Evaluation of the immunomodulatory activity of thalidomide on tumor-associated macrophages in the 4T1 murine metastatic breast cancer model / [Avaliação do efeito imunomodulador da talidomida em macrófagos associados a tumores no modelo murino de câncer de mama metastático 4T1]

The present work evaluated the immunomodulatory effect of thalidomide (Thal) at different doses on tumor-associated macrophages (TAMs) using a mouse model of human breast cancer. Mice were inoculated with 4T1 cells in the left flank and treated with Thal once a day at concentrations of 50, 100, and 150mg/kg body weight from the 5th day until the 28th day of tumor inoculation. The tumors were sized, proliferation index and TAMs count were evaluated in primary tumors and metastatic lungs. In addition, the metastasis rate was evaluated in the lungs. Thal at 150mg/kg significantly decreased tumor growth, proliferation index, and TAMs infiltration in primary tumors. Conversely, a higher number of TAMs and lower proliferation index were observed in metastatic lungs in mice treated with 150mg/kg of Thal. Furthermore, Thal at 150mg/kg significantly decreased the metastatic nodules in the lungs. Our findings demonstrated that Thal treatment considerably decreased the primary tumor and lung metastasis in mice associated with different TAM infiltration effects in these sites.(AU)

No presente trabalho, foi avaliado o efeito imunomodulador de diferentes doses de talidomida em macrófagos associados ao tumor (TAMs), em um modelo murino de câncer de mama. Camundongos foram inoculados com células 4T1, na região do flanco esquerdo, e tratados com talidomida, uma vez ao dia, nas doses de 50, 100 e 150mg/k, por massa corporal, do quinto dia ao 28º dia de inoculação tumoral. Os tumores foram medidos, o índice de proliferação celular e a contagem de TAMs foram avaliados nos tumores primários e nos pulmões com metástases. Além disso, a taxa de metástases pulmonares também foi avaliada. A talidomida na dose de 150mg/kg diminuiu significativamente o crescimento tumoral, o índice de proliferação celular e a infiltração de TAMs nos tumores primários. Por outro lado, maior número de TAMs e menor índice de proliferação celular foram observados nos pulmões metastáticos, em camundongos tratados com 150mg/kg de talidomida. Ademais, a talidomida na dose de 150mg/kg diminuiu significativamente os nódulos metastáticos nos pulmões. Os resultados demonstraram que o tratamento com talidomida diminuiu o crescimento tumoral e as metástases pulmonares em camundongos, associado com diferentes efeitos na infiltração de TAMs nesses locais.(AU)

Synbiotic supplementation attenuates the promoting effect of indole-3-carbinol on colon tumorigenesis.

Indole-3 carbinol (I3C) has shown dual effects on the promotion and progression stages of colon carcinogenesis while synbiotics (Syn) have exerted anti-carcinogenic activities in most rodent studies. This study aimed to investigate the effects of I3C given alone or together with a Syn intervention on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis. All animals were given four subcutaneous DMH injections (4×40 mg/kg bodyweight, twice a week for two weeks) and then received either basal diet (G1), basal diet containing I3C (1g/kg chow) (G2) or basal diet containing I3C+Syn (I3C + inulin 50g/kg chow + Bifidobacterium lactis BB-12®), 2.5×1010 cfu/g of basal diet), (G3) for 21 weeks. Dietary I3C (G2) significantly increased tumour volume and cell proliferation when compared to the DMH control group (G1). Syn intervention (G3) significantly reduced tumour volume and cell proliferation when compared to I3C (G2). The colon tumours found were classified into well-differentiated tubular adenomas or adenocarcinomas. Dietary I3C or I3C+Syn did not significantly affect the incidence and the multiplicity of tumours in comparison with the DMH control group. Furthermore, Syn intervention (G3) increased Gstm1 and reduced Mapk9 gene expression in colonic tumours. The findings of the present study show that the dietary I3C shows a weak promoting activity, while the combination with Syn ameliorates I3C effects.

Antimicrobial alumina nanobiostructures of disulfide- and triazole-linked peptides: Synthesis, characterization, membrane interactions and biological activity.

