Decolorization of the benzidine-based azo dye Congo red by the new strain Shewanella xiamenensis G5-03 / Descoloração do corante azo derivado da benzidina vermelho Congo por uma nova linhagem de Shewanella xiamenensis G5-03

Shewanella xiamenensis G5-03 was observed to decolorize the azo dye Congo red in synthetic wastewater. The influence of some factors on the dye decolorization efficiency was evaluated. The optimal decolorization conditions were temperature 30-35 °C, pH 10.0, incubation time 10 h, and static condition. The kinetic of Congo red decolorization fitted to the Michaelis–Menten model (Vmax = 111.11 mg L-¹ h-¹ and Km = 448.3 mg L-¹). The bacterium was also able to degrade benzidine, a product of azo bond breakage of the Congo red, which contributed to reduce the phytotoxicity. The ability of S. xiamenensis G5-03 for simultaneous decolorization and degradation of Congo red shows its potential application for the biological treatment of wastewaters containing azo dyes.

Shewanella xiamenensis G5-03 foi capaz de descolorir o corante azo vermelho Congo em água residuária sintética. A influência de alguns fatores na eficiência da descoloração do corante foi avaliada. As condições ótimas de descoloração foram temperatura de 30-35 °C, pH 10,0 e condições estáticas. A cinética de descoloração do vermelho Congo se ajustou ao modelo de Michaelis–Menten (Vmax = 111,11 mg L-¹ h-¹ and Km = 448,3 mg L-¹). A bactéria também foi capaz de degradar a benzidina, um produto da quebra da ligação azo do vermelho Congo, o que contribuiu para a redução da fitotoxicidade. A habilidade da S. xiamenensis G5-03 em simultaneamente descolorir e degradar o vermelho Congo demostra seu potencial de aplicação no tratamento de águas residuárias contendo corantes azo.

Decolorization of the benzidine-based azo dye Congo red by the new strain Shewanella xiamenensis G5-03 / Descoloração do corante azo derivado da benzidina vermelho Congo por uma nova linhagem de Shewanella xiamenensis G5-03

Shewanella xiamenensis G5-03 was observed to decolorize the azo dye Congo red in synthetic wastewater. The influence of some factors on the dye decolorization efficiency was evaluated. The optimal decolorization conditions were temperature 30-35 °C, pH 10.0, incubation time 10 h, and static condition. The kinetic of Congo red decolorization fitted to the Michaelis­Menten model (Vmax = 111.11 mg L-1 h-1 and Km = 448.3 mg L-1). The bacterium was also able to degrade benzidine, a product of azo bond breakage of the Congo red, which contributed to reduce the phytotoxicity. The ability of S. xiamenensis G5-03 for simultaneous decolorization and degradation of Congo red shows its potential application for the biological treatment of wastewaters containing azo dyes.

Shewanella xiamenensis G5-03 foi capaz de descolorir o corante azo vermelho Congo em água residuária sintética. A influência de alguns fatores na eficiência da descoloração do corante foi avaliada. As condições ótimas de descoloração foram temperatura de 30-35 °C, pH 10,0 e condições estáticas. A cinética de descoloração do vermelho Congo se ajustou ao modelo de Michaelis­Menten (Vmax = 111,11 mg L-1 h-1 and Km = 448,3 mg L-1). A bactéria também foi capaz de degradar a benzidina, um produto da quebra da ligação azo do vermelho Congo, o que contribuiu para a redução da fitotoxicidade. A habilidade da S. xiamenensis G5-03 em simultaneamente descolorir e degradar o vermelho Congo demostra seu potencial de aplicação no tratamento de águas residuárias contendo corantes azo.

