulatory EHR system, but gaps in key concept coverage were detected; coverage rates for relative information in free-text clinical documents were 95.2% and 98.6%, respectively. Compared to structured entries, richer family history information was only available in free-text documents. Using a comprehensive lexicon that included concepts and terms of relative information from different sources, we expanded the MTERMS NLP system to extract and encode relative information in clinical documents and achieved a corresponding precision of 100% and recall of 97.4%. CONCLUSIONS: Comprehensive assessment and user guidance are critical to adopting standards into EHR systems in a meaningful way. A significant portion of patients' family history information is only documented in free-text clinical documents and NLP can be used to extract this information.

[Peculiarities of health condition of Tbilisi metro workers accordind to age and length of service].

The purpose of the study was to evaluate the influence of professional factors on health condition of Tbilisi metro workers. Working conditions were analyzed; paraclinic study was conducted; health status according to age and length of service was investigated. It was found that age and length of service affects the health condition of the worker; cardiovascular and nervous system diseases are the most frequent.

Analysis of ß-blockers in groundwater using large-volume injection coupled-column reversed-phase liquid chromatography with fluorescence detection and liquid chromatography time-of-flight mass spectrometry.

Atenolol, nadolol, metoprolol, bisoprolol and betaxolol were simultaneously determined in groundwater samples by large-volume injection coupled-column reversed-phase liquid chromatography with fluorescence detection (LVI-LC-LC-FD) and liquid chromatography-time-of-flight mass spectrometry (LC-TOF-MS). The LVI-LC-LC-FD method combines analyte isolation, preconcentration and determination into a single step. Significant reductions in costs for sample pre-treatment (solvent and solid phases for clean up) and method development times are also achieved. Using LC-TOF-MS, accurate mass measurements within 3 ppm error were obtained for all of the ß-blockers studied. Empirical formula information can be obtained by this method, allowing the unequivocal identification of the target compounds in the samples. To increase the sensitivity, a solid-phase extraction step with Oasis MCX cartridge was carried out yielding recoveries of 79-114% (n=5) with RSD 2-7% for the LC-TOF-MS method. SPE gives a high purification of ß-blockers compared with the existing methods. A 100% methanol wash was allowed for these compounds with no loss of analytes. Limit of quantification was 1-7 ng/L for LVI-LC-LC-FD and 0.25-5 ng/L for LC-TOF-MS. As a result of selective extraction and effective removal of coextractives, no matrix effect was observed in LVI-LC-LC-FD and LC-TOF-MS analyses. The methods were applied to detect and quantify ß-blockers in groundwater samples of Almería (Spain).

Stress fiber formation is required for matrix reorganization in a corneal myofibroblast cell line.

Corneal wound healing fibroblasts (myofibroblasts) develop a muscle-like contractile apparatus composed of prominent microfilament bundles (stress fibers) and express alpha-smooth muscle actin (alpha-SMA). In this study, gelsolin, an actin filament-severing protein, was overexpressed in a alpha-SMA-expressing corneal myofibroblast cell line (TRK43) to assess whether intact stress fibers are required for in vitro matrix organization and wound contraction. Stably integrated gelsolin was introduced by electroporation of an expression construct (pREPCG8) into cultured cells. Thirty-seven clones were isolated with half of the clones showing a fibroblastic phenotype while the remaining half appeared epithelioid. One fibroblastic clone, GS56, and one epithelioid clone, GS44, were selected for detailed characterization. The GS56 cells appeared highly elongated and spindle-shaped and had prominent stress fibers and focal adhesions. GS44 cells showed disruption of stress fibers and a cortical f-actin organization as well as the down regulation of alpha-SMA expression by immunocytochemistry and Western blotting. Both phenotypes showed enhanced gelsolin expression; however, fractionation of cell extracts demonstrated differences in the subcellular distribution of gelsolin with GS44 cells having markedly reduced and GS56 cells having markedly increased cytoskeletal gelsolin. In an in vitro wound contraction assay, epithelioid GS44 cells showed a significantly impaired ability to contract a collagen matrix compared to that of TRK43 cells, CT9 or GS56 transfectants. Loss of stress fibers in GS44 cells also correlated with enhanced cell motility. Together, these results demonstrate that the ability to form microfilament bundles or stress fibers is required for matrix organization and contraction by corneal myofibroblasts. Although no clear explanation is available, we suspect that differences in gene insertion of the gelsolin overexpression vector may have led to differential intercellular localization of gelsolin and its effect on stress fiber formation in the two cell lines.

