Complicatedness associated to whole blood discard in the Hemotherapy Service in the National Institute of Cancer / Complicações associadas ao descarte de sangue total no Serviço de Hemoterapia do Instituto Nacional de Cancer

Objective: The aim of this study is to minimize the whole blood discharge in the donor collection room to increase viable total blood units. Methods: The data were extracted from the computer systems of Hemotherapy Service in the National Institute of Cancer from 2012 to 2015. The bivariate analysis in Excel® was used. Results: 46,478 (100%) whole blood unit were collected, out of which 44,686 (96.14%) were used, 1,792 (3.86%) were discarded at donor collection room. The discard indicators were: slow flow 576 (32.14%), difficult venous access 438 (24.44%), interrupted flow 293 (16.35%), reaction during collection 198 (11.05%) and high volume 142 (7.92%). A discard of 2,722 blood bag inputs and 14 complications were noticed. Conclusion: In donor reactions, we observed psychological and physiological factors that must be considered during puncture. The research suggests it is possible to minimize the discard and optimize the quality of service with the implementation of a theoretical/practical training on complications related to non-compliances, as well as the review and updating of documents. A trained, cohesive and stable collection team helps to minimize the discard of whole blood unit due to complications.

Objetivo: Minimizar o descarte de sangue total na sala de coleta de doadores para aumentar as unidades de sangue total viáveis. Métodos: Os dados foram extraídos dos sistemas informatizados do Serviço de Hemoterapia do Instituto Nacional de Câncer de 2012 a 2015. A análise bivariada Excel® foi usada. Resultados: Foram coletadas 46.478 (100%) unidades de sangue total, das quais 44.686 (96,14%) foram utilizadas, 1.792 (3,86%) foram descartadas na sala de coleta do doador. Os indicadores de descarte foram: fluxo lento 576 (32,14%), acesso venoso difícil 438 (24,44%), fluxo interrompido 293 (16,35%), reação durante a coleta 198 (11,05%) e alto volume 142 (7,92%). Descartes de 2.722 insumos de bolsa de sangue e 14 complicações foram notados. Conclusão: Nas reações dos doadores, observamos fatores psicológicos e fisiológicos que devem ser considerados durante a punção. A pesquisa sugere que é possível minimizar o descarte e otimizar a qualidade do serviço com a implementação de um treinamento teórico/prático sobre complicações relacionadas a não conformidades, bem como a revisão e atualização de documentos. Uma equipe de coleta treinada, coesa e estável ajuda a minimizar o descarte de sangue total devido a complicações.

Sintomas osteomusculares, absenteísmo e fatores associados em citotecnologistas: estudo transversal / Musculoskeletal complaints, absenteeism and associated factors among cytotechnologists: cross-sectional study

Introdução: Citotecnologistas realizam leitura de lâminas citológicas utilizando o microscópio óptico. Postura estática, velocidade e repetição dos movimentos são fatores que acarretam o aparecimento de sintomas osteomusculares. Objetivos: Identificar os principais sintomas osteomusculares que afetam os citotecnologistas do Instituto Nacional de Câncer (INCA). Estudar possíveis associações entre absenteísmo de citotecnologistas por sintomas osteomusculares e variáveis individuais e profissionais. Caracterizar o absenteísmo por doenças dos citopatologistas do INCA, entre 2016 e 2017, no que tange o grupo de doenças do sistema osteomuscular. Método: Estudo transversal, baseado na aplicação do Questionário Nórdico de Sintomas Osteomusculares. Por meio dos dados do questionário, realizou-se a associação entre as variáveis de exposição e o absenteísmo. Além disso, foram analisados os registros de doenças na Divisão de Saúde do Trabalhador (DISAT) para verificar as principais doenças que resultaram em absenteísmo. As associações foram testadas por meio do Teste de Fisher utilizando o SPSS, versão 20.0, e a significância estatística considerada foi de p<0,05. Resultados: Do total, 34,4% relataram afastamento do trabalho por sintomas osteomusculares. As principais queixas musculoesqueléticas são na coluna cervical (18%). De acordo com os registros da DISAT, os principais motivos de absenteísmo foram por doenças do sistema osteomuscular e do tecido conjuntivo (25%). Observou-se relação estatisticamente significativa entre o tempo de trabalho, Índice de Massa Corporal (IMC) e o absenteísmo por sintomas osteomusculares. Conclusão: As doenças do sistema osteomuscular e do tecido conjuntivo foram os principais motivos de afastamento dos citotecnologistas, existindo associação do absenteísmo com tempo de serviço e IMC elevado.

