Detección rápida de Enterobacterias productoras de carbapenemasas en hisopados rectales de pacientes neonatos colonizados / Rapid detection of carbapenemase-producing Enterobacteriaceae in rectal swabs of colonized neonatal patients

Resumen Objetivo: Detectar la presencia de Enterobacterias productoras de carbapenemasas en hisopados rectales de neonatos mediante técnica de nefelometría láser y caracterización del tipo de carbapenemasa mediante test inmunocromatográfico. Materiales y Métodos: Estudio descriptivo de corte transversal. Fueron incluidos 57 neonatos, tamizados al ingreso a UCI, mediante hisopado rectal, procesado por nefelometría laser HB&L Carbapenemase (Alifax®) y caracterización del tipo de carbapenemasa por inmunocromatografía rápida RESIST-3 (Coris BioConcept®). Resultados: Encontramos un alto porcentaje de colonización rectal (22.9%) correspondiente a 13 hisopados positivos y 44 (77.1%) fueron negativos por nefelometría láser. Por VITEK 2® se obtuvo identificación de Klebsiella pneumoniae resistente a carbapenémicos en los 13 aislamientos y el test inmunocromatográfico reveló la presencia de carbapenemasas blaKPC en estos aislamientos. Discusión: Estudios evidencian el aumento de la colonización por microorganismos productores de carbapenemasas en neonatos. Los resultados de este estudio demuestran que un porcentaje significativo de neonatos que ingresan a las Unidades de Cuidado Neonatal se encuentran colonizados con Enterobacterias productoras de carbapenemasas en tracto intestinal. Lo anterior constituye un riesgo potencial para su diseminación y posterior desarrollo de brotes, en donde surge la importancia de implementar estrategias de vigilancia activa como la tamización rectal para la detección oportuna de neonatos colonizados.

Abstract Objective: To detect the presence of carbapenemase-producing Enterobacteriaceae in rectal swabs of neonates by means of laser nephelometry technique and characterization of the type of carbapenemase by immunochromatographic test. Materials and Methods: Descriptive cross-sectional study. 57 neonatal patients were included; They underwent rectal screening upon admission to the ICU, using swabs which were processed by HB&L Carbapenemase laser nephelometry (Alifax®) and characterization of the type of carbapenemase by RESIST-3 rapid immu nochromatography (Coris BioConcept®). Results: We found a high percentage of rectal colonization (22.9%) corresponding to 13 positive swabs and 44 samples (77.1%) were negative by laser nephelome try. Identification of carbapenem-resistant Klebsiella pneumoniae was obtained by VITEK 2® in the 13 isolates and the immunochromatographic test revealed the presence of blaKPC carbapenemases in these isolates. Discussion: Studies show increased colonization by carbapenemase-producing microorganisms in neonates. The results of this study demonstrate that a significant percentage of neonates who enter Neonatal Care Units are colonized with Enterobacteriaceae that produce carbapenemases in the intestinal tract. This constitu tes a potential risk for its spread and subsequent development of outbreaks, where the importance of implementing active surveillance strategies such as rectal screening for the timely detection of colonized neonates arises.

Role of Salivary Immune Parameters in Patients With Primary Sjögren's Syndrome

BACKGROUND: Several factors, including clinical manifestations and laboratory data, have been used to evaluate the disease activity of Sjögren's syndrome (SS). We investigated saliva indicators of disease activity in primary SS patients. METHODS: We enrolled 138 Taiwanese patients with primary SS and 100 Taiwanese normal controls. Interleukin (IL)-6, IL-17A, tumor necrosis factor-alpha (TNF-α), and rheumatoid factor (RF)-IgA levels in saliva samples were measured using ELISA or fluorescent enzyme-linked immunoassay. Serum IgG, IgA, and IgM levels were measured by nephelometry. Erythrocyte sedimentation rate (ESR) was measured with an automatic ESR analyzer. The t-test and Pearson correlation test were used. RESULTS: IL-6 level was higher in primary SS patients than in normal controls (14.23±14.77 vs 9.87±7.32, P=0.012), but there were no significant differences in IL-17A, TNF-α, and RF-IgA levels. In primary SS patients, IL-6 level correlated weakly with ESR and IgG levels (r=0.252, P=0.015, and r=0.248, P=0.017, respectively), and TNF-α level correlated weakly with IgG level (r=0.231, P=0.024). CONCLUSIONS: IL-6 may play a role in SS pathogenesis. Saliva IL-6 might be an indicator of disease activity in primary SS patients.

