Regorafenib monotherapy as second-line treatment of patients with RAS-mutant advanced colorectal cancer (STREAM): an academic, multicenter, single-arm, two-stage, phase II study.

BACKGROUND: Maintaining angiogenesis inhibition and switching the chemotherapy backbone represent the current second-line therapy in patients with RAS-mutant metastatic colorectal cancer (mCRC). Regorafenib, an oral multikinase inhibitor, prolonged overall survival (OS) in the chemorefractory setting. MATERIALS AND METHODS: STREAM was an academic, multicenter, single-arm phase II trial, evaluating the activity of regorafenib in RAS-mutant mCRC, in terms of the rate of patients who were progression-free after 6 months from study entry (6mo-PF). Patients were pretreated with fluoropyrimidine, oxaliplatin, and bevacizumab. According to Simon's two-stage design, ≥18 patients 6mo-PF were needed in the overall population (N = 46). Secondary endpoints were safety, objective response rate (ORR), progression-free survival (PFS), and OS. Early metabolic response by [18F]2-fluoro-2-deoxy-D-glucose-positron emission tomography/computed tomography ([18F]-FDG PET/CT) scan was an exploratory endpoint. EudraCT Number: 2015-001105-13. RESULTS: The number of patients 6mo-PF was 8/22 at the first stage and 14/46 in the overall population. The ORR was 10.9%, disease control rate was 54.6%, median (m)PFS was 3.6 months [95% confidence interval (CI) 1.9-6.7 months], mOS was 18.9 months (95% CI 10.3-35.3 months), and mPFS2 (from study entry to subsequent-line progression) was 13.3 months (95% CI 8.4-19.7 months). Long benefiter patients (>6mo-PF) significantly more often had a single metastatic site and lung-limited disease. No unexpected toxicity was reported. Grade ≥3 events occurred in 39.1% of patients, with hand-foot syndrome (13%), fatigue, and hyperbilirubinemia (6.5%) occurring mostly. Baseline metabolic assessment was associated with OS in the multivariate analysis, while early metabolic response was not associated with clinical outcomes. CONCLUSIONS: The study did not meet its primary endpoint. However, regorafenib was well tolerated and did not preclude subsequent treatments. Patients with good prognostic features (single metastatic site and lung-limited disease) reported clinical benefit with regorafenib. The exploratory metabolic analysis suggests that baseline [18F]-FDG PET/CT might be useful to select patients with a favorable outcome. A chemotherapy-free interval with regorafenib was associated with durable disease control in a selected group of patients with favorable clinical characteristics.

Viability of human fibroblasts in coconut water as a storage medium.

AIM: To evaluate the effectiveness of a new storage medium for avulsed teeth, coconut water, in maintaining the viability of human fibroblasts. METHODOLOGY: Cell viability after different time periods was evaluated in the following storage media: coconut water, coconut water with sodium bicarbonate, milk, saline and still mineral water. Human fibroblasts were seeded in Eagle's minimal essential medium (EMEM) supplemented with 7.5% foetal calf serum. After trypsinisation, 100 microL of culture medium containing approximately 10(4) cells mL(-1) were collected and pipetted into the wells of 96-well plates, which were incubated overnight in 5% CO(2) and 95% air mixture at 37 degrees C. EMEM was then replaced by the storage media and the plates were incubated at 37 degrees C for 1, 2 and 4 h. Cell viability was determined using the neutral red assay. The proportions of viable cells after exposure to the storage media were analysed statistically by anova and the least significant difference (LSD) test (alpha = 5%). RESULTS: Milk had the greatest capacity to maintain cell viability (P < 0.05), followed by coconut water with sodium bicarbonate and saline. Coconut water was significantly worse at maintaining cell viability compared to milk, coconut water with sodium bicarbonate and saline. The smallest number of viable cells was observed for mineral water (P < 0.05). CONCLUSION: Coconut water was worse than milk in maintaining human fibroblast cell viability.

