Two cases of neural tube defects with dolutegravir use at conception in south Brazil

ABSTRACT Dolutegravir (DTG) is amongst the most prescribed antiretrovirals worldwide and is recommended as first line regimen in most HIV treatment guidelines. Its use, although infrequently, had been associated to an increased chance of neural tube defects (NTD) in Botswana, Africa. Herein we describe two cases of NTD in women who conceived while taking DTG as part of their antiretroviral treatment in the city of Porto Alegre, Brazil.

Which antiretroviral regimen is associated with higher adherence in Brazil? A comparison of single, multi, and dolutegravir-based regimens / Qual esquema antirretroviral está associado à maior adesão no Brasil? Uma comparação dos esquemas de comprimido único, múltiplos comprimidos e com dolutegravir / ¿Qué tratamiento antirretroviral está asociado con adherencia más alta en Brasil? Una comparación de tratamientos únicos, múltiples y basados en el dolutegravir

Abstract: We evaluated adherence to highly active antiretroviral therapy (HAART) and its associated factors according to the type of regimen in patients initiating treatment in Belo Horizonte, Minas Gerais State, Brazil. We measured adherence using the eight items Morisky Therapeutic Adhesion Scale (MMAS-8) and compared the use of "backbone" tenofovir/lamivudine plus efavirenz one tablet once-daily (STR) or dolutegravir in multi-tablet once-daily (MTR-DTG), or other multi-tablet regimens (MTR-other). We conducted a multivariate logistic regression analysis to address factors associated with adherence. A total of 393 patients were included, 254 used STR, 106 MTR-DTG, and 33 MTR-other. The overall adhesion rate was 44.8% (95%CI: 39.4; 50.1), 50% for MTR-DTG, 43.3% for STR and 39.4% for MTR-other. Multivariate analysis showed a higher chance of adherence among patients using MTR-DTG, those who received and understood counseling about their treatment and with a higher quality of life. Prior use of illicit drugs in the lifetime was associated with poorer adherence. Overall adherence was low, highlighting the need for strategies focusing on counseling about medicines and substance use. Pill burden was not an issue for patients using MTR-DTG once-daily, who achieved better results.

Resumo: Avaliamos a adesão à terapia antirretroviral (TARV) e fatores associados de acordo com o tipo de esquema em pacientes no início do tratamento em Belo Horizonte, Minas Gerais, Brasil. Mensuramos a adesão com a Escala de Adesão Terapêutica de Morisky, de oito itens (MMAS-8), e comparamos o uso de tenofovir/lamivudina com efavirenz, um comprimido uma vez ao dia (STR), ou dolutegravir em múltiplos comprimidos uma vez ao dia (MTR-DTG), com outros esquemas com múltiplos comprimidos ao dia (MTR-outros). Conduzimos uma análise de regressão logística multivariada para avaliar os fatores associados à adesão. Foram incluídos 393 pacientes: 254 em uso de STR, 106 MTR-DTG e 33 MTR-outros. A taxa global de adesão foi 44,8% (IC95%: 39,4; 50,1), sendo 50% para MTR-DTG, 43,3% para STR e 39,4% para MTR-outros. A análise multivariada mostrou chances maiores de adesão em pacientes em uso de MTR-DTG, pacientes que haviam recebido e compreendido o aconselhamento sobre o tratamento e pacientes com melhor qualidade de vida. Uso anterior de drogas ilícitas em qualquer período da vida está associada à pior adesão. A adesão global foi baixa, enfatizando a necessidade de estratégias focadas no aconselhamento sobre medicamentos e uso de drogas. A quantidade de comprimidos não foi um problema para pacientes em uso de MTR-DTG uma vez ao dia, os quais alcançaram melhores taxas de adesão.