Method for determining o-tolidine in workplace air by gas chromatography / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To establish a method for determining o-tolidine in workplace air by gas chromatography.</p><p><b>METHODS</b>o-tolidine in workplace air was collected with a glass fiber filter, desorbed with methanol, and determined by gas chromatography-flame ionization detector.</p><p><b>RESULTS</b>The concentration of o-tolidine showed a linear relationship with peak area within 0.04∼9.00 µg/ml; the detection limit was 0.04 µg/ml; the minimum detectable concentration was 0.0002 mg/m(3) (calculated by 375 L air sample); the sampling efficiency was 93%∼100%; the elution efficiency was 94%∼96%; the relative standard deviation was 0.8-2.5%. Sample could be stored at 4 °C for at least 8 days and at room temperature for as long as 6 days.</p><p><b>CONCLUSION</b>This determination method meets the requirements of Guide for establishing occupational heath standards-Part 4 Determination methods of air chemicals in workplace (GBZ/T 210.4-2008) and is suitable for determination of o-tolidine in workplace air.</p>

Clinical Characteristics in Attention-Deficit Hyperactivity Disorder with/or without Developmental Coordination Disorder Patients

OBJECTIVES: This study explored the clinical differences in attention-deficit hyperactivity disorder (ADHD) patients with and without developmental coordination disorder (DCD). METHODS: Participants were 49 children and adolescents with ages between 6 and 18 years. These subjects were placed into 2 groups: ADHD without DCD (24) and ADHD with DCD (25). We used several evaluation tools on both groups: the Affective Disorders and Schizophrenia-Present and Lifetime Version- Korean Version (K-SADS-PL), Wechsler Intelligence Scale for Children-III (WISC-IIII), Child Behavior Check List (CBCL), Korean Personality Rating Scale for Children (K-PRC), and Bruininks-Osretsky Test of Motor (BOT-2). RESULTS: Patients with both ADHD and DCD had a lower performance intelligence quotient and more internal and external behavioral symptoms than patients with ADHD but not DCD. It is possible that patients with ADHD and motor coordination problems should be noticed earlier and given intensive treatment.

Clinical Characteristics in Attention-Deficit Hyperactivity Disorder with/or without Developmental Coordination Disorder Patients

OBJECTIVES: This study explored the clinical differences in attention-deficit hyperactivity disorder (ADHD) patients with and without developmental coordination disorder (DCD). METHODS: Participants were 49 children and adolescents with ages between 6 and 18 years. These subjects were placed into 2 groups: ADHD without DCD (24) and ADHD with DCD (25). We used several evaluation tools on both groups: the Affective Disorders and Schizophrenia-Present and Lifetime Version- Korean Version (K-SADS-PL), Wechsler Intelligence Scale for Children-III (WISC-IIII), Child Behavior Check List (CBCL), Korean Personality Rating Scale for Children (K-PRC), and Bruininks-Osretsky Test of Motor (BOT-2). RESULTS: Patients with both ADHD and DCD had a lower performance intelligence quotient and more internal and external behavioral symptoms than patients with ADHD but not DCD. It is possible that patients with ADHD and motor coordination problems should be noticed earlier and given intensive treatment.

Programando o software Imagej para a seleção automática de áreas marcadas por imuno-histoquímica e coradas com o cromógeno dab e contra-coradas pela hematoxilina / Programming Imagej software for automatic selection of immunohistochemistry marked areas stained by dab chromogen and counter-stained by hematoxylin

Introdução: A quantificação dos parâmetros de imunomarcação na análise de imagens de imuno-histoquímica depende da adequada delimitação das regiões marcadas, sendo a seleção destas regiões realizada manualmente ou através de métodos computacionais automáticos ou semi-automáticos. Objetivos: Viabilizar um método de seleção automática de áreas marcadas por imuno-histoquímica por meio do software ImageJ, apresentando a programação envolvida e analisando quantitativamente as imagens. Métodos: Para a aplicação e avaliação da programação proposta neste trabalho foram utilizadas imagens de imuno-histoquímica para localização de alfa-actina de músculo liso (α-SMA) provenientes de estudo de modelo animal experimental de fibrose renal em ratos Wistar machos. Foram capturadas imagens de áreas distintas da região do córtex renal contendo regiões positivas coradas pelo 3,3'-Diaminobenzidina (DAB) em uma lâmina com imunomarcação extensa (n=10) e em outra lâmina com Imunomarcação focal (n=10), provenientes de dois animais. A contra-coloração foi feita com hematoxilina. Resultados: A diferença da extensão da imunomarcação foi qualitativamente evidente entre os dois grupos de imagens. As áreas reveladas pelo DAB nas lâminas dos animais I e II foram selecionadas automaticamente pelo programa, ajustadas pelo operador, e quantificadas, apresentando fração de área ocupada de 23,48 ± 6,52% e de 0,10 ± 0,04% respectivamente, valores expressos em média ± desvio padrão com diferença estatística significativa (p < 0,0001).Conclusões: O software ImageJ mostrou-se adequado para a aplicação da seleção automática de áreas marcadas por imuno-histoquímica, embora a intervenção do operador tenha sido necessária para a correta identificação dos pontos de corte do "threshold", supervisionando a correta delimitação das áreas.

