Effect of Curcumin on Apoptosis of Acute T-Lymphoblastic Leukemia Cells induced by UMI-77 and Its Related Mechanism / 中国实验血液学杂志

AbstractObjective: To explore the effect and mechanism of curcumin on human T-cell acute lymphoblastic leukemia (T-ALL) cell apoptosis induced by Mcl-1 small molecule inhibitors UMI-77.@*METHODS@#T-ALL cell line Molt-4 was cultured, and the cells were treated with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77 for 24 h. The MTT method was used to detect the cell survival rate after different treatment; According to the results of curcumin and UMI-77, the experimental settings were divided into control group, curcumin group (20 μmol/L curcumin treated cells), UMI-77 group (15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells) and curcumin+ UMI-77 group (20 μmol/L curcumin and 15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells), MTT method was used to detect cell proliferation inhibition rate, Annexin V-FITC/PI double staining method and TUNEL staining were used to detect cell apoptosis, DCFH-DA probe was used to detect cell reactive oxygen species, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, Western blot was used to detect the expression levels of apoptosis-related proteins and Notch1 signaling pathway-related proteins.@*RESULTS@#After the treatment of Molt-4 cells with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77, the cell survival rate was decreased (P<0.05); Compared with the control group, the cell proliferation inhibition rate of the curcumin group and the UMI-77 group were increased, the apoptosis rate of cell was increased, the level of ROS was increased, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, and the protein expression of Bcl-2 was reduced (P<0.05); Compared with the curcumin group or UMI-77 group, the cell proliferation inhibition rate and apoptosis rate of the curcumin+UMI-77 group were further increased, and the level of ROS was increased. At the same time, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, the protein expression of Bcl-2 was reduced (P<0.05); In addition, the mitochondrial membrane potential of the cells after curcumin treatment was decreased, and the proteins expression of Notch1 and HES1 were reduced (P<0.05).@*CONCLUSION@#Curcumin can enhance the apoptosis of T-ALL cells induced by Mcl-1 small molecule inhibitor UMI-77 by reducing the mitochondrial membrane potential, the mechanism may be related to the inhibition of Notch1 signaling pathway.

Erdosteine ameliorates the harmful effects of ischemia-reperfusion injury on the liver of rats

Abstract Purpose: To investigate the potential protective effects of erdosteine against the harmful effects of ischemia-reperfusion injury on the liver in an experimental rat model. Methods: Forty rats were divided into 4 groups. In the sham group, only the hepatic pedicle was mobilized. No other manipulation or treatment was performed. In the other groups, ischemia was achieved by clamping the hepatic pedicle for 60 min. After that, 90 min reperfusion was provided. In the control group, no treatment was given. In the perioperative treatment group, 100 mg/kg erdosteine was administered 2 hours before ischemia induction. In the preoperative treatment group, 100 mg/kg/day erdosteine was administered daily for ten days before the operation. At the end of the procedures, blood and liver samples were obtained for biochemical and histopathological assessment. Results: Treatment with erdosteine ameliorated the histopathological abnormalities when compared with the control group. Furthermore, this treatment significantly decreased the serum liver function test values. It was also found that erdosteine ameliorated the oxidative stress parameters in both the perioperative and preoperative treatment groups. Conclusion: The current study is the first to have shown the favorable effects of erdosteine on the harmful effects of experimental hepatic ischemia-reperfusion injury.

Perkinsus sp. infecting the oyster Crassostrea rhizophorae from estuaries of the septentrional Northeast, Brazil / Perkinsus sp. infectando a ostra Crassostrea rhizophorae de estuários do Nordeste setentrional, Brasil