Due to the its physical-chemical properties, alumina nanoparticles have potential applications in several areas, such as nanobiomaterials for medicinal or orthodontic implants, although the introduction of these devices poses a serious risk of microbial infection. One convenient strategy to circumvent this problem is to associate the nanomaterials to antimicrobial peptides with broad-spectrum of activities. In this study we present two novel synthesis approaches to obtain fibrous type alumina nanoparticles covalently bound to antimicrobial peptides. In the first strategy, thiol functionalized alumina nanoparticles were linked via disulfide bond formation to a cysteine residue of an analog of the peptide BP100 containing a four amino acid spacer (Cys-Ala-Ala-Ala). In the second strategy, alumina nanoparticles were functionalized with azide groups and then bound to alkyne-decorated analogs of the peptides BP100 and DD K through a triazole linkage obtained via a copper(I)-catalyzed cycloaddition reaction. The complete physical-chemical characterization of the intermediates and final materials is presented along with in vitro biological assays and membrane interaction studies, which confirmed the activity of the obtained nanobiostructures against both bacteria and fungi. To our knowledge, this is the first report of aluminum nanoparticles covalently bound to triazole-peptides and to a disulfide bound antimicrobial peptide with high potential for biotechnological applications.

Tumour-associated macrophages: Relation with progression and invasiveness, and assessment of M1/M2 macrophages in canine mammary tumours.

Macrophages represent a major component of the overall leucocyte population within neoplasms and are important for tumour behaviour in several cancers in human beings. However, little information regarding their role in canine mammary tumours (CMTs) is available. The aim of this study was to address the potential role of tumour-associated macrophages (TAMs) in CMTs. TAMs in CMTs excised from 82 female dogs were quantified at high power (400× magnification) and categorised as low (≤50) or high (>50) TAM counts. Higher TAM counts were associated with clinical stage (P<0.001), tumour type (P=0.016), tumour size (P=0.013), vascular invasion (P=0.031), lymph node metastasis (P=0.003), high proliferation rates (P=0.009), vascular microdensity (P=0.008), invasive tumour profile (P=0.002) and worse prognosis (P=0.018; hazard ratio=0.283). Almost all macrophages infiltrating malignant tumours with high TAM counts expressed CD206 (macrophage mannose receptor 1), while all benign tumours were infiltrated by macrophages expressing inducible nitric oxide synthase (NOS2), suggesting a phenotypic shift from classically activated macrophage (M1) subpopulations towards alternatively activated macrophage (M2) subpopulations in malignant tumours. A triple staining pattern revealed mixed M1/M2 profiles in some tumours, thus characterising an intermediate state. The results indicate that TAMs are associated with more aggressive types of mammary cancer in dogs.

Development of asymmetric resorbable membranes for guided bone and surrounding tissue regeneration.

Membranes design for guided tissue engineering have been studied to aid in cell viability and function as tissue barriers. Two asymmetric resorbable membranes for guided bone regeneration (GBR) were produced: chitosan/pectin/poly-caprolactone (PECm) and poly(vinyl alcohol)/polyethylenimine/poly(ethylene glycol) (PVAm). Both membranes were characterized by physical, chemical, mechanical, degradation rate, and in vitro biological assessment. Scanning electron microscopy (SEM) confirmed the membranes asymmetry, in which PECm asymmetry is given by roughness and chemical composition, while PVAm's only by differences in porosity. Fourier transform infrared spectroscopy (FTIR) identified chemical groups and bonds between polymers. Both sides of PVAm revealed to be hydrophobic, whereas the PECm presented one side with higher hydrophobicity than the other. In vitro biological assessment disclosed that PECm presented a higher cell adhesion growth pattern than PVAm, where it seemed to occur a delay in proliferation due to initial low cell adhesion. Both developed membranes are suitable for GBR, since both membranes fulfil the requirements to be used as a tissue barrier. The PECm has an additional role in cell viability that was not observed in the PVAm. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2141-2150, 2018.

Migraine, hormones and the menopausal transition.

Migraine is a common, disabling and incapacitating headache disorder that may be triggered by many factors, such as hormones especially during the perimenopausal period, where large alterations in estradiol levels can occur. The evidence implies that hormonal fluctuations are one of the important triggers of migraine. During reproductive life and during hormonal contraception, the course of migraine can be impacted. Different types of migraine with and without aura can be variously influenced by hormones. Migraine can constitute a risk factor for stroke and this must be taken in account for menopause hormone therapy. Hormone therapy is a possible approach to prevent migraine that happens during the menopause transition. Scarce data on the various regimens and types of hormone therapy are available. Transdermal estradiol displays a more favorable profile on migraine than oral estrogens because it may provide more constant levels of estrogens.