Hereditary angioedema with C1 inhibitor (C1-INH) deficit: the strength of recognition (51 cases)

Hereditary angioedema (HAE) is a rare autosomal dominant disease due to C1 esterase inhibitor deficiency (C1-INH). The disease is characterized by subcutaneous and submucosal edema in the absence of urticaria due to the accumulation of bradykinin. This descriptive study aimed to evaluate the clinical characteristics of patients with a confirmed diagnosis of HAE referred to our Outpatient Clinic between December 2009 and November 2017. Fifty-one patients (38 F, 13 M) with a mean age of 32 years (range: 7-70 y) were included. Family history of HAE was reported in 70% (36/51) of the cases; 33/46 patients became symptomatic by 18 years of age. The median time between onset of symptoms and diagnosis was 13 years (3 mo-50 y). The most frequent triggering factors for attacks were stress (74.4%), trauma (56.4%), and hormonal variations (56%). The main symptoms were subcutaneous edema in 93.5% (43/46) of patients, gastrointestinal symptoms in 84.8% (39/46), and obstruction in the upper airways in 34.8% (16/46). Hospitalization occurred in 65.2%, of whom 13.3% had to be transferred to the Intensive Care Unit. Prophylactic treatment was instituted in 87% (40/46) of patients, and 56.5% (26/46) required additional treatment to control attacks. Owing to our data collection over a period of 8 years, a significant number of patients were identified by this HAE reference center. Despite early recognition and prophylactic treatment, a high percentage of patients were hospitalized. HAE is still diagnosed late, reinforcing the need for more reference centers specialized in diagnosis and educational projects for health professionals.

Quality of bevacizumab (Avastin®) repacked in single-use glass vials for intravitreal administration / Avaliação da qualidade do bevacizumabe (Avastin®) fracionado em frascos de vidro de dose única para administração intravítrea

ABSTRACT Purpose: Avastin® (bevacizumab) is an anti-vascular endothelial growth factor (VEGF) monoclonal antibody given as an off-label drug by intravitreal administration for treatment of ocular diseases. The drug's clinical application and its cost-benefit profile has generated demand for its division into single-use vials to meet the low volume and low-cost doses necessary for intraocular administration. However, the safety of compounding the drug in single-use vials is still under discussion. In this study, the stability and efficacy of Avastin® repacked in individual single-use glass vials and glass ampoules by external compounding pharmacies were evaluated. Methods: Polyacrylamide gel electrophoresis (PAGE), size-exclusion chromatography (SEC), dynamic light scattering (DLS), and turbidimetry were selected to detect the formation of aggregates of various sizes. Changes in bevacizumab biological efficacy were investigated by using an enzyme-linked immunosorbent assay (ELISA). Results: Repacked and reference bevacizumab showed similar results when analyzed by PAGE. By SEC, a slight increase in high molecular weight aggregates and a reduction in bevacizumab monomers were observed in the products of the three compounding pharmacies relative to those in the reference bevacizumab. A comparison of repacked and reference SEC chromatograms showed that the mean monomer loss was ≤1% for all compounding pharmacies. Protein aggregates in the nanometer- and micrometer-size ranges were not detected by DLS and turbidimetry. In the efficacy assay, the biological function of repacked bevacizumab was preserved, with <3% loss of VEGF binding capacity relative to that of the reference. Conclusion: The results showed that bevacizumab remained stable after compounding in ampoules and single-use glass vials; no significant aggregation, fragmentation, or loss of biological activity was observed.

RESUMO Objetivos: Avastin® (bevacizumabe) é um anticorpo monoclonal inibidor do fator de crescimento endotelial de vasos (VEGF) utilizado "off-label" por meio de administração intravítrea para o tratamento de doenças oculares. A sua aplicação clínica associada ao custo-benefício do medicamento gerou uma demanda para seu fracionamento em frascos de dose única para utilização pela via intraocular. No entanto, a segurança do fracionamento do anticorpo em frascos de dose única ainda é alvo de discussão. Neste trabalho, a estabilidade e a eficácia do Avastin® fracionado em frascos ou ampolas de vidro de dose unitária por farmácias de manipulação do mercado foram avaliadas. Métodos: As técnicas de eletroforese em gel de poliacrilamida (PAGE), cromatografia por exclusão de tamanho (SEC), espalhamento dinâmico da luz (DLS) e turbidimetria foram empregadas para avaliar a formação de agregados de diferentes tamanhos. Alterações na atividade biológica do bevacizumabe foram estudadas utilizando ELISA. Resultados: Amostras referência e do bevacizumabe fracionado apresentaram resultados semelhantes quando analisado por gel de poliacrilamida. Por cromatografia por exclusão de tamanho, um pequeno aumento na quantidade de agregados de alta massa molar seguido de uma redução nos monômeros do bevacizumabe foram observados para as amostras das três farmácias de manipulação quando comparado ao referência. A comparação dos cromatogramas mostrou uma quantidade de redução do monômero inferior a 1% para todas as amostras fracionadas. Por espalhamento dinâmico da luz e turbidimetria, não foram detectados agregados de proteína na faixa de tamanho de micrômetro e nanômetro. No ensaio de eficácia, o bevacizumabe fracionado preservou sua função biológica pois apresentou menos de 3% de perda na capacidade de ligação ao VEGF quando comparado ao referência. Conclusão: Este estudo sugere que o bevacizumabe se mantem estável após fracionamento em ampolas e frascos de vidro de dose unitária pois não foram observadas agregação e/ou fragmentação de proteínas e perda de atividade biológica em quan tidades significativas.

