A ten-color tube with dried antibody reagents for the screening of hematological malignancies.

INTRODUCTION: The workflow in clinical flow cytometry laboratories must constantly be reviewed to develop technical procedures that improve quality and productivity and reduce costs. Using the Beckman Coulter dry coating technology, we customized a ten-color tube with dried antibody reagents, designated the Duraclone screening tube (DST), for screening hematological malignancies. Here, we compared the applicability, clinical and numerical equivalence, and cost and time required for the technical procedures between the liquid reagents and the DST. METHODS: The DST contains CD4 + Kappa-FITC, CD8 + Lambda-PE, CD3 + CD14-ECD, CD33-PE-Cy5.5, CD20 + CD56-PE-Cy7, CD34-APC, CD19-APC-AlexaFluor700, CD10-APC-AlexaFluor750, CD5-Pacific Blue, and CD45-Krome Orange. We evaluated 20 bone marrow samples, 13 peripheral blood samples, 6 lymph node biopsy samples, 5 fine-needle aspirate samples, 5 cerebrospinal fluid samples, and 1 pleural fluid sample. RESULTS: The DST was useful for more than 60% of our samples. It was able to enumerate the majority of the populations in all types of samples with a statistically acceptable correlation with the liquid reagents. The use of the DST translated into significant time and cost savings of 15.8% and 12.3%, respectively, compared with the use of the liquid reagent. The cost was reduced by $14.36 per sample. CONCLUSIONS: The DST is an efficient solution for screening hematological malignancies with improved quality, productivity, standardization, and sustainability. These improvements could benefit patients by providing faster diagnoses using a higher quality and lower cost reagent.

Prevalence and etiology of buffalo mastitis and milk somatic cell count in dry and rainy seasons in a buffalo herd from Analândia, São Paulo State, Brazil / Prevalência e etiologia da mastite bubalina e contagem de células somáticas no leite, nas estações seca e chuvosa, em um rebanho bubalino do município de Analândia, Estado de São Paulo, Brasil

The aim of this study was to evaluate somatic cell count (SCC), prevalence and etiology of mastitis in a dairy buffalo herd from Analândia, São Paulo State, Brazil, in the dry and rainy seasons. Additionally, antimicrobial susceptibility profile of microorganisms isolated from milk samples was also evaluated. 1,042 milk samples from female Murrah buffaloes in a dairy farm located in Analândia, São Paulo State, Brazil, collected between May 2011 and November 2012 were analyzed. After the mammary gland physical examination, strip cup test and California Mastitis Test (CMT) were performed. Afterwards, 50mL of milk samples from each mammary quarter were collected aseptically for SCC in automatic equipment and microbiological examination. The antimicrobial sensitivity profile to ampicillin, cefoperazone, ceftiofur, enrofloxacin, gentamicin, neomycin, oxacillin, penicillin, and sulfamethoxazole/trimethoprim was evaluated by disk diffusion method. The monthly average temperature and pluviometric index were obtained from "Centro Integrado de Informações Agrometeorológicas" (CIIAGRO) of "Instituto Agronômico de Campinas" (IAC). Milk samples with positive results in the microbiological test showed average SCC of 137,720 cells/mL in the dry period and 190,309 cells/mL in the rainy period. Although a higher number of isolated microorganisms was observed in buffalo milk samples during the rainy period (69/600) compared to the dry period (50/442), the season had no significant effect on the frequency of isolation of microorganisms. The main genera of microorganisms isolated were coagulase-negative Staphylococcus (38.4%), Streptococcus agalactiae (28.8%), and Bacillus spp. (7.56%) during the dry season and Corynebacterium sp. (23.5%), Streptococcus spp. (32.3%), and Streptococcus agalactiae (9.24%) during the rainy period. Multidrug resistance was observed in 30.1% of the isolated microorganisms...

O objetivo do presente estudo foi avaliar a contagem de células somáticas, a prevalência e a etiologia da mastite bubalina nas estações seca e chuvosa em um rebanho de bubalinos do município de Analândia, estado de São Paulo, Brasil. Adicionalmente, verificou-se o perfil de sensibilidade antimicrobiana dos micro-organismos isolados nas amostras de leite das búfalas. Foram avaliadas 1.042 amostras de leite de búfalas da raça Murrah pertencentes a uma propriedade rural localizada no município de Analândia-SP, obtidas no período de maio de 2011 e novembro de 2012. Após o exame físico da glândula mamária, foram realizados o teste da caneca de fundo escuro e o California Mastits Test (CMT); em seguida, foram colhidas, de forma asséptica, amostras de 50mL de leite de cada quarto mamário, para a contagem de células somáticas (CCS) em aparelho automático e exame microbiológico. Também, foi avaliado o perfil de sensibilidade antimicrobiana a ampicilina, cefoperazona, ceftiofur, enrofloxacina, gentamicina, neomicina, oxacilina, penicilina e sulfametoxazol/trimetoprim, pelo método de difusão em disco. A temperatura média e o índice pluviométrico mensais foram obtidos no Centro Integrado de Informações Agrometeorológicas (CIIAGRO) do Instituto Agronômico de Campinas (IAC). Notou-se que as amostras de leite com resultado positivo no exame microbiológico apresentaram CCS média de 137.720 células/mL, no período seco, e 190.309 células/mL, no período chuvoso. Embora tenha se constatado maior índice de isolamentos de micro-organismos nas amostras de leite obtidas no período chuvoso (69/600) do que no período seco (50/442), a frequência de isolamentos não foi influenciada significativamente pela estação do ano...

