Use of the dried blood spot sampling process coupled with fast gas chromatography and negative-ion chemical ionization tandem mass spectrometry: application to fluoxetine, norfluoxetine, reboxetine, and paroxetine analysis.

The objective of this work was to combine the advantages of the dried blood spot (DBS) sampling process with the highly sensitive and selective negative-ion chemical ionization tandem mass spectrometry (NICI-MS-MS) to analyze for recent antidepressants including fluoxetine, norfluoxetine, reboxetine, and paroxetine from micro whole blood samples (i.e., 10 microL). Before analysis, DBS samples were punched out, and antidepressants were simultaneously extracted and derivatized in a single step by use of pentafluoropropionic acid anhydride and 0.02% triethylamine in butyl chloride for 30 min at 60 degrees C under ultrasonication. Derivatives were then separated on a gas chromatograph coupled with a triple-quadrupole mass spectrometer operating in negative selected reaction monitoring mode for a total run time of 5 min. To establish the validity of the method, trueness, precision, and selectivity were determined on the basis of the guidelines of the "Société Française des Sciences et des Techniques Pharmaceutiques" (SFSTP). The assay was found to be linear in the concentration ranges 1 to 500 ng mL(-1) for fluoxetine and norfluoxetine and 20 to 500 ng mL(-1) for reboxetine and paroxetine. Despite the small sampling volume, the limit of detection was estimated at 20 pg mL(-1) for all the analytes. The stability of DBS was also evaluated at -20 degrees C, 4 degrees C, 25 degrees C, and 40 degrees C for up to 30 days. Furthermore, the method was successfully applied to a pharmacokinetic investigation performed on a healthy volunteer after oral administration of a single 40-mg dose of fluoxetine. Thus, this validated DBS method combines an extractive-derivative single step with a fast and sensitive GC-NICI-MS-MS technique. Using microliter blood samples, this procedure offers a patient-friendly tool in many biomedical fields such as checking treatment adherence, therapeutic drug monitoring, toxicological analyses, or pharmacokinetic studies.

Estudo das variáveis de processamento das matérias-primas da região do crato - ce na fabricação de produtos cerâmicos por extrusão e por prensagem

The limits to inform is about the character stico of basic, quimica, mineralogical and mechaniques of matlaughed material used in the manufacturing process the product certified in economic region the Cariri, specifically in the city of Crato, Ceará state, motivated the development of this work, since in this region the exist ing economic context that a general appear as important in the production chains. Were made twentyfive soils-test specimen collection and the study was performed to differentiate the mat laugh materials of variaveis processing of mathing raw materials in the factory The product mica monkeys by extrusion and pressing. The results were obtained ap s as analyzes: grain size, index of plasticity, fluoresce incidence X-ray difration the X-ray, and analyzes thermicals and properties technological. through s of curves gresifica returned to was a comparison between the retro the linear, absorb to water, porosity and bulk density. the results show that the excellent distribution and character acceptable available for the processing of the structure color dark red. needing, therefore, of the mixture of a less plastic clay with thick granulation, that works as plasticity reducer. In spite of the different resignation forms for prensagem and extrusion, the characteristics of absorption of water and rupture tension the flexing was shown inside of the patterns of ABNT