Abstract The mangrove oyster Crassostrea rhizophorae is an estuarine resource exploited by riverside communities in Northeast Brazil. Despite its socioeconomic importance, studies on the health status of this bivalve are scanty in this region. The purpose of the present study was to investigate the presence of the protozoan Perkinsus sp. in C. rhizophorae collected in August and September 2011 in three estuaries of the septentrional Northeast, Brazil: Jaguaribe (Ceará), Camurupim (Piauí) and Carnaubeiras (Maranhão) (n= 150 specimens/site). The samples were submitted to Ray’s fluid thioglycollate medium (RFTM), PCR and histology assays. The RFTM assay revealed spherical, blue or bluish-black hypnospores of the genus Perkinsus in 50 specimens (Jaguaribe= 17.3%, Camurupim= 5.3%, Carnaubeiras= 10.6%). The intensity of the infection ranged from very light (1-10 cells per slide) to severe (more than 40 cells in each of 10 fields of the slide) for Jaguaribe; very light for Camurupim and very light to moderate (at least 40 cells observed in each of 10 fields of the slide) for Carnaubeiras. When submitted to confirmatory PCR analysis, 6 cases were confirmed (Jaguaribe=3, Camurupim=1, Carnaubeiras=2). The histology confirmed 21 cases of infection in specimens from the three estuaries. Although local collectors have reported no mortality in oyster populations that might be attributed to infection by Perkinsus, health surveillance of oyster populations in the septentrional region of Northeast Brazil is advisable.

Resumo A ostra-do-mangue Crassostrea rhizophorae é um recurso estuarino explorado por comunidades ribeirinhas do Nordeste do Brasil. Apesar de sua importância socioeconômica, estudos sobre o estado de saúde deste bivalve são escassos na região. O objetivo deste estudo foi investigar a presença do protozoário Perkinsus sp. em C. rhizophorae coletada em agosto e setembro de 2011, em três estuários da região setentrional do Nordeste brasileiro: Jaguaribe (Ceará), Camurupim (Piauí) e Carnaubeiras (Maranhão) (n = 150 espécimes/local). As amostras foram submetidas ao meio líquido de tioglicolato de Ray (RFTM), PCR e ensaios histológicos. A análise em RFTM revelou hipnósporos esféricos azuis ou preto-azulados do gênero Perkinsus em 50 espécimes (Jaguaribe= 17,3%, Camurupim= 5,3%, Carnaubeiras= 10,6%). A intensidade de infecção variou de muito leve (1-10 células por lâmina) a severa (mais de 40 células em cada um dos 10 campos da lâmina) para o Rio Jaguaribe; muito leve para o Rio Camurupim e muito leve a moderada (pelo menos 40 células observadas, em cada um dos 10 campos da lâmina) para o Rio Carnaubeiras. Quando submetidos à análise confirmatória por PCR, foram confirmados 6 casos (Jaguaribe= 3, Camurupim= 1, Carnaubeiras= 2). A histologia confirmou 21 casos de infecção em espécimes dos três estuários. Embora os coletores locais não tenham relatado nenhuma mortalidade em populações de ostras que pudesse ser atribuída à infecção por Perkinsus, é aconselhável um monitoramento sobre o estado de saúde de populações de ostras da região.

Avaliação in vitro do efeito de diferentes processos de alisamento químico/térmico na fibra capilar / In vitro evaluation of the straightening effect by different chemical/thermal processes in the hair fiber

A aparência dos cabelos é de fundamental importância na sociedade atual. Estando em moda, cabelos mais lisos e com menos volume, os consumidores que antes os alisavam com produtos químicos e força mecânica, passaram a utilizar um tratamento térmico, além do secador de cabelos: as piastras ("chapinhas") que atuam em valores de temperatura ao redor de 230°C. Esse procedimento ocasiona além dos danos mecânicos e químicos também dano térmico, tornando os cabelos ainda mais fragilizados. O escopo deste estudo foi avaliar o dano na fibra capilar, de amostras não tratadas e nas que receberam aplicação de alisantes/relaxantes tradicionais e alternativos. O estudo foi dividido em cinco capítulos que avaliam: aplicação dos alisantes/relaxantes com ingredientes ativos distintos; danos mecânicos, perda Protéica; análise térmica e microscopia eletrônica de varredura. As amostras de cabelo utilizadas em todos os estudos foram tratadas como descrito no primeiro capítulo. Foram aplicados produtos comerciais contendo os seguintes ingredientes ativos: Hidróxido de Sódio, Tioglicolato de Amônio, Hidróxido de Guanidina (reação de hidróxido de cálcio com carbonato de guanidina), formaldeído e ácido glioxílico isolado e em combinação com carbocisteína. O uso de formaldeído e ácido glioxilico em formulações de alisantes/relaxantes está proibido pela Agência Nacional de Vigilância Sanitária. Todos os produtos aplicados alisaram os cabelos; os procedimentos que utilizaram a piastra tornaram os fios mais lisos. Os alisantes/relaxantes à base de ácido glioxilico e formaldeído reduziram de forma expressiva a tensão de ruptura dos cabelos tornando-os mais frágeis. A maior perda protéica foi observada na amostra tratada com carbocisteína (1,74 mg/g cabelo). Nos estudos de análise térmica, na fase de desidratação a amostra tratada com carbocisteína apresentou maior perda de massa (15,17%); na fase de denaturação da proteína, a tratada com hidróxido de sódio (51,06%); e na fase de eliminação do material carbonáceo, todas as amostras apresentaram perda de massa maior que a amostra não tratada; as menores temperaturas de pico foram as das amostras sem tratamento alisante (630°C) e ácido glioxílico (640°C). Observando-se as imagens de microscopia eletrônica nota-se modificação nas bordas das cutículas das amostras indicando que sofreram agressão; o hidróxido de guanidina deixou adicionalmente resíduo; as amostras tratadas com ácido glioxílico e formaldeído apresentaram a formação de filme superficial como um "envelopamento" da fibra. Os resultados sugerem que não há predominância de um procedimento mais danoso que os demais; porém os que utilizaram a piastra (alisamentos/relaxamento ácidos) acentuaram os danos