Mutational profile of TP53 in esophageal squamous cell carcinoma associated with chagasic megaesophagus.

Chaga's disease is an important communicable neglected disease that is gaining wider attention due to its increasing incidence worldwide. Achalasia due to chagasic megaesophagus (CM), a complication of this disease, is a known-yet, poorly understood-etiological factor for esophageal squamous cell carcinoma (ESCC) development. In this study, we aimed to perform the analysis of TP53 mutations in a series of Brazilian patients with ESCC that developed in the context CM (ESCC/CM), and to compare with the TP53 mutation profile of patients with benign CM and patients with nonchagasic ESCC. Additionally, we intended to correlate the TP53 mutation results with patient's clinical pathological features. By polymerase chain reaction (PCR) followed by direct sequencing of the hotspot regions of TP53 (exon 5 to 8), we found that TP53 mutations were present in 40.6% (13/32) of the ESCC/CM group, 45% (18/40) of the nonchagasic ESCC group, and in only 3% (1/33) of the benign CM group. Missense mutations were the most common in the three groups, yet, the type and mutated exon mutation varied significantly among the groups. Clinically, the groups exhibited distinct features, with both cancer groups (ESCC and ESCC/CM) been significantly associated higher consumption of alcohol and tobacco, older age, worse Karnofsky performance status, poor outcome than the patients with benign CM. No significant association was found between TP53 mutation profile and clinical-pathological features in any of the three groups. We describe first the time the analysis of TP53 mutations in ESCC that developed in the context of CM, and the observed high frequency of mutations, suggest that TP53 also plays an important role in the tumorigenic process of this unexplored etiological condition.

Sentinel node biopsy using blue dye and technetium99 in advanced gastric cancer: anatomical drainage and clinical application.

Lymph node metastases are an independent prognosis factor in gastric carcinoma (GC) patients. Radical lymphadenectomy can improve survival but it can also increase surgical morbidity. As a principle, sentinel node (SN) navigation surgery can avoid unnecessary lymphadenectomy without compromising prognosis. In this pilot study, 24 patients with untreated GC were initially screened for SN navigation surgery, of which 12 were eligible. Five patients had T2 tumors, 5 had T3 tumors and 2 had T1 tumors. In 33% of cases, tumor diameter was greater than 5.0 cm. Three hundred and eighty-seven lymph nodes were excised with a median of 32.3 per patient. The SN navigation surgery was feasible in all patients, with a median of 4.5 SNs per patient. The detection success rate was 100%. All the SNs were located in N1 and N2 nodal level. In 70.9% of cases, the SNs were located at lymphatic chains 6 and 7. The SN sensitivity for nodal staging was 91.6%, with 8.3% of false negative. In 4 patients who were initially staged as N0, the SNs were submitted to multisection analyses and immunohistochemistry, confirming the N0 stage, without micrometastases. In one case initially staged as negative for nodal metastases based on SN analyses, metastases in lymph nodes other than SN were found, resulting in a 20% skip metastases incidence. This surgery is a reproducible procedure with 100% detection rate of SN. Tumor size, GC location and obesity were factors that imposed some limitations regarding SN identification. Results from nodal multisection histology and immunohistochemistry analysis did not change initial nodal staging.

Optimized automated data analysis for the cytokinesis-block micronucleus assay using imaging flow cytometry for high throughput radiation biodosimetry.

The cytokinesis-block micronucleus (CBMN) assay is a well-established technique that can be employed in triage radiation biodosimetry to estimate whole body doses of radiation to potentially exposed individuals through quantitation of the frequency of micronuclei (MN) in binucleated lymphocyte cells (BNCs). The assay has been partially automated using traditional microscope-based methods and most recently has been modified for application on the ImageStream(X) (IS(X) ) imaging flow cytometer. This modification has allowed for a similar number of BNCs to be automatically scored as compared to traditional microscopy in a much shorter time period. However, the MN frequency measured was much lower than both manual and automated slide-based methods of performing the assay. This work describes the optimized analysis template which implements newly developed functions in the IDEAS(®) data analysis software for the IS(X) that enhances specificity for BNCs and increases the frequency of scored MN. A new dose response calibration curve is presented in which the average rate of MN per BNC is of similar magnitude to those presented in the literature using automated CBMN slide scoring methods. In addition, dose estimates were generated for nine irradiated, blinded samples and were found to be within ±0.5 Gy of the delivered dose. Results demonstrate that the improved identification accuracy for MN and BNCs in the IS(X) -based version of the CBMN assay will translate to increased accuracy when estimating unknown radiation doses received by exposed individuals following large-scale radiological or nuclear emergencies. © 2016 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of ISAC.

