Método microespectrofotométrico para la determinación de valores de referencia de la fosfomanosa isomerasa / A Microspectrophotometric Method for the Determination of Reference Values of Phosphomannose Isomerase / Método microespectrofotométrico para determinar valores de referência da fosfomanose isomerase

Resumen: Los desórdenes congénitos de glicosilación son un conjunto de defectos genéticos de tipo multisistémico que afectan la función de la proteína. Se han descrito cerca de 75 enfermedades desde sus primeros estudios. En el presente estudio se desarrolló un método microespectrofotométrico para el diagnóstico de la enzima citosólica fosfomanosa isomerasa EC 5.3.1.8 (PMI), se analizaron 32 muestras de individuos con rango de edad de 0,6 a 27 años y se estableció el intervalo y el valor de referencia de actividad enzimática específica. Este estudio permitirá iniciar el diagnóstico de pacientes deficientes de la PMI de forma temprana y oportuna, lo cual la convierte en una posible enzima candidata para pruebas de tamizaje neonatal, ya que esta patología tiene un tratamiento fácil y de bajo costo, que consiste en la suplementación de manosa en forma oral. El diagnóstico clínico de este desorden metabólico beneficiará al paciente y a su familia al mejorar su calidad de vida, como también al sistema de salud colombiano.

Abstract: Congenital glycosylation disorders are a set of multi-systemic genetic defects affecting protein function. About 75 diseases have been described since early studies. This study developed a microspectrophotometric method for the diagnosis of the cytosolic enzyme phosphomannose isomerase (PMI) (EC 5.3.1.8), analyzed 32 samples of individuals ranging between 0.6 and 27 years old, and established the interval and reference value of specific enzyme activity. This study will allow early and timely diagnosis of PMI deficient patients, which makes this enzyme a potential candidate for neonatal screening tests since this pathology has an easy, low-cost treatment (oral administration of mannose supplements). Clinical diagnosis of this metabolic disorder will benefit the patient and his family by improving his quality of life, as well as the Colombian healthcare system.

Resumo: Os defeitos congénitos de glicosilação são um conjunto de defeitos genéticos de tipo mul-tissistêmico que afetam a função da proteína. Foram descritas 75 doenças desde seus primeiros estudos. Neste estudo, foi desenvolvido um método microespectrofotométrico para diagnosticar a enzima citosólica fosfomanose isomerase EC 5.3.1.8 (PMI); foram analisadas 32 amostras de indivíduos entre 0,6 e 27 anos e estabelecidos o intervalo e o valor de referência de atividade enzimática específica. Este estudo permitirá iniciar o diagnóstico de pacientes deficientes da PMI de forma precoce e oportuna, o que a converte em uma possível enzima candidata para testes genéticos de rastreio pré-natal, já que essa patologia tem um tratamento fácil e de baixo custo, que consiste na suplementação de manose por via oral. O diagnóstico clínico desse defeito metabólico beneficiará o paciente e sua família ao melhorar a qualidade de vida e o sistema de saúde colombiano.

Fourier Transform Infrared Microspectroscopy of Rat Kidney with Regard to Fa- tal Hyperthermia / 法医学杂志

OBJECTIVE@#To observe the chemical groups changing in rat kidney with regard to fatal hyperthermia by Fourier transform infrared microspectroscopy (FTIR-MSP) and to provide a new method to diagnose fatal hyperthermia.@*METHODS@#Rats were sacrificed by hyperthermia, brainstem injury, massive hemorrhage and asphyxiation and divided into groups. The renal samples were dissected immediately after death. The data of infrared spectroscopy in glomerulus were measured by FTIR-MSP.@*RESULTS@#The absorbances of 3290, 3070, 2850, 1540 and 1396 cm(-1) significantly increased (P < 0.05), and the ratios of Al650/A3290 and A1650/A1540 significantly decreased (P < 0.05) in group of hyperthermia.@*CONCLUSION@#FTIR-MSP can analyze the changes of chemical groups of kidney as an auxiliary diagnosis for discriminating hyperthermia with other causes of death.

assessment of the accuracy of visual diagnosis of meconium-stained amniotic fluid