Resumen: Evaluamos la adherencia a la terapia antirretroviral altamente activa (TARAA) y sus factores asociados, según el tipo de tratamiento en pacientes que comenzaron su tratamiento en Belo Horizonte, Minas Gerais, Brasil. La adherencia se mensuró por la Escala de Adhesión Terapéutica de Morisky, de ocho ítems (MMAS-8), y se comparó el uso del "eje" tenofovir/lamivudina, además de un comprimido de efavirenz una vez al día (STR) o dolutegravir con varios comprimidos una vez al día (MTR-DTG), u otros tratamientos con múltiples comprimidos (MTR-otros). Se realizó un análisis multivariado de regresión logística para evaluar los factores asociados a la adherencia. Se incluyeron un total de 393 pacientes, 254 usaron STR, 106 MTR-DTG, y 33 MTR-Otros. La tasa de adherencia general fue de un 44,8% (95%CI: 39,4; 50,1), 50% en el MTR-DTG, 43,3% en el STR y 39,4% en el MTR-otros. El análisis multivariado mostró una probabilidad más alta de adherencia entre pacientes usando MTR-DTG, quienes recibieron y comprendieron las orientaciones acerca de sus tratamientos y los que disfrutaban de una calidad mejor de vida. El consumo previo de drogas ilícitas a lo largo de la vida estuvo asociado con una adherencia más escasa. La adherencia general fue baja y resalta la necesidad de estrategias que se enfoquen en brindar orientación sobre el uso de la medicación y de sustancias. El número de comprimidos no fue un problema para los pacientes que tomaban MTR-DTG una vez al día, que obtuvieron mejores resultados.

Detection of carboxylesterase and esterase activity in culturable gut bacterial flora isolated from diamondback moth, Plutella xylostella (Linnaeus), from India and its possible role in indoxacarb degradation

Abstract Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n = 13) and adults (n = 12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32 µmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus – KC985225 and Pantoea agglomerans – KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26 µmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.

Dormancy models for Mycobacterium tuberculosis: A minireview

Dormancy models for Mycobacterium tuberculosis play important roles in understanding various aspects of tuberculosis pathogenesis and in the testing of novel therapeutic regimens. By simulating the latent tuberculosis infection, in which the bacteria exist in a non-replicative state, the models demonstrate reduced susceptibility to antimycobacterial agents. This minireview outlines the models available for simulating latent tuberculosis both in vitro and in several animal species. Additionally, this minireview discusses the advantages and disadvantages of these models for investigating the bacterial subpopulations and susceptibilities to sterilization by various antituberculosis drugs.

Neonatal screening program for hemoglobinopathies in the city of São Carlos, state of São Paulo, Brazil: analysis of a series of cases / Triagem neonatal de hemoglobinopatias no município de São Carlos, São Paulo, Brasil: análise de uma série de casos

OBJECTIVE: To analyze the neonatal screening program for hemoglobinopathies in São Carlos, Southeast Brazil, by investigating a series of cases in which the screening test was abnormal. More specifically, it was aimed to know the information regarding the neonatal screening received by mothers at the hospital and at primary health care, in addition to information related to genetic counseling. METHODS: A descriptive study that enrolled 119 mothers, accounting for 73% of all children born between 2010 and 2011 with abnormal results of neonatal screening for hemoglobinopathies. The mothers completed a questionnaire that assessed the information received at hospital and primary health care, and issues related to genetic counseling. Descriptive statistics was performed. RESULTS: Of the 119 participating mothers, 69 (58%) had children with sickle cell trait, 22 (18.5%) with hemoglobin C trait, 18 (15.1%) with alpha thalassemia trait and, in 10 cases (8.4%), the result was inconclusive. At the hospital, 118 mothers (99.2%) received information about where to go to collect the test and 115 (96.6%) were informed about the correct time to collect the test. Only 4 mothers (3.4%) were informed about which diseases are investigated and the risks of not performing the screening. Seventeen mothers (14.3%) recognized the difference between trait and disease, and 42 (35.3%) considered that a positive screening test could have implications for future pregnancies. In 70 cases (58.8%), the child's physician was not informed about the screening test results. CONCLUSIONS: The neonatal screening program needs further improvement. In both scenarios investigated, health professionals demonstrated a lack of training in providing information to mothers and families. .