The significance of serum IgD quantitation for evaluation of clinical efficacy in IgD multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the significance of serum IgD quantitation in evaluation of clinical efficacy in IgD myeloma.</p><p><b>METHODS</b>Serum IgD and free light chain (sFLC) levels were determined by immune scatter turbidimetry with SPA plus analysis machine in 29 patients with IgD multiple myeloma (MM) achieving VGPR or better response following previous treatments. The concurrent immunofixation electrophoresis (IFE) results were also incorporated and analyzed.</p><p><b>RESULTS</b>Increased IgD levels were detected in 1 of 12 patients achieving sCR, 2 of 5 patients achieving CR and 4 of 12 patients achieving VGPR, respectively. The median progression-free survival (PFS) was 38.5 months, 34.1 months and 15.5 months for patients achieving sCR, CR and VGPR, respectively, with a significant difference between sCR and VGPR groups (P=0.022), and between CR and VGPR groups (P=0.018). There was no difference in overall survival (OS) among sCR, CR and VGPR groups (P>0.05). The median PFS were 7.8, 33.7 and 43.9 months, respectively for the patients with both abnormal sFLC ratios and IgD levels (6 cases, Group A), with either abnormal sFLC ratios or increased IgD levels (10 cases, Group B) or with normal sFLC ratios and IgD levels (13 cases, Group C). A significant PFS benefit of Group A over Group C was found (P=0.033), and no differences in terms of OS among three groups (P>0.05).</p><p><b>CONCLUSION</b>IgD levels may remain abnormal in IgD MM patients who have achieved VGPR or better response, and IgD quantitation represented a useful assay complementary to the current lab examinations. IgD quantitation assay was of significance in clinical efficacy evaluation and survival judgement, and should be incorporated into the evaluation parameters used for IgD MM in addition to sFLC and IFE assays.</p>

Efecto del ejercicio físico de hidrogimnasia sobre la concentración sérica de inmunoglobulina A en mujeres adultas mayores / Effect of hydrogymnastics physical exercise on serum level of immunoglobulin A in elderly women

Background: Decreases in function of the immune system with age contribute to the increase the risk of infectious diseases especially the upper respiratory tract. On the other hand, physical activity has been widely recommended for health. However, more studies are needed to support the benefit effect of exercise on immune system in elderly. Aim: To evaluate the effect of the hydrogymnastics on the serum level of immunoglobulin A (IgA) in elderly women. Methods: Twenty-six participants were randomly assigned to an experimental (n: 16) or a control group (n: 10). IgA concentrations were determined by nephelometry (BN2 Analyzer, Dade Behring). The hydrogymnastics training protocol was performed 5 times a week during 12 weeks. The intensity of the aerobic exercise was 50-60% of the maximum heart rate monitored by telemetry (Polar-FT7). Results: No significant differences (p: 0.797) resulted by ANOVA on serum IgA levels; the percentage of change in experimental group was Δ% = -6.7 mg/dL compared to the control group, Δ%= -7.4 mg/dL. Conclusion: After three months of hydrogymnastics the IgA level did not show significant change on elderly women; However, positive improvement in percentage of change Δ% was found.

Introducción: La disminución de la función del sistema inmune con la edad es asociada a la incidencia de enfermedades infecciosas del tracto respiratorio superior; por otro lado, el ejercicio físico ha sido ampliamente recomendado para la salud. Sin embargo se requiere aún fundamentar con mayor exactitud los efectos del ejercicio físico en el sistema inmunológico en adultos mayores. Objetivo: Evaluar el efecto de un programa de hidrogimnasia sobre la concentración sérica de Inmunoglobulina A (IgA) en adultas mayores. Métodos: 26 participantes fueron asignadas aleatoriamente a un grupo experimental (n: 16) y un grupo control (n: 10). Se determinó la concentración sérica de IgA por nefelometría (BN2 Analyser, DADE Behring). Las sesiones de hidrogimnasia se realizaron cinco veces por semana durante 12 semanas; el ejercicio fue aeróbico a una intensidad de 50-60% de la frecuencia cardíaca máxima controlada por telemetría (Polar-FT7). Resultados: Se utilizó la prueba de ANOVA, indicando no interacción significativa (p: 0,797). El porcentaje de cambio resultó de Δ% = -6,7 mg/dL del grupo experimental en comparación con el grupo control Δ% = -7,4 mg/dL. Conclusiones: Un programa de tres meses de hidrogimnasia en mujeres no mejora significativamente los niveles de IgA. Sin embargo, los datos refieren una mejora positiva el porcentaje de cambio A%.

