An ELISA-like time-resolved fluorescence immunoassay for microcystin detection

Background: A time-resolved fluorescence immunoassay (TRFIA), based on anti-microcystin-LR (MCLR) monoclonal antibodies (MAbs) and europium-labeled antimouse IgG conjugate, was first developed for microcystin detection. Methods: Anti-MCLR MAbs were prepared by a standard method, and the attained MAbs showed a good cross reactivity with MCLR, MCRR and MCYR. The TRFIA was performed in an indirect competitive mode. The detection method of TRFIA was compared with indirect competitive enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC). Results: The TRFIA exhibited a typical sigmoidal response for MCLR at concentrations of 0.005-50 ng/ml, with a wide quantitative range between 0.01 and 10 ng/ml, indicating the broadest detective range and the most sensitive of all the methods for microcystins (MCs) detection. Additionally, the TRFIA maintained good reliability through its quantitative range, as evidenced by low coefficients of variation (1.6-12.2%). The toxin data of algal samples assayed from TRFIA were in the same range as those with ELISA and HPLC, implying that the method was reliable and practical for the detection of MCs. Conclusions: The TRFIA may offer a valuable alternative or a substitute for conventional ELISA for microcystin detection. (C) 2004 Elsevier B.V. All rights reserved.

A new method for screening alpha-glucosidase inhibitors and application to marine microorganisms

A new enzyme assay method for screening alpha-glucosidase inhibitors with rapidity and simplicity was developed. The enzyme-substituted alpha-glucosidases for this assay was glucoamylase. Samples were spotted or developed on the silica gel plate. The agar solution containing substrate was poured on the plate, and paper impregnated with enzyme was layered on the agar. After incubation, an inhibitory circle would appear around the inhibitor. By using this method, more than 200 strains of marine microorganisms were screened. Among them, three active strains were found to secrete inhibitors in the culture medium.