Introduction: The immunostaining quantitative parameters of immunohistochemistry image analysis depend on the proper delimitation of marked regions. The selection of those marked regions is carried out manually, or by computed semi-automatic or automatic methods.Objectives: to carry out a feasible automatic method on selection of immunohistochemistry stained areas applying the Image J software (NIH), presenting the computed program and the quantitative analysis of some images.Methods: For application and evaluation of the proposed program of the study, we used immunehistochemistry images of alpha-smooth muscle actin (α-SMA), a marker of epithelial dedifferentiation, coming from an experimental study of renal fibrosis in Wistar male rats. The images have been captured from non-overlapping regions of the renal cortex, comprising positive 3,3'-Diaminobenzidine (DAB)-stained fields of a strong immunostained slide (N=10, animal I) and another slide with focal immune pattern (N=10, animal II), obtained from two different animals. The slides were counter-stained by Hematoxylin.Results: The difference by immunostaning between the two specimens was qualitatively evident. The areas revealed as DAB-positive in animals I and II slides were automatically selected by the proposed program, visually adjusted by the operator, and finally, quantified, resulting in an occupied area fraction of 23.48 ± 6.52% and 0.10 ± 0.04%, respectively, p<0.0001. Conclusion: The software Image J was able to perform a feasible automatic selection of immunohistochemistry DAB stained areas, although the operator intervention has been necessary to identify the correct threshold cut-off points, supervising if the delimitation of marked areas was precise.

Clinical Features and Effects of Occupational Therapy in Children with Developmental Coordination Disorder

OBJECTIVE: To assess the clinical features of children with developmental coordination disorder (DCD) and the effects of occupational therapy. METHOD: Seventeen children were diagnosed with DCD using by Bruininks-Osteretsky test of motor proficiency (BOTMP). To investigate the clinical features, neurological examinations such as soft neurological signs, speech evaluation, cognitive assessment, and psychiatric evaluation were performed on the 17 participants. Among the participants, 8 children had occupational therapy. BOTMP and cognitive function were evaluated both prior to and 6 months after the intervention. RESULTS: Children with DCD showed various soft neurological signs and co-morbidities such as attention-deficit hyperactivity disorder (ADHD), developmental language disorder, and emotional problems. The cognitive assessment revealed disproportionately low performance IQ for all subjects. After occupational therapy, scores for BOTMP and performance IQ increased significantly. CONCLUSION: The clinical features of DCD are heterogeneous and occupational therapy focused on motor proficiency was effective. This study supports raising interest in and attention on children with motor coordination difficulties.

Occupational Reproductive Function Abnormalities and Bladder Cancer in Korea

The purpose of this study was to review occupational reproductive abnormalities and occupational bladder cancer in Korea and to discuss their toxicological implications. Reproductive dysfunction as a result of 2-bromopropane poisoning was first reported in Korean workers. In 1995, 23 of the 33 workers (25 female and 8 male workers) who were exposed to 2-bromopropane during the assembly of tactile switch parts developed reproductive and/or hematopoietic disorders. A total of 17 (68%) workers were diagnosed with ovarian failure. Two of the eight male workers experienced azoospermia and four workers experienced some degree of oligospermia or reduced sperm motility. In summary, 2-bromopropane poisoning caused severe reproductive effects in Korean workers. The prognosis was poor for reproductive dysfunction. A few cases of occupational bladder cancer have been reported in Korea, whereas other cancers of the urinary tract have not been reported after occupational exposure. A few cases of benzidine-induced cancer have been reported in Korea and 592 workers in Japan have received compensation for benzidine and beta-naphthylamine-induced cancer. In conclusion, a few cases of benzidine-induced occupational bladder cancer have been reported in Korea. However, benzidine-induced bladder cancer will likely be an important occupational health issue in Korea in the coming years.

