CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Os agrotÃxicos estÃo entre os mais importantes fatores de risco para a saÃde dos trabalhadores. No PiauÃ, a expansÃo agrÃcola implementada nos cerrados, por intermÃdio da intensificaÃÃo do plantio de soja, a partir da dÃcada de 1990, consistiu em um forte fator para o aumento do uso de agrotÃxicos. A utilizaÃÃo indiscriminada dos agrotÃxicos, geralmente, està associada a intoxicaÃÃes agudas e crÃnicas. A dosagem de colinesterase plasmÃtica e eritrocitÃria sÃo parÃmetros relevantes para diagnÃstico de casos cuja exposiÃÃo se deu, principalmente, de forma aguda. A genotoxicidade associada ao uso destes produtos vem sendo estudada, sendo fortes as evidÃncias que estes produtos sÃo carcinogÃnicos. No estado do PiauÃ, em setembro de 2005, no municÃpio de Ribeiro GonÃalves, ocorreram mortes cuja suspeita principal era de intoxicaÃÃo por uso dos pesticidas agrÃcolas. Este fato desencadeou uma investigaÃÃo nos municÃpios produtores de soja (UruÃuÃ, Ribeiro GonÃalves e Baixa Grande do Ribeiro). O objetivo deste trabalho foi avaliar o impacto do uso de agrotÃxicos sobre a saÃde dos trabalhadores rurais dos municÃpios de UruÃuÃ, Ribeiro GonÃalves e Baixa Grande do Ribeiro. Dividiu-se, metodologicamente, o estudo em trÃs etapas. Na primeira, aplicou-se questionÃrio sÃcio epidemiolÃgico de acordo com OrganizaÃÃo Mundial de SaÃde (OMS) e coletaram-se amostras de material biolÃgico para anÃlises bioquÃmicas, hematolÃgicas e enzimÃticas (acetil colinesterase plasmÃtica e eritrocitÃria) de 120 trabalhadores agrÃcolas da regiÃo sul do PiauÃ. Na segunda etapa, estudou-se 75 indivÃduos. Estes se submeteram a uma segunda avaliaÃÃo, tendo como ferramenta os mesmos exames anteriormente realizados. Como terceira etapa, realizou-se o teste de micronÃcleo em 31 indivÃduos que encontravam-se com os parÃmetros alterados. Fez-se coleta de cÃlulas da mucosa oral, para avaliaÃÃo do nÃvel de lesÃo de DNA atravÃs de contagem de micronÃcleos. Quanto ao questionÃrio sÃcio epidemiolÃgico os resultados encontrados indicaram que 100% da populaÃÃo estudada pertencia ao sexo masculino, com idade mÃdia de 35 anos. Ainda sobre o questionÃrio, 66% eram casados, 76,67% trabalhavam em fazendas (lavradores, operadores de mÃquinas e tÃcnicos agrÃcolas), 16,67% eram agentes de endemias ou servidores pÃblico, sendo que 21,3% tomavam medicamento sem prescriÃÃo mÃdica e que 71,67% nÃo realizaram exames mÃdicos nos Ãltimos 06 meses. A investigaÃÃo evidenciou ainda que, quanto ao estilo de vida e trabalho, 33,33% dos trabalhadores possuem o hÃbito de fumar, 55,83% de consumir bebidas alcoÃlicas, 29,9% entraram em contato direto com o agrotÃxico durante o preparo do produto, 38% atravÃs da aplicaÃÃo do produto por pulverizaÃÃo, 10% atravÃs da limpeza e manutenÃÃo dos equipamentos, 17,5% nÃo utilizavam nenhuma proteÃÃo, sendo que a grande maioria utilizavam apenas 01 tipo de EPI. Outro achado relatado refere-se que os trabalhadores utilizaram mais de 34 diferentes formulaÃÃes, sendo as classes mais utilizadas os inseticidas (45,18%), herbicidas (27,68%) e fungicidas (6,21%). Quanto ao tempo de exposiÃÃo dos trabalhadores aos pesticidas, 50,84% estavam expostos a menos de 2 anos e 10% estavam expostos a mais de 10 anos. Dentre os sintomas mais relatados, os efeitos no SNC destacaram-se com 230 queixas relatadas durante a aplicaÃÃo do questionÃrio sÃcio-epidemiolÃgico. Sobre o destino final das embalagens, 84,21% dos mesmos eram queimados, enterrados e deixados a cÃu aberto, sendo 15,79% entregues na central de recebimento. Em relaÃÃo Ãs anÃlises bioquÃmicas e hematolÃgicas dos indivÃduos submetidos à primeira coleta, os resultados indicaram que 11,67%; 4,17%; 10,83%; 0,83% ; 5,83%; 5% e 9,17% encontravam-se com nÃveis respectivamente alterados de Gama GT, Fosfatase Alcalina e TGO/ TGP, UrÃia, Creatinina componentes protÃicos, acetilcolinesterase plasmÃtica e eritrocitÃria . Na segunda etapa, os achados indicaram que os indivÃduos estavam com os nÃveis alterados de: gama GT ( 4,00%); fosfatase alcalina e TGO/TGP ( 4,00%); urÃia (4,00%); creatinina (5,33%); proteÃnas totais (17,33%); acetilcolinesterase plasmÃtica (4,00%) e acetilcolinesterase eritrocitÃria (2,67%). Com relaÃÃo à terceira etapa, dos 31 indivÃduos que realizaram teste de micronÃcleo 84,00% encontravam-se com danos citogenÃticos. Os danos citogenÃticos evidenciados nÃo apresentaram correlaÃÃes com idade, fumo, bebidas, equipamentos de proteÃÃo, medicamentos e doenÃas, confirmando a correlaÃÃo dos danos citogenÃticos evidenciados, com a exposiÃÃo aos agrotÃxicos (P<0,05) / The pesticides are some of the main risky factors for agricultural laborersâ health. In the state of PiauÃ, the agricultural expansion which was implemented in the cerrados through the growth of the soy crops in the 90âs was a relevant factor that increased the use of pesticides. The unbalanced usage of pesticides is often related to acute and chronic poisonings. The dosage of plasma cholinesterase and erythrocyte cholinesterase are relevant parameters to diagnose mainly cases in which an acute contact has occurred. The genotoxicity related to the use of these kinds of products has been studied and there are strong evidences that they are carcinogenic. Deaths which the main suspicion was poisoning by the misuse of agricultural pesticides happened in September of 2007 in the city of Ribeiro GonÃalves, state of PiauÃ. The fact unchained an investigation in the cities that produce soy (UruÃuÃ, Ribeiro GonÃalves and Baixa Grande do Ribeiro). The purpose of this study was evaluating the impact of the use of pesticides on agricultural laborerâs health in the cities of UruÃuÃ, Ribeiro GonÃalves and Baixa Grande do Ribeiro. The study was divided in three steps. Firstly, a social epidemiologic questionnaire Word Health Organization (WHO) was applied and samples and collected material to biochemical, hematological and enzymatic plasma acetylcholinesterase and erythrocyte acetylcholinesterase analysis of 120 agricultural laborers from the region. The second step was a study of these workers. These ones submitted salf of Piauà to a second evaluation which used, as a tool, the tests which had been previously discribed. In the third step 31 individuals with irregular parameters were submitted to a micronucleus test. Cells from the oral mucosa were collected to evaluate the level of DNA injury through micronucleus counting. The results of the social epidemiologic questionnaire pointed that 100% of the subjects who had benn studied were males with an average age of 35 years old. 66% were married, 76.67% worked in farms (farmers, machinists and agricultural technicians), 16.67% worked as endemic diseases agents or civil servants. 