Condição microbiológica dos leitos hospitalares antes e depois de sua limpeza

INTRODUÇÃO: O meio ambiente hospitalar guarda uma íntima relação com as infecções hospitalares, podendo proporcionar focos de contato e de transmissão. Como a higiene representa uma das formas de controlar a contaminação ambiental, realizou-se estudo para avaliar as condições microbiológicas dos colchões hospitalares antes e depois de sua limpeza. MÉTODOS: Utilizaram-se, para a colheita dos espécimes, placas de contato -- Rodac-plate ¾ preparadas com meio de cultura ágar-sangue. Selecionaram-se os leitos de acordo com critérios previamente estabelecidos, e os locais de colheita sob o colchão foram escolhidos por sorteio aleatório. Aplicou-se o teste estatístico de Goodman para o estudo das alterações numéricas quanto a positividade das placas. RESULTADOS: Foram investigados 52 colchões, totalizando 520 placas, das quais 514 (98,8%) resultaram em culturas positivas, sendo que 259 corresponderam ao período anterior à limpeza e 255 ao período posterior ao procedimento. Houve redução de culturas positivas em apenas 4 placas. CONCLUSÕES: Os resultados obtidos sugerem que a limpeza, da forma como vem sendo conduzida, provoca o deslocamento da carga microbiana para outros pontos do colchão em vez de diminuí-la, resultando na manutenção da quantidade de microorganismos que existia anteriormente à limpeza.

Eritrograma e morfometria eritrocitária de avestruzes (struthio camelus) criados na região de São José do Rio Preto, São Paulo, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Stability of coagulation assays performed in plasma from citrated whole blood transported at ambient temperature

Many preanalytical variables affect the results of coagulation assays. A possible way to control some of them would be to accept blood specimens shipped in the original collection tube. The aim of our study was to investigate the stability of coagulation assays in citrated whole blood transported at ambient temperature for up to two days after specimen collection. Blood samples from 59 patients who attended our haematology outpatient ward for thrombophilia screening were transported at ambient temperature (outdoor during the day, indoor overnight) for following periods of time: <1 hour, 4-6, 8-12, 24-28 and 48-52 hours prior to centrifugation and plasma-freezing. The following coagulation tests were performed: PT, aPTT, fibrinogen, FII:C, FV:C, FVII:C, FVIII:C, FIX:C, FX:C, FXI:C, VWF:RCo, VWF:Ag, AT, PC activity, total and free PS antigen, modified APC-sensitivity-ratio, thrombin-antithrombin-complex and D-dimer. Clinically significant changes, defined as a percentage change of more than 10% from the initial value, were observed for FV:C, FVIII:C and total PS antigen starting at 24-28 hours, and for PT, aPTT and FVII:C at 48-52 hours. No statistically significant differences were seen for fibrinogen, antithrombin, or thrombin-antithrombin complexes (Friedman repeated measures analysis of variance). The present data suggest that the use of whole blood samples transported at ambient temperature may be an acceptable means of delivering specimens for coagulation analysis. With the exception of factor V and VIII coagulant activity, and total PS antigen all investigated parameters can be measured 24-28 hours after specimen collection without observing clinically relevant changes.

A study of the cost and effect of newborn screening for Congenital Adrenal Hyperplasia in Texas

Congenital Adrenal Hyperplasia (CAH), due to 21-Hydroxylase deficiency, has an estimated incidence of 1:15,000 births and can result in death, salt-wasting crisis or impaired growth. It has been proposed that early diagnosis and treatment of infants detected from newborn screening for CAH will decrease the incidence of mortality and morbidity in the affected population. The Texas Department of Health (TDH) began mandatory screening for CAH in June, 1989 and Texas is one of fourteen states to provide neonatal screening for the disorder.^ The purpose of this study was to describe the cost and effect of screening for CAH in Texas during 1994 and to compare cases first detected by screen and first detected clinically between January 1, 1990 and December 31, 1994. This study used a longitudinal descriptive research design. The data was secondary and previously collected by the Texas Department of Health. Along with the descriptive study, an economic analysis was done. The cost of the program was defined, measured and valued for four phases of screening: specimen collection, specimen testing, follow-up and diagnostic evaluation.^ There were 103 infants with Classical CAH diagnosed during the study and 71 of the cases had the more serious Salt-Wasting form of the disease. Of the infants diagnosed with Classical CAH, 60% of the cases were first detected by screen and 40% were first detected because of clinical findings before the screening results were returned. The base case cost of adding newborn screening to an existing program (excluding the cost of specimen collection) was $357,989 for 100,000 infants. The cost per case of Classical CAH diagnosed, based on the number of infants first detected by screen in 1994, was \$126,892. There were 42 infants diagnosed with the more benign Nonclassical form of the disease. When these cases were included in the total, the cost per infant to diagnose Congenital Adrenal/Hyperplasia was $87,848. ^