OBJETIVO: Fazer uma análise do programa de triagem neonatal de hemoglobinopatias no município de São Carlos, São Paulo, Brasil, por meio da investigação de série de casos cujo resultado do teste de rastreio foi alterado. Objetivou-se conhecer as informações a respeito da triagem neonatal recebidas pelas mães na maternidade e na atenção primária à saúde, além das informações relacionadas à orientação genética. MÉTODOS: Estudo descritivo, no qual participaram 119 mães cujos filhos apresentaram teste de triagem de hemoglobinopatia alterado, o que correspondeu a 73% das crianças nascidas entre 2010 e 2011 com resultado de triagem neonatal para hemoglobinopatia anormal. As mães responderam um questionário que avaliou informações recebidas na maternidade e na atenção primária à saúde, além de aspectos relacionados à orientação genética. Foi feita estatística descritiva dos dados. RESULTADOS: Das 119 mães participantes, 69 (58%) tinham filhos com traço falciforme, 22 (18,5%) traço C, 18 (15,1%) traço alfatalassêmico e 10 (8,4%) resultado inconclusivo. Na maternidade, 118 mães (99,2%) receberam informação sobre onde ir e 115 (96,6%) foram orientadas sobre o momento correto para coleta do teste. Somente quatro mães (3,4%) foram informadas sobre quais doenças seriam investigadas e os riscos de não fazer o rastreio. Das 119 mães participantes, 17 (14,3%) reconheceram a diferença entre traço e doença e 42 (35,3%) consideraram que um teste alterado poderia ter implicações para futuras gestações. Em 70 casos (58,8%), o médico da criança não foi informado sobre o resultado da triagem. CONCLUSÕES: O programa de triagem neonatal necessita de aperfeiçoamento. Nos dois cenários investigados, os profissionais de saúde carecem de treinamento para orientar mães e famílias. .

In vitro antimycobacterial activity of six Cameroonian medicinal plants using microplate alamarBlue assay

Objective/background: The latest incidence of tuberculosis [TB] [per 100,000 people] in Cameroon was 243.00 as of 2011. Over the past 21 years, the value for this indicator has fluctuated between 112.00 in 1990 and 320.00 in 2003. Worldwide, this incidence has also increased, bringing back TB as a reemerging disease. On the same note, resistance to anti-TB drugs has increased, urging the search for new molecules

Methods: This study was carried out to evaluate the antimycobacterial activity of six medicinal plants on the virulent strain, H37Rv, using the microplate alamarBlue assay. Mycobacterium tuberculosis [H37Rv strain] was incubated with decreased concentrations of six plant extracts, ranging from 250 microg/mL to 31.25 microg/mL. After 7 days of incubation at 37 degree C, the effects of these plant extracts on the viability of the mycobacteria were evaluated. For each plant extract, the minimal inhibitory concentration was determined

Results: The results showed that the compounds MBC1, MBC24, MBC68, MBC81, MBC117, and MBC118 were the best candidates with minimal inhibitory concentrations of 31.25, 62.5, 125, 62.5, and 125 microg/mL, respectively

Conclusion: These results confirm and validate the traditional use of these plants to treat respiratory diseases, which could be good sources and alternatives of plant metabolites for anti-TB-drug development

Study on standard of safe application of thiamethoxam on GAP of Lonicera japonica / 中国中药杂志

The paper is aimed to establish a method of residue analysis for thiamethoxam and to study its degradation dynamic and final residue and its standard of safe application of thiamethoxam on Lonicera japonica. Samples extracted with methanol by ultrasonication were purified with dichloromethane by liquid-liquid extraction and SPE column and analysed by HPLC-UV. The results showed that average rate was 84.91%-94.44% and RSD 1.74%-4.96% with addition of thiamethoxam in respectively diverse concentration, which meets inspection requirement of pesticide residue. Two kinds of dosages of thiamethoxam were treated- varying from recommended dosage (90 g x hm(-2)) to high dosage (135 g x hm(-2)), Results of two years test showed that thiamethoxam was degraded more than 90% seven days after application and the half - life period of thiamethoxam was 1.54-1.66 d. The digestion rate of thiamethoxam was fast in the L. japonica. The recommended MRL of thiamethoxam in the L. japonica is 0.1 mg x kg(-1), the dosage of 25% thiamethoxam WDG from 90-135 g x hm(-2) is sprayed less than three times a year on L. japonica and 14 days is proposed for the safety interval of the last pesticide application's and harvest's date.

Evaluation of some 1H-pyrazole derivatives as a dual acting antimalarial and anti-leishmanial agents