Effect of immunoglobulin heavy/light chain detection on the minimal residual disease monitoring in IgG multiple myeloma patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the specificity and sensitivity of immunoglobulin heavy/light chain (HLC) and serum free light chain (FLC) level in minimal residual disease monitoring of IgG type multiple myeloma (MM) patients during complete remission (CR).</p><p><b>METHODS</b>Immunoglobulin HLC was assessed in 20 IgG myeloma patients by immune turbidimetry using SPAplus Analyzer. The serum level of HLC and FLC was detected at same time. Combine with those obtained by serum protein electrophoresis (SPE) and immune fixation electrophoresis (IFE), the specificity and sensitivity of HLC in detection of serum immunoglobulin were analyzed. Combined with the clinical efficacy, kappa/lambda ratios of HLC (rHLC) and FLC (rFLC) were compared between the patients and normal controls.</p><p><b>RESULTS</b>Among 20 patients, there were 10 male and 10 female, the median age was 56 years (35-70). There were 6 patients with abnormal rHLC but normal rFLC; 3 patients with abnormal rFLC but normal HLC; and 11 patients with both normal rHLC and rFLC. During the mean follow-up time of 18 months, 4 of the6 patients with abnormal rHLC accepted intervention therapies, 1 case relapsed in 9 months,the other 2 untreated patients relapsed in 3 months. Among the 3 cases with abnormal rFLC, 2 patients are still in remission after intervention therapies,the other untreated patient relapsed in 1.5 months. Among the 11 untreated patients with both normal rHLC and rFLC, 3 relapsed with the disease free survival time of 3.5 months, 5.0 months and 5.5 months respectively.</p><p><b>CONCLUSION</b>The combined detection of HLC and FLC is helpful to assess the curative efficacy and the accuracy of minimal residual disease monitoring, and more effectively evaluate the prognosis of MM patients. Abnormal rHLC and rFLC are correlated with poor prognosis, while early intervention therapies can help to improve disease free survival.</p>

Expression of soluble programmed death-1, soluble programmed death ligand 1 proteins and immune status in patients with oral lichen planus / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the possible role and significance of soluble programmed death-1 (sPD-1)/soluble programmed death ligand 1 (sPD-L1) in the pathogenesis of oral lichen planus (OLP).</p><p><b>METHODS</b>Thirty-six patients with OLP (20 cases of reticular OLP and 16 cases of erosive OLP) were enrolled in this study, and 18 healthy people served as controls. Lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺, CD19⁺, CD16⁺ + 56⁺) were examined by flow cytometric analysis and humoral immunity indexes (IgG, IgA, IgM, C3, C4) tested by nephelometry immunoassay. The levels of sPD-1 and sPD-L1 proteins in serum of patients with OLP were determined by enzyme-linked immunosorbent assay. The correlations between the level of sPD-1, sPD-L1 proteins and the immune status and clinical characteristics of patients with OLP were analyzed by SPSS 19.0.</p><p><b>RESULTS</b>CD3⁺, CD4⁺, CD8⁺, CD16⁺ + 56⁺ in patients with OLP were decreased compared with the normal value, while CD19⁺ in patients with OLP was increased compared with the normal value (P < 0.05). C3 and C4 in patients with OLP were decreased compared with the normal value, but IgM in patients with OLP was increased (P < 0.05). The levels of sPD-1 and sPD-L1 proteins in patients with OLP were significantly higher than that in control group [26.10(8.81, 40.00) ng/L vs 17.65(0.00, 26.10) ng/L, 29.53(21.47, 36.76) ng/L vs 22.79(1.19, 28.29) ng/L] (P < 0.05), but the expression of sPD-1 and sPD-L1 was not related with clinical characteristics of OLP. There were negative correlations between the levels of sPD-1 protein and CD4⁺ T cells or CD16⁺ + 56⁺ cells (r1 = -0.378, P1 = 0.007; r2 = -0.365, P2 = 0.009), while there was a positive correlation between the levels of sPD-1 and CD19⁺ B cells (r = 0.482, P = 0.000). There was a negative correlation between sPD-L1 expression level and CD4⁺ and a positive correlation between sPD-L1 expression level and IgG (r¹ = -0.286, P1 = 0.044; r² = 0.365, P₂= 0.029).</p><p><b>CONCLUSIONS</b>In patients with OLP, the cellular immune function is low with humoral immunity function disorder. PD-1/PD-L1 signaling pathway, which might be influenced by the involvement of sPD-1 and sPD-L1 proteins in a certain extent, may play an important role in the immune pathogenesis of OLP.</p>

Are bovine teeth a suitable substitute for human teeth in in vitro studies to assess endotoxin load in root canals?

The present study aimed to determine the feasibility of using bovine teeth as a suitable alternative for human teeth, in experiments involving in vitro endotoxin contamination. Twenty bovine central incisors and 20 human single-root premolars had their dental crowns removed and root lengths set at 16 mm. Root canals were prepared up to #60 K-file size and sterilized with cobalt-60 gamma irradiation (20 kGy, 6 h). The teeth were randomly divided into four groups: G1-bovine teeth (bovine negative control, n = 10), G2-human teeth (human negative control, n = 10), G3-bovine teeth, inoculated withEscherichia coli (055:B55) LPS, and G4-human teeth inoculated with E. coli LPS. The G1 and G2 groups were exposed to apyrogenic water. After the teeth had been incubated at 37 °C and atmospheric humidity for 24 h, the samples of solutions in the main canals were collected with apyrogenic absorbent paper tips. LPS levels were quantified using Limulus Amebocyte Lysate assay. The data obtained were statistically analyzed using one-way ANOVA, with a significance level of 5%. A high amount of endotoxin was detected in the inoculated human teeth (G4) when compared to the sterilized teeth (G2), as well as in the inoculated bovine teeth (G3) when compared to the inoculated human teeth (G4). However, there was no statistical difference between bovine teeth before and after the E. coli endotoxin inoculation. Therefore, under the mentioned experimental conditions, the use of bovine teeth should not be a choice for laboratory research on endotoxin contamination.