The appearance of the hair is of fundamental importance in today's society. Being in fashion, hair straight and with less volume, consumers that before straighted hair with chemicals products and mechanical strength began to use a heat treatment, in addition to hair dryers: the hot plates ("chapinhas") acting on temperature values around 230°C. This procedure causes not only mechanical and chemical damage but also thermal one, making the hair more fragile. The scope of this study was to evaluate the damage to the hair fiber, in untreated samples and these receiving straighteners/relaxers application of traditional and alternative products.The study was divided into five chapters that evaluated: application of straighteners/relaxers with different active ingredients; mechanical damage, protein loss; thermal analysis and scanning electron microscopy. The hair samples used in all studies were treated as described in the chapter one. Commercial products containing the following active ingredients were used: Sodium Hydroxide, Ammonium Thioglycolate, Guanidine Hydroxide (calcium hydroxide reaction with guanidine carbonate), Formaldehyde and Glyoxylic Acid alone and in combination with Carbocysteine. The use of Formaldehyde andGlyoxylicAcid in straightening/relaxing formulations are prohibited by the National Agency for Sanitary Vigilance. All applied products, straight the hair samples; the procedures that used the hot plates become the hair more straight. The straightening/relaxing based on Glyoxylic Acid and Formaldehyde reduced significantly the hair break point making them more fragile. Most protein loss was observed in the sample treated with Carbocysteine (1.74mg/g hair).In the thermal analysis studies at the dewatering stage, Carbocystein treated samples showed greater weight loss(15.17%), at the protein denaturation stage this treated with Sodium Hydroxide (51.06%) and in the carbonaceous material elimination phase all samples showed mass loss greater than the untreated sample;. The lower peak temperatures were observed in the samples without treatment (630°C) and with Glyoxylic Acid (640°C). Observing the images of electron microscopy is noted the change in the cuticle aspect of the samples showing that the edges were damaged, Guanidine Hydroxide, left further residue: the samples treated with Glyoxylic Acid and Formaldehyde showed the formation of surface film as an "enveloping" fiber. The results suggest that there is not a predominance of a more harmful treatment than other, but those using hot plates(straightening/relaxing acids) emphasize the damage

Determination of thiodiglycolic acid in urine with gas chromatography-mass spectrometer / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To establish a rapid determination method with gas chromatography-mass spectrometer (GC-MS) for thiodiglycolic acid (TDGA), a vinyl chloride (VCM) biomarker.</p><p><b>METHODS</b>A high- sensitivity determination method was established using a moderate methyl esterification instead of methyl esterification of highly toxic diazo reaction.</p><p><b>RESULTS</b>The standard curve regression linear equation of the method was: y=8460.5x-4758.2, the linear coefficient was 0.999 7, the minimum quantity concentration was 2.0 µg/L, the range of precision value was 0.81%-2.38%, and the average recovery of standard addition was 99.0%-102.9%.</p><p><b>CONCLUSION</b>This method reduces the risk of traditional methyl esterification, improves the determination sensitivity compared with the GC-FPD method, and meets the determination requirement of TDGA.</p>