Background: Cytotechnologists are laboratory professionals who analyze cytology slides under optical microscopes. Static postures, speed and repetitive movements are associated with occurrence of musculoskeletal complaints. Objective: To establish the main musculoskeletal complaints among cytotechnologists at the National Cancer Institute, Brazil, test associations between absenteeism due to musculoskeletal complaints and individual and occupational variables, and characterize absenteeism related to diseases of the musculoskeletal system in 2016 and 2017. Method: Cross-sectional study in which we administered the Nordic Musculoskeletal Questionnaire and tested associations between exposure variables and absenteeism. We also analyzed morbidity records kept at the Occupational Health Division (OHD) to establish the main disorders related to absenteeism. Associations were investigated by means of Fisher's test using SPSS version 20.0. The significance level was set to p<0.05. Results: 34.4% of the sample required sick leave due to musculoskeletal complaints. The most affected body site was the neck (18%). As per the OHD records, sickness absenteeism was mainly due to diseases of the musculoskeletal system and connective tissue (25%). We found statistically significant association of absenteeism with length in the job and body mass index. Conclusion: Diseases of the musculoskeletal system and connective tissue were the main reason for missing work days. Absenteeism was associated with length in the job and high body mass index.

Detection of multidrug-resistant Enterobacteriaceae isolated from river waters flowing to the Guanabara Bay and from clinical samples of hospitals in Rio de Janeiro, Brazil / Detección de enterobacterias multirresistentes aisladas en aguas de los ríos que desembocan en la bahía de Guanabara y en muestras de hospitales de Río de Janeiro, Brasil

Abstract Introduction: The use of antibiotics in humans, animal husbandry and veterinary activities induces selective pressure leading to the colonization and infection by resistant strains. Objective: We evaluated water samples collected from rivers of the Guanabara Bay, which have suffered minor and major environmental degradation, and clinical samples of hospital origin to detect evidence of the presence of resistance genes to aminoglycosides, beta-lactam antibiotics and fluoroquinolones in strains of Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae and Escherichia coli. Materials and methods: For isolation of the water strains we employed culture media containing 32 μg/ml cephalotin and 8 μg/ml gentamicin. The strains from clinical materials were selected using culture media containing 8 μg/ml gentamicin. The strains were identified and subjected to antimicrobial susceptibility testing (AST), plasmid DNA extraction and polymerase chain reaction (PCR) to detect genes encoding enzymes modifying aminoglycosides (EMA), extended-spectrum beta-lactamases (ESBL) and plasmid mechanisms of quinolone resistance (PMQR). Results: The AST of the isolates recovered from water samples showed multidrug-resistance profiles similar to those found in isolates recovered from clinical materials. All isolates from water samples and 90% of the isolates from clinical samples showed at least one plasmid band. In the PCR assays, 7.4% of the isolates recovered from water samples and 20% of those from clinical materials showed amplification products for the three antimicrobial classes. Conclusion: We believe that the detection of microorganisms presenting genetic elements in environments such as water is necessary for the prevention and control of their dissemination with potential to infect humans and other animals in eventual contact with these environments.

Resumen Introducción. El uso de antibióticos en seres humanos, en la industria pecuaria y en las actividades veterinarias induce una presión selectiva que resulta en la colonización e infección con cepas resistentes. Objetivo. Determinar la presencia de genes de resistencia a aminoglucósidos, betalactámicos y fluoroquinolonas en cepas de Klebsiella pneumoniae subsp. pneumoniae, K. pneumoniae subsp. ozaenae y Escherichia coli, obtenidas de muestras de agua de los ríos que desembocan en la bahía de Guanabara y de muestras clínicas de hospitales de Río de Janeiro. Materiales y métodos. En la selección de las cepas resistentes obtenidas de las muestras de agua de los ríos, se emplearon medios de cultivo que contenían 32 μg/ml de cefalotina y 8 μg/ ml de gentamicina. En el caso de las muestras de especímenes clínicos, se usaron medios de cultivo que contenían 8 μg/ml de gentamicina. Las cepas se identificaron y se sometieron a pruebas de sensibilidad antimicrobiana, extracción de ADN plasmídico y pruebas de reacción en cadena de la polimerasa (PCR) para detectar los genes que codifican aquellas enzimas que modifican los aminoglucósidos, las betalactamasas de espectro extendido (BLEE) y los mecanismos de resistencia a las quinolonas mediados por plásmidos. Resultados. Se encontraron perfiles de resistencia a los antimicrobianos similares en los dos grupos. En todas las bacterias obtenidas de las muestras de agua y en 90 % de las muestras clínicas, se evidenciaron bandas de plásmidos asociados con la transferencia de genes de resistencia. En las pruebas de PCR, se obtuvieron productos de amplificación de los genes de resistencia para las tres clases de antimicrobianos analizados, en el 7,4 % de las bacterias recuperadas de las muestras de agua y en el 20 % de aquellas recuperadas de las muestras clínicas. Conclusión. La detección de microorganismos con elementos genéticos que confieren resistencia a los antibióticos en ambientes como el agua, es una estrategia necesaria para prevenir y controlar la diseminación de estos agentes patógenos con potencial para infectar a humanos y a otros animales en dichos ambientes.