21.3% of them used non-prescribed medication and 71.67% had not did medical tests for the last 6 months. About life and work style the investigation showed that 33.33% of the workers is smokers, 55.83% drink alcohol, 29.9% has direct contact with the pesticide while it was being prepared to be use, 38% during the pulverization process, 10% during the equipment cleaning and maintenance, 17.5% did not take any safety protection measure, while the most used only a single kind of EPI. Another related discovery about the workers showed that they used more than 34 different formulations. Insecticides (45.18%), herbicides (27.68%) and fungicides (6.21%) were the most used products. Concerning the time of exposure the workers had to the pesticides, 50.84% were exposed to them for less than 2 years and 10% for more than 10 years. The effects on the SNC detached themselves among the most related symptoms with a 230 complaints reported during the application of the social epidemiologic questionnaire. Concerning the final destination of the containers, 84.21% were burned, buried and left outdoors while 15.79% were brought to the appropriate organ headquarter. The results from the biochemical and hematological analysis indicated that 11.67%; 4.17%; 10.83%; 0.83%; 5.83%; 5% e 9.17% of the individuals who were submitted to the first sample collection had, respectively, irregular levels of Gamma-Glutamyl Transferase (GGT); Alkaline Fosfatase and Glutamic-Oxaloacetic Transaminase/ Glutamic Pyruvic Transaminase; Urea; Creatinine; Proteic components; Plasma acetylcholinesterase and Erythrocyte acetylcholinesterase. The findings in the second step indicated that the individuals had irregular levels of: Gamma GT (4.00%); Alkaline Fosfatase and GOT/GPT (4.00%); Urea (4.00%); Creatinine (5.33%); Total Protein (17.33%); Plasma acetylcholinesterase (4.00%) and Erythrocyte acetylcholinesterase (2.67%). In the third step, 84.00% of the 31 subjects who had a micronucleus test presented cytogenetic injuries. The cytogenetic injuries which could be observed were not related to age, smoke, alcohol consumption; safety equipment, medications and diseases which confirms the co-relation between the presented cytogenetic injuries and the exposure to pesticides ( P<0.05)

Recentemente, foi descrito um grupo de alterações no metabolismo da creatina denominado Síndromes de Deficiência de Creatina. Há três formas da doença geneticamente determinadas que cursam com deficiência de creatina, seja por comprometimento de sua síntese ou por defeito na proteína transportadora. O espectro de apresentação clínica dessa condição é inespecífico e inclui atraso ou estagnação do desenvolvimento neuromotor, hipotonia muscular, movimentos involuntários do tipo coreoatetose, retardo ou ausência do desenvolvimento da fala, retardo mental de grau variável, comportamento autista e epilepsia. Neste trabalho, foi desenvolvida e validada uma alternativa metodológica àquelas disponíveis na literatura, com a utilização de extração por troca catiônica forte e separação e detecção por cromatografia líquida de interação hidrofílica acoplada a espectrometria de massas em tandem em que foram exploradas as características químicas das moléculas de creatina e guanidinoacetato, metabólito intermediário da síntese de creatina. Os valores de referência para o método foram definidos pela sua aplicação a 150 amostras de urina e 197 amostras de soro de indivíduos de ambos os sexos e idades entre 0 e 16 anos. Foram também analisadas amostras de urina, soro e plasma de 54 pacientes com clínica compatível com a síndrome de deficiência de creatina sendo que 3 deles apresentaram perfil bioquímico característico de uma das formas dessa condição / Recently, a new group of inborn errors of metabolism, collectively named as creatine deficiency syndrome, was identified. Three genetically determined presentations are currently known, affecting both creatine synthesis and transport. Clinical presentation spectrum is non-specific and includes developmental delay, hypotonia, involuntary movements as choreoathetosis, delay or lack of speech acquisition, mental retardation of variable severity, autistic behavior, and epilepsy. Herein, we developed and validated an innovative method for determination of creatine and of its metabolic intermediate, guanidinoacetate, based on cation-exchange solid-phase extraction and hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry. Reference values for the method were defined testing 150 urine and 197 serum samples in males and females with age ranging from 0 to 16 years. Urine and serum samples from 54 patients with some clinical features that might be attributable to creatine deficiency were also evaluated, and in three, biochemical profile characteristic of one of the disorders was detected