Development and validation of a rapid turbidimetric assay to determine the potency of norfloxacin in tablets

Norfloxacin is one of the first commercially available (and most widely used) fluoroquinolone antibiotics. This paper reports the development and validation of a simple, sensitive, accurate and reproducible turbidimetric assay method to quantify norfloxacin in tablets formulations in only 4 hours. The bioassay is based on the inhibitory effect of norfloxacin upon the strain ofStaphylococcus epidermidis ATCC 12228 IAL 2150 used as test microorganism. The assay was performed 3x3 parallel lines like, three tubes for each concentration of reference substance and three tubes for each sample concentration. The results were treated statistically by analysis of variance and were found to be linear (r2 = 0.9999) in the selected range of 25-100 μg mL-1; precise (intra-assay: relative standard deviation (RSD) = 1.33%; inter-assay: RSD = 0.21%), accurate (100.74%) and robust with RSD lower than 4.5%. The student's t-test showed no statistically significant difference between the proposed turbidimetric method and an HPLC method previously validated. However the turbidimetric assay can be used as a valuable alternative methodology for the routine quality control of this medicine, complementary to other physical-chemical methods.

O norfloxacino foi a primeira fluorquinolona (e mais utilizada) disponível no mercado. Este trabalho divulga um novo desenvolvimento e validação de um método turbidimétrico simples, sensível, preciso e reprodutível para a quantificação de norfloxacino em comprimidos em apenas 4 horas. O bioensaio é baseado no efeito inibitório de norfloxacino sobre a cepa Staphylococcus epidermidis ATCC 12228 IAL 2150, utilizada como micro-organismo teste. O bioensaio foi efetuado através do delineamento de linhas paralelas 3x3, em que três tubos foram utilizados para a solução padrão e três tubos para as concentrações da amostra. Os resultados foram tratados estatisticamente pela análise de variância, apresentando coeficiente de correlação linear der2 = 0,9999, na faixa de 20 a 100 μg mL-1; precisão (intra-ensaio: desvio padrão relativo (RSD) 1,33%; inter-ensaio: RSD=0,21%), exatidão (100,74%) e robustez com RSD menor que 4,5%. O teste-tmostrou não haver diferença estatisticamente significativa entre o método turbidimétrico proposto e um método por HPLC previamente validado. No entanto, o ensaio turbidimétrico pode ser utilizado como método alternativo para o controle de qualidade de rotina para este antimicrobiano, como um complemento de outros métodos físico-químicos.

La formación y el trabajo del agente indígena de salud en el Subsistema de Salud Indígena en Brasil / Professional education and work of indigenous health agents in the Brazilian health system

Objetivo. Reflexionar sobre la figura del agente indígena de salud en Brasil y sobre el papel que éste ejerce en el modelo de atención diferenciada o intercultural. Material y métodos. Se revisó la bibliografía de investigaciones existentes en el área del trabajo y la formación de los agentes indígenas de salud, del subsistema de salud indígena en Brasil. Resultados. Existe subordinación del agente al modelo médico hegemónico. Los agentes carecen de procesos formativos iniciales, los cursos ocurren con irregularidad y los contenidos se enfocan en la biomedicina. Hay conflictos con el equipo y con la comunidad, lo que genera su desvalorización. El agente no ejerce la función de mediación que se espera entre saberes y prácticas. Conclusiones. La discusión sobre la atención diferenciada debe partir de la relación entre el sector salud y la autoatención.

Objective. To discuss the role of indigenous health agents in the implementation of the model of differentiated attention or intercultural health in Brazil. Materials and methods. We revised the scientific literature about the work and professional education of indigenous health agents in the Brazilian indigenous health system. Results. There is a subordination of the agents to the hegemonic medical model. With regards to professional education, we observe the absence and irregularity of these processes, with a general emphasis the biomedicine. There are conflicts with the health team and community, with devaluation of the agents. The agent does not plays the role of mediator between the different health knowledge and practices. Conclusions. We suggest that the discussion of the model of differentiated attention should strengthen the relationship between the health system and the selfcare.