Determinação da resistência de Staphylococcus aureus: um desafio? / Identification of drug resistance in Staphylococcus aureus: a challenge?

O objetivo deste estudo foi avaliar o perfil de resistência de cepas de Staphylococcus aureus, com diferentes padrões de eletroforese em campo pulsado (PFGE), em relação à resistência induzida à clindamicina e caracterizar cepas resistentes à oxacilina por testes fenotípicos. Do total de 18 cepas diferenciadas por PFGE, isoladas dos sítios nasais ou linguais de portadores adultos saudáveis, sem doença de base, sem histórico de uso de antibióticos e internações hospitalares, quatro (22,2%) apresentaram sensibilidade à clindamicina no antibiograma convencional, mas demonstraram resistência no D-teste; uma cepa (5,6%) foi caracterizada como BORSA (borderline) em relação à resistência a oxacilina e outra (5,6%) CA MRSA (S.aureus meticilina/oxacilina resistente associado à comunidade), ambas sensíveis à cefoxitina pelo teste de disco difusão. A caracterização molecular pela reação em cadeia para polimerase (PCR) da cepa identificada fenotipicamente como CA MRSA não revelou a presença do gene mecA, indicando tratar-se de cepa BORSA. Estes resultados apontam a importância do emprego rotineiro do D-teste como ferramenta para a determinação da resistência do tipo induzida à clindamicina, bem como para a importância da inclusão do teste de resistência à cefoxitina entre os métodos fenotípicos para caracterização de MRSA.

The aim of this study was to identify the resistance profile of Staphylococcus aureus strains, in relation to induced clindamycin resistance, and to detect oxacillin resistance by the routine phenotypic methods. The strains were isolated from nasal or lingual swabs taken from healthy adult carriers with no medical history of hospitalization or antibiotic treatment. Eighteen strains were distinguished by the different patterns generated by pulsed gel electrophoresis (PFGE). Four (22.2%) of these showed sensitivity to clindamycin by the conventional antibacterial susceptibility test, but demonstrated inducible resistance to it by the D-test. One strain (5.6%) was characterized as borderline oxacillin-resistant S. aureus (BORSA), and another (5.6%) as CA MRSA (community-associated methicillin-resistant Staphylococcus aureus). Both of these strains were shown to be cefoxitin susceptible by the disk diffusion test. The polymerase chain reaction (PCR) failed to detect the mecA gene in this last strain and it was thus classified as BORSA. These results show the importance of incorporating the D-test into the routine lab tests for S. aureus inducible clindamycin resistance and also of including the cefoxitin resistance test among the phenotypic methods for MRSA characterization.

Influência de fluídos biológicos na sobrevivência de Staphylococcus aureus sobre diferentes superfícies secas / Influence of biological fluids on survival of Staphylococcus aureus on various dried surfaces

Sabe-se que a transmissão de infecções hospitalares, via cruzada ou ambiental, é facilitada pela sobrevivência de microrganismos em superfícies secas que pode ser favorecida pela presença de fluídos biológicos. Visando demonstrar o cuidado com substâncias corporais narotina de limpeza, esse estudo avaliou a influência de alguns fluídos biológicos (sangue, urina e saliva artificial) na sobrevivência de Staphylococcus aureus (ATCC 25923), depositados de modo similar, sobre diferentes superfícies após secagem. O sangue foi capaz de preservar a viabilidade bacteriana por até 72 diasquando depositado sobre piso cerâmico. O tecido em fibra de algodão permitiu maior tempo de sobrevivência em comparação ao tecido sintético. Esses resultados demonstram que a composição do fluído biológico e o tipo de suporte altera o tempo da sobrevivência bacteriana em condições ambientais.