Helicobacter pylori infection is high in paediatric nonulcer dyspepsia but not associated with specific gastrointestinal symptoms.

AIM: The association between Helicobacter pylori infection and gastrointestinal symptoms is debatable in childhood. We examined the potential relationship between H. pylori infection and gastrointestinal symptoms in Brazilian children with nonulcer dyspepsia. METHODS: This prospective observational study analysed 240 Brazilian children and adolescents (68.7% girls) with chronic nonulcer dyspepsia, who underwent upper gastrointestinal endoscopy and biopsy. Their mean age was 9.8 years (range 4-17). Upper gastrointestinal symptoms, including abdominal pain, nausea, burning, early satiety, belching and weigh loss, were evaluated by a questionnaire and H. pylori infection was determined by histopathology of gastric biopsies. RESULTS: H. pylori infection was identified in 123/240 patients (52%). There was no significant association between the H. pylori infection and gastrointestinal symptoms and no relationship between the infection and abdominal pain or pain characteristics. However, nausea was significantly associated with the H. pylori infection, with an odds ratio of 1.76 and 95% confidence interval of 1.1-2.94 p < 0.03. Symptoms lasting longer than 12 months were significantly more frequent in children with pangastritis than in those with antral gastritis (p < 0.05). CONCLUSION: The prevalence of H. pylori infection was high in Brazilian children with nonulcer dyspepsia, but was not associated with specific signs and symptoms, except for nausea.

Sentinel node biopsy using blue dye and technetium99 in advanced gastric cancer: anatomical drainage and clinical application

Lymph node metastases are an independent prognosis factor in gastric carcinoma (GC) patients. Radical lymphadenectomy can improve survival but it can also increase surgical morbidity. As a principle, sentinel node (SN) navigation surgery can avoid unnecessary lymphadenectomy without compromising prognosis. In this pilot study, 24 patients with untreated GC were initially screened for SN navigation surgery, of which 12 were eligible. Five patients had T2 tumors, 5 had T3 tumors and 2 had T1 tumors. In 33% of cases, tumor diameter was greater than 5.0 cm. Three hundred and eighty-seven lymph nodes were excised with a median of 32.3 per patient. The SN navigation surgery was feasible in all patients, with a median of 4.5 SNs per patient. The detection success rate was 100%. All the SNs were located in N1 and N2 nodal level. In 70.9% of cases, the SNs were located at lymphatic chains 6 and 7. The SN sensitivity for nodal staging was 91.6%, with 8.3% of false negative. In 4 patients who were initially staged as N0, the SNs were submitted to multisection analyses and immunohistochemistry, confirming the N0 stage, without micrometastases. In one case initially staged as negative for nodal metastases based on SN analyses, metastases in lymph nodes other than SN were found, resulting in a 20% skip metastases incidence. This surgery is a reproducible procedure with 100% detection rate of SN. Tumor size, GC location and obesity were factors that imposed some limitations regarding SN identification. Results from nodal multisection histology and immunohistochemistry analysis did not change initial nodal staging.

Degradation and inactivation of adenovirus in water by photo-electro-oxidation.

The present study analyzed the efficiency of the photo-electro-oxidation process as a method for degradation and inactivation of adenovirus in water. The experimental design employed a solution prepared from sterile water containing 5.107 genomic copies/L (gc/L) of a standard strain of human adenovirus type 5 (HAdV-5) divided into two equal parts, one to serve as control and one treated by photo-electro-oxidation (PEO) for 3 hours and with a 5A current. Samples collected throughout the exposure process were analyzed by real-time polymerase chain reaction (qPCR) for viral genome identification and quantitation. Prior to gene extraction, a parallel DNAse treatment step was carried out to assess the integrity of viral particles. Integrated cell culture (ICC) analyses assessed the viability of infection in a cell culture. The tested process proved effective for viral degradation, with a 7 log10 reduction in viral load after 60 minutes of treatment. The DNAse-treated samples exhibited complete reduction of viral load after a 75 minute exposure to the process, and ICC analyses showed completely non-viable viral particles at 30 minutes of treatment.