Decision support for clinical trial eligibility determination in breast cancer.

We have developed a system for clinical trial eligibility determination where patients or primary care providers can enter clinical information about a patient and obtain a ranked list of clinical trials for which the patient is likely to be eligible. We used clinical trial eligibility information from the National Cancer Institute's Physician Data Query (PDQ) database. We translated each free-text eligibility criterion into a machine executable statement using a derivation of the Arden Syntax. Clinical trial protocols were then structured as collections of these eligibility criteria using XML. The application compares the entered patient information against each of the eligibility criteria and returns a numerical score. Results are displayed in order of likelihood of match. We have tested our system using all phase II and III clinical trials for treatment of metastatic breast cancer found in the PDQ database. Preliminary results are encouraging.

Characterization of novel phorbol ester- and serum-responsive sequences of the rat ornithine decarboxylase gene promoter.

Ornithine decarboxylase (ODC), the key regulatory enzyme in mammalian polyamine biosynthesis, is rapidly induced by mitogens and tumor promoters. We used transient expression assays and DNA-protein binding studies to examine the regulation of ODC promoter activity by phorbol esters and serum growth factors. A fragment of the ODC 5' flanking region (nt-1156 to +13) was sufficient to confer 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive expression to a luciferase reporter gene when transfected into H35 cells. However, induction by TPA was not observed in Rat2 fibroblasts, although refeeding of serum-starved Rat2 cells with fresh serum-containing medium rapidly induced a fivefold to sixfold increase in ODC promoter activity, maximal about 8 h after refeeding. Deletion analysis demonstrated that several sequences contributed to basal ODC promoter activity but that nt -92 to +13 was sufficient for induction by TPA or by serum. This sequence lacked canonical TPA-responsive elements, and an activator protein-1 (AP-1) consensus oligonucleotide failed to compete effectively for proteins binding to this region. Two of four protein complexes observed by gel-shift analysis of nt -92 to +13 were competitively inhibited by wild-type but not mutant oligonucleotides encompassing a variant cyclic AMP-response element (CRE) (ODC nt -50 to -42); however, a consensus CRE did not compete. Mutagenesis of this site demonstrated that it contributes to basal expression of the ODC promoter but not to TPA or serum responsiveness. Thus, we conclude that the proximal ODC promoter (nt -92 to +13) responds to TPA and serum stimulation in a cell-type-specific manner that is not mediated by canonical AP-1 elements.

Regulation of rat ornithine decarboxylase promoter activity by binding of transcription factor Sp1.

Ornithine decarboxylase (ODC) is the rate-limiting enzyme of polyamine biosynthesis. We investigated the transcriptional regulation of the rat ODC gene using transient expression assays. The 5'-flanking region (-1156 to +13) of the ODC gene was sufficient to mediate strong basal expression of a luciferase reporter. Sequences between -345 and -93 contributed to basal promoter activity. This region, containing five potential Sp1 binding sites, was analyzed by electrophoretic mobility shift assays. Three specific DNA-protein complexes were identified using H35 nuclear extracts and the -345/-93 ODC probe. Binding to all three was eliminated by competition with an oligonucleotide containing an Sp1 binding site, but not by a mutant Sp1 oligonucleotide. Preincubation with an antibody against Sp1 supershifted complexes associated with one or more of Sp1 binding sites 1-4 as well as with site 5. DNase I footprinting revealed two protected regions: PR-I (-92 to -130) and PR-II (-304 to -332). PR-I contains a putative binding site for Sp1 that was protected by recombinant Sp1 protein. Transfection studies in Schneider SL2 cells demonstrated that the ODC promoter is transactivated up to 350-fold by Sp1 and that this transactivation is dependent on the presence of Sp1 binding sites 1-4. Thus, although the ODC promoter binds multiple nuclear proteins, Sp1 or a related protein appears to be a critical determinant of ODC transcription, possibly through cooperative interactions between Sp1 and additional transcription factors.