Simultaneous determination of erdosteine and its active metabolite in human plasma by liquid chromatography-tandem mass spectrometry with pre-column derivatization / 药学学报

A sensitive, rapid and accurate liquid chromatography-tandem mass spectrometric (LC-MS/MS) method with pre-column derivatization was developed for the simultaneous determination of erdosteine and its thiol-containing active metabolite in human plasma. Paracetamol and captopril were chosen as the internal standard of erdosteine and its active metabolite, respectively. Aliquots of 100 microL plasma sample were derivatized by 2-bromine-3'-methoxy acetophenone, then separated on an Agilent XDB-C18 (50 mm x 4.6 mm ID, 1.8 microm) column using 0.1% formic acid methanol--0.1% formic acid 5 mmol x L(-1) ammonium acetate as mobile phase, in a gradient mode. Detection of erdosteine and its active metabolite were achieved by ESI MS/MS in the positive ion mode. The linear calibration curves for erdosteine and its active metabolite were obtained in the concentration ranges of 5-3 000 ng x mL(-1) and 5-10 000 ng x mL(-1), respectively. The lower limit of quantification of erdosteine and its active metabolite were both 5.00 ng x mL(-1). The pharmacokinetic results of erdosteine and its thiol-containing active metabolite showed that the area under curve (AUC) of the thiol-containing active metabolite was 6.2 times of that of erdosteine after a single oral dose of 600 mg erdosteine tables in 32 healthy volunteers, The mean residence time (MRT) of the thiol-containing active metabolite was (7.51 +/- 0.788) h, which provided a pharmacokinetic basis for the rational dosage regimen.

Pannexin-1 influences peritoneal cavity cell population but is not involved in NLRP3 inflammasome activation

Pannexin-1 (Panx1) forms nonselective large channel in cell plasma membrane and has been shown to be associated with NLRP3 inflammasome activation, ATP release and phagocytes recruitment. In the current study, by manipulation of Panx1 expression in human myeloid cells and application of Panx1 deficient mice, we failed to find a correlation between Panx1 and NLRP3 inflammasome activation, although an interaction between these two proteins was evident. However, in thioglycollate induced peritonitis, Panx1 deficient mice showed much more phagocytes infiltration. Further analyses showed that mice deficient for Panx1 exhibited enlarged F4/80(low)Gr1(-)Ly6C(-)cell population in the peritonea. Our study thus reveals an important role for Panx1 in regulation of peritoneal cell population and peritonitis development.

Comparative study of the effect of LPS on the function of BALB/c and C57BL/6 peritoneal macrophages

Macrophages influence their environment and surrounding immune cells as soon as stimulators affect them. Different sources of macrophages induce different reactions in their neighboring immune cells,which result in non-uniform immunologic outcomes. In this experimental research, we compare the behavior of peritoneal macrophages to lipopolysaccharide [LPS] stimulation from BALB/cmice as an indicator of a type 2 immune response and from C57BL/6 mice as an indicator of a type 1 immune response. In this experimental study, peritoneal macrophages prepared from thioglycolate stimulated BALB/c and C57BL/6 micewere treated with 1microg/ml LPS. At different time points after LPS treatment, nitric oxide [NO], interferon gamma [IFN-gamma]. interleukin 4 [IL-4],transforming growth factor beta[1] [TGF-beta[1]], interleukin 17 [IL-17], and interleukin 10[IL-10] production were measured in the supernatants of all macrophage cultures.Indoleamine 2, 3 dioxygenase [IDO] and phagocytic activitywere analyzed in the different experimental groups. The supernatant effects of LPS-treated macrophages on splenocyte proliferation was assessed by the colorimetric method using a 3-[4,5-Dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide [MTT] reagent. According to cytokine analysis, different mouse strains show different cytokine patterns in response to LPS. C57BL/6 macrophages produced more IL-17, IL-10, and IFN-gamma while BALB/c macrophages produced more TGF-beta[1]., and IL-4. There was no significant difference in IDO activity between strains [p