The new generation of giant ground-based telescopes will see their first light this decade. These state-of-the-art facilities will significantly surpass the resolving power of modern space-based observatories such as the James Webb telescope, thanks to their enormous aperture size and adaptive optics (AO) facilities. Without AO, atmospheric turbulence would degrade the image quality of these enormous telescopes to that of a 50 cm amateur one. These extremely large telescopes (ELTs) will further benefit from a particular branch of AO called multi-conjugate adaptive optics (MCAO), which provides an extremely high resolving power over a much wider field of view as compared to classical AO systems. The design and fabrication of such systems, as well as their optimal use for science operation, pose a great challenge as they are an order of magnitude more complicated than current AO systems. To face such a challenge, the combined knowledge of MCAO system design and fabrication, working in tandem with scientific insights into new astronomy science cases, is an extremely valuable and essential pairing. This thesis is an effort to not only contribute to the design and fabrication of ELT MCAO facilities, but also provide guidance on the optimal method to utilize these giant telescopes to achieve unprecedented astrometric measurements. On the instrumentation side, in partnership with the National Research Council of Canada's - Herzberg Astronomy and Astrophysics Institute as well as W.M. Keck Observatory in Hawaii, I was involved in the design and fabrication of a cutting edge new wavefront sensor, which is the eye of an AO system. I performed opto-mechanical design and verification studies for components of the Keck infrared pyramid wavefront sensor (IR-PWFS) as well as the Keck Planet Imager and characterizer (KPIC) instrument, which have both been commissioned and are in science operation. Furthermore, I designed the alignment plan and participated in the modification and alignment operation of a few components on the Keck II adaptive optics bench on the summit of Mauna Kea. To pave the way for the design verification of future MCAO systems for ELTs, I proposed a new method for an old challenge in the path of AO system design and verification: a flexible method for precise intensity pattern injection into laboratory AO benches. AO benches are the backbone of instrument design and modeling. One of the challenges especially important for the future generation of MCAO systems for ELTs is the verification of the effect of shadowed regions on the primary mirror. During my PhD, I successfully demonstrated the feasibility of a new proposed method to accurately model the telescope pupil. This work was done in partnership with the Laboratoire d'Astrophysique de Marseille (LAM) in France. The method I developed at LAM will be implemented in the AO Lab at NRC Herzberg Astronomy and Astrophysics. As an observational astronomer, I focused on developing methods for making optimal astrometric measurements with MCAO-enabled telescopes. The expected unparalleled astrometric precision of ELTs comes with many unprecedented challenges that if left unresolved, would jeopardize the success of these facilities as they would not be able to reach their science goals. I used observations with the only available MCAO system in science operation, the Gemini MCAO system on the 8-meter Gemini South telescope in Chile, to develop and verify a pipeline specifically designed for very high-precision astrometric studies with MCAO-fed imagers. I successfully used the pipeline to provide the precise on-sky differential distortion of the Gemini South telescope and its MCAO facilities by looking deep into the core of globular cluster NGC~6723. Using this pipeline, I produced high quality proper motions with an uncertainty floor of $\sim 45$\,$\mu$as~yr$^{-1}$ as well as measured the proper motion dispersion profile of NGC~6723 from a radius of $\sim 10$ arcseconds out to $\sim 1$\,arcminute, based on $\sim 12000$ stars. I also produced a high-quality optical-near-infrared color magnitude diagram which clearly shows the extreme horizontal branch and main-sequence knee of this cluster. / Graduate

Since the Institute of Medicine highlighted the devastating impact of medical errors in their seminal report, “To Err is Human” (2000), efforts have been underway to improve patient safety. A portion of medical errors are due to medication errors, and a large portion of these can be attributed to inadequate laboratory monitoring. In this thesis, I attempt to address this small but important corner of this patient safety endeavor. Why are patients not getting their laboratory monitoring tests? Do they fail to complete them or do doctors not order the tests in the first place? Which prescribers and which patients are least likely to do what is needed for testing to happen and what interventions would be most promising? To address these questions, I conducted a systematic review of existing interventions. I then proceeded with three aims: 1) To identify reasons that patients give for missing monitoring tests; 2) To identify patient and provider factors associated with monitoring test ordering; and 3) To identify patient and provider factors associated with completion of ordered testing. To achieve these aims, I worked with patients and data at the Fallon Clinic. For aim 1, I conducted a qualitative analysis of their reasons for missing tests as well as reporting completion and ordering rates. For aims 2 and 3, I used electronic medical record data and conducted a regression with patient and provider characteristics as covariates to identify factors contributing to test ordering and completion. Interviews revealed that patients had few barriers to completion, with forgetting being the most common reason for missing a test. The quantitative studies showed that: older patients with more interactions with the health care system were more likely to have tests ordered and were more likely to complete them; providers who more frequently prescribe a drug were more likely to order testing for it; and drug-test combinations that were particularly dangerous, indicated by a black box warning, were more likely to have appropriate ordering, though for these combinations, primary care providers were less likely to order tests appropriately, and patients were less likely to complete tests. Taken together, my work can inform future interventions in laboratory monitoring and patient safety.