Nurse case management: collaborative beyond the hospital walls.

In 1989, St. Joseph Medical Center initiated a community-based nursing case management program with two nurses providing care to high-risk, chronically ill, frail elderly patients. This program has expanded to five registered nurses actively following over 120 patients. Target populations now served have expanded to include: frail elderly, high-risk pregnant women, premature infants, AIDS patients, and those with chronic physical and mental illness. The nurse manages and coordinates the care for patients through all settings (community and hospital), brokering services, acting as a patient advocate, and giving traditional hands-on care as needed. Outcomes analysis has shown that, after nursing case management intervention, the patients demonstrated a 71% decrease in admissions to the medical center, a 21% decrease in length of stay, and a 64% decrease in Emergency Room usage. Nursing case management helps to prevent patients' health problems from becoming more complex. Consequently, managing their health effectively requires fewer, less costly resources while achieving improved patient outcomes.

Regulation of ornithine decarboxylase mRNA by phorbol esters and insulin in normal and C-kinase-deficient rat hepatoma cells.

Tumor-promoting phorbol esters and insulin produce similar effects in Reuber H35 rat hepatoma cell proliferation, including increased ornithine decarboxylase (ODC) enzyme activity, DNA synthesis, and mitogenesis. We investigated ODC mRNA accumulation in cells treated with either insulin or 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Both agents caused rapid accumulation of ODC mRNA: for TPA, it was maximal 3 hr after treatment (4-6-fold greater than control cells) and returned quickly to control levels; for insulin, it was significantly longer, continuing to increase for at least 6 hr. Simultaneous treatment with TPA and insulin led to additive effects on ODC mRNA. Induction of ODC by TPA was blocked by down-regulation or inhibition of protein kinase C (PKC), consistent with a PKC-mediated mechanism. In contrast, PKC down-regulation had little effect on ODC induction by insulin. Furthermore, although both agents stimulated ribosomal S6 protein phosphorylation in cells containing normal amounts of PKC, the response to TPA was abolished in PKC-depleted cells; the effect of insulin was only slightly inhibited. TPA caused a rapid redistribution of essentially all of the PKC activity from the cytosolic to the membrane fraction of the cells, whereas insulin had no effect on PKC distribution. These results suggest that although insulin and TPA share some common cytoplasmic signalling pathways, their effects on phosphorylation of nuclear proteins and transcription of ODC may be mediated by distinct factors.

Involvement of protein kinase C in the regulation of ornithine decarboxylase mRNA by phorbol esters in rat hepatoma cells.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulates a rapid increase in ornithine decarboxylase (EC 4.1.1.17; ODC) activity in target cells. Here we demonstrate that this process involves a rapid accumulation of ODC mRNA, which is maximal 3 h after treatment (three- to eightfold greater than control cells) and decays to control levels within 18 h. Stimulation of ODC mRNA by TPA is blocked by phorbol dibutyrate down-regulation of protein kinase C (PKC). ODC mRNA was also induced by the PKC activators, phospholipase C and 1-oleoyl-2-acetyl-rac-glycerol, and blocked by kinase inhibitors (trifluoroperazine, H7, and palmitoyl-L-carnitine), consistent with a requirement for PKC activation in the induction mechanism. However, the non-PKC-specific protein kinase inhibitor HA1004 also suppressed expression of ODC mRNA in response to TPA, under conditions where it did not inhibit PKC, suggesting that additional kinases may be involved in the intracellular signalling process. The stability of the ODC mRNA (control value = 6.2 +/- 1.6 h) is not significantly changed by either TPA (5.7 +/- 0.8 h) or by cycloheximide (6.0 h). These results are inconsistent with any contribution from altered mRNA half-life towards the accumulation of ODC mRNA following treatment with phorbol ester tumor promoters.