The synthesis of a novel series of 1H-pyrazole derivatives was achieved by condensation of pyrazole aldehyde 1 with hydrazine hydrate to give hydrazone 7. On the other hand, cyclization of alpha, beta-unsaturated ketone counterpart 2 using hydrazine hydrate in liquid aliphatic acids rendered compounds 4-6 and hydrazine hydrate in ethanol afforded compound 3. The later was allowed to react with aroyl chloride giving rise to compounds 8, 9. All compounds were tested for their in vivo anti-malarial and in vitro antileishmanial activities. The anti-malarial activity was performed using Plasmodium berghei infected mice, while the anti-leishmanial activity of the compounds was determined against Leishmania aethiopica promastigotes using alamar blue reduction assay. Compound 3, 1-[4-methylphenyl]-3-phenyl-4-[3-[2-thienyl]-2-pyrazolin-5-yl]-1H-pyrazole, possessed the highest anti-malarial activity with suppression of 70.26%. The highest anti-leishmanial activity was exhibited by compound 2, 1-[4-methylphenyl]-3-phenyl-4-[1-[2-thienyl]-prop-2-en-1-one]-1H-pyrazole, with an IC[50] of 0.079 microg/ml. Hydrazone 7 showed appreciable dual anti-malarial [suppression = 62.30%] and anti-leishmanial activity [IC[50] = 1.823 microg/ml]

Developmental competence of bovine oocytes selected based on follicle size and using the brilliant cresyl blue [BCB] test

Many studies reported that follicle size has an essential role in developmental potential of oocytes. Also, the brilliant cresyl blue [BCB] test is one of the most important criteria in selection of more competent oocytes. Selection of developmentally competent bovine oocytes. A total of 1730 bovine cumulus oocyte complexes [COCs] were recovered from the ovaries by follicles isolation and classified into 3 categories according to the diameters of the follicles [small, <3 mm; medium 3-6 mm and large >6 mm]. Oocytes were exposed to the BCB stain, diluted in Dulbecco's phosphate-buffered saline, modified with 0.4% bovine serum albumin [BSA] for 90 min. Oocytes with or without blue coloration of the cytoplasm were designated as BCB[+] and BCB[-], respectively. The BCB[+] and control oocytes originated from large and medium follicles exhibited a higher [p<0.0001] cleavage and blastocyst rate than BCB[2] oocytes. Furthermore, the BCB[+] oocytes from large and medium follicles had the highest [p<0.0001] proportion of blastocyst than other treatment groups. In contrast, the BCB[-] oocytes from small follicles had the lowest [p<0.0001] proportion of blastocyst than other treatment groups. Interestingly, the percentage of the BCB[+] oocytes from the large and medium ovarian follicles was significantly higher [p<0.0001], than the BCB[+] oocytes from the small follicles. Current results confirmed that each BCB[+] oocyte could not lead to perfect embryo development and the BCB test is not sufficient enough for the identification of oocytes that are competent for in vitro embryo development

effect of melatonin on maturation, glutathione level and expression of HMGB1 gene in brilliant cresyl blue [BCB] stained immature oocyte

Nutrients and antioxidants in the medium of immature oocyte have a profound effect on maturation, fertilization and development of resulting embryos. In this study the effects of melatonin as an antioxidant agent on maturation, glutathione level and expression of high mobility group box-1 [HMGB1] gene were evaluated in immature oocytes of mice stained with brilliant cresyl blue [BCB]. In this experimental study, immature oocytes were harvested from ovaries of Naval Medical Research Institute [NMRI] mice. Oocytes were stained with 26 Micro M BCB for 90 minutes and transferred to in vitro maturation medium containing varying doses of melatonin [10[-12], 10[-9], 10[-6], 10[-3] M] and without melatonin, for 22-24 hours. Maturation was monitored using an inverted microscope. Glutathione was assessed by monochlorobimane [MCB] staining and HMGB1 expression in mature oocyte was analyzed using real-time polymerase chain reaction [PCR]. Melatonin in the concentration of 10[-6] M had the most effect on maturation and HMGB1 expression of BCB+ oocytes [p<0.05]. Meanwhile melatonin had no effects on glutathione levels. Additionally in immature BCB- oocytes, compared to the control group, melatonin did not affect cytoplasm maturation [p>0.05]. In vitro treatment with melatonin increases the maturation and HMGB1 expression in BCB+ immature oocytes and has no significant effect on glutathione levels

Comparison of the in-vitro effects of bevacizumab, mitomycin-C, 5-fluorouracil,and triamcinolone acetonide on the viability of cultured human tenon's fibroblasts