Degradation and inactivation of adenovirus in water by photo-electro-oxidation / Degradação e inativação de adenovírus na água por fotoeletrooxidação

The present study analyzed the efficiency of the photo-electro-oxidation process as a method for degradation and inactivation of adenovirus in water. The experimental design employed a solution prepared from sterile water containing 5.107 genomic copies/L (gc/L) of a standard strain of human adenovirus type 5 (HAdV-5) divided into two equal parts, one to serve as control and one treated by photo-electro-oxidation (PEO) for 3 hours and with a 5A current. Samples collected throughout the exposure process were analyzed by real-time polymerase chain reaction (qPCR) for viral genome identification and quantitation. Prior to gene extraction, a parallel DNAse treatment step was carried out to assess the integrity of viral particles. Integrated cell culture (ICC) analyses assessed the viability of infection in a cell culture. The tested process proved effective for viral degradation, with a 7 log10 reduction in viral load after 60 minutes of treatment. The DNAse-treated samples exhibited complete reduction of viral load after a 75 minute exposure to the process, and ICC analyses showed completely non-viable viral particles at 30 minutes of treatment.

Resumo O presente estudo analisou a eficiência do processo de fotoeletrooxidação como metodologia para a degradação e inativação de adenovírus em água. A concepção experimental emprega uma solução preparada a partir de água estéril contendo 5,107 cópias genômicas/L (gc/L) de uma amostra padrão de adenovírus humano tipo 5 (HAdV-5), dividida em duas partes iguais, uma para servir como controle e outra tratada por fotoeletrooxidação (PEO) durante 3 horas e com uma corrente de 5A. As amostras recolhidas durante o processo de exposição foram analisadas por PCR quantitativo em tempo real (qPCR) para identificação e quantificação do genoma viral. Antes da extração de ácidos nucleicos, um passo de tratamento com DNAse paralelo foi realizado para avaliar a integridade das partículas virais. Um ensaio de qPCR integrado à cultura de células (ICC-qPCR) permitiu analisar a viabilidade de infecção em uma cultura de células. O processo mostrou-se eficaz testada para a degradação viral, com uma redução de 7 log10 da carga viral após 60 minutos de tratamento. As amostras tratadas com DNAse exibiram redução completa da carga viral após uma exposição de 75 minutos ao processo, e a análise de ICC-qPCR mostrou partículas virais completamente não-viáveis ​​em 30 minutos de tratamento.

Cytotoxicity assays to evaluate tannery effluents treated by photoelectrooxidation / Ensaios de citotoxicidade na avaliação de efluentes de curtume tratados por fotoeletrooxidação

The advanced oxidation process (AOP) is used to increase the treatment efficiency of effluents however, it is necessary to compare the toxicity of treated and untreated effluents to evaluate if the decontamination process does not cause any biological harm. Cultured cells have been previously used to assess the genotoxic and cytotoxic potential of various compounds. Hence, the aim of this work was to assess the applicability of cytotoxicity assays to evaluate the toxicity related to the AOP treatment. Samples of an industrial effluent were collected after their treatment by a conventional method. Cytotoxicity of standard and AOP treated effluents was assessed in CRIB and HEp-2 cell line using the MTT and neutral red assays. We observed decrease at cell viability in the both assays (50% MTT and 13% NRU) when cells were exposed to the AOP treatment in the highest concentration. Thus, cytotoxic assays in cultured cells can be explored as an useful method to evaluate toxicity as well as to optimize effluents treatment process.