Association between clinical indications and coproparasitological tests carried out at the Pedro Ernesto University Hospital (HUPE/UERJ) in Rio de Janeiro, Brazil

Enteroparasitosis is a public health problem in Brazil. Clinical indications and the appropriate stool examination are essential to obtain an adequate result. This study aims to evaluate whether the clinical indications and the choice of coproparasitological tests requested by the medical services may influence the diagnosis of enteroparasitosis. The data was obtained from the records in the Laboratory of Parasitology at the Pedro Ernesto University Hospital (HUPE/ UERJ) of the State University of Rio de Janeiro (UERJ) from 2009 to 2014. The qualitative variables were grouped in medical services (medical surgery, infectious and parasitic diseases, gastroenterology, pediatrics and rheumatology); types of tests requested (parasitological stool examination (PSE), merthiolate-iodine-formaldehyde (MIF), and sodium-acetate acetic acidformaldehyde (SAF)) and clinical indications (anemia, diarrhea, abdominal pain, eosinophilia, routine tests, HTLV patients, HIV patients, parasitosis and transplantation research). The chi square (X²) and the Spearman coefficient correlation tests were performed to calculate the association between the clinical indications and the coproparasitological tests. A significant association was evident in the clinical indication: parasitosis found among the MIF tests and Trichrome Wheatley (ρ = 0.980). In other clinical indications such as anemia, surgery/ transplant, diarrhea, patients with HIV, HTLV and eosinophilia (despite the PSE tests and MIF having presented a strong link (ρ = 0.802), there was no significant association among the tests. Clinical indications are essential and they have a great influence on the parasitological diagnosis, requiring a combination of diagnostic methods for the detection of protozoa and helminths of medical interest.

Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents

During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins.

Detection and differentiation of Cryptosporidium by real-time polymerase chain reaction in stool samples from patients in Rio de Janeiro, Brazil

This study reports the first genetic characterisation of Cryptosporidium isolates in Brazil using real-time polymerase chain reaction (RT-PCR). A total of 1,197 faecal specimens from children and 10 specimens from human immunodeficiency virus-infected patients were collected between 1999-2010 and screened using microscopy. Forty-eight Cryptosporidium oocyst-positive isolates were identified and analysed using a generic TaqMan assay targeting the 18S rRNA to detect Cryptosporidium species and two other TaqMan assays to identify Cryptosporidium hominis and Cryptosporidium parvum. The 18S rRNA assay detected Cryptosporidium species in all 48 of the stool specimens. The C. parvum TaqMan assay correctly identified five/48 stool samples, while 37/48 stool specimens were correctly amplified in the C. hominis TaqMan assay. The results obtained in this study support previous findings showing that C. hominis infections are more prevalent than C. parvum infections in Brazil and they demonstrate that the TaqMan RT-PCR procedure is a simple, fast and valuable tool for the detection and differentiation of Cryptosporidium species.

β-lactamase producing enterobacteria isolated from surveillance swabs of patients in a cardiac intensive care unit in Rio de Janeiro, Brazil