@#<p style="text-align: justify;"><strong>Objective:</strong> To evaluate and compare the effects of bevacizumab, mitoinycin-C (MMC), 5-fluorouracil (5-FU), and triamcinolone acetonide (TA) on the viability of cultured human Tenon's capsule fibroblasts (cHTF) in vitro.</p> <p style="text-align: justify;"><strong>Methods:</strong> Human Tenon's fibroblasts (HTF) were harvested and cultured in a Roswell-Park-Memorial 1-Institute (RPMI) media. MMC, 5-FU, bevaciz. umab, and TA were administered to the cHTF at 3-fold decreasing concentrations starting from 20 ug, 5 mg, 25 mg, and 4 mg respectively. A negative control/untreated group containing RPMI media only was included in the study. Fibroblast cell viability was assessed using resazurin fluorim etric assay. Half¬maximal inhibitory concentration (IC50) was computed for agents which showed significant decrease in cHTF viability compared to the untreated group.</p> <p style="text-align: justify;"><strong>Results:</strong> There was no significant difference in cH IF viability between the untreated control group compared to 5-FU (p=0.97), bevacizumab (p=0.10), and TA (p=0.06) groups. Mitomycin-C showed a significant decrease in cHTF viability (p<0.001) which was dose dependent. The IC50 of MMC was computed at 12.16 ug using the prism software.</p> <p style="text-align: justify;"><strong>Conclusion:</strong> Mitomycin-C demonstrated dose-dependent decrease in viability of cultured human Tenon's fibroblasts. 5-FU, bevacizumab, and triamcinolone did not show this effect.</p> <p style="text-align: justify;"><strong>Key Words:</strong> Mitomycin-C, 5-fluorouracil, Bevaciz. umab, Tria. mcinolone acetonide, Fibroblast, Trabeculectomy</p>

Early responses of the STAT3 pathway to platinum drugs are associated with cisplatin resistance in epithelial ovarian cancer

Cisplatin resistance remains one of the major obstacles when treating epithelial ovarian cancer. Because oxaliplatin and nedaplatin are effective against cisplatin-resistant ovarian cancer in clinical trials and signal transducer and activator of transcription 3 (STAT3) is associated with cisplatin resistance, we investigated whether overcoming cisplatin resistance by oxaliplatin and nedaplatin was associated with the STAT3 pathway in ovarian cancer. Alamar blue, clonogenic, and wound healing assays, and Western blot analysis were used to compare the effects of platinum drugs in SKOV-3 cells. At an equitoxic dose, oxaliplatin and nedaplatin exhibited similar inhibitory effects on colony-forming ability and greater inhibition on cell motility than cisplatin in ovarian cancer. Early in the time course of drug administration, cisplatin increased the expression of pSTAT3 (Tyr705), STAT3α, VEGF, survivin, and Bcl-XL, while oxaliplatin and nedaplatin exhibited the opposite effects, and upregulated pSTAT3 (Ser727) and STAT3β. The STAT3 pathway responded early to platinum drugs associated with cisplatin resistance in epithelial ovarian cancer and provided a rationale for new therapeutic strategies to reverse cisplatin resistance.

Effect of Dipyridamole on the Reactive Oxygen Species and Oxidative Stress in Trabecular Meshwork Cells

PURPOSE: To investigate the effects of dipyridamole (DPD) on the production of reactive oxygen species (ROS) and oxidative stress in cultured human trabecular meshwork cells (HTMC). METHODS: Antioxidant activity of DPD was determined by DPPH assay. Primarily cultured HTMC were exposed to 0, 20, and 50 microm DPD using serum-deprived media. The effect of DPD on the production of ROS was assessed with the DCHFDA assay. The effect of DPD on the t-butyl hydroperoxide (tBHP)-induced oxidative stress was assessed with resazurin assay. RESULTS: DPD showed significant antioxidant activity. DPD significantly decreased the production of ROS (p < 0.05) and improved cellular activity significantly after treatment with t-BHP (p < 0.05). DPD did not affect the generation of nitric oxides. CONCLUSIONS: DPD suppressed the formation of ROS and possessed cytoprotective activity against the oxidative stress in HTMC.