Parâmetros ósseos de frangos de corte alimentados com rações contendo fitase e baixos níveis de fósforo disponível e proteína bruta / Bone parameters for broilers fed diets with phytase and low phosphorus and crude protein levels

Foram utilizados 504 pintos de linhagem comercial (Ag Ross 308) para frangos de corte de um dia de idade, distribuídos em 12 tratamentos com seis repetições. O delineamento experimental utilizado foi de blocos ao acaso em esquema fatorial 2x2x3, com dois níveis de fósforo disponível (0,45 e 0,34%), dois níveis de fitase (0 e 1200 FTU/kg) e três níveis de proteína bruta (22,5; 20,5 e 18,5%). A porcentagem de cálcio e fósforo nas tíbias foi influenciada significativamente pelos níveis de proteína e de fósforo na dieta com o uso da fitase. Os teores de matéria mineral nas tíbias apresentaram efeito linear com o uso da enzima e efeito quadrático com a ausência dela, apresentando maior valor com o nível de 22,5% de proteína na dieta. Já com os maiores níveis de fósforo houve efeito linear entre os níveis de proteína bruta na dieta e os pesos da matéria mineral nas tíbias, ou seja, quanto maior o nível de proteína, menor o peso da matéria mineral.

504 (five hundred four) 1 day old male chicks from a commercial broiler line (Ag Ross 308) were used, distributed in 12 treatments, with 6 replicates per treatment. The experimental design was casually blocked and treatments were organized in a 2x2x3 factorial arrangement: two available phosphorus levels (0.45 and 0.34%), two phytase inclusion levels (0 and 1200 FTU/kg) and three crude protein levels (22.5; 20.5 and 18.5%). Tibia calcium percentage was influenced by protein and phosphorus levels in the diet, when using phytase. Tibia ash levels showed a linear effect when enzymes were added and showed a quadractic effect without it, and the higher value was observed with 22.5% crude protein in the diet. With higher phosphorus levels a linear effect was observed between crude protein in the diet and tibia ash weights, as a higher protein level had lower tíbia ash weight.

[Reduced and oxidized glutathione of the placenta in pregnancy complicated by pre-eclampsia]. / Glutatião reduzido e oxidado da placenta na gravidez complicada com preeclâmpsia.

Significative enhancement of free radical formation (FRO) in vivo is an important feature of hypertensive disorders of pregnancy (HDP), namely preeclampsia (PIH). The latest investigations about the pathology of HDP, showed the contribution of placental circulation to the development and evolution of such disease. The placental bed can be a potential source of FRO or activation of cells that can produce FRO. Glutathione, is an important molecule for cellular protection against damage, is a cofactor of many enzymes, in particular, for the glutathione peroxidase of the placental tissue; this enzyme in the placenta bed prevent the production of thromboxan and lipoperoxides; the latter are potentially damaging to the endothelium cells and can cause vasoconstriction, the most important feature of PIH. The activity of that enzyme is deficient in PIH. We studied, by fluorometric assay, the concentrations of the two states of glutathione in placental homogenates (PLH) from pregnant women without pathology (PWN) and from pregnant women with PIH (PWPIH). The data showed significant low concentrations in the PLH of the two states of glutathione in the PWN against high concentrations of this molecule in the PLH from PWPIH. This feature can result from a deficient user of the glutathione by the cellular mechanism for prevention against oxidative factors. In addition, our study shows a biochemical marker that is suggestive that the placental bed is a potential source of FRO production in PIH.

[Determination of HBSAg in pregnancy. Epidemiologic study of 100 cases]. / Determinação do AgHBs na grávida. Estudo epidemiológico de 100 casos.