Resumo O processo de oxidação avançada (POA) tem sido usado para aumentar a eficiência do tratamento de efluentes; no entanto, é necessário comparar a toxicidade de efluentes tratados e não tratados para avaliar se o processo de descontaminação não é capaz de causar algum risco biológico. Cultivos celulares têm sido utilizados para avaliar o potencial genotóxico e citotóxico de vários compostos. Assim, o objetivo deste trabalho foi avaliar a aplicabilidade de ensaios de citotoxicidade para avaliar a toxicidade relacionada ao tratamento com POA. As amostras de um efluente industrial foram recolhidas após o tratamento por um método convencional. A citotoxicidade dos efluentes padrão e tratado com POA foi avaliada nas linhagens celulares CRIB e HEp-2 usando os ensaios do MTT e do vermelho neutro. Observou-se diminuição da viabilidade celular em ambos os ensaios (50% MTT e 13% VN) quando as células foram expostas à concentração mais elevada do efluente tratado com POA. Assim, os ensaios de citotoxicidade em cultivos celulares podem ser explorados como um método útil para avaliar a toxicidade, bem como para otimizar os processos de tratamento de efluentes.

Concentrations of PM2.5-10 and PM2.5 and metallic elements around the Schmidt Stream area, in the Sinos River Basin, southern Brazil / Concentrações de MP2,5-10 e MP2,5 e elementos metálicos junto à área do Arroio Schmidt, na bacia do Rio dos Sinos, sul do Brasil

This research aimed to evaluate the air quality, by determining the concentrations of PM2.5-10, PM2.5 and the metallic elements Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn and Hg in the leaf part of ryegrass (Lolium multiflorum) in an area close to Schmidt Stream, at the lower section of Sinos River Basin (SRB), in a research campaign of six months, from October 2013 to March 2014. The particles collected in the PM sampling were analyzed by Scanning Electron Microscopy (SEM) combined with Energy Dispersive X-ray Spectrometry (EDS), in order to study their morphology and chemical composition. The mean concentration of PM2.5-10 was 9.1 µg m–3, with a range of 2.2 µg m–3 to 15.4 µg m–3 and the mean concentration of PM2.5was 4.7 µg m–3, with a range of 1.9 µg m–3 to 8.2 µg m–3. Concentrations of metallic elements, especially Pb, Cr and Zn, were classified as Class 4 (very high pollution levels), according to the classification proposed by Klumpp et al. (2004). Chemical and morphological analysis of PM revealed the presence of particles of biological origin, soot (Cr, Fe, Ni, Zn, Cd, Hg and Pb), salts (KCl) and soil resuspension (Al and Si). The integrated study methodology, employing environmental variables, such as PM and ryegrass, can be of help in the preparation of wide-ranging environmental diagnoses, in addition providing information needed to develop precautionary measures designed to minimize the effects of atmospheric pollution that takes into consideration the environment’s supportive capacity and environmental quality.

Resumo O objetivo desta pesquisa foi avaliar a qualidade do ar, por meio da determinação das concentrações do MP2,5-10, MP2,5 e dos elementos metálicos Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn e Hg na folha do azevém (Lolium multiflorum) em uma área próxima ao Arroio Schmidt, no trecho inferior da Bacia do Rio dos Sinos (BRS), em uma campanha de seis meses de pesquisa, de outubro de 2013 a março de 2014. As partículas coletadas na amostragem do MP foram analisadas por Microscopia Eletrônica de Varredura (MEV), combinada com Espectrometria de Energia Dispersiva de Raios X (EDS), a fim de estudar sua morfologia e composição química. As concentrações médias de MP2,5-10 e MP2,5 foram de 9,1 µg m–3 e 4,7 µg m–3, respectivamente. As concentrações médias dos elementos metálicos Pb, Cr e Zn na parte foliar do azevém foram de 13,58 mg kg–1, 5,26 mg kg–1 e 88,80 mg kg–1, respectivamente, caracterizando a área como Classe 4 (nível muito elevado de poluição), conforme classificação proposta por Klumpp et al. (2004). A caracterização química e morfológica das partículas coletadas revelou a presença de fuligem (Cr, Fe, Ni, Zn, Cd, Hg e Pb), materiais biológicos, cristais salinos (KCl) e partículas ressuspensas de poeira do solo (Al e Si). Estudos integrados empregando variáveis ambientais como o MP e o biomonitoramento com azevém podem auxiliar na elaboração de diagnósticos ambientais robustos, além de fornecer informações para o desenvolvimento de medidas de precaução que considerem a capacidade de suporte do meio ambiente e que visem à minimização dos efeitos prejudiciais da poluição atmosférica.