Clinical Characteristics of Patients with Indoxacarb Insecticide Poisoning

PURPOSE: Indoxacarb insecticide poisoning causes methemoglobinemia, which is occasionally life-threatening. However, there is limited data on indoxacarb effects after human ingestion. The purpose of this study was to examine the clinical features, complications, management, and medical outcome of patients with indoxacarb insecticide poisoning. METHODS: We retrospectively reviewed the medical records of 10 patients with indoxacarb insecticide poisoning who had visited our emergency centers from January 2008 to December 2011. We collected data on the general characteristics of the patients, their clinical symptoms and signs, laboratory data, management of their condition, and clinical results. RESULTS: Among the 10 patients, 8 were diagnosed with methemoglobinemia. The clinical manifestations of indoxacarb insecticide poisoning were hypotension (3 patients), altered mentality (5 patients), cyanosis (5 patients), dyspnea (2 patients), seizure (3 patients), and cardiac arrest (2 patients). Four patients had a poisoning severity score of 3 and 2 patients had a poisoning severity score of 2. Four patients were treated with methylene blue for methemoglobinemia and one patient was treated with a high dose (150 mg/kg) of ascorbic acid. The serum methemoglobin saturation of five patients who were treated with methylene blue or a high dose of ascorbic acid was nearly normalized. Four patients experienced rhabdomyolysis, pneumonia, hemolytic anemia, acute pancreatitis, and heart failure as a complication of indoxacarb insecticide poisoning. CONCLUSION: We observed a variety of clinical features, complications, management, medical outcome, and clinical course of patients with indoxacarb insecticide poisoning. We could also ascertain the efficacy of methylene blue and high dose ascorbic acid for indoxacarb-induced metheglobinemia.

Determination of pazufloxacin concentration in saliva, gingival crevicular fluid and serum of healthy adults using high-performance liquid chromatography / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a rapid and sensitive high-performance liquid chromatography (HPLC) method for detecting pazufloxacin concentrations in the saliva, gingival crevicular fluid and serum of healthy adults.</p><p><b>METHODS</b>Samples of saliva, gingival crevicular fluid and serum were obtained from healthy adults receiving intravenous infusion of pazufloxacin. The concentrations of pazufloxacin in the samples were quantified by HPLC equipped with a reversed-phase column (Agilent Zorbax SB-C18 5 µm, 250 mm×4.6 mm). The mobile phase for pazufloxacin was a mixture of acetonitrile and 0.5% phosphoric acid containing 1% triethylamine (155:850), and 20 µl of the resulting solution was injected into the HPLC system at a flow rate of 1.0 ml/min. The detection wavelength was set at 245 nm. The samples were first deproteinized by precipitation with methanol followed by supernatant drying; the residue was reconstituted with the mobile phase and centrifuged, and the supernatants were directly injected into the HPLC system.</p><p><b>RESULTS</b>Pazufloxacin in the samples were totally separated without interference by any endogenous substances. The calibration curves showed a good linear regression (r>0.999). The detection limit was 10 ng/ml with within-day and between-day coefficients of variation performance all below 5% and recovery rates all above 91%.</p><p><b>CONCLUSION</b>HPLC is both sensitive and selective for quantification of pazufloxacin in saliva, gingival crevicular fluid and serum.</p>

Comparative evaluation of the microplate nitrate reductase assay and the rezasurin microtitre assay for the rapid detection of multidrug resistant Mycobacterium tuberculosis clinical isolates

The microplate nitrate reductase assay (MNRA) and the rezasurin microtitre assay (REMA) were used for the susceptibility testing of 73 clinical isolates and the results were compared with those that were obtained using the Bactec 460 TB and Bactec MGIT 960 systems. The REMA and the MNRA were performed in 96-well plates. For the REMA, the concentrations of isoniazid (INH) and rifampicin (RIF) ranged from 1.0-0.01 µg/mL and 2.0-0.03 µg/mL, respectively. For the MNRA, the INH concentration was between 1.0-0.03 µg/mL and the RIF concentration was between 2.0-0.06 µg/mL. For the MNRA, the sensitivity, specificity, positive predictive value, negative predictive value and INH/RIF agreement were 100/95.6, 97.6/100, 96.8/100, 100/98 and 98.6/98.6, respectively, and for the REMA, they were 100/91.3, 90.4/100, 88.5/100, 100/96.1 and 94.5/97.2, respectively. Our data suggest that these two rapid, low-cost methods may be inexpensive, alternative assays for the rapid detection of multidrug resistant tuberculosis in low-income countries.