<p>UVOD: Tuberkuloza je odavno poznata bolest koja i danas u 21. veku jo&scaron; uvek predstavlja veliki javnozdravstveni problem, uprkos primeni moćnih antituberkuloznih lekova. Trećina svetske populacije inficirana je bacilom tuberkuloze. Svake godine oboli oko osam miliona, a umre oko dva miliona ljudi, zbog čega je tuberkuloza i dalje infektivno oboljenje sa najvećom stopom smrtnosti. Kasna dijagnoza, multirezistentna tuberkuloza i udruženost sa HIV infekcijom predstavljaju jednu od najvećih prepreka za efikasnu kontrolu ove bolesti u svetu. Rano otkrivanje se oslanja na kvalitetnu bakteriolo&scaron;ku dijagnostiku koja je kamen temeljac svakog nacionalnog programa za kontrolu tuberkuloze. Brza i tačna mikrobiolo&scaron;ka dijagnostika predstavlja osnovu programa kontrole tuberkuloze i zbog toga je uvođenje novih i brzih laboratorijskih testova od veoma velikog značaja. Razvijen je novi komercijalno dostupni imunohromatografski test koji se zasniva na detekciji antigena MPT64 glavnog sekretovanog proteina M. tuberculosis. Test je brz i pouzdan u identifikaciji izolovanih sojeva M. tuberculosis i jeftiniji je od konvencionalnih biohemijskih i molekularnih testova. CILJ: Ciljevi istraživanja su bili da se evaluiraju karakteristike novog brzog imunohromatografskog testa u identifikaciji mikobakterija izolovanih iz respiratornih uzoraka bolesnika sa tuberkulozom pluća i referentnih sojeva klinički značajnih vrsta netuberkuloznih mikobakterija (NTM). MATERIJAL I METODE: Istraživanje je sprovedeno u periodu od 1.1.2010. do 31.12.2013. i obuhvatilo je 43563 respiratornih uzoraka dobijenih od bolesnika hospitalizovanih u Institutu za plućne bolesti Vojvodine. Iz obrađenih respiratornih uzoraka izolovano je 3469 izolata mikobakterija. Identifikacija do nivoa vrste urađena je primenom standardnih biohemijskih testova, molekularnog testa (GenoType&reg; Mycobacterium) i imunohromatografskog testa (BDMGIT Tbc). U istraživanje je uključeno 100 sojeva Gram pozitivnih i Gram negativnih bakterija (n = 19 vrsta) izolovanih iz respiratornih kliničkih uzoraka. Identifikacija do nivoa vrste je potvrđena komercijalnim identifikacionim sistemima. REZULTATI: U toku četvorogodi&scaron;njeg istraživanja izolovano je 3469 izolata mikobakterija iz respiratornih uzoraka. U ispitivanom periodu ne postoji opadajući trend izolacije mikobakterija &scaron;to potvrđuje i koeficijent korelacije (r = 0,31). Svi izolati mikobakterija su identifikovani konvencionalnim biohemijskim ispitivanjima koja pokazuju da je 89% od svih izolata identifikovano kao Mycobacterium tuberculosis (M. tuberculosis), a 11% izolata kao NTM. Mycobacterium xenopi je bila najzastupljenija NTM vrsta identifikovana kod 55,3% izolata. Nakon biohemijske identifikacije kod 300 izolata M. tuberculosis i 100 izolata NTM, identifikacija je potvrđena komercijalno dostupnim molekularnim i imunohromatografskim testom. Na osnovu rezultata testiranja mikobakterija imunohromatografskim testom, senzitivnost, specifičnost, pozitivne i negativne prediktivne vrednosti bile su: 99,7%, 100%, 100% i 99%. U poređenju imunohromatografskog testa sa konvencionalnim biohemijskim ispitivanjima nije nađena statistički značajna razlika (p&gt; 0,5). Kappa vrednost testa je iznosila 0,993, a interval poverenja CI =0,98 &ndash; 1,00. U poređenju imunohromatografskog sa molekularnim testom vrednost kappa je iznosila 0,993, a interval poverenja CI = 0,98 &ndash; 1,00. Slaganje rezultata je potvrđeno i McNemar testom sa vredno&scaron;ću 0,99. Utvrđena je stabilnost sekretovanog antigena MPT64 i posle 5 godina od prvog testiranja. ZAKLJUČAK: Visoka senzitivnost i specifičnost imunohromatografskog testa omogućuju tačnu i preciznu identifikaciju M. tuberculosis kao i pouzdanu diferencijaciju M.tuberculosis od NTM &ndash; a. Imunohromatografski test može da predstavlja zamenu za konvencionalne biohemijske i molekularne testove u identifikaciji M. tuberculosis. Jeftiniji je, jednostavniji za izvođenje i brže se dobijaju rezultati čime seskraćuje vreme za postavljanje dijagnoze.</p> / <p>INTRODUCTION: Tuberculosis (TB) has been known as a disease for a long time, but nevertheless it represents a major public health issue even nowadays in the 21st century, despite potent antituberculous drugs applied. One third of the world population is infected by the TB bacillus. About eight million people get infected and two million die of tuberculosis in a year, so tuberculosis is still an infectious disease with the greatest mortality rate. Late diagnosis, multiresistant tuberculosis and concomitant HIV infection interfere mostly with an efficient control of the disease all over the world. Early TB detection largely depends on the high-quality bacteriological diagnostics, which is the corner stone of each national TB control programme. A fast and accurate microbiological TB diagnosis plays a crucial role in any TB control programme. It is therefore very important to introduce new and fast laboratory tests. A novel commercially available immunochromatographic test has been designed, based on the MPT64 antigen of the major M. tuberculosis &ndash; secreted protein. This is a rapid and reliable test to identify the isolated strains of M. tuberculosis, which is not expensive as conventional biochemical and molecular tests. OBJECTIVE: The objective of the investigation was to evaluate the new immunochromatographic rapid test to identify mycobacteria isolated from respiratory samples from pulmonary TB patients, and referential strains of clinically relevant species of nontuberculous mycobacteria (NTM). MATERIAL AND METHODS: The research was carried out in the period from 1st January, 2010 to 31st December, 2013. It included 43 563 respiratory samples obtained from the patients hospitalized in the Institute for Pulmonary Diseases of Vojvodina, Sremska Kamenica (Serbia). There were 3 469 mycobacterial isolates obtained from the processed respiratory samples. The species &ndash; level identification was performed by standard biochemical tests, the molecular test (GenoType&reg;Mycobacterium), and the immunochromatographic test (BD MGIT Tbc). The study included one hundred (100) of Gram positive and Gram negative bacteria (n = 19 species) isolated from respiratory clinical samples. The species &ndash; level identification was confirmed by commercial identification systems. RESULTS: During the four &ndash; year investigation, 3 469 mycobacterial isolates were obtained from respiratory samples. No declining tendency of mycobacterial isolation was registered in the examined period, as confirmed by the correlation coefficient (r = 0.31). All mycobacterial isolates were identified by conventional biochemical tests showing that 89% of all isolates were identified as M. tuberculosis, and 11% of the isolates as NTM. Mycobacterium xenopi was the most common NTM species identified in 55.3% of the isolates. Following the biochemical identification in 300 M. tuberculosis isolates and 100 NTM isolates, the identification was confirmed by commercially available molecular and immunochromatographic tests. Based on immunochromatographic testing of mycobacteria, the sensitivity, specificity, positive and negative predictive values of the test were 99.7%, 100%, 100% and 99% respectively. There is no statistically significant difference (p&gt; 0.5) when comparing features of immunochromatographic test with conventional biochemical assay. The kappa test value was 0.993, and the confidence interval CI = 0.98 &ndash; 1.00. Comparing the immunochromatographic with the molecular test, the kappa value was 0.993, and the confidence interval CI = 0.98 &ndash; 1.00. The congruence of the tests findings was also confirmed by the McNemar test, estimated to 0.99. The stability of the secreted MPT64 antigen was registered even five years after the first testing episode. CONCLUSION: The high sensitivity and specificity of the imunochromatographic test enable an accurate and precise identification of M. tuberculosis, as well as a reliable differentiation of M. tuberculosis from NTM. The immunochromatographic test may substitute conventional biochemical and molecular tests to identify M. tuberculosis. It is easier to perform and provides faster test results, thus reducing the time of establishing the diagnosis.</p>