Protein expression of 5-lipoxygenase and activation and cytotoxicity of Benzidine in human bronchial epithelial cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effect of intracellular 5-lipoxygenase on oxidation of benzidine in HBE cells and to provide further evidence that lipoxygenase is an alternative pathway for the oxidation of xenobiotics mediated by cytochrome P450.</p><p><b>METHODS</b>Enzyme system test: Soybean lipoxygenase (SLO), substrate (benzidine) and other components reacted in the enzyme system, followed by detection of the reaction products by spectrophotometry. In vitro test: After HBE cells were exposed to benzidine, the protein levels of 5-lipoxygenase in HBE cells were assessed by Western-blot, and the DNA damage by the single cell gel electrophoresis. At last, the effect of the specific inhibitor of 5-lipoxygenase (AA861) on 5-lipoxygenase protein expression and DNA damage in HBE cells were detected.</p><p><b>RESULTS</b>SLO could catalyze the co-oxidation of benzidine to generate benzidine diimine in the presence of hydrogen peroxide. Under optimal condition, numax value of the oxidation of benzidine catalyzed by SLO was 1.42 nmol*min(-1) SLO, and the Km value of benzidine was 1.48 mmol/L. EGCG could inhibit the oxidation of benzidine by SLO. Benzidine could induce 5-lipoxygenase protein expression in HBE cells, but AA861 was invalid. Benzidine caused DNA damage in HBE cells, which could be significantly inhibited by AA861.</p><p><b>CONCLUSION</b>5-LOX protein expression in HBE cells can be regulated by benzidine, which suggests that the co-oxidation of benzidine by 5-LOX could produce into electrophile that could covalently bind to DNA and induce DNA damage, which could be one of the mechanisms for carcinogenesis of BZD. 5-LOX inhibitor AA861 can inhibit this effect.</p>

Expression of protein p53 in workers occupationally exposed to benzidine and bladder cancer patients / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study expression of mutant p53 protein in workers occupationally exposed to benzidine and bladder cancer patients.</p><p><b>METHODS</b>Mutant p53 protein in serum from the workers occupationally exposed to benzidine and bladder cancer patients were determined with Immuno-PCR, while exfoliated urothelial cells in the urine samples were classified with Papanicolau grading.</p><p><b>RESULTS</b>Positive rate of mutant p53 protein increased with the exposed intensity index in workers occupationally exposed to benzidine. The positive rate of mutant p53 protein in bladder cancer patients (83.3%) was significantly higher than that in the group 1 of exposed intensity index. The average scanning integrals of PCR amplified band in the group of bladder cancer patients and group 2 of exposed intensity index were both higher than that in the group 1 significantly. Workers in the groups of different exposed intensity indices were further stratified according to Papanicolau grades. In the group 2 of exposed intensity index, the average scanning integrals of PCR amplified band in the stratum of Papanicolau grade II and III were significantly higher than that in the strata of Papanicolau grade I. And in the group 3 of exposed intensity index, the positive rate of mutant p53 protein in the strata of Papanicolau grade III was higher than that in the strata of Papanicolau grade I significantly.</p><p><b>CONCLUSION</b>The increase of exposed intensity may not only result in the positive rate of mutant p53 protein, but also the quantity of mutant p53 protein in serum within the low range of benzidine exposure. Once the exposed intensity was beyond that spectrum, the positive rate of mutant p53 protein in serum and the average scanning integrals of PCR amplified band were no longer enhanced with the increase of exposed intensity. There was tight correlation between Papanicolau grade of exfoliated urothelial cells and the positive rate or the quantity of mutant p53 protein for the higher benzidine exposure intensity.</p>