Antigen receptor rearrangement and expression in acute leukemias.

Twenty-two leukemias, 11 of which were undifferentiated with respect to surface antigen markers, were investigated for their antigen receptor gene rearrangement, transcription products of these antigen receptor genes, and surface antigen pattern of the cells. Among the three less-differentiated groups rearrangement was observed in 2/10 cases for the TCR beta-chain and in 4/11 cases for the heavy-chain gene. TCR beta-mRNA, however, was expressed in seven out of eight cases and the mu heavy-chain mRNA in eight out of ten cases investigated. Also mRNA of TCR alpha, the rearrangement of which could not be detected with our probes, was expressed as frequently as TCR beta. Although rearrangement of the appropriate gene was found regularly in the more mature leukemias, transcription of these genes was lower or even lacking. These findings indicate that expression of antigen receptor mRNA in undifferentiated leukemias can be activated by events other than maturational rearrangement.

Quantitative tracing of mRNAs for T- and B-lymphocyte receptor genes in individual cells by in situ hybridization with fluorochrome-labeled gene probes. I. Expression in malignancies carrying B-lineage associated antigens.

Acute and chronic lymphatic leukemias were investigated on the single-cell level for the activity of genes coding for the IgM heavy chain and the alpha and beta chains of the T-cell antigen receptor (TCR). We used a new method for preparing highly fluorochrome-labeled gene probes for in situ hybridization, which allowed rapid and quantitative detection of mRNA at the individual cell level. Leukemic cell populations classified as belonging to the B lineage according to their surface antigenic patterns revealed increasing expression of mRNA for the IgM heavy chain (mu mRNA) in a maturation-dependent fashion, which was not correlated to rearrangement of the immunoglobulin mu chain gene--only 66% of the leukemias with rearranged mu gene also transcribed it. TCR mRNA was detected in B-antigen positive leukemic cells. High levels of both TCR and mu mRNA expression in all cells of some of these leukemias allowed the conclusion that these cells simultaneously transcribed the genes for T and B cell antigen receptors. TCR mRNA was also found in what are considered relatively mature B leukemias, lineage cross-over on the mRNA level being observed at a frequency of 23% (five of 22 cases), comparable with that of "inappropriate" receptor gene rearrangement. The quantitation of mRNA with fluorochrome-labeled gene probes in situ may allow determining the degree of gene activation in individual antigenically defined cells and may thus contribute a new tool for characterization of normal and malignant cells.

[Antibodies to lipoid A in the treatment of septic shock]. / Antikörper gegen Lipoid A in der Behandlung des septischen Schocks.

The protective effect of high-titer anti-lipid A hyperimmune globulin with respect to the course of the disease and the mortality rate was studied in patients with septicemia verified by positive blood cultures. Six patients were treated with anti-lipid A in an open study. Dramatic improvement in fever curves and clinical condition in some of the patients encouraged us to start a randomized double blind study. So far, 17 patients have entered the study, 16 of whom were evaluable. Immediately after a positive blood culture was found, patients received either high doses of anti-lipid A or placebo (saline solution) on two subsequent days. Before and after each infusion blood samples were taken in order to assess serum bactericidal activity and anti-lipid A titers. Because of the still small numbers of patients the results of both studies were summarized. In all patients treated with anti-lipid A clear-cut increases in anti-lipid A titers were shown. Patients with repeated gram-negative infections showed higher median anti-lipid A titers than patients without such a history. The patients treated with anti-lipid A immune globulin ran a significantly milder course than the placebo group. The severe signs of septic shock were reversed in seven of 15 patients on anti-lipid A compared to two of seven patients treated with placebo. In the anti-lipid A-treated group, three of 15 patients died, and in the placebo group two of seven. This difference is not statistically significant.