An ethnographic study of home remedy use for African-American children

Background and significance. The use of herbs and other remedies by adult and elderly African-Americans has been documented. However, little is understood regarding the use of herbs for African-American children. The purpose of this study was to document and describe the historical and present day uses of herbal and other remedies, specifically for the health and illness of African-American children. This information will provide health care providers with a better understanding of their African-American patients. This information may also contribute to the emerging appreciation of indigenous uses of phytotherapeutics. ^ Methods. A focused ethnographic approach was used to describe the cultural context, including the beliefs, values, customs, and behaviors of a particular culture. The use of intensive fieldwork, including participant observation, audiotaped formal interviews, photographs, and specimen collection of plants helped to describe herb use in this population. Information on the growing, harvesting, preparation, and storage of these plant remedies, as well as the amount and dosage of these compounds was collected in a typology. Detailed information was gathered to discern how, when, and under what conditions these remedies were used and their expected results. Further data collection focused on the history of herbal use, and explanations for how and why informants thought the herbs work. ^ Setting and participants. The setting for this study was in East Texas and field work extended over the period of one year. Thirty African-Americans, age 38 to 98, were interviewed for the study. The African-American population in this area has been relatively stable, with roots dating back prior to the reconstruction period, which allowed excellent historical information. Informants were chosen by a nominated sampling technique starting with two key informants knowledgeable about the use of home remedies for children. ^ Findings. The findings of this study suggest that African-American children in East Texas have a long history of receiving herbs and home remedies for health promotion and illness. Data further suggests that there is a strong connection between spirituality and the health beliefs and practices of this community. This spiritual component underlies the accuracy of oral recall for remedies that have been used over many generations and the use of natural folk remedies. A typology of the herbal remedies was developed with folk and Latin name, herb place of origin, known scientific properties, and informant folk usage and dosage information. ^

Manual of services /

"February 2006."

FOOTNOTES : S.C. DHEC NEWBORN SCREENING NEWSLETTER

These publications will include helpful tips for specimen collection, submission, follow-up contact information for DHEC NBS staff, and an educational section on specific disorders found on the newborn screening panel.

Collection and evaluation of semen from the three-toed sloth (Bradypus tridactylus)

Sloths (Bradypus sp.) are extremely sensitive animals that suffer with the destruction and fragmentation of forests. They present a low population growth rate and need to be further studied for the preservation of the specie. Thus, the aim of this study was to establish an efficient semen collection protocol as well as characterize sperm concentration, motility and morphology in order to contribute with information about the reproductive traits of this specie, which has never been described in the literature before. For that, nine Bradypus tridactylus males were captured during the wet season and six during the dry season, in Manaus (AM), located in the north region of Brazil, semen was collected by electroejaculation with shocks given in sequences of progressive intensities (minimum 20 mA and maximum 60 mA). All animals ejaculated small volumes of semen and in some of them, the volume ejaculated was not enough for a complete spermiogram. Physical characteristics observed on the collections of the wet season were different from those seen in the specimen collected in the dry season. Motility an vigor was very low and did not show forward progression, only oscillatory movement. After Spermac stain, spermatozoa presented a wide variety of defects; however, the differences in morphology were not significant between seasons. The morphology assessed by scanning electron microscopy shows that the head in both groups could be elongated, short or could have a base narrower than the apex and the midpiece narrowed abruptly, forming a nip in its transition to the tail. Although further studies are necessary to verify our preliminary findings concerning seasonal variation in sperm quality, these results demonstrate that semen can be safely collected from sloths by electroejaculation and provide the first reports of semen characteristics in this species. (C) 2008 Elsevier Ltd. All rights reserved.