Validation of a modified fluorimetric assay for the screening of trichomonacidal drugs

A fluorimetric microassay that uses a redox dye to determine the viability of the flagellate Trichomonas vaginalis has been optimised to provide a more sensitive method to evaluate potential trichomonacidal compounds. Resazurin has been used in recent years to test drugs against different parasites, including trichomonadid protozoa; however, the reproducibility of these resazurin-based methods in our laboratory has been limited because the flagellate culture medium spontaneously reduces the resazurin. The objective of this work was to refine the fluorimetric microassay method previously developed by other research groups to reduce the fluorescence background generated by the media and increase the sensitivity of the screening assay. The experimental conditions, time of incubation, resazurin concentration and media used in the microtitre plates were adjusted. Different drug sensitivity studies against T. vaginalis were developed using the 5-nitroimidazole reference drugs, new 5-nitroindazolinones and 5-nitroindazole synthetic derivatives. Haemocytometer count results were compared with the resazurin assay using a 10% solution of 3 mM resazurin dissolved in phosphate buffered saline with glucose (1 mg/mL). The fluorimetric assay and the haemocytometer counts resulted in similar percentages of trichomonacidal activity in all the experiments, demonstrating that the fluorimetric microtitre assay has the necessary accuracy for high-throughput screening of new drugs against T. vaginalis.

Antifungal susceptibility testing of Candida tropicalis biofilms against fluconazole using calorimetric indicator resazurin.

Background: C.tropicalis is an important cause of nosocomial infections particularly in immunocompromised patients. Infections caused by Candida spp. are often associated with biofilm formation on implanted medical devices or on epithelial cell surfaces. Phenotypic characteristics of sessile cells in biofilms are known to be different from those of their free-living, planktonic counterparts. Biofilm forming strains often show increased resistance to antimicrobial agents. Materials and Methods : We measured susceptibility to fluconazole of fifty C.tropicalis isolates from immunocompromised (29) and immunocompetent (21) patients by minimum inhibitory concentration (MIC) and minimum biofilm inhibitory concentration (MBIC) assays. MBIC was done using the calorimetric indicator resazurin, to measure the metabolically active cells. Results : Biofilm forming cells showed increased resistance to fluconazole. Conclusion : The resazurin dye test was found to be a good method for determining MBIC.

Methemoglobinemia and Seizure Following Indoxacarb Poisoning

Indoxacarb is an oxadiazine insecticide with selective lethality through blockade of neuronal voltage-dependent sodium channels. It has a low mammalian toxicity, and few cases of human toxicity after indoxacarb ingestion can be found in the literature. A 36 year-old male patient visited our ED after a generalized tonic clonic seizure, which was witnessed by his mother. His past medical history was nonspecific. On initial presentation, he showed a decreased level of consciousness with a Glasgow coma score of 5/15 (E1V1M3), unprotected airway, hypoxia, and cyanosis. The saturation gap and cyanosis after intubation and mechanical ventilation was strongly suggestive of methemoglobinemia due to poisoning. Finally, the methemogobin (metHb) level was 27.4%. Therefore, the patient received 100 mg of methylene blue (2 mg/kg, 1% solution) and 50 g of charcoal. The insecticide was found to be ingested xenobiotic (Steward Gold(R); 5% indoxacarb; 95% inert ingredients and other components). On the second hospital day, the patient became alert. The patient's metHb level was 0.1%. The endotracheal tube was removed. On the fifth hospital day, he was discharged in good condition. Herein we present a case of indoxacarb poisoning with methemoglobiemia and seizure, which are unusual presentations.

Small Molecule Inhibitors in Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease predominantly affecting diarthroidal joints. Following the successful application of biologic agents, several small molecule inhibitors are currently under clinical trials. Small molecule inhibitors have several strengths compared with biologics. First, they can target several inflammatory cytokines together by blocking common signal transduction pathways. Second, they can be taken orally. Third, the price can be made flexible. Among the several small molecule inhibitors in the development process, fostamatinib and tofacitinib are the closest to the clinics at the moment. Fostamatinib, which is a Syk inhibitor, showed superior efficacy over placebo with tolerable safety signals. Diarrhea, hypertension and infection are representative adverse events. Tofacitinib, which is JAK inhibitor, is now finishing phase 3 clinical trials. It showed clinical efficacy comparable to Adalimumab and similar adverse effect profiles to the biologics, which include opportunistic infections. For laboratory abnormalities, leukopenia, anemia, increase of LDL and serum Cr were reported, which, however, were stabilized with prolonged use. Other classes of small molecule inhibitors did not show impressive efficacy as these small molecule inhibitors. In conclusion, small molecule inhibitors are promising novel therapeutic agents for the treatment of RA. They will be able to change the treatment paradigm of RA if they can show long-term safety.