Aplicabilidade do TTC para a detecção de bacteriúria / Applicability of TTC test to bacteriuria detection

Exames laboratoriais para detecção da infecção urinária com custo menor que da urocultura têm sidobuscados. O cloridrato de trifeniltetrazólio foi avaliado em 342 amostras de urina paralelamente à urocultura objetivando a detecção de bacteriúria significativa. Os resultados demonstraram que o teste apresenta boa sensibilidade (91,3%) e baixa especificidade (64,3%), com valor preditivo negativo de 99,0%. Apesar do teste não substituir a urocultura como método diagnóstico, pode ser recomendado para a triagem de bacteriúria, eliminando a realização da cultura em amostras negativas.

Ocorrência de infecção urinária em pacientes de um hospital universitário / Occurrence of urinary infection in patients at an university hospital

Infecção Hospitalar é um grande problema de saúde pública, sendo responsável pelo aumento da morbimortalidade dos pacientes, bem como do período de internação e custos assistenciais. Dentre as infecções hospitalares, a infecção do trato urinário (ITU) é a mais comum, sendo a presença de cateter urinário o principal fator de risco. O presente estudo teve como objetivo avaliar as ITUs em pacientes internados num Hospital Universitário, no período de outubro a dezembro de 2003. Das 271 amostras de urina analisadas, 51 foram positivas, sendo 27 de pacientes com infecção comunitária do trato urinário e 24 de origem hospitalar. As ITUs comunitárias foram mais comuns em pacientes do sexo feminino (63%), com idade entre 0 e 15 anos (37%), sendo Escherichia coli o agente mais frequente (74,1%). Os episódios de ITU de origem hospitalar ocorreram, na sua maioria, em pacientes que faziam uso de sonda vesical de demora, do sexo masculino (68%) e com idade acimade 50 anos (68%), e tiveram como agentes etiológicos mais frequentes Escherichia coli (29,1%) e Klebsiella spp. (29,1%). Quanto à sensibilidade/resistência aos antimicrobianos, E. coli e Klebsiella pneumoniae apresentaram sensibilidade elevada (62,5%) ao sulfametoxazol/trimetoprim e a ampicilina, sugerindo a não utilização desses medicamentos nessa instituição.

[Preponderance of Mycobacterium tuberculosis strains of the family Beijing and risk factors of their transmission in the Samara Region].

Current molecular and epidemiological studies could reveal individual families of Mycobacterium tuberculosis strains. The purpose of the present study was to reveal the spread of Mycobacterium tuberculosis strains of the family Beijing in the Samara Region and to define risk factors for their transmission. This was a cross-sectional populational molecular epidemiological study that showed that the Beijing genotype prevailed among the obtained isolates (66.6%; 586/880) and it was encountered among convicts and young persons (RR 1.3; 95% CI 1.2-1.5 and RR 1.2; 95% CI 1.0-1.3, respectively), which is indicative of active and recent transmission. Multifactorial analysis indicated that male sex (OR 1.5; 95% CI 1.1-1.9), younger age (OR 1.3; 95% CI 1.1-1.7), homelessness (OR 5.6; 95% CI 1.1-6.3), and prior or current confinement (OR 2.0; 95 CI 1.5-2.7) were substantially associated with the risk of contamination with the strain of the Beijung family. Drug resistance, including multidrug resistance, was twice higher among the strains of this family.

In vitro basal and metabolism-mediated cytotoxicity of flavonoids.

The purpose of this study was to compare the basal cytotoxicity and metabolism-mediated cytotoxicity of kaempferol, quercetin and rutin. McCoy cells were exposed to various concentrations of the flavonols with and without the S9 system. The neutral red uptake assay was used to determine viability after 24 h at 35-37 degrees C. Dose-response curves were established for each flavonol in the presence and absence of external metabolizing systems. Kaempferol and quercetin were cytotoxic and provoked a dose-dependent decrease in cell viability, without the S9 system. The hepatic S9 microsomal fraction metabolized these compounds to less cytotoxic metabolites. In contrast, rutin at 500 microg/ml failed to produce any overt signs of toxicity in either assay.

[Analysis of risk factors of the occurrence of drug resistance in patients with tuberculosis from civil and penitentiary sectors in the Samara Region].