Aislamiento de trichomonas tenax en pacientes con periodontitis crónica al medio de cultivo de tioglicolato modificado / Isolation of trichomonas tenax in patients with chronic periodontitis in the modified thioglycolate culture médium

Objetivo. Establecer la asociación entre la presencia del protozoario flagelado de la cavidad bucal Trichomonas tenax y la periodontitis crónica en los pacientes atendidos en la clínica especializada en odontología de la universidad de San Martín de Porres (USMP). Material y métodos. Fueron seleccionados 53 pacientes con periodontitis crónica y 41 pacientes periodontalmente sanos, los cuales acudieron a la clínica entre los meses de setiembre a diciembre de 2011, tomándose muestras de cálculo dental y placa dental subgingival respectivamente. Las muestras fueron depositadas en un tubo de vidrio conteniendo el medio de cultivo de Tioglicolato modificado para Trichomonas tenax, los cuales fueron incubados a 35ºC por 4 a 5 días en el laboratorio. Resultados. Se encontró presencia del parásito de Trichomonas tenax en 9 pacientes con periodontitis crónica (17,0%) y 10 pacientes periodontalmente sanos (24,4 %), con un valor de p =0,781, por ser un estudio de casos y controles, el OR= 0,634. Conclusiones. Se determinó que no existe asociación entre la presencia de Trichomonas tenax y la periodontitis crónica. Asimismo, la presencia del parásito no se vio condicionada por la edad, el sexo y el índice de higiene oral de los pacientes atendidos en el mencionado recinto.

Objective. To establish the association between the presence of the flagellated protozoan of the oral cavity Trichomonas tenax and chronic periodontitis in patients treated at the specialized clinic in dentistry of San Martin de Porres University (SMPU). Material and methods. 53 patients with chronic periodontitis and 41 periodontal healthy patients were selected, who attended to the clinic during the months of September to December 2011, taking of them samples of dental calculus and subgingival dental plaque, respectively. The samples were placed in a glass tube containing the culture medium of modified thioglycollate to Trichomonas tenax, which were incubated at 350 grades centigrades for 4 or 5 days in the laboratory. Results. We found the presence of Trichomonas tenax in 9 patients with chronic periodontitis (17,0%) and in 10 periodontal healthy patients (24,4%), with a p-value =0,781. Conclusions. We conclude that there is no association between the presence of Trichomonas tenax and chronic periodontitis. Also the presence of the parasite was not influenced by age, sex and oral hygiene index of patients seen in the clinic.

HPLC determination of the related substances in erdosteine / 药学学报

An HPLC method was established for the determination of the related substance in erdosteine. Waters ODS-SunFire (250 mm x 4.6 mm ID, 5 microm) column was used, the mobile phase was composed of methanol-acetonitrile-0.01 mol x L(-1) citric acid (20:4:76, the pH value was adjusted by triethylamine to 2.5). The flow rate was 1 mL x min(-1). The detection wavelength was 254 nm. The related substances in the sample of erdosteine taken were calculated by self control with or without the response factor of impurity relative to that of erdosteine. Under the chromatographic condition developed, the impurities in erdosteine were isolated well. The detection limit was 0.2 microg x mL(-1) (signal/noise = 3) by principal component calculated. The method can be adopted to control the related substances in erdosteine.

Acute aerocistitis induced by thioglycolate, lipopolysaccharide and inactivated Aeromonas hydrophila in Piaractus mesopotamicus: hematological effects / Aerocistite aguda induzida por tioglicolato, lipolisacarídeo e Aeromona hydrophila inativada em Piaractus mesopotamicus: efeitos hematológicos

The effects of swim bladder injection with thioglycolate, Escherichia coli lipopolysaccharide (LPS) and heat-inactivat ed Aeromonas hydrophila were assessed on hematological responses in pacu, Piaractus mesopotamicus (Characidae). A quantitative assessment was done on erythrocytes, thrombocytes e leucocytes at 6, 24, and 48 h pos-injection of the inflammatory agents and compared with fish injected with saline solution (control). Fish injected with inactivated A. hydrophila showed a reduction of erythrocytes and hemoglobin, whereas the hematocrit increased 6 h pos-injection. The results show that thioglycolate and LPS also induced a reduction on hemoglobin and an increase on the hematocrit. The thrombocytes count decreased 6 h post A. hydrophila injection, whereas increased 48 hours post LPS injection. The leukocytes count increased after 6 h post A. hydrophila injection, while the lymphocytes and PAS-positive granularleukocytes (PAS-LG) count decreased after 24 h post injection. In fish injected with thioglycolate or with LPS showed an increase in the LG-PAS counts when compared to A. hydrophila or control groups. The monocytes count was not affected by the different inflammatory agents.