军队医学科技人员身心健康与影响因素的研究

To study the relationship between mental-physical health and coping behavior,job stress and job motivation in medical laboratory scientists and technicians. A cross-section survey was conducted,using the anxiety rating scale(SAS), depression rating scale(SDS), coping behavior rating scale,self-rating subhealth scale,self-rating stress scale,and self-rating job motivation scale among 289 medical laboratory scientists and technicians with analysis of ANOVA and Regression. The study was carried out in three steps:step1 is to analyze the general situation of the mental-physical health.Step 2 is to analyze the features of the main influencial facotors.Step 3 is to study the influence of the relative factors on mental-physical health.The following are the results: 1.The anxiety and depression scores of the medical researchers and technicians were 30±6 and 37±8 respectively,both significantly lower than those of the national norm（34±6 and 42±11 respectively,both P<0.01）,which are of significant difference. The depression score of the associate chief technician was 42±7, significantly higher than those of the medical researchers and technicians with any other professional titles (all P<0.05）, and the anxiety score of the associate chief technicians was 32±7, significantly higher than that of the research fellows（28±4, P<0.05），with no significant difference to the score of the medical researchers and technicians with other professional titles. The depression score,anxiety score and the subhealth score of the age group of 36～49 were 39±9,31±7 and 32±9 respectively, both higher than that of the age group of 20～35 （36±8,29±6 and 29±7 respectively,both P<0.05）,while there is much diference among other age groups. And the subhealth status has nothing to do with marital status,education background and professional titles. 2.The coping behaviors,job stress and motivation of the military medical laboratory scientists and technicians. 1）Coping behaviors:The seeking help score of the associate chief technicians was 2.8±0.5,not significantly different from that of the associate research fellows（2.8±0.5）,but significantly lower than those of the medical researchers and technicians with other professional titles(all P<0.01),and whose self-blame score was higher than that of the research fellows,which is of significant difference.The self-blame score and the imagination score of the associate research fellows were 2.0±0.5 and 2.4±0.5 respectively, significantly higher than that of the research fellows(1.6±0.4, 2.1±0.4,both P<0.01). The seeking help score of the females was 3.1±0.5, significantly higher than that of the males(2.9±0.5, both P<0.01),and the solving problem score was 3.4±0.6,significantly lower than the male(3.6±0.7, both P<0.01).There is no any significant difference in coping behaviors among researchers and technicians of different marital status,education background and age groups. 2)Job stress: The score of job stress of the age group of 35~49 and 50~60 were 23±8 and 25±6 respectively,significantly higher than that of the 20~35(21±7,both P<0.05). There is no any significant difference in job stress among researchers and technicians of different marital status,gender,professional titles and education background. 3)The score of meeting personal desire for reputation and interests of the males was 19.6±5.4，significantly higher than that of the females(18.4±5.0, both P<0.05), while there is no significant difference in job motivation among researchers and technicians of different titles,education,age and marital status. 3.The relationship of the mental-physical health of the military medical laboratory scientists and technicians with their coping behaviors,job stress and motivations 1) Coping behaviors:Regression analysis showed that, the best predictors of subhealth were anxiety,depression and escaping (β=-0.40,β=0.23, β=0.14, both P<0.01).the best predictors of anxiety and depression were active coping behaviors and negative coping behaviors (β=-0.40,β=0.40, both P<0.01).The moderate(rational) coping behaviors is the best predictor only of anxiety and of significant difference. 2)job stress : Regression analysis demonstrated that anxiety,depression,job stress 2 and job stress 4 are the best predictor for subhealth;job stress 12 is the best retro-predictor for depression; job stress 3 and 8 are the best predictor for depression;and job stress 2 and 9 are the best predictor for anxiety ,which are of significant difference. 3)Motivation:Regression analysis demonstrated that depression is the best predictor for subhealth while motivation itself doesn't predict subhealth;with respect to the specific contents of motivation,"probing unknown rules and making contributions to the human society "is the best retro-predictor for depression and has nothing to do with anxiety prediction. 4) The combined influences of coping behaviors,job stress and motivations: Regression analysis showed that, the best predictors for subhealth were anxiety,depression and job stress(β=0.41,β=0.24, β=0.19, both P<0.01).the best predictors for anxiety and depression were active coping behaviors and negative coping behaviors (β=-0.40,β=0.40, both P<0.01).The moderate(rational) coping behaviors is the best predictor only for anxiety and of significant difference. CONCLUSIONS: Regression analysis showed that the best predictors for anxiety and depression were active coping behaviors and negative coping behaviors, the best predictors for subhealth were anxiety,depression and job stress.Coping behaviors, job stress and motivations have significant influences on the mental-physical health of the military medical researchers and and technicians. Among them,coping behavior is the most important factor while job stress and motivation follow. Seeking help more often,less self-blame ,imagination and job stress could help release the anxiety, depression and improve the subhealth of the medical researchers and technicians. The findings of this study indicate we should address the physical and mental health of the military laboratory researchers and technicians.

Desmoplastic Small Round Cell Tumor of Major Salivary Glands: Report of 1 Case and a Review of the Literature

Desmoplastic small round cell tumor is a rare malignant neoplasm mostly occurring in the vicinity of or within the peritoneal cavity, and is uncommon in the head and neck region. Tumor location within a major salivary gland is exceptional. We report a case of a 41-year-old Chinese man with a history of diabetes mellitus and end-stage renal failure on peritoneal dialysis with a desmoplastic small round cell tumor occurring in the left submandibular gland. Fine-needle aspiration cytology showed variably cohesive clusters of small cells with hyperchromatic nuclei and fine granular chromatin. On histology the neoplasm displayed classic features of a desmoplastic small round cell tumor with angulated nests of small round blue cells in a fibromyxoid/desmoplastic stroma. Neoplastic cells were immunoreactive for cytokeratins (AE1/3), desmin (paranuclear dot-like), WT-1 (nuclear), epithelial membrane antigen, and CD56. EWS gene translocation and EWS-WT1 gene fusion were detected by fluorescence in situ hybridization and reverse transcriptase polymerase chain reaction, respectively. The case presented is the sixth case of and the oldest reported patient with a desmoplastic small round cell tumor occurring in a major salivary gland to date. Desmoplastic small round cell tumor should be considered in the differential diagnosis of a salivary gland neoplasm with a basaloid or small cell pattern on fine-needle aspiration cytology.