The true prevalence rates of multidrug-resistant tuberculosis (MDRT) are unknown for most regions of Russia. This study was conducted in the Samara Region that differs from other regions in the rapid spread of HIV infection. The purpose of this study was to determine the primary and acquired resistance of Mycobacterium tuberculosis (MBT) to first-line antituberculous drugs in patients from civil and penitentiary sectors and to reveal risk factors of drug resistance of MBT. Six hundred patients (309 civilians and 291 prisoners who had been bacteriologically diagnosed as having tuberculosis. The authors have established the following:--in new cases, primary drug resistance is as follows: to isoniazid [38.9% (95% CI, 31.3-36.9%)], to rifampicin [25.9% (95% CI, 19.4-33.4%)] and to MDRT [23.0% (95% CI, 16.7-30.3%)];--in prisoners, the primary resistance of MBT was statistically more significant than in civilians;--male sex, in adequate prior or current treatment for tuberculosis for more than 4 weeks, the presence of fibrocavernous tuberculosis and previous prison stay are essential risk factors of the development of resistance of MBT to both any first-line drug and MDRT;--HIV infection is unassociated with resistance.

Eficiência de máscaras cirúrgicas como equipamento de proteção respiratória contra aerossóis bacterianos / Efficiency of surgical masks as respiratory protective equipment against bacterial aerosols

O uso de máscaras é aceito como método de proteção individual contra agentes microbianos presentes em ambientes que possam ser inalados. O objetivo desse estudo foi avaliar a performance de máscaras descartáveis do tipo cirúrgica, comercializadas no Brasil, na filtração de bioaerossóis emitidos por aeradores de uma estação de tratamento de esgoto sanitário. A eficiência na filtração variou de 50,9% a 99,9% para seis produtos de diferentes procedências

Bioaerossóis bacterianos em um hospital / Bacterial bioaerosols in hospital

A contaminação microbiológica de ambientes internos é afetada pela presença de bioaerossóis do ambiente externo e as geradas no próprio ambiente. O objetivo deste trabalho é relacionar a concentração de bioaerossóis encontrados em ambientes internos de um hospital em relação ao ar exterior, levando em consideração alguns parâmetros ambientais que influenciam na qualidade do ar, como número de ocupantes e tipo de ventilação. Quinhentos litros de ar foram coletados, por impacto, durante cinco minutos, utilizando-se um amostrador de ar de um estágio sobre placas de meios de cultura. A concentração média de bioaerossóis bacterianos viáveis no ambiente externo de um hospital do interior do Estado de São Paulo foi de 77 ± 4 UFC/m3 e no ambiente interior de 302 ± 260 UFC/m3 de ar. O centro cirúrgico, após cirurgia ortopédica, único ambiente climatizado amostrado, apresentou a maior concentração (867 ± 482 UFC/m3). Nos ambientes interiores nove espécies bacterianas foram identificadas. Apesar da dificuldade em se estabelecer valores para a concentração de bioaerossóis bacterianos em ambientes hospitalares, os resultados demonstram a necessidade de programas apropriados para a manutenção da concentração microbiana baixa nesses ambientes e ausência de microrganismos que possam significar risco para os seus ocupantes. O monitoramento do ar deveria ser recomendado objetivando definir quantidades aceitáveis de contaminantes bacterianos

Cytotoxicity of chlorhexidine digluconate to murine macrophages and its effect on hydrogen peroxide and nitric oxide induction.