Discrepancies between Aedes aegypti identification in the field and in the laboratory after collection with a sticky trap

Currently, sticky traps are regularly employed to assist in the surveillance of Aedes aegypti infestation. We tested two alternative procedures for specimen identification performed by local health agents: directly in the field, as recommended by certain manufacturers, or after transportation to the laboratory. A total of 384 sticky traps (MosquiTRAP) were monitored monthly during one year in four geographically representative Brazilian municipalities. When the same samples were inspected in the field and in the laboratory, large differences were noted in the total number of mosquitoes recorded and in the number of specimens identified as Ae. aegypti by both procedures. Although field identification has the potential to speed vector surveillance, these results point to uncertainties in the evaluated protocol.

Effect of collection and preprocessing methods on neutrophil elastase plasma concentrations.

OBJECTIVES: Elevated plasma levels of the elastase alpha 1-proteinase inhibitor complex (E-alpha 1 PI) have been proposed as a marker of bacterial infection and neutrophil activation. Liberation of elastase from neutrophils after collection of blood may cause falsely elevated results. Collection methods have not been validated for critically ill neonates and children. We evaluated the influence of preanalytical methods on E-alpha 1 PI results including the recommended collection into EDTA tubes. DESIGN AND METHODS: First, we compared varying acceleration speeds and centrifugation times. Centrifugation at 1550 g for 3 min resulted in reliable preparation of leukocyte free plasma. Second, we evaluated all collection tubes under consideration for absorption of E-alpha 1 PI. Finally, 12 sets of samples from healthy adults and 42 sets obtained from critically ill neonates and children were distributed into the various sampling tubes. Samples were centrifuged within 15 min of collection and analyzed with a new turbidimetric assay adapted to routine laboratory analyzers. RESULTS: One of the two tubes containing a plasma-cell separation gel absorbed 22.1% of the E-alpha 1 PI content. In the remaining tubes without absorption of E-alpha 1 PI no differences were observed for samples from healthy adult patients. However, in samples from critically ill neonates or children, significantly higher results were obtained for plain Li-heparin tubes (mean = 183 micrograms/L), EDTA tubes (mean = 93 micrograms/L), and citrate tubes (mean = 88.5 micrograms/L) than for the Li-hep tube with cell-plasma separation gel and no absorption of E-alpha 1 PI (mean = 62.4 micrograms/L, p < 0.01). CONCLUSION: Contrary to healthy adults, E-alpha 1 PI results in plasma samples from critically ill neonates and children depend on the type of collection tube.

Recovery of DNA for the detection and genotyping of human papillomavirus from clinical cervical specimens stored for up to 2 years in a universal collection medium with denaturing reagent

The recovery and stability of DNA for the detection and genotyping of HPV in UCM-containing specimens, after exposure to denaturing reagents and stored for up to 2 years were evaluated. Samples were collected from 60 women who had cervical cytology specimens harboring cervical intraepithelial neoplasia (CIN) 2 or 3. All samples were stored in UCM and had been frozen at -20 degrees C following the addition of the denaturing reagent (sodium hydroxide) and the removal of the aliquot required for Hybrid Capture 2 testing for the identification of HPV DNA. The samples had been stored for 6, 12 and 24 months (20 samples for each storage time). HPV DNA extraction was performed according to a protocol designed specifically and the presence and quality of DNA was confirmed by human P-globin detection using the consensus primers G73 and G74. HPV DNA was amplified using the consensus primers PGMY09 and PGMY11, and reverse line-blot hybridization was used to detect type-specific amplicons for 37 HPV types. The DNA extracted from the denatured specimen was recovered in 57/60 (95%) of the samples. HPV DNA was detected in 56/57 (98%) of the recovered samples. Twenty-six of the 56 samples recovered (48%) were genotyped successfully. (c) 2007 Elsevier B.V. All rights reserved.