Polymorphism of N-acetyltransferase 2 (NAT2) gene polymorphism in shanghai population: occupational and non-occupational bladder cancer patient groups / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Arylamine N-acetyltransferases (NATs) are involved in the detoxification of aromatic amines and hydrazine. In order to explore the possible association of NAT2 polymorphism with bladder cancer risk in benzidine exposed or non-exposed Chinese individuals, healthy subjects, subjects with bladder cancer of a former benzidine exposed cohort in Shanghai dyestuff industry and a group of bladder cancer patients without known occupational exposure to aromatic amines were genotyped for NAT2 gene polymorphism.</p><p><b>METHODS</b>NAT2 genotyping was performed with a set of RFLP procedures at seven major polymorphic loci of gene coding area: G191A, C282T, T341C, C481T, G590A, A803G and G857A.</p><p><b>RESULTS</b>The wild allele NAT2 *4 was the most prevalent allele (59%) in healthy individuals. The alleles NAT2*6A and NAT2*7B were also frequently observed (21% and 17%, respectively). In contrast to Caucasians, the percentage of slow acetylators was lower (12% in Chinese vs. 58% in Caucasians, P < 0.001). No relevant differences were observed for homogenous rapid, heterogeneous rapid/slow and homogeneous slow acetylation genotypes between the healthy subjects and both groups of bladder cancer patients.</p><p><b>CONCLUSION</b>The present work did not support the association of slow acetylating genotypes of NAT2 gene with elevated risk of bladder cancer in Chinese whereas it was documented as an important genetically determined risk factor in Caucasians. Different mechanisms might play a role in individual susceptibility to bladder cancer related with aromatic amine exposure in various races or ethnic groups.</p>

Stimulatory response to the oxidation of benzidine and other xenobiotics mediated by lipoxygenase in the presence of phenothiazines / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To test the possibility that certain efficient substrates for lipoxygenase (LOX) produce shuttle oxidants that stimulate the generation of reactive oxygen species from other chemicals.</p><p><b>METHODS</b>Metabolic interactions were conducted in vitro between chlorpromazine (BZ) or other phenothiazines and benzidine or other xenobiotics mediated by soybean lipoxygenase (SLO) or purified human term placental lipoxygenase (HTPLO) in the presence of hydrogen peroxide, and the rates of xenobiotics oxidation were measured by spectroscopic analysis.</p><p><b>RESULTS</b>Chlorpromazine cation radical (CPZ(*+)) generated by lipoxygenase triggered a rapid oxidation of benzidine to benzidine diimine cation. Under the experimental conditions used, the metabolic interaction resulted in a 42-fold greater stimulation than BZ alone in the rate of BZ oxidation. The magnitude of stimulation of benzidine oxidation exhibited a dependence on the pH of the reaction medium, amount of the enzyme, and concentration of chlorpromazine, BZ, and hydrogen peroxide. A number of other phenothiazines were also found to stimulate BZ oxidation, albeit to a lesser degree. Chlorpromazine cation radical stimulated the oxidation of all six other xenobiotics tested. The highest stimulation (94-fold) was noted in the oxidation of tetramethyl phenylenediamine to Wursters blue radical, while the lowest stimulatory response (2-fold) was observed in guaiacol. Preliminary data showed that purified HTPLO also displayed a similar stimulatory response to benzidine oxidation in the presence of chlorpromazine.</p><p><b>CONCLUSION</b>Both soybean lipoxygenase and purified human term placental lipoxygenase can mediate stimulatory response to the oxidation of benzidine and other xenobiotics in the presence of phenothiazines.</p>

Biodegradation of benzidine based azodyes Direct red and Direct blue by the immobilized cells of Pseudomonas fluorescens D41.

Benzidine based azodyes are proven carcinogens, mutagens and have been linked to bladder cancer of human beings and laboratory animals. The textile and dyestuff manufacturing industry are the two major sources that released azodyes in their effluents. The dye, Direct blue contains two carcinogenic compounds namely benzidine (BZ), 4-amino biphenyl (4-ABP), while the dye Direct red has benzidine (BZ). Among 40 isolates of Pseudomonas fluorescens screened, one isolate designated as D41 was found to be capable of extensively degrading the dyes Direct blue and Direct red. Immobilized cells of P. fluorescens D41 efficiently degraded Direct red (82%) and Direct blue (71%) in the presence of glucose.