Chlorhexidine, even at low concentrations, is toxic for a variety of eukaryotic cells; however, its effects on host immune cells are not well known. We evaluated in vitro chlorhexidine-induced cytotoxicity and its effects on reactive oxygen/nitrogen intermediate induction by murine peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 ml of 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/ml) in appropriate medium for each test. Cell preparations contained more than 95% macrophages. The cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. The midpoint cytotoxicity values for 1- and 24-h exposures were 61.12 +/- 2.46 and 21.22 +/- 2.44 microg/ml, respectively. Chlorhexidine did not induce synthesis or liberation of reactive oxygen/nitrogen intermediates. When macrophages were treated with various sub-toxic doses for 1 h (1, 5, 10, and 20 microg/ml) and 24 h (0.5, 1, and 5 microg/ml) and stimulated with 200 nM phorbol myristate acetate (PMA) solution, the H2O2 production was not altered; however, the NO production induced by 10 microg/ml lipopolysaccharide (LPS) solution varied from 14.47 +/- 1.46 to 22.35 +/- 1.94 micromol/l and 13.50 +/- 1.42 to 20.44 +/- 1.40 micromol/l (N = 5). The results showed that chlorhexidine has no immunostimulating activity and sub-toxic concentrations did not affect the response of macrophages to the soluble stimulus PMA but can interfere with the receptor-dependent stimulus LPS.

Cytotoxicity of chlorhexidine digluconate to murine macrophages and its effect on hydrogen peroxide and nitric oxide induction

Chlorhexidine, even at low concentrations, is toxic for a variety of eukaryotic cells; however, its effects on host immune cells are not well known. We evaluated in vitro chlorhexidine-induced cytotoxicity and its effects on reactive oxygen/nitrogen intermediate induction by murine peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 ml of 10 mM phosphate-buffered saline, washed twice and resuspended (10(6) cells/ml) in appropriate medium for each test. Cell preparations contained more than 95 percent macrophages. The cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. The midpoint cytotoxicity values for 1- and 24-h exposures were 61.12 ± 2.46 and 21.22 ± 2.44 æg/ml, respectively. Chlorhexidine did not induce synthesis or liberation of reactive oxygen/nitrogen intermediates. When macrophages were treated with various sub-toxic doses for 1 h (1, 5, 10, and 20 æg/ml) and 24 h (0.5, 1, and 5 æg/ml) and stimulated with 200 nM phorbol myristate acetate (PMA) solution, the H2O2 production was not altered; however, the NO production induced by 10 æg/ml lipopolysaccharide (LPS) solution varied from 14.47 ± 1.46 to 22.35 ± 1.94 æmol/l and 13.50 ± 1.42 to 20.44 ± 1.40 æmol/l (N = 5). The results showed that chlorhexidine has no immunostimulating activity and sub-toxic concentrations did not affect the response of macrophages to the soluble stimulus PMA but can interfere with the receptor-dependent stimulus LPS.

Mutagenicity of paepalantine dimer and glycoside derivatives from Paepalanthus bromelioides.

The first isocoumarin isolated from the methylene chloride extract of Paepalanthus bromelioides, named paepalantine (isocoumarin 1), was found to have antimicrobial activity; but, it is mutagenic clastogenic and cytotoxic. Two other isocoumarins, paepalantine-9-O-beta-D-glucopyranoside (isocoumarin 2) and paepalantine-9-O-beta-D-allopyranosyl(1 --> 6) glucopyranoside (isocoumarin 3) were isolated from the ethanolic extract. A fourth new isocoumarin, also isolated from the methylene chloride extract of the capitula of P. bromelioides, was characterized as an 8-8' dimer of paepalantine and denominated isocoumarin 4. The abilities of isocoumarins 2, 3 and 4 to induce mutations in Salmonella typhimurium strains TA97a, TA98, TA100 and TA102 were investigated. Mutagenic activity was observed in strain TA97a treated with isocoumarin 2 in the presence of S9 mixture. The substitution of H at position 9 by glucose or glucose-allose caused reductions in the mutagenic activities of paepalantine, indicating this to be an important site for these properties.