Modelling the risk of mortality of Corbicula fluminea (Müller, 1774) (Bivalvia: Corbiculidae) exposed to different turbidity conditions / Modelagem do risco de mortalidade de Corbicula fluminea (Müller, 1774) (Bivalvia: Corbiculidae) exposta a diferentes condições de turbidez

The provision of sediment in rivers, due to erosion processes that occur in the environment, consists of a major source of pollution and alteration of the physicochemical conditions of water resources. In addition, the increase in water turbidity may cause siltation, dramatically impacting aquatic communities. Specifically considering the bivalve Corbicula fluminea (Müller, 1774), the aim of this study was to analyse the effect of exposure to different turbidity conditions of sediments, as a risk factor for the animals. For this purpose, a docking device was designed to ensure water circulation in a closed system and to maintain the desired levels of turbidity. Although C. fluminea can generally tolerate environmental changes in aquatic systems, an intolerance to high turbidity levels was experimentally observed, expressed by the mortality rate of the animals when exposed to conditions above 150 nephelometric turbidity units (NTU). This value was similar to the one recorded at study sites in the rivers Pardo (Serrana-SP-Brazil) and Mogi Guaçu (Porto Ferreira-SP-Brazil) during the rainy season. Using a logistic regression model, the experimental results were analysed and the observed mortality rates indicate that the exposure of the animals to turbidity levels above 150 nephelometric turbidity units (NTU), for periods longer than 120 hours, may be considered a probable cause of mortality for the species.

A disponibilização de sedimentos nos cursos d'água, em função dos processos erosivos que ocorrem no entorno, constitui-se e uma das principais fontes de poluição e alteração das condições físico-químicas dos recursos hídricos. Além disso, o aumento da turbidez da água pode causar assoreamento, impactando drasticamente as comunidades aquáticas. Especificamente para o bivalve Corbicula fluminea (MÜLLER, 1774), este estudo teve por objetivo analisar o efeito da exposição a diferentes condições de turbidez, como fator de risco para os animals. Para esta finalidade, um dispositivo de acoplamento foi concebido para assegurar a circulação da água, em um sistema fechado mantendo os níveis desejados de turbidez. Embora C. fluminea possa tolerar mudanças ambientais em sistemas aquáticos, foi observadao experimentalmente, uma intolerância a turbidez acima de 150 unidades nefelométricas de turbidez (NTU), um valor próximo ao registrado em pontos de estudo nos rios Pardo (Serrana-SP-Brasil) e Mogi Guaçu (Porto Ferreira-SP-Brasil) durante a estação chuvosa. Por intermédio da utilização de um modelo de regressão logística, os resultados experimentais foram analisados. As taxas de mortalidade observadas indicam ser a exposição dos animais a níveis de turbidez acima desse limite e por períodos mais longos do que 120 horas uma provável causa de mortalidade desta espécie.

Efficacy of Intravitreal Anti-vascular Endothelial Growth Factor or Steroid Injection in Diabetic Macular Edema According to Fluid Turbidity in Optical Coherence Tomography

PURPOSE: To determine if short term effects of intravitreal anti-vascular endothelial growth factor or steroid injection are correlated with fluid turbidity, as detected by spectral domain optical coherence tomography (SD-OCT) in diabetic macular edema (DME) patients. METHODS: A total of 583 medical records were reviewed and 104 cases were enrolled. Sixty eyes received a single intravitreal bevacizumab injection (IVB) on the first attack of DME and 44 eyes received triamcinolone acetonide treatment (IVTA). Intraretinal fluid turbidity in DME patients was estimated with initialintravitreal SD-OCT and analyzed with color histograms from a Photoshop program. Central macular thickness and visual acuity using a logarithm from the minimum angle of resolution chart, were assessed at the initial period and 2 months after injections. RESULTS: Visual acuity and central macular thickness improved after injections in both groups. In the IVB group, visual acuity and central macular thickness changed less as the intraretinal fluid became more turbid. In the IVTA group, visual acuity underwent less change while central macular thickness had a greater reduction (r = -0.675, p = 0.001) as the intraretinal fluid was more turbid. CONCLUSIONS: IVB and IVTA injections were effective in reducing central macular thickness and improving visual acuity in DME patients. Further, fluid turbidity, which was detected by SD-OCT may be one of the indexes that highlight the influence of the steroid-dependent pathogenetic mechanism.

Análisis de concordancia entre el dosaje de inmunoglobulinas y la fracción gamma del proteinograma sérico / Analysis of agreement between the dosage of immunoglobulins and gamma fraction of serum protein electrophoresis / Análise de concordancia entre a dosagem de imunoglobulinas e a fragao gamma do proteinograma sérico

En este trabajo se proponen dos criterios que evalúan el acuerdo existente entre densitometría y nefelometría, en la evaluación de los niveles de inmunoglobulinas en suero, basados en la propia población de pacientes y su posible aplicación en un algoritmo de autoverificación. Se construyó la variable Diferencia GAMgamma = Suma Igs - gamma; se efectuó el análisis según el método de Bland y Altman. Se estimó el IC95% empleando como referencia una cohorte de 497 pacientes concurrentes al laboratorio del Hospital A. Posadas en octubre y noviembre de 2011 y que tenían un dosaje de inmunoglobulinas G, A y M normal para población adulta. Estos resultados fueron contrastados con 2 poblaciones: a) 977 sujetos concurrentes en el mismo período y b) 111 pacientes con presencia de componente monoclonal (CM). En la población de referencia el IC95% Diferencia (mg/dL) fue de 115-653 y el DE para los límites ± 21 mg/dL. Utilizando los criterios Valores de referencia seguido por Diferencia se obtuvieron los siguientes resultados: Población de pacientes: 470/977 (48,1%) pasaron las dos alarmas y serán autovalidados. Población con CM: el 75,6% dio al menos una alarma. No obstante, si bien se puede considerar aceptable esta sensibilidad, hubo 27 CM que pasaron las dos alarmas. Se concluye que luego de una inspección visual previa, casi el 50% de los resultados serán autovalidados.