Lymphocytic follicles and aggregates are a determinant of mucosal damage and duration of diarrhea

Nonspecific changes (nonspecific chronic inflammation) in patients with chronic diarrhea represent the commonest diagnosis in colorectal biopsy interpretation, but these changes are of little clinical significance.

Mammalian triokinase and dihydroxyacetone kinase are the same enzyme

It is widely accepted that the ATP-dependent phosphorylation of D-glyceraldehyde in the fructokinase pathway of fructose metabolism requires the enzyme “triokinase”. However, experimental data on this enzyme are remarkably scarce. The enzyme has been purified from a variety of sources and peptides derived from the pig kidney enzyme show high similarity to human dihydroxyacetone kinase – an enzyme which also has FMN cyclase activity in high manganese ion concentrations. The properties of the two enzymes are also highly similar. Therefore it is proposed that mammalian triokinase and dihydroxyacetone kinase are, in fact, the same enzyme. This has consequences for investigations of normal and aberrant fructose metabolism and for the teaching of biochemistry in medical and science courses.

Evaluación del nivel de seguridad ocupacional en laboratorios universitarios de ingeniería en Bogotá

Antecedentes: Los laboratorios estudiantiles ofrecen riesgos para los estudiantes, la normatividad específica vigente no es de carácter obligatorio y en Bogotá aproximadamente hay 387 laboratorios. Objetivo: Evaluar el nivel de seguridad referente a salud, higiene y seguridad industrial en las prácticas que se desarrollan en los laboratorios de pregrado de Ingeniería en universidades de Bogotá. Métodos: Estudio descriptivo con un muestreo estratificado por conglomerado en ocho (8) universidades de la ciudad de Bogotá, con una confiabilidad del 95% y una precisión absoluta del 6% para una proporción máxima esperada; para evaluar los laboratorios se utilizó el Cuestionario de seguridad para laboratorios de secundaria (NTP 921) del Instituto Nacional de Seguridad e Higiene del Trabajo de España Resultados: Se evaluaron 166 laboratorios de 8 universidades de Bogotá, 65 de los laboratorios evaluados presentaron un riesgo bajo de los aspectos evaluados. Los aspectos con un porcentaje mayor o igual al 80% de cumplimiento de condiciones fueron aspectos generales del laboratorio, trabajo de los alumnos, manipulación de material, manipulación del producto, protección contra Incendios, eliminación de residuos, seguridad eléctrica y formación. Los ítems con un porcentaje menor del 60% de cumplimiento son los siguientes: sistemas de alarma contra incendios o emergencias; mantenimiento preventivo de la red eléctrica del laboratorio, capacitaciones anuales sobre seguridad en los laboratorios. aspectos generales del laboratorio, trabajo de los alumnos, manipulación de material, manipulación del producto, protección contra Incendios, eliminación de residuos, seguridad eléctrica y formación. Se realizó una prueba de asociación entre el tipo de laboratorio y cada ítem de las condiciones de seguridad del laboratorio. Los laboratorios que cumplen con un mayor número de condiciones de seguridad fueron los laboratorios químicos, el aspecto que obtuvo un porcentaje bajo de cumplimiento fue protección personal (40%); los aspectos que se cumplieron son aspectos generales del laboratorio, trabajo de los alumnos, manipulación de material, manipulación del producto, protección contra Incendios, eliminación de residuos, seguridad eléctrica y formación.

Validation of a novel ELISA for measurement of electronegative low-density lipoprotein

Background: Oxidative modification of low-density lipoprotein (LDL) plays a key role in the pathogenesis of atherosclerosis. LDL(-) is present in blood plasma of healthy subjects and at higher concentrations in diseases with high cardiovascular risk, such as familial hypercholesterolemia or diabetes. Methods: We developed and validated a sandwich ELISA for LDL(-) in human plasma using two monoclonal antibodies against LDL(-) that do not bind to native LDL, extensively copper-oxidized LDL or malondialdehyde-modified LDL. The characteristics of assay performance, such as limits of detection and quantification, accuracy, inter- and intra-assay precision were evaluated. The linearity, interferences and stability tests were also performed. Results: The calibration range of the assay is 0.625-20.0 mU/L at 1: 2000 sample dilution. ELISA validation showed intra- and inter- assay precision and recovery within the required limits for immunoassays. The limits of detection and quantification were 0.423 mU/L and 0.517 mU/L LDL(-), respectively. The intra- and inter- assay coefficient of variation ranged from 9.5% to 11.5% and from 11.3% to 18.9%, respectively. Recovery of LDL(-) ranged from 92.8% to 105.1%. Conclusions: This ELISA represents a very practical tool for measuring LDL(-) in human blood for widespread research and clinical sample use. Clin Chem Lab Med 2008; 46: 1769-75.