O Laboratório de Análises Clínicas (LAC) vem buscando, no decorrer dos anos, alternativas quanto à sua capacidade em gerar receita. A busca por melhores resultados incrementou a parceria com as organizações chamadas de \"convênios médicos\" ou \"medicina de grupo\" que, em geral, remuneram as análises laboratoriais com base em tabelas de procedimentos criadas pela Associação Médica Brasileira AMB. Destas, a mais utilizada é a Tabela AMB 92, devido a utilizar valores de Coeficiente de Honorários (CH), que convertidos em reais, são mais interessantes para os LAC. Este estudo teve como base o método recomendado pelo \"National Committee for Clinical Laboratory Standards\" - NCCLS, que normatiza a apuração do custo baseado na atividade desempenhada durante a sua realização. O objetivo deste trabalho foi estabelecer o custo das análises laboratoriais e verificar se, em comparação à tabela AMB 92, a opção pelos contratos com os \"convênios médicos\" realmente gera lucro a um laboratório de pequeno porte. Para tanto foram selecionadas as análises mais solicitadas no mês de agosto de 2004 em um laboratório de pequeno porte, que presta atendimento a pacientes conveniados a planos de saúde e particulares. O ressarcimento aos laboratórios prestadores de serviço é efetuado com base no valor do CH, que pode variar de acordo com o contrato firmado com os convênios médicos. Os resultados mostraram que na comparação dos custos unitários das 69 análises apuradas com a Tabela AMB-92 houve lucro em 52% das análises quando o valor de CH foi de R$ 0,2610 e quando atribuído um valor de CH de R$ 0,1800, houve lucro em apenas 28% das análises. / The Clinical Laboratory (CL) has been seeking, throughout these years, alternative forms of increasing budget. The search for better results has flourished partnerships with organizations named \"prepaid group practice\" or \"group medicine\" which usually reward the clinical analysis based on tables of procedures established by the Brazilian Medicai Association - BMA. Among them, the most employed is the BMA 92 Table, due to the use of payment coefficient values (PC) which, expressed in Brazilian currency (reais), are in CLs interests. This study was carried out based on the method recommended by the National Committee for Clinical Laboratory Standards - NCCLS, who draws up the regulation of the cost estimate of a procedure while it is carried out. The objective of this study was to establish the cost of clinical analysis and to verify if, comparing to the BMA 92 Table, the contracts with prepaid group practices are actually profitable for a small size laboratory. In order to achieve it, most requested analysis during August 2004 in a small size laboratory which offers services to patients affiliated with prepaid health plans and private health plans. The payment to the laboratories which offer services is made regarding the PC values, which may change according to the contract with the health plans. The results demonstrated that in comparison to the cost of each one of 69 analysis verified according to the BMA 92 Table, 52% of the analysis were profitable when the PC value was R$ 0.2610 and for the PC value of R$ 0.1800 only 28% of the analysis were profitable.

O Laboratório de Análises Clínicas (LAC) vem buscando, no decorrer dos anos, alternativas quanto à sua capacidade em gerar receita. A busca por melhores resultados incrementou a parceria com as organizações chamadas de \"convênios médicos\" ou \"medicina de grupo\" que, em geral, remuneram as análises laboratoriais com base em tabelas de procedimentos criadas pela Associação Médica Brasileira AMB. Destas, a mais utilizada é a Tabela AMB 92, devido a utilizar valores de Coeficiente de Honorários (CH), que convertidos em reais, são mais interessantes para os LAC. Este estudo teve como base o método recomendado pelo \"National Committee for Clinical Laboratory Standards\" - NCCLS, que normatiza a apuração do custo baseado na atividade desempenhada durante a sua realização. O objetivo deste trabalho foi estabelecer o custo das análises laboratoriais e verificar se, em comparação à tabela AMB 92, a opção pelos contratos com os \"convênios médicos\" realmente gera lucro a um laboratório de pequeno porte. Para tanto foram selecionadas as análises mais solicitadas no mês de agosto de 2004 em um laboratório de pequeno porte, que presta atendimento a pacientes conveniados a planos de saúde e particulares. O ressarcimento aos laboratórios prestadores de serviço é efetuado com base no valor do CH, que pode variar de acordo com o contrato firmado com os convênios médicos. Os resultados mostraram que na comparação dos custos unitários das 69 análises apuradas com a Tabela AMB-92 houve lucro em 52% das análises quando o valor de CH foi de R$ 0,2610 e quando atribuído um valor de CH de R$ 0,1800, houve lucro em apenas 28% das análises. / The Clinical Laboratory (CL) has been seeking, throughout these years, alternative forms of increasing budget. The search for better results has flourished partnerships with organizations named \"prepaid group practice\" or \"group medicine\" which usually reward the clinical analysis based on tables of procedures established by the Brazilian Medicai Association - BMA. Among them, the most employed is the BMA 92 Table, due to the use of payment coefficient values (PC) which, expressed in Brazilian currency (reais), are in CLs interests. This study was carried out based on the method recommended by the National Committee for Clinical Laboratory Standards - NCCLS, who draws up the regulation of the cost estimate of a procedure while it is carried out. The objective of this study was to establish the cost of clinical analysis and to verify if, comparing to the BMA 92 Table, the contracts with prepaid group practices are actually profitable for a small size laboratory. In order to achieve it, most requested analysis during August 2004 in a small size laboratory which offers services to patients affiliated with prepaid health plans and private health plans. The payment to the laboratories which offer services is made regarding the PC values, which may change according to the contract with the health plans. The results demonstrated that in comparison to the cost of each one of 69 analysis verified according to the BMA 92 Table, 52% of the analysis were profitable when the PC value was R$ 0.2610 and for the PC value of R$ 0.1800 only 28% of the analysis were profitable.