Os efeitos da injeção de tioglicolato, lipolissacarídio de Escherichia coli e Aeromonas hydrophila inativada na bexiga natatória de pacus, Piaractus mesopotamicus (Characidae) foram avaliados quanto às respostas de células vermelhas, leucócitos e trombócitos do sangue. Ensaios quantitativos de eritrócitos, leucócitos e trombócitos foram realizados 6, 24 e 48 h após os estímulos e comparados com peixes que receberam solução salina 0,65% pela mesma via. Peixes inoculados com A. hydrophila apresentaram redução do número de eritrócitos e da taxa de hemoglobina enquanto o hematócrito aumentou 6 h após o estímulo. Os resultados mostraram que o tioglicolato e o LPS também induziram redução da hemoglobina e aumento do hematócrito. A contagem de trombócitos diminuiu 6 h após a inoculação de A. hydrophila inativada e aumentou 48 horas após a injeção de LPS. A contagem de leucócitos aumentou 6 h após a inoculação de A. hydrophila enquanto a de linfócitos a leucócitos granulares PAS positivos (PAS_LG) diminuiu 24 h depois. Peixes injetados com tioglicolato o LPS apresentaram aumento do número de LG_PAS em relação aos inoculados com A. hydrophila inativada ou grupo controle. A contagem de monócitos não foi afetada pelos diferentes agentes.

Thioglycolic acid esterified into rice straw for removing lead from aqueous solution

Thiol rice straw [TRS] was prepared by esterifying thioglycolic acid onto rice straw in the medium of acetic anhydride and acetic acid with sulfuric acid as catalyst. The sorption of lead [Pb] on TRS from aqueous solution was subsequently investigated. The batch experiments showed that Pb removal was dependent on initial pH, sorbent dose, Pb concentration, contact time, and temperature. The maximum value of Pb removal appeared at pH 5. For 100 mg/L of Pb solution, a removal ratio of greater than 98% could be achieved with 2.0 g/L or more of TRS. The isothermal data of Pb sorption conformed well to the Langmuir model, and the maximum sorption capacity [Q[m]] of TRS for Pb was 104.17 mg/g. The equilibrium of Pb removal was reached within 120 min. The Pb removal process could be described by the pseudo-first-order kinetic model. The thermodynamic study indicated that the Pb removal process was spontaneous and endothermic

Characterization of the acute inflammatory response in the hybrid tambacu (Piaractus mesopotamicus male × Colossoma macropomum female) (Osteichthyes) / Caracterização da resposta inflamatória aguda no híbrido tambacu (Piaractus mesopotamicus macho × Colossoma macropomum fêmea) (Osteichthyes)

This work evaluated the acute inflammatory response induced by injections of 0.5 mL saline solution (control), 500 µg carrageenin and 0.5 mL thioglycollate 3 percent in the swim bladder of juvenile tambacu hybrid. Fish were distributed in three treatments, three replications and acclimated for a period of 10 days before assay. The cell characterization from the inflammatory exudate was performed in Giemsa and PAS stained smears. Carrageenin, injected in fish, showed an increase on the total number of cells in the inflammatory exudate when compared to saline and thioglycollate injected. Whereas, for carrageenin-injected fish, the percentage of thrombocyte was higher than thioglycollate. On the other hand, granulocyte percentage in thioglycollate-injected fish was higher than the ones injected using carrageenin. Carrageenin provoked the highest migration of macrophage to the inflammatory site. The PAS method confirmed the presence of three types of granulocytes: eosinophilic granular cell (EGC) type 1 with the characteristics of a special granulocytic cell commonly found in the circulating blood; EGC type 2 shorter than the last one and neutrophil. This study contributes to a better understanding of the inflammatory response and infectious processes in native fish.