Polymorphism of glutathione S-transferase T1, M1 and P1 genes in a Shanghai population: patients with occupational or non-occupational bladder cancer / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Glutathione S-transferases are involved in the conjugation of xenobiotics. To explore whether GSTs polymorphisms are involved in the development of occupational or non-occupational bladder cancer, polymorphism frequencies of GSTT1, M1 and P1 were investigated in a normal population, which had been settled in a rural area in Shanghai suburb for at least 5 generations as well as in a group of patients with benzidine exposure related occupational bladder cancer in Shanghai dyestuff industry and a group of patients with non-occupational bladder cancer.</p><p><b>METHODS</b>PCR based procedures were performed in the study populations to confirm the genotypes of GSTT1, M1 and P1.</p><p><b>RESULTS</b>The polymorphisms at locus of GSTP1-A1578G in the normal population differed significantly from those in Caucasians or African Americans. All the subjects genotyped so far (n = 118) bore only homogenous wild genotype (C2293/C2293) at GSTP1-C2293T locus. This locus seemed to be a monomorphic in Shanghai population. No significant difference in GSTT1 and GSTM1 polymorphic form frequencies could be confirmed among three groups of subjects. An overrepresentation of GSTP1 AG or GG genotype corresponding a less stable and less effective isozyme protein was detected in patients with benzidine related occupational bladder cancer, compared with that in the normal population though a statistical significance was not yet reached (P = 0.09, OR = 1.96, 95% CI 0.89-4.32).</p><p><b>CONCLUSION</b>This study suggests that GSTM1 or GSTT1 homozygous deficiency genotypes and their combination do not have a clear impact on bladder cancer incidence in a Shanghai population. It seems that GSTP1 polymorphism is not associated with non-occupational bladder cancer. GSTP1 AG or GG genotype has a higher frequency in the patients with benzidine related occupational bladder cancer, and further work is needed to confirm if GSTP1 AG or GG genotype plays a role in the development of occupational bladder cancer.</p>

Pueden usarse las tiras reactivas para orina en la detección de sangre oculta en heces? / Can diagnostic urine strips be used to search blood in stool?

Introducción. En diversos hospitales se emplean las tiras reactivas para orina (Labstix) para demostrar sangre oculta en heces (SOH). El objetivo de este estudio fue valorar la efectividad de las tiras reactivas para orina en la detección de SOH versus bencidina como estándar de oro. Material y métodos. Se realizó un estudio de investigación prospectivo, comparativo, transversal y observacional en el laboratorio del Hospital Infantil de México Federico Gómez. Doscientas sesenta muestras se obtuvieron de pacientes con posibilidad de sangrado por tubo digestivo; en cada una de ellos se realizaron las técnicas de bencidina y tiras reactivas para orina. La observación en las tiras reactivas se realizó en dos formas: introduciéndola directamente en las heces formadas y la otra impregnándola con heces previamente hidratadas. Resultados. Del total de muestras, sólo 54 resultaron positivas con la técnica de bencidina y 206 negativas. Para la tira reactiva en directo la sensibilidad fue de 88.8 por ciento (44/54) y la especificidad de 22.8 por ciento; con la tira reactiva en directo la sensibilidad fue de 100 por ciento (54/54) y 0 por ciento, respectivamente. Conclusión. Se demostró estadísticamente que no existe utilidad diagnóstica con las tiras reactivas para orina para detectar SOH

Effect of age on susceptibility of carcinogen benzidine-hydrochloride.

Susceptibility of carcinogen (benzidine hydrochloride) was checked by degranulation technique in rats of three different (young, adult and old) age groups. The dose response curves of these three different groups showed different per cent degranulation. Comparative data of dose response of benzidine hydrochloride observed on the basis of RNA/Protein ratio basis showed that old animals were more susceptible to carcinogens than young and adult animals.