Dengue fever (DF) and dengue hemorrhagic fever (DHF) are emerging infectious diseases which are endemic in many regions of the globe, many of which are resource-poor areas. DHF and DF impose a severe economic health burden in tropical and subtropical areas. Dengue virus causes an acute febrile illness that can be a self-limited febrile illness, as seen in most cases of DF, or a life-threatening illness with plasma leakage and shock, as seen in cases of DHF. A systematic review of the literature revealed gaps in the knowledge base of clinical laboratory findings of dengue illness with regards to longitudinal dynamics and classification and predictive modeling of disease severity. The objective of this thesis was to investigate the utility of clinical laboratory variables for classification and prediction of disease outcomes. The data used in this investigation was derived from a prospective study of Thai children presenting to either of two study hospitals within 72 hours of onset of an acute febrile illness. Systematic data collection, including clinical laboratory parameters, and routine clinical management continued each day until 24 hours after the fever had subsided. A final diagnosis of DHF, DF, or other febrile illness (OFI) was assigned by an expert physician after chart review. The first research objective of this study was to describe the temporal dynamics of clinical laboratory parameters among subjects with DHF, DF, or OFI. Data were analyzed using lowess curves and population-average models. Quadratic functions of clinical variables over time were established and demonstrated significantly divergent patterns between the various diagnostic groups. The second research objective was to establish and validate tools for classification of illness severity using easily obtained clinical laboratory measures. Bivariate logistic regression models were established using data from one hospital in an urban area of Thailand as a training data set and validated with a second data set from a hospital in a rural area of Thailand. The validated models maintained a high sensitivity and specificity in distinguishing severe dengue illnesses without using the hallmark indicators of plasma leakage. The third research objective used classification and regression tree (CART) analysis to established diagnostic decisions trees using data obtained on the day of study enrollment, within the first 3 days of acute illness. Decision trees with high sensitivity were established for severe dengue defined either as: 1) DHF with evidence of shock (dengue shock syndrome, DSS); or 2) DSS or dengue with significant pleural effusion. This study expands existing knowledge of the potential utility of clinical laboratory variables during different phases of dengue illness. The application of the results of these studies should lead to promising opportunities in the fields of epidemiological research and disease surveillance to reduce the health burden, and improve the clinical management, of dengue illness. Future directions involve application of these algorithms to different study populations and age groups. Additionally, other analytical techniques, such as those involving CART analysis, can be explored with these data.

INTRODUÇÃO: As infecções por bactérias anaeróbias são geralmente de origem endógena, polimicrobianas e mistas. Devido a sua natureza fastidiosa, essas bactérias necessitam de uma prévia incubação em meios líquidos enriquecidos, como o caldo Thioglicolato (CT) para serem recuperadas, o isolamento desses microrganismos é trabalhoso e o tempo de resposta - TAT (turn around time) estendido desse exame pode estar associado a falhas terapêuticas e ao aumento da resistência bacteriana. A cultura de anaeróbios (CANA) ainda é um desafio para os laboratórios clínicos de rotina e novas estratégias para diminuir o TAT são fundamentais para que esse exame forneça um impacto clínico significativo. OBJETIVO: Otimizar o processo de triagem da CANA pela modificação do CT; comparar a identificação dos anaeróbios pelas metodologias fenotípicas ANC (Vitek 2- bioMérieux, France) e MALDI-TOF (Vitek MS - bioMérieux, France) e verificar o impacto econômico das ações propostas MÉTODOS: O caldo de triagem CT foi modificado eluindo individualmente discos comerciais de antibióticos (em concentrações fixas) selecionados por apresentarem baixa ou nenhuma ação contra microrganismos anaeróbios e com um bom espectro de ação para os principais aeróbios associados em culturas mistas e foram escolhidos aqueles que após uma bateria de testes frente a 15 cepas dos principais anaeróbios envolvidos em infecções humanas mantiveram a viabilidade inicial. O caldo Thioglicolato modificado (CTM) foi composto pela adição dos antibióticos que apresentaram a melhor \"performance\" acima descrita. A sensibilidade e especificidade do CTM foram avaliadas paralelamente com CT na rotina de CANA do HCFMUSP. Para a avaliar a identificação fenotípica, 421 anaeróbios isolados no período de seis meses foram submetidos a identificação pelo ANC (Vitek 2) e MALDI-TOF (Vitek MS). Os resultados discordantes ou com baixa discriminação da espécie foram avaliados pelo sequenciamento 16S rRNA. O impacto econômico da introdução do CTM bem como os custos diretos da identificação pelo MALDI-TOF foram avaliados. RESULTADOS: O CTM foi composto por amicacina, gentamicina e aztreonam. Das 159 amostras clínicas triadas pelo CT e CTM, 11 (7%) foram positivas para CANA com as mesmas espécies isoladas em ambos os meios. Utilizando o CTM, foi obtida uma redução dos falsos positivos de 97 (61%) para 69 (43%) quando comparado ao CT (p < 0,05). O TAT do resultado negativo da CANA com o CTM foi reduzido de 14 para sete dias em 28 (18%) amostras; o CTM permitiu a liberação do resultado positivo da CANA 48 horas à frente do CT. A sensibilidade do CTM foi igual ao CT, porém a especificidade foi superior em 19%. Das 421 cepas avaliadas, 35 foram identificadas