The assessment of meconium content in the amniotic fluid depends on visual observation by clinicians at the bedside. The aim of the present study was to compare visual evaluation of meconium-stained amniotic fluid with spectrophotometer evaluation. Study Design: Ten gram of meconium was added to 100 ml of amniotic fluid and mixed. The solution was serially two-fold diluted with amniotic fluid. The serially diluted tubes' absorbance spectrum was measured at 420 nm and thus a standard scale was established. Ninetyfive samples of meconium-stained amniotic fluid were collected from labouring women and the grade of meconium was determined visually at the bedside. The samples' absorbance spectrum was measured at 420 nm and recorded. Spectrophotometer was considered gold standard and the ranges of optical density in the standard scale was used to test the accuracy of visual categorization of the samples. In the statistical analysis chi-square test was used and significance was p<0.05. The accuracy rate of visual diagnosis of meconium-stained amniotic fluid were found as statistically significant [accuracy rate=54.74%, p<0.001]. Visual evaluation was correct in 19.4% of thin, 53.1% of moderate and 90.6% of thick meconium samples when examined with spectrophotometer. Visually diagnosed thin meconium can be moderate or thick meconium when examined objectively. The visual diagnosis at bedside is not always reliable and should be replaced with an objective method like spectrophotometery

Pharmaceutical study on cinnarizine floating systems for oral sustained release drug delivery

In this study, cinnarizine was formulated in three different floating systems to control its release in gastric fluid. Four formulae containing different concentrations of Eudragit L100 [15-30% [w/w]] were prepared using sodium bicarbonate as CO2 generating agent. Three formulae containing methyl cellulose, hydroxypropyl methyl cellulose [HPMC] and sodium alginate of different viscosity grades [low, medium and high] were studied. Six formulae were also prepared using mixed polymers containing constant Eudragit L100 concentration and two concentrations of sodium alginate [of different viscosity grades]. All prepared formulae were evaluated through determining floating time, in vitro release rate of cinnarizine and in vivo bioavailability assessment of cinnarizine from selected floating systems. The obtained results in this study proved the effectiveness of the prepared cinnarizine floating systems in increasing its bioavailability. It also indicated the superiority of Eudragit L100 system over the alginate-cellulose system, which in turn has a higher bioavailability than the conventional cinnarizine capsule [197.57% and 176%, respectively]

Bilirrubinometría transcutánea y egreso hospitalario temprano en neonatos de bajo riesgo con ictericia / Transcutameous bilirubinometry and aerly hospital discharge in low risk newborns with jaundice

Introducción. El egreso y el inicio de intervenciones terapéuticas tempranas en recién nacidos ictéricos, dependen de la cuantificación de la bilirrubina sérica. Objetivo. Comparar las cifras de bilirrubina sérica total con las obtenidas mediante la bilirrubinometría transcutánea y evaluar la utilidad como herramienta diagnóstica para decidir el alta hospitalaria temprana. Material y métodos. Estudio transversal y prolectivo. Se realizaron determinaciones casi simultáneas de bilirrubina sérica total y bilirrubinometría transcutánea en neonatos con peso > 2, 000 g, edad gestacional > 36 semanas y edad extrauterina < 72 horas de vida. Resultados. 100 neonatos cumplieron los criterios de inclusión, con peso de 3,135 ñ 499.9 g, edad gestacional de 38.85 ñ 1.4 semanas y edad extrauterina al momento del estudio de 45.46 ( 1.75 horas. El coeficiente de correlación fue de 0.81 (p< 0.0001). Discusión. La bilirrubinometría transcutánea debe realizarse en todo neonato incluido en el programa de puerperio de bajo riesgo. Cifras < 4.9 mg/dL durante las primeras 24 horas de vida y niveles menores de 7.9 mg/dL entre las 48 y 72 horas, se consideran seguras para permitir el egreso hospitalario temprano en los neonatos estudiados.

Effects of Blood-Brain Barrier Disruption on Cerebral Oxygen Balance / 대한구급학회지

BACKGOUND: Disruption of the blood-brain barrier (BBB) can alter the internal milieu and may increase the release of excitatory amino acid neurotransmitters or catecholamines, which may affect metabolic rate or coupling. This study was performed to evaluate whether disruption of BBB by unilateral intracarotid injection of hyperosmolar mannitol would alter oxygen supply/consumption balance in the ipsilateral cortex. METHODS: Rats were anesthetized with 1.4% isoflurane using mechanical ventilation via tracheostomy. 25% mannitol was administered at a rate of 0.25 mlxkg-1s-1 for 30 s through unilateral internal carotid artery. The BBB transfer coefficient (Ki) of 14C-alpha-aminoisobutyric acid was measured in one group (N=7) after administering mannitol. Regional cerebral blood flow (rCBF), regional arterial and venous O2 saturation and O2 consumption were measured in another group using a 14C-iodoantipyrine and microspectrophotometry (N=7). RESULTS: Vital signs were similar before and after administering mannitol. Ki was significantly higher in the ipsilateral cortex (IC) than in the contralateral cortex (CC), (22.3+/-8.4 vs 4.4+/-1.1 microliterxg-1min-1). rCBF was similar between IC (105+/-21 mlxg-1min-1) and the CC (93+/-20). Venous O2 saturation was lower in the IC (43+/-7%) than in the CC (55+/-4). O2 consumption was higher in the IC (9.6+/-3.0 mlx100 g-1min-1) than in the CC (6.7+/-1.5). CONCLUSIONS: Our data suggested that increasing permeability of the BBB increased cerebral O2 consumption and deteriorated cerebral oxygen balance.