Este estudo avaliou a resposta inflamatória aguda induzida por injeções de 0,5 mL de solução salina (controle), 500 µg de carragenina e 0,5 mL de tioglicolato a 3 por cento na bexiga natatória de juvenis do híbrido tambacu. Os peixes foram distribuídos em três tratamentos, três repetições e aclimatados durante 10 dias antes do ensaio. A caracterização das células do exsudato inflamatório foi feita após coloração com Giemsa e PAS. Peixes injetados com carragenina apresentaram maior número de células no exsudato inflamatório do que com salina e tioglicolato. A porcentagem de trombócitos no exsudato foi maior nos injetados com carragenina quando comparada com a dos injetados com tioglicolato. Por outro lado, o percentual de granulócitos foi maior em animais injetados com tioglicolato do que em animais injetados com carragenina. A carragenina provocou maior migração de macrófagos para o foco inflamatório. O método de PAS confirmou a presença de três tipos de granulócitos: célula granular eosinofílica (CGE) tipo 1 com as características da célula granulocítica especial encontrada no sangue, CGE tipo 2, menor do que esta última, e de neutrófilos. Este estudo contribui para o melhor entendimento da resposta inflamatória e dos processos infecciosos em peixes nativos.

Polymerase chain reaction versus conventional methods in the diagnosis of vaginal trichomoniasis

A total of 200 females of whom 120 had manifestations of vaginal trichomoniasis and 80 asymptomatic ones were studied. In 54/120 symptomatic female [45%] and in 28/80 asymptomatic ones [35%], T. vaginalis was diagnosed by wet mount of bedside vaginal swab samples of 120 samples from symptomatic females, T. vaginalis was detected in 93 [77.5%] when cultured onto InPouch and 95 [79.16%] in modified thioglycolate media. Culturing 80 samples of asymptomatic females showed T. vaginalis in 35 [43.75%] onto either media. T. vaginalis genomic DNAs was amplified by PCR from 130 [65%] by using TVA5-TVA6 primer pair in 95 [79.16%] samples of 120 symptomatic females, and in 35 [43.75%] samples of 80 asymptomatic ones. Difference between groups was statistically significant. The motile trichomonads was detected by wet mount in 82/130 positive cultures giving 63.07% sensitivity and 100% positive predictive value [PPV]. Flagellates were not detected by wet mount in any negative culture, giving 100% specificity and 59.32% negative predictive value [NPV]. The wet mount diagnostic accuracy [DA] was 76%, without false-positive, but false negative was 48/130 [36.93%]. DNA was amplified from 129/130 positive culture by TVA5-TVA6 primer pair, giving 99.23% sensitivity. No amplification was detected from one positive culture. DNA was not amplified from 69/70 negative culture using TVA5-TVA6 primer pair, giving 98.57% specificity, 99.23% PPV, 98.57% NPV and 99% DA

Effect of ascorbic acid on stimulatory status of activated mouse peritoneal phagocytes.

Mouse peritoneal macrophages (MPM) when elicited by the antioxidant ascorbic acid have been found to be significantly stimulatory, exhibiting marked alteration at the cellular and enzyme levels. Alterations recorded were as follows--cellular yield per mouse, their protein content, lysosomal acid hydrolase levels and capability to phagocyte, all were significantly enhanced. The new stimulant was observed to produce no synergistic action on MPM when thioglycollate, BCG or endotoxin along with the same stimulated the latter. Levels of antioxidants like ascorbic acid and glutathione were found to be enhanced in elicited macrophages whereas superoxide dismutase levels varied when the three above stimulators were administered. However, the ascorbic acid elicited cells showed an increase in glutathione levels and a decrease in SOD levels but no change in total intracellular ascorbic acid levels. Further, though ascorbic acid interaction enhanced the phagocytic capability of MPM as compared to resident cells, no significant boosting of phagocytic process could be observed when each of three stimulators coupled with ascorbic acid was used for macrophage elicitation.