somente pelo MALDI-TOF (Vitek MS) sendo que uma (Clostridium innocum) foi identificada somente pelo sequenciamento 16S rRNA. Das 386 avaliadas por ambas as metodologias, houve uma concordância de 97% e os resultados das 13 (3%) cepas submetidas ao sequenciamento foram concordantes em 92% com o MALDI-TOF (Vitek MS) que promoveu a redução do TAT do resultado positivo em cinco dias. A implementação do CTM possibilitou uma redução de custos nessa amostragem, de R$ 2.240,00 e a identificação pelo MALDI-TOF proporcionou uma economia de R$ 7.786,00. Considerando os valores econômicos encontrados nesse estudo e projetando-os nas estatísticas de CANA do HCFMUSP em 2017, o CTM poderia proporcionar uma economia de R$ 132.560,00 /ano e o MALDI-TOF uma redução nos gastos de R$ 13.579,00/ ano CONCLUSÕES: A padronização e implementação do CTM permitiu uma um aumento significativo de especificidade da cultura anaeróbia com redução do TAT e dos custos. A utilização do MALDI-TOF diminuiu o TAT das identificações aliado a uma melhor performance de forma custo efetiva / INTRODUCTION: Anaerobic bacterial infections are usually of endogenous origin, polymicrobial and mixed. Because of their fastidious nature, these bacteria require prior incubation in enriched liquid media, such as Thioglycolate broth (TB) to be recovered, the isolation of these microorganisms is laborious, and the TAT (turn around time) extended time of this examination may be associated with therapeutic failures and increased bacterial resistance. Anaerobic culture (AC) is still a challenge for routine clinical laboratories, and new strategies for lowering TAT are critical to provide a significant clinical impact. OBJECTIVE: To optimize the AC screening process by modifying the TB; Compare anaerobical identification between (Vitek 2- bioMérieux, France) and MALDI-TOF (Vitek MS - bioMérieux, France) and to verify the economic impact of the proposed actions. METHODS: TB broth was modified by eluting individually antibiotic commercial discs (at fixed concentrations) selected for low or no action against anaerobic microorganisms and with a good action spectrum for the main associated aerobes in mixed cultures. Those who maintained the initial viability after a battery of tests against 15 strains of the major anaerobes involved in human infections were selected. Modified Thioglycolate Broth (MTB) was composed of the antibiotics that presented the best performance described above. The sensitivity and specificity of MTB were evaluated in parallel with TB in the HCFMUSP AC routine. To evaluate the phenotypic identification, 421 anaerobes isolated in the six-month period were submitted to identification by ANC (Vitek 2) and MALDI-TOF (Vitek MS). Discordant results or those with low discrimination of the species were submitted to 16S rRNA sequencing. The economic impact of the introduction of MTB as well as the direct costs of MALDI-TOF identification were assessed. RESULTS: MTB was composed of amikacin, gentamicin and aztreonam. Of the 159 clinical samples screened by TB and MTB, 11 (7%) were positive for AC with the same species isolated in both media. Using MTB, a reduction of false positives was obtained from 97 (61%) to 69 (43%) when compared to TB (p < 0.05). The TAT of the negative result of the AC with the MTB was reduced from 14 to 7 days in 28 (18%) samples; the MTB allowed the release of the AC positive result 48 hours ahead of the TB. The sensitivity of MTB was equal to TB, but the specificity was higher in 19%. Of the 421 strains evaluated, 35 were identified only by MALDI-TOF (Vitek MS) and one (Clostridium innocum) was identified only by 16S rRNA sequencing. Of the 386 evaluated by both methodologies, there was a concordance of 97% and the results of the 13 (3%) strains submitted to the sequencing were concordant in 92% with the MALDI-TOF (Vitek MS) that promoted TAT of the positive result reduction in five days. The implementation of the MTB made possible a reduction of costs in this sampling, of US $ 677,00 and the identification by MALDI-TOF provided a saving of US $ 2354,00. Considering the economic values found in this study and projecting them in the HCFMUSP AC statistics in 2017, the MTB could provide savings of US $40,070.00 / year and MALDI-TOF a reduction in expenses of US $ 4,100.00 / year. CONCLUSIONS: Standardization and implementation of MTB allowed a significant increase of anaerobic culture specificity with TAT and costs reduction. The use of MALDI-TOF reduced the TAT of the identifications and also resulted in a better performance in a cost effective way

INTRODUÇÃO: As infecções por bactérias anaeróbias são geralmente de origem endógena, polimicrobianas e mistas. Devido a sua natureza fastidiosa, essas bactérias necessitam de uma prévia incubação em meios líquidos enriquecidos, como o caldo Thioglicolato (CT) para serem recuperadas, o isolamento desses microrganismos é trabalhoso e o tempo de resposta - TAT (turn around time) estendido desse exame pode estar associado a falhas terapêuticas e ao aumento da resistência bacteriana. A cultura de anaeróbios (CANA) ainda é um desafio para os laboratórios clínicos de rotina e novas estratégias para diminuir o TAT são fundamentais para que esse exame forneça um impacto clínico significativo. OBJETIVO: Otimizar o processo de triagem da CANA pela modificação do CT; comparar a identificação dos anaeróbios pelas metodologias fenotípicas ANC (Vitek 2- bioMérieux, France) e MALDI-TOF (Vitek MS - bioMérieux, France) e verificar o impacto econômico das ações propostas MÉTODOS: O caldo de triagem CT foi modificado eluindo individualmente discos comerciais de antibióticos (em concentrações fixas) selecionados por apresentarem baixa ou nenhuma ação contra microrganismos anaeróbios e com um bom espectro de ação para os principais aeróbios associados em culturas mistas e foram escolhidos aqueles que após uma bateria de testes frente a 15 cepas dos principais