The Effects of Nitric Oxide on Oxygen Balance in Cerebral Ischemia / 대한구급학회지

Bockground: Nitric oxide (NO) is an important regulator of blood flow and also works as a neuronal messenger via cyclic GMP. Recent studies regarding the therapeutic utility of nitric oxide synthase (NOS) inhibitors in reducing ischemia-induced neuronal damage are very controversial. The possible neuroprotective effect of NO or NOS inhibitors in ischemic neuronal damage could occur at the vascular and or neuronal level. This study investigated whether the NOS inhibitor, NG-nitro-L-arginine-methyl ester (L-NAME) would alter oxygen balance in ischemic cerebrocortex of isoflurane-anesthetized rats. METHODS: Fifteen minutes after middle cerebral artery occlusion, L-NAME (1.5 mgxmin-1kg-1) was infused intravenously to the L-NAME group (n=14), and normal saline was given to the control group (n=14) for 45 minutes. Regional cerebral blood flow was determined with [14C]iodoantipyrine, and arterial and venous oxygen saturations were determined by microspectrophotometry. RESULTS: Regional cerebral blood flow of the ischemic cortex was significantly lower than that of the contralateral cortex in both groups. In the control group, ischemic cortex; 55+/-13, contralateral cortex; 110+/-29 mlxmin-1100 g-1, and in the L-NAME group, ischemic cortex; 35+/-13, contralateral cortex; 90+/-24 mlxmin-1100 g-1. Compared with the blood flow in the ischemic cortex of the control group, L-NAME significantly reduced ischemic blood flow by 36%. Venous oxygen saturation was significantly increased in the ischemic cortex (41+/-1% in control, 44+/-3% in L-NAME) but decreased in the contralateral cortex (65+/-3% in control, 61+/-3% in L-NAME) by L-NAME. Ischemic cortical oxygen consumption in the L-NAME group was 39% lower than that in the corresponding control group, whereas the difference was only 11% in the contralateral sides between groups. The ratio of oxygen supply to consumption was lower in the ischemic than in the nonischemic regions in both groups. In the ischemic cortex, this ratio was significantly lower in the control group (1.7+/-0.1) than in the L-NAME group (1.9+/-0.1). In contrast, the ratio tended to be decreased by L-NAME in nonischemic regions. CONCLUSIONS: These observations suggest that despite a decrease in cerebral blood flow, inhibition of nitric oxide synthesis mildly improves the oxygen supply and consumption balance in the ischemic cortex.

Human renal carcinoma: Feulgen-DNA, area and chromatin condesation determined by scanning cytometry

Säo reportadas neste trabalho as variaçöes em conteúdo de DNA, as correspondentes variaçöes das áreas cobertas por cromatina-Feulgen reativa e o grau de empacotamento (condensaçäo) cromatínica em núcleos de células de carcinomas renais. O método eleito, para levar a efeito o trabalho, foi o de análise de imagem a partir de dados obtidos com microespectrofotometria de varredura. Controles de muita precisäo foram levados a efeito usando-se núcleos de eritrócitos de frango, neutrófilos humanos e núcleos epiteliais de rim humano normal. Os núcleos tumorais apresentaram uma populaçäo poliplóide, tendo como origem provável uma populaçäo hipodiplóide ("stem cell"). Detectou-se nos núcleos tumorais, um aumento de área coberta por material Feulgen-positivo, tal fato correspondendo a um fenômeno geral de descondensaçäo cromatínica. Este achado, contudo, foi acompanhado por um aumento dos valores da razäo das absorbâncias médias (AAR) significando que as regiöes de cromatina condensada, nos casos de tumores renais, säo mais condensadas do que as correspondentes das células normais