The effect of sixteen medicinal plants used in the Brazilian pharmacopoeia on the expression and activity of glutathione S-transferase in hepatocytes and leukemia cells

Glutathione S-transferase (GST) is a family of enzymes involved in the detoxification of electrophilic compounds. Different classes of GST are expressed in various organs, such as liver, lungs, stomach and others. Expression of GST can be modulated by diet components and plant-derived compounds. The importance of controlling GST expression is twofold: increasing levels of GST are beneficial to prevent deleterious effects of toxic and carcinogenic compounds, while inhibition of GST in tumor cells may help overcoming tumor resistance to chemotherapy. A screening of 16 plants used in the Brazilian pharmacopoeia tested their effects on GST expression in hepatocytes and Jurkat (leukemia) T-cells. The methanol extracts of five plants inhibited GST expression in hepatocytes. Three plants significantly inhibited and four others induced GST expression in Jurkat cells. Among these, the extracts of Bauhinia forficata Link. (Leguminosae) and Cecropia pachystachya Trec. (Urticaceae) inhibited GST expression at relatively low concentrations. With the exception of B. forficata, all plants were cytotoxic when administered to Jurkat cells at high doses (1 mg/mL) and some extracts were considerably cytotoxic even at lower concentrations.

Correlation Between Etest (R), Disk Diffusion, and Microdilution Methods for Antifungal Susceptibility Testing of Candida Species From Infection and Colonization

The correlation between the microdilution (MD), Etest (R) (ET), and disk diffusion (DD) methods was determined for amphotericin B, itraconazole and fluconazole. The minimal inhibitory concentration (MIC) of those antifungal agents was established for a total of 70 Candida spp. isolates from colonization and infection. The species distribution was: Candida albicans (n = 27), C. tropicalis (n = 17), C. glabrata (n = 16), C. parapsilosis (n = 8), and C. lusitaniae (n = 2). Non-Candida albicans Candida species showed higher MICs for the three antifungal agents when compared with C. albicans isolates. The overall concordance (based on the MIC value obtained within two dilutions) between the ET and the MD method was 83% for amphotericin B, 63% for itraconazole, and 64% for fluconazole. Considering the breakpoint, the agreement between the DD and MD methods was 71% for itraconazole and 67% for fluconazole. The DD zone diameters are highly reproducible and correlate well with the MD method, making agar-based methods a viable alternative to MD for susceptibility testing. However, data on agar-based tests for itraconazole and amphotericin B are yet scarce. Thus, further research must still be carded out to ensure the standardization to other antifungal agents. J. Clin. Lab. Anal. 23:324-330, 2009. (C) 2009 Wiley-Liss, Inc.

Anti-inflammatory activity of essential oil from leaves of Myrciaria tenella and Calycorectes sellowianus

The GC-MS analysis revealed that the leaf essential oils of Myrciaria tenella (DC.) Berg and Calycorectes sellowianus O. Berg (Myrtaceae) were composed of 34 and 37 compounds, respectively. The main constituents of M. tenella oil were beta-caryophyllene (25.1%), and spathulenol (9.7%), while for C. sellowianus were guaiol (13.1%) and beta-caryophyllene (8.6%). The anti-inflammatory effect of both essential oils was investigated in vitro and in vivo. Both oils reduced significantly (p < 0.005) the treated neutrophils chemotaxis with 93% and 91% inhibition for M. tenella and C. sellowianus, respectively. However, in the systemic treatment with the essential oils (50 mg/kg p.o.) only the M. tenella oil was able to significantly reduce the carrageenan-induced paw edema with a similar effect to that observed for indomethacin (10 mg/kg), the positive control.