Dispersão de bioaerossóis por aeradores em uma estação de tratamento de esgoto sanitário / Bioaerosols dispersion from aerators of a sanitary sewage treatment station

O objetivo desse trabalho foi investigar o alcance de bioaerossóis emitidos por aeradores da Estaçãdo de Tratamento de Esgoto Sanitário de Araraquara, Brasil, utilizando coliformes totais como microrganismos indicadores. A concentração de coliformes viáveis no ar foi determinada a partir de várias distâncias dos aeradores, utilizando-se impactador de ar de um estágio em Chromocult Coliform Agar (MERCK), tendo como padrão referencial a contagem obtida em outros pontos da cidade. Parâmetros meteorológicos foram determinados simultaneamente com a amostragem bacteriológica. ANOVA foi utilizada para avaliação estatística dos resultados. A concentração de coliformes totais mostrou diferença significativa em distâncias de até 100 m dos aeradores em relação ao padrão referencial. Os fatores meteorológicos não influenciaram na preservação desses microrganismos a partir de 100 m das fontes emissoras. A dispersão de bioaerossóis pelos aeradores da Estação de Tratamento de Esgoto Sanitário de Araraquara não é fonte de contaminação para áreas que circundam essa estação

A new cytotoxic naphthopyrone dimer from Paepalanthus bromelioides.

A new naphthopyrone dimer was isolated from the capitula of Paepalanthus bromelioides by chromatographic procedures. It structure was deduced from spectrometric data. On colorimetric assay for cytotoxicity the new dimer showed IC50 of 55.9 microM.

Mutagenic and cytotoxic activity of an isocoumarin (Paepalantine) isolated from Paepalanthus vellozioides.

A new isocoumarin with antimicrobial activity was isolated from Paepalanthus vellozioides (a native Brazilian plant) and called paepalantine. This study was carried out to assess the mutagenic activity of this new agent in assays with Salmonella typhimurium TA100, TA98, and TA102 and in Chinese hamster ovary (CHO) cell cultures, as well as cytotoxicity to McCoy cells. paepalantine caused a significant dose-dependent increase in the frequency of revertants in the three strains used in the assay, both with and without S9 mix, in concentrations varying from 2 to 128 micrograms/plate. The mutagenicity was confirmed in assays with CHO cells treated in the G1, S, and G2 phases of the cell cycle. There was an increase in the chromosomal aberration frequency, mainly in the G2 phase. Furthermore, the mitotic index of the treated cultures (40,80, and 160 micrograms/ml) was significantly lower, indicating cytotoxicity. The midpoint cytotoxicity values of McCoy cells by the neutral red (NR) and microculture tetrazolium (MTT) techniques resulted in a NR50 and MTT50 of 30 and 38 micrograms/ml, respectively. Alterations to the paepalantine structure are suggested to reduce its mutagenic and cytotoxic activity in investigations for its antineoplastic potential.

Characteristics of gram-stained cervical smear from patients with Chlamydia trachomatis infection.

Cervical discharges from 142 women attending the Public Gynecologic Service of Araraqura (SESA), Brazil were cultured for Chlamydia trachomatis. Gram-smears and plating on semiquantitative sheep blood agar and chocolate agar were also carried out. An isolation rate of 18% was reported. The presence of purulent cervical secretion was observed in 8 (32%) out of 25 women. It was also observed that a substantial proportion of culture-positive women had no symptoms. Our data demonstrate that screening tests should be based on specific diagnostic techniques for Chlamydia trachomatis since the majority of infected women we examined were asymptomatic.

Estudo comparativo do espermograma em portadores e näo portadores de Ureaplasma urealyticum no líquido seminal / Seminal parameters in patients with and without Ureaplasma urealyticum

Foram analisadas 47 amostras de líquido seminal com o objetivo de determinar se alteraçöes nos valores de referência do espermograma poderiam estar associadas à presença de Ureaplasma urealyticum na amostra. Observamos positividade em 24(51,1%) dos materiais examinados. Os valores de referência para vitalidade foram considerados satisfatórios em ambos os grupos. Portadores de ureaplasmas apresentaram contagem menor do número de espermatozóides em relaçäo ao grupo näo portador. A presença de títulos elevados de U. urealyticum (> ou = 10**3 CFU/ml)näo altera significativamente os valores de referência dos testes do espermograma