There is a high incidence of infections caused by betalactamase-producing Gram-negative microorganisms in Brazil. These organisms are of clinical and epidemiological importance, since their mobile genetic elements facilitate cross-infection. The present study was conducted in sentinel rectal swabs from patients admitted to a cardiac surgery hospital in Rio de Janeiro, from January through December 2007, in a consecutive manner. The aim of the study was to characterize the genotype and phenotype of these isolates from colonized patients. Biochemical tests, antimicrobial susceptibility tests, a confirmatory test for the expression of extended spectrum betalactamase (ESBL) production and polymerase chain reaction for the blaTEM, blaSHV, CTX-M1, Toho-1 and AmpC genes were performed at the University Hospital of Universidade do Estado do Rio de Janeiro (UERJ). The most frequently isolated bacteria were Escherichia coli 9/41 (21.95 percent) and Klebsiella pneumoniae 14/41 (34.1 percent). In 24/41 (58 percent), the ESBL genotype was confirmed. The most prevalent genes in samples that expressed ESBL were blaTEM 13/24 (54 percent), AmpC 12/24 (50 percent), blaSHV 6/24 (25 percent), CTX-M1 7/24 (29 percent), and Toho-1 6/24 (25 percent). Of these, 14/24 (58 percent) presented more than one genotype for the tested primers. In nine (37 percent) samples other than E. coli, K. pneumoniae or Proteus spp., the phenotype for ESBL was found and confirmed by PCR. The most sensitive substrate in the approximation test in ESBL positive samples was ceftriaxone (83 percent). Fifty percent of the samples expressed AmpC were associated with other genes. Intermediate susceptibility to ertapenem was found in 2/41 (5 percent).

Detection of Toxoplasma gondii DNA by polymerase chain reaction in experimentally desiccated tissues

Despite toxoplasmosis being a common infection among human and other warm-blooded animals worldwide, there are no findings about Toxoplasma gondii evolutionary forms in ancient populations. The molecular techniques used for amplification of genetic material have allowed recovery of ancient DNA (aDNA) from parasites contained in mummified tissues. The application of polymerase chain reaction (PCR) to paleoparasitological toxoplasmosis research becomes a promising option, since it might allow diagnosis, acquisition of paleoepidemiological data, access to toxoplasmosis information related origin, evolution, and distribution among the ancient populations.Furthermore, it makes possible the analysis of parasite aDNA aiming at phylogenetic studies. To standardize and evaluate PCR applicability to toxoplasmosis paleodiagnostic, an experimental mummification protocol was tested using desiccated tissues from mice infected with the ME49 strain cysts, the chronic infection group (CIG), or infected with tachyzoites (RH strain), the acute infection group (AIG). Tissues were subjected to DNA extraction followed by PCR amplification of T. gondii B1 gene. PCR recovered T. gondii DNA in thigh muscle, encephalon, heart, and lung samples. AIG presented PCR positivity in encephalon, lungs, hearts, and livers. Based on this results, we propose this molecular approach for toxoplasmosis research in past populations.

Random amplified polymorphic DNA analysis of DNA extracted from Trichuris trichiura (Linnaeus, 1771) eggs and its prospective application to paleoparasitological studies

Random amplified polymorphic DNA analysis was applied to DNAs extracted from Trichuris trichiura eggs recovered from human fecal samples. Four out of 6 primers tested displayed 18 distinct and well defined polymorphic patterns, ranging from 650 to 3200 base pairs. These results, upon retrieval and DNA sequencing of some of these bands from agarose gels, might help in establishing T. trichiura specific genetic markers, not available yet, and an important step to design primers to be used in molecular diagnosis approaches

Evidences of gentamicin resistance amplification in Klebsiella pneumoniae isolated from faeces of hospitalized newborns

The intestinal microbiota, a barrier to the establishment of pathogenic bacteria, is also an important reservoir of opportunistic pathogens. It plays a key role in the process of resistance-genes dissemination, commonly carried by specialized genetic elements, like plasmids, phages, and conjugative transposons. We obtained from strains of enterobacteria, isolated from faeces of newborns in a university hospital nursery, indication of phenothypical gentamicin resistance amplification (frequencies of 10-3 to 10-5, compatible with transposition frequencies). Southern blotting assays showed strong hybridization signals for both plasmidial and chromossomal regions in DNA extracted from variants selected at high gentamicin concentrations, using as a probe a labeled cloned insert containing aminoglycoside modifying enzyme (AME) gene sequence originated from a plasmid of a Klebsiella pneumoniae strain previously isolated in the same hospital. Further, we found indications of inactivation to other resistance genes in variants selected under similar conditions, as well as, indications of co-amplification of other AME markers (amikacin). Since the intestinal environment is a scenario of selective processes due to the therapeutic and prophylactic use of antimicrobial agents, the processes of amplification of low level antimicrobial resistance (not usually detected or sought by common methods used for antibiotic resistance surveillance) might compromise the effectiveness of antibiotic chemotherapy