Thioglycollate-elicited murine macrophages are cytotoxic to Mycoplasma arginini-infected YAC-1 tumor cells

Macrophages are important components of natural immunity involved in inhibition of tumor growth and destruction of tumor cells. It is known that these cells can be activated for tumoricidal activity by lymphokines and bacterial products. We investigated whether YAC-1 tumor cells infected with Mycoplasma arginini stimulate nitric oxide (NO) release and macrophage cytotoxic activity. Thioglycollate-elicited macrophages from male BALB/c mice were co-cultured for 20 h with YAC-1 tumor cells infected or not with Mycoplasma arginini. The cytotoxic activity was evaluated by MTT assay and nitrite levels were determined with the Griess reagent. Thioglycollate-elicited macrophages co-cultured with noninfected YAC-1 cells showed low cytotoxic activity (34.7 ñ 8.6per cent) and low production of NO (4.7 ñ 3.1 µM NO2-). These macrophages co-cultured with mycoplasma-infected YAC-1 cells showed significantly higher cytotoxic activity (61.4 ñ 9.1 per cent; P=0.05) and higher NO production (48.5 ñ 13 µM NO2-; P=0.05). Addition of L-NAME (10 mM), an inhibitor of NO synthesis, to these co-cultures reduced the cytotoxic activity to 37.4 ñ 2per cent (P=0.05) and NO production to 3 ñ 4 µM NO2- (P=0.05). The present data show that Mycoplasma arginini is able to induce macrophage cytotoxic activity and that this activity is partially mediated by NO.

Evaluación de tres métodos de laboratorio para la cuantificación de hierro sérico y capacidad total fijación al hierro / Evaluation of three laboratory methods for the determination of serum iron and iron binding capacity

Se evaluaron tres métodos de laboratorio para la determinación de hierro sérico (FeS) y capacidad total de fijación de hierro sérico (CTFFeS): el método de Beale (método de referencia) y un método comercial tanto con sus reactivos originales como con reactivos preparados en el laboratorio, con el fin de sustituir el método utilizado como referencia ya que es muy tedioso y consume mucho tiempo. Se realizaron los espectros de absorción, intervalos analíticos, estabilidad de los complejos coloreados, precisión en un mismo día y día a día durante 20 días consecutivos, recuperación, interferentes como hemoglobina, bilirrubina y cobre y un estudio de correlación entre estos métodos. Se concluye que a pesar de que el método de Beale no posee la mejor aplicabilidad analítica, sí presenta un desempeño analítico superior a los otros dos métodos evaluados, por lo tanto sigue siendo el método de elección para este medio

Effect of dithiothreitol and sodium thioglycollate on protoplast production of saccharomyces cerevisae

Investigou-se a influência do ditiotreitol e do tioglicolato de sódio na produçäo de protoplastos de S. cerevisae. Resultados favoráveis foram obtidos a partir de 30 min. de digestäo, usando-se o ditioteitrol como pré-tratamento e adicionado à mesma soluçäo enzimática, inibiu a produçäo de protoplastos

Canatoxin induces activation on mice peritoneal macrophages

Canatoxin (CNTX), the toxic protein from Canavalia ensiformis seeds, injected into the peritoneal cavities of mice (10 micrograms/cavity) induced a significant neutrophil migration (10.5 +/- 0.5 x 10(6) cells/cavity) after 4 h. A later migratory effect (48 h) on mononuclear cells, predominantly macrophages, was also observed (controls: 7 +/- 0.9; CNTX: 17 +/- 2.0 x 10(6) cells/cavity). These CNTX-elicited macrophages, when compared to resident cells (R) or cells elicited by thioglycollate (TG), had an increased content of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase (R: 4.5 +/- 0.5; TG: 7.2 +/- 1.0; CNTX: 20.2 +/- 3.0 mU/10(6) cells) and a greater (> or = 100%) phagocytic activity. The data suggest that CNTX-stimulated macrophages presented some characteristics of activated cells

Comportamiento de los receptores de insulina de macrofagos inducidos con tioglicolato y en condiciones de cultivo / Behaviour of insulin receptors in macrophages induced with thioglycate and under culture conditions

Se estudian los receptores de insulina en macrófagos peritoneales de ratón, inducidos con medio tioglicolato y en macrófagos residentes cultivados in vitro durante distintos tiempos. Los macrófagos inducidos presentan una disminución del número de receptores de insulina (1,5 x 10 4/célula con respecto a las residencias (5,2 x 10 4/célula) El número de receptores aumenta en los macrófagos peritoneales de 3,7 x 10 4/célula hasta 2,1 x 10 5/célula durante 72 horas de cultivo in vitro