There are some arguments concerning prenatal Hepatitis B testing: universal screening versus testing of risk groups. A thousand parturients were evaluated in order to assess the sensitivity of risk factors in the identification of Hepatitis B surface antigen (HBsAg) positive women. To achieve that purpose they were questioned regarding risk factors and blood samples were collected for HBsAg testing. Thirty two HBsAg positive parturients were identified. A larger number of HBsAg positive was found amongst women included in the class 5 of the Graffard scale (p = 0.005), or referring contact with patients with hepatitis (p = 0.01). These risk factors identified no more than 17 of the 32 positive mothers (sensitivity = 53%). None of the other risk factors proved significant. The entire questionnaire allows a sensitivity of 87.5% but presents a specificity of 22.5%. In this population, risk factors are not a good method of screening HBsAg positive women. A universal screening must be undertaken in order to prevent vertical transmission of the virus and to detect families in risk.

Role of neuropeptide Y (NPY) in cardiovascular responses to stress.

Neuropeptide Y (NPY), a putative sympathetic neurotransmitter and neuromodulator, is released during sympathetic nerve stimulation and causes vasoconstriction and cardiodepression. Whether the release of NPY contributes to stress-induced cardiovascular responses was assessed by studying i) plasma levels of circulating NPY-immunoreactivity (NPY-ir) during various stress paradigms and ii) mechanisms of action of NPY in the cardiovascular system of the rat. Plasma NPY-ir was increased by all stress situations, such as transfer to a new environment (by 52%), exposure to cold water (4 degrees C) (by 117%) and hemorrhage (4 ml/300 g body weight) (by 231%). The cold water, stress-induced, maximal increase in circulating plasma NPY-ir was delayed in relation to the peak pressor response by 10-20 min. Administration of NPY caused dose-dependent pressor responses that were greater in pithed rats--which have all centrally mediated circulatory reflexes removed--than in conscious rats. Infusion of a low pressor (8.5 +/- 1.5 mm Hg) dose of norepinephrine into conscious rats potentiated NPY-mediated pressor responses 2-fold and also tended to increase bradycardic effect of a higher dose of NPY (by 19%). Thus hypertensive and bradycardiac actions of NPY appear to depend on the level of adrenergic activity and on the interactions at the level of vascular smooth muscle, heart, and baroreceptor reflexes. During a hyperadrenergic state such as stress, cardiovascular effects of NPY may be greatly accentuated. NPY may enhance and prolong the stress-induced hypertensive responses while antagonizing adrenergic cardiostimulation.

Estudo multicêntrico aberto, näo comparativo, com etomidato em procedimentos cirúrgicos de curta duraçäo / Open multicenter study, non-comparative, with etomidate in short-term surgical procedures

Realizou-se um estudo multicêntrico em 155 pacientes de ambos os sexos, ASA 1, 2, e 3, submetidos a anestesia venosa com etomidato (0,3 mg.k-1) para procedimentos cirúrgicos de curta duraçäo. Foram administrados, como medicaçäo pré-anestésica, 10 minutos antes da induçäo, 100 micron-grama de fentanil por via venosa. Ao final do procedimento anestésico, verificou-se aumento das pressöes sistólica e diastólica, enquanto que a freqüência cardíaca manteve-se estável. A freqüência respiratória apresentou aumento, porém sem significado clínico. Em 113 pacientes (72,9%) houve necessidade de se administrar doses complementares da droga. O tempo médio de perda da consciência foi de 48 + ou - 22s do início da injeçäo. O tempo de recuperaçäo da consciência foi de 8,26 + ou - 5,16 min. A média do tempo para possível alta hospitalar foi de 61,7 + ou - 30,5 min. Com relaçäo aos efeitos secundários observou-se que em 15 (9,7%) pacientes houve dor no local da injeçäo, em 79 (51%) surgiram mioclonias, na maioria das vezes de grau leve, e em 36 (23,2%) pacientes ocorreu ereçäo pilosa. Concluíram ser o etomidato uma droga adequada para uso venoso em procedimentos de curta duraçäo

Complemento hemolitico total, C3 e taxa de conversao de C3 nas formas cardiaca e indeterminada da Doenca de Chagas. / Total hemolytic complement, C3 and C3 convertase levels in cardiac and indeterminate forms of Chagas' disease

Os valores de complemento hemolitico total C3 total (nativo + produtos de degradacao) e o grau de conversao de C3 nativo foram estudados em dois subgrupos de pacientes chagasicos, nas formas cardiaca e indeterminada, e em um subgrupo de individuos nao chagasicos, clinicamente sadios.Os niveis de C3 total e as taxas de conversao de C3 em seus produtos de degradacao foram semelhantes nos tres subgrupos. Os valores de complemento hemolitico total foram estatisticamente diferentes nos tres subgrupos (nivel de significancia descritivo p= 0,0757), tendo sido observada media aritmetica mais baixa no subgrupo de cardiacos e mais elevada no subgrupo de controles.Maior amplitude de variacao dos niveis de complemento hemolitico total foi notada no subgrupo de cardiacos, no qual se encontraram os valores extremos (maximo e minimo), considerando-se todos os subgrupos