Two criteria are proposed to evaluate the agreement between densitometry and nephelometry of serum immunoglobulin level, based on own patient population and its possible application in a self-validation algorithm. A difference GAMgamma = Sum Igs -gamma variable was constructed; analysis was performed according to the Bland and Altman method. CI 95% was estimated using as reference a cohort of 497 patients attending Posadas Hospital's laboratory in October and November 2011 who had a normal dosage of G, A and M immunoglobulins for an adult population. These results were compared with two populations: a) 977 patients attending the hospital in the same period and b) 111 patients with monoclonal component (MC). In the reference population, the CI 95% Difference (mg/dL) was 115-653 and the limits were ± 21 mg/dL. Using Reference Values criteria followed by Difference, the following results were obtained: in the patient population, 470/977 (48.1%) passed both alarms and will be validated. In the MC population, 75.6% gave at least one alarm. However, although this may be considered acceptable sensitivity, there were 27 MC who passed both alarms. It can be concluded that after a visual inspection, almost 50% of the results will be self-validated.

Neste trabalho sáo propostos dois critérios que avaliam o acordo existente entre densitometria e nefelometría, na avaliagáo dos níveis de imunoglobulinas em soro, com base na pròpria populagáo de doentes e a sua possível aplicagáo num algoritmo de autovalidagáo. Foi construida a variável Diferenga GAMgamma = Soma Igs - gama, a análise foi realizada de acordo com o método de Bland & Altman. IC95% foi estimada utilizando-se como referencia uma coorte com 497 doentes concorrentes ao laboratòrio do hospital Posadas em outubro e novembro de 2011 e que tinham uma dosagem normal de imunoglobulinas G, A e M para populagáo adulta. Estes resultados foram comparados com duas populagdes-. a) 977 doentes concorrentes no mesmo período e b) 111 doentes com presenga de componente monoclonal (CM). Na populagáo de referencia, IC95% Diferenga (mg/dL): (115-653) e o DE para os limites ± 21 mg/dL foram os resultados. Utilizando os critérios Valores de Referencia seguidos de Diferenga foram obtidos os seguintes resultados: A populagáo de doentes de 470/977 (48,1%) passou os dois alarmes e seráo autovalidados. A populagáo com CM que deu pelo menos um alarme foi de 75,6%. No entanto, embora esta sensibilidade possa ser considerada aceitável, houve 27 CM que passaram os dois alarmes. A conclusáo é que depois de uma inspegáo visual prévia, cerca de 50% dos resultados seráo autovalidados.

Validation and development of an immunonephelometric assay for the determination of alpha-1 antitrypsin levels in dried blood spots from patients with COPD / Desenvolvimento e validação de um método de imunonefelometria em amostras de sangue em papel-filtro para a dosagem da alfa-1 antitripsina em pacientes com DPOC

OBJECTIVE: To validate and develop an immunonephelometric assay for the determination of alpha-1 antitrypsin (AAT) levels in dried blood spots from COPD patients in Brazil. METHODS: We determined AAT levels in serum samples and dried blood spots from 192 COPD patients. For the preparation of dried blood spots, a disk (diameter, 6 mm) was placed into a tube, eluted with 200 µL of PBS, and stored overnight at 4ºC. All of the samples were analyzed by immunonephelometry in duplicate. We used the bootstrap resampling method in order to determine a cut-off point for AAT levels in dried blood spots. RESULTS: The correlation coefficient between the AAT levels in serum samples and those in dried blood spots was r = 0.45. For dried blood spots, the cut-off value was 2.02 mg/dL (97% CI: 1.45-2.64 mg/dL), with a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 95.7%, 27.2%, and 100%, respectively. CONCLUSIONS: This method for the determination of AAT levels in dried blood spots appears to be a reliable screening tool for patients with AAT deficiency. .

OBJETIVO: Validar e desenvolver um método de dosagem de alfa-1 antitripsina (AAT) por imunonefelometria em amostras de sangue em papel-filtro em pacientes com DPOC no Brasil. MÉTODOS: Amostras de soro e de sangue em papel-filtro de 192 pacientes com DPOC foram utilizadas para a dosagem de AAT. Para a preparação das amostras de sangue em papel-filtro, um disco do papel com diâmetro de 6 mm foi colocado em um tubo e eluído com 200 µL de PBS, permanecendo por toda a noite a 4ºC. Todas as amostras foram analisadas em duplicata por imunonefelometria. O método de reamostragem bootstrap foi utilizado para a determinação de um ponto de corte para o nível de AAT nas amostras de sangue em papel-filtro. RESULTADOS: O coeficiente de correlação entre os níveis de AAT em soro e em sangue em papel-filtro foi de r = 0,45. Para as amostras em papel-filtro, o ponto de corte foi de 2,02 mg/dL (IC97%: 1,45-2,64 mg/dL), com sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 100%, 95,7%, 27,2% e 100%, respectivamente. CONCLUSÕES: Este método de determinação dos níveis de AAT em sangue em papel-filtro se mostrou uma ferramenta confiável para o rastreamento de pacientes com deficiência de AAT. .