Water quality assessment of the Sinos River – RS, Brazil / Qualidade da água do Rio dos Sinos – RS, Brasil

Worldwide environmental pollution is increasing at the same rate as social and economic development. This growth, however, is disorganized and leads to increased degradation of water resources. Water, which was once considered inexhaustible, has become the focus of environmental concerns because it is essential for life and for many production processes. This article describes monitoring of the water quality at three points along the Sinos River (RS, Brazil), one in each of the upper, middle and lower stretches. The points were sampled in 2013 and again in 2014. The water samples were analyzed to determine the following physical and chemical parameters plus genotoxicity to fish: metals (Cr, Fe, Al), chemical oxygen demand, biochemical oxygen demand, chlorides, conductivity, total suspended solids, total phosphorous, total and fecal coliforms, pH, dissolved oxygen, turbidity, total Kjeldahl nitrogen nitrate and ammoniacal nitrogen. Genotoxicity was tested by exposing individuals of the species Astyanax jacuhiensis to water samples and then comparing them with a control group exposed to water from the public water supply. The results confirmed the presence of substances with genotoxic potential at the sample points located in the middle and lower stretches of the river. The results for samples from the upper stretch, at P1, did not exhibit differences in relation to the control group. The physical and chemical analyses did not detect reductions in water quality in the lower stretch, as had been expected in view of the large volumes of domestic and industrial effluents discharged into this part of the river.

Resumo A poluição ambiental aumenta mundialmente, no mesmo ritmo que o desenvolvimento social e econômico. Este crescimento é, muitas vezes, desorganizado e proporciona um aumento da degradação dos recursos hídricos. Água, que já foi considerado inesgotável, tornou-se o foco das preocupações ambientais, pois é essencial para a vida e para muitos processos de produção. Este artigo aborda o monitoramento da qualidade da água, em três pontos do Rio dos Sinos, compreendendo o trecho superior, médio e inferior. Os pontos foram monitorados entre 2013 e 2014. As amostras de água foram analisadas para determinar os parâmetros físico-químicos complementarmente a genotoxicidade em peixes: metais (Cr, Fe, Al), demanda química de oxigenio, demanda bioquímica de oxigênio, cloretos, condutividade, sólidos suspensos totais, fósforo total, coliformes totais e fecais, pH, oxigênio dissolvido, turbidez, nitrogênio total Kjeldahl, nitrato e nitrogênio amoniacal. A avaliação de genotoxicidade foi conduzida pela exposição à água de diferentes pontos do rio de indivíduos da espécie Astyanax jacuhiensis. O grupo controle foi exposto à água de abastecimento público. Os resultados confirmam a presença de substâncias com potencial genotóxico nos pontos localizados nos trechos médio e inferior. No trecho superior, ponto 1, não houve diferenças em relação ao grupo controle. A diminuição da qualidade da água do trecho inferior quando comparada com o superior também foi confirmada através das análises físico-químicas, as quais indicaram uma redução na qualidade da água nos trechos inferiores, como o esperado devido ao grande escoamento de esgotos domésticos e industriais neste trecho do rio.

Degradation and inactivation of adenovirus in water by photo-electro-oxidation

The present study analyzed the efficiency of the photo-electro-oxidation process as a method for degradation and inactivation of adenovirus in water. The experimental design employed a solution prepared from sterile water containing 5.107 genomic copies/L (gc/L) of a standard strain of human adenovirus type 5 (HAdV-5) divided into two equal parts, one to serve as control and one treated by photo-electro-oxidation (PEO) for 3 hours and with a 5A current. Samples collected throughout the exposure process were analyzed by real-time polymerase chain reaction (qPCR) for viral genome identification and quantitation. Prior to gene extraction, a parallel DNAse treatment step was carried out to assess the integrity of viral particles. Integrated cell culture (ICC) analyses assessed the viability of infection in a cell culture. The tested process proved effective for viral degradation, with a 7 log10 reduction in viral load after 60 minutes of treatment. The DNAse-treated samples exhibited complete reduction of viral load after a 75 minute exposure to the process, and ICC analyses showed completely non-viable viral particles at 30 minutes of treatment.