anaeróbios envolvidos em infecções humanas mantiveram a viabilidade inicial. O caldo Thioglicolato modificado (CTM) foi composto pela adição dos antibióticos que apresentaram a melhor \"performance\" acima descrita. A sensibilidade e especificidade do CTM foram avaliadas paralelamente com CT na rotina de CANA do HCFMUSP. Para a avaliar a identificação fenotípica, 421 anaeróbios isolados no período de seis meses foram submetidos a identificação pelo ANC (Vitek 2) e MALDI-TOF (Vitek MS). Os resultados discordantes ou com baixa discriminação da espécie foram avaliados pelo sequenciamento 16S rRNA. O impacto econômico da introdução do CTM bem como os custos diretos da identificação pelo MALDI-TOF foram avaliados. RESULTADOS: O CTM foi composto por amicacina, gentamicina e aztreonam. Das 159 amostras clínicas triadas pelo CT e CTM, 11 (7%) foram positivas para CANA com as mesmas espécies isoladas em ambos os meios. Utilizando o CTM, foi obtida uma redução dos falsos positivos de 97 (61%) para 69 (43%) quando comparado ao CT (p < 0,05). O TAT do resultado negativo da CANA com o CTM foi reduzido de 14 para sete dias em 28 (18%) amostras; o CTM permitiu a liberação do resultado positivo da CANA 48 horas à frente do CT. A sensibilidade do CTM foi igual ao CT, porém a especificidade foi superior em 19%. Das 421 cepas avaliadas, 35 foram identificadas somente pelo MALDI-TOF (Vitek MS) sendo que uma (Clostridium innocum) foi identificada somente pelo sequenciamento 16S rRNA. Das 386 avaliadas por ambas as metodologias, houve uma concordância de 97% e os resultados das 13 (3%) cepas submetidas ao sequenciamento foram concordantes em 92% com o MALDI-TOF (Vitek MS) que promoveu a redução do TAT do resultado positivo em cinco dias. A implementação do CTM possibilitou uma redução de custos nessa amostragem, de R$ 2.240,00 e a identificação pelo MALDI-TOF proporcionou uma economia de R$ 7.786,00. Considerando os valores econômicos encontrados nesse estudo e projetando-os nas estatísticas de CANA do HCFMUSP em 2017, o CTM poderia proporcionar uma economia de R$ 132.560,00 /ano e o MALDI-TOF uma redução nos gastos de R$ 13.579,00/ ano CONCLUSÕES: A padronização e implementação do CTM permitiu uma um aumento significativo de especificidade da cultura anaeróbia com redução do TAT e dos custos. A utilização do MALDI-TOF diminuiu o TAT das identificações aliado a uma melhor performance de forma custo efetiva / INTRODUCTION: Anaerobic bacterial infections are usually of endogenous origin, polymicrobial and mixed. Because of their fastidious nature, these bacteria require prior incubation in enriched liquid media, such as Thioglycolate broth (TB) to be recovered, the isolation of these microorganisms is laborious, and the TAT (turn around time) extended time of this examination may be associated with therapeutic failures and increased bacterial resistance. Anaerobic culture (AC) is still a challenge for routine clinical laboratories, and new strategies for lowering TAT are critical to provide a significant clinical impact. OBJECTIVE: To optimize the AC screening process by modifying the TB; Compare anaerobical identification between (Vitek 2- bioMérieux, France) and MALDI-TOF (Vitek MS - bioMérieux, France) and to verify the economic impact of the proposed actions. METHODS: TB broth was modified by eluting individually antibiotic commercial discs (at fixed concentrations) selected for low or no action against anaerobic microorganisms and with a good action spectrum for the main associated aerobes in mixed cultures. Those who maintained the initial viability after a battery of tests against 15 strains of the major anaerobes involved in human infections were selected. Modified Thioglycolate Broth (MTB) was composed of the antibiotics that presented the best performance described above. The sensitivity and specificity of MTB were evaluated in parallel with TB in the HCFMUSP AC routine. To evaluate the phenotypic identification, 421 anaerobes isolated in the six-month period were submitted to identification by ANC (Vitek 2) and MALDI-TOF (Vitek MS). Discordant results or those with low discrimination of the species were submitted to 16S rRNA sequencing. The economic impact of the introduction of MTB as well as the direct costs of MALDI-TOF identification were assessed. RESULTS: MTB was composed of amikacin, gentamicin and aztreonam. Of the 159 clinical samples screened by TB and MTB, 11 (7%) were positive for AC with the same species isolated in both media. Using MTB, a reduction of false positives was obtained from 97 (61%) to 69 (43%) when compared to TB (p < 0.05). The TAT of the negative result of the AC with the MTB was reduced from 14 to 7 days in 28 (18%) samples; the MTB allowed the release of the AC positive result 48 hours ahead of the TB. The sensitivity of MTB was equal to TB, but the specificity was higher in 19%. Of the 421 strains evaluated, 35 were identified only by MALDI-TOF (Vitek MS) and one (Clostridium innocum) was identified only by 16S rRNA sequencing. Of the 386 evaluated by both methodologies, there was a concordance of 97% and the results of the 13 (3%) strains submitted to the sequencing were concordant in 92% with the MALDI-TOF (Vitek MS) that promoted TAT of the positive result reduction in five days. The implementation of the MTB made possible a reduction of costs in this sampling, of US $ 677,00 and the identification by MALDI-TOF provided a saving of US $ 2354,00. Considering the economic values found in this study and projecting them in the HCFMUSP AC statistics in 2017, the MTB could provide savings of US $40,070.00 / year and MALDI-TOF a reduction in expenses of US $ 4,100.00 / year. CONCLUSIONS: Standardization and implementation of MTB allowed a significant increase of anaerobic culture specificity with TAT and costs reduction. The use of MALDI-TOF reduced the TAT of the identifications and also resulted in a better performance in a cost effective way