Dosagens séricas de proteína C-reativa na fase inicial da sepse abdominal e pulmonar / Serum C-reactive protein concentrations in early abdominal and pulmonary sepsis

OBJETIVO: Avaliar os níveis séricos de proteína C-reativa em pacientes com sepse pulmonar e abdominal nos primeiros 5 dias de progressão da sepse. MÉTODOS: Estudo de coorte retrospectivo em hospital universitário. Foram selecionados 345 pacientes admitidos em unidade de terapia intensiva e diagnosticados com sepse de origem pulmonar ou abdominal. A dosagem sérica de proteína C-reativa foi realizada por imunoensaio turbidimétrico. Para análises da proteína C-reativa, o dia 1 foi definido como o do diagnóstico clínico da sepse. RESULTADOS: Foram avaliados 34 pacientes com sepse (9,8%), 114 com sepse grave (33,0%) e 197 com choque séptico (57,2%). A idade dos pacientes foram 56,4±19,8 anos. Concentrações séricas de proteína C-reativa foram mais elevadas no dia do diagnóstico de sepse no grupo com infecção de origem abdominal em comparação ao grupo com sepse pulmonar (17,8±10,1 mg/dL versus 14,9±11,1 mg/dL; p=0,025) e mantiveram-se significativamente mais elevadas nos primeiros 5 dias de evolução da sepse. CONCLUSÃO: As concentrações séricas de proteína C-reativa foram significativamente mais elevadas nos pacientes com sepse de origem abdominal do que em pacientes com sepse de origem pulmonar nos 5 primeiros dias de evolução da sepse.

OBJECTIVES: To evaluate the C-reactive protein serum levels in patients with pulmonary and abdominal sepsis during the first five days of sepsis progression. METHODS: The present investigation was a retrospective cohort study conducted at the university hospital with 345 patients who were admitted to the intensive care unit and diagnosed with sepsis of pulmonary or abdominal origin. Serum C-reactive protein concentrations were measured by the turbidimetric immunoassay. For analysis of C-reactive protein, day 1 was defined as the day on which the patient was clinically diagnosed with sepsis. RESULTS: Thirty-four patients with sepsis (9.8%), 114 patients with severe sepsis (33.0%), and 197 patients with septic shock (57.2%) were evaluated. The age of the patients was 56.4±19.8 years. The serum C-reactive protein concentrations were higher on the day of sepsis diagnosis in the group with abdominal infection compared with the group with pulmonary sepsis (17.8±10.1 mg/dL versus 14.9±11.1 mg/dL, p=0.025) and remained significantly higher during the first five days of sepsis progression. CONCLUSION: The serum C-reactive protein concentrations were significantly higher in the patients with abdominal sepsis compared with the patients with pulmonary sepsis during the first five days of sepsis progression.

Comparative study between immunoturbidimetric and latex agglutination methods for the detection of rheumatoid factor / Estudo comparativo entre as técnicas de aglutinação em látex e de imunoturbidimetria para a detecção de fator reumatoide

INTRODUCTION: The rheumatoid factor (RF) is the most common antibody found in patients with rheumatoid arthritis. It is an inflammatory chronic disease characterized by articular involvement, inflammation of synovial fluid, tissue infiltration by leucocytes and joint destruction, which ultimately determine articular deformities. The rheumatoid factor is found in 70%-80% of the adult population and in 10% of the young population. OBJECTIVE: The aim of this research was to compare immunoturbidimetric and latex agglutination methods for the detection of RF in serum. RESULTS: The immunoturbidimetric method displayed sensitivity (95.2%), specificity (89.4%) and high positive correlation (R² = 0,8077) with the latex agglutination method in positive serum samples. CONCLUSION: The study allowed to demonstrate that both immunoturbidimetric and latex agglutination methods equally discriminate between negative and positive serum samples for RF.

INTRODUÇÃO: O fator reumatoide (FR) é o autoanticorpo mais comum encontrado em pacientes com artrite reumatoide, uma doença crônica inflamatória caracterizada pelo envolvimento articular com inflamação do líquido sinovial, infiltração de tecido por leucócitos e destruição das articulações, que acaba por determinar deformidades articulares. O FR é encontrado em 70%-80% da população adulta e em 10% da população juvenil. OBJETIVO: Comparar os métodos de imunoturbidimetria e aglutinação (prova do látex) para a determinação de FR em soro. RESULTADO: Foi possível observar que o método imunoturbidimétrico apresenta sensibilidade (95,2%), especificidade (89,4%) e correlação positiva elevada (R² = 0,8077) com o método de aglutinação pelo látex em amostras de soro positivas. CONCLUSÃO: O estudo permitiu demonstrar que o método imunoturbidimétrico e o método de aglutinação pelo látex são igualmente capazes de discriminar amostras negativas e positivas para FR.