Resumo O presente estudo analisou a eficiência do processo de fotoeletrooxidação como metodologia para a degradação e inativação de adenovírus em água. A concepção experimental emprega uma solução preparada a partir de água estéril contendo 5,107 cópias genômicas/L (gc/L) de uma amostra padrão de adenovírus humano tipo 5 (HAdV-5), dividida em duas partes iguais, uma para servir como controle e outra tratada por fotoeletrooxidação (PEO) durante 3 horas e com uma corrente de 5A. As amostras recolhidas durante o processo de exposição foram analisadas por PCR quantitativo em tempo real (qPCR) para identificação e quantificação do genoma viral. Antes da extração de ácidos nucleicos, um passo de tratamento com DNAse paralelo foi realizado para avaliar a integridade das partículas virais. Um ensaio de qPCR integrado à cultura de células (ICC-qPCR) permitiu analisar a viabilidade de infecção em uma cultura de células. O processo mostrou-se eficaz testada para a degradação viral, com uma redução de 7 log10 da carga viral após 60 minutos de tratamento. As amostras tratadas com DNAse exibiram redução completa da carga viral após uma exposição de 75 minutos ao processo, e a análise de ICC-qPCR mostrou partículas virais completamente não-viáveis em 30 minutos de tratamento.

Concentrations of PM2.5-10 and PM2.5 and metallic elements around the Schmidt Stream area, in the Sinos River Basin, southern Brazil

This research aimed to evaluate the air quality, by determining the concentrations of PM2.5-10, PM2.5 and the metallic elements Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn and Hg in the leaf part of ryegrass (Lolium multiflorum) in an area close to Schmidt Stream, at the lower section of Sinos River Basin (SRB), in a research campaign of six months, from October 2013 to March 2014. The particles collected in the PM sampling were analyzed by Scanning Electron Microscopy (SEM) combined with Energy Dispersive X-ray Spectrometry (EDS), in order to study their morphology and chemical composition. The mean concentration of PM2.5-10 was 9.1 µg m3, with a range of 2.2 µg m3 to 15.4 µg m3 and the mean concentration of PM2.5was 4.7 µg m3, with a range of 1.9 µg m3 to 8.2 µg m3. Concentrations of metallic elements, especially Pb, Cr and Zn, were classified as Class 4 (very high pollution levels), according to the classification proposed by Klumpp et al. (2004). Chemical and morphological analysis of PM revealed the presence of particles of biological origin, soot (Cr, Fe, Ni, Zn, Cd, Hg and Pb), salts (KCl) and soil resuspension (Al and Si). The integrated study methodology, employing environmental variables, such as PM and ryegrass, can be of help in the preparation of wide-ranging environmental diagnoses, in addition providing information needed to develop precautionary measures designed to minimize the effects of atmospheric pollution that takes into consideration the environments supportive capacity and environmental quality.

Resumo O objetivo desta pesquisa foi avaliar a qualidade do ar, por meio da determinação das concentrações do MP2,5-10, MP2,5 e dos elementos metálicos Al, Ba, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn e Hg na folha do azevém (Lolium multiflorum) em uma área próxima ao Arroio Schmidt, no trecho inferior da Bacia do Rio dos Sinos (BRS), em uma campanha de seis meses de pesquisa, de outubro de 2013 a março de 2014. As partículas coletadas na amostragem do MP foram analisadas por Microscopia Eletrônica de Varredura (MEV), combinada com Espectrometria de Energia Dispersiva de Raios X (EDS), a fim de estudar sua morfologia e composição química. As concentrações médias de MP2,5-10 e MP2,5 foram de 9,1 µg m3 e 4,7 µg m3, respectivamente. As concentrações médias dos elementos metálicos Pb, Cr e Zn na parte foliar do azevém foram de 13,58 mg kg1, 5,26 mg kg1 e 88,80 mg kg1, respectivamente, caracterizando a área como Classe 4 (nível muito elevado de poluição), conforme classificação proposta por Klumpp et al. (2004). A caracterização química e morfológica das partículas coletadas revelou a presença de fuligem (Cr, Fe, Ni, Zn, Cd, Hg e Pb), materiais biológicos, cristais salinos (KCl) e partículas ressuspensas de poeira do solo (Al e Si). Estudos integrados empregando variáveis ambientais como o MP e o biomonitoramento com azevém podem auxiliar na elaboração de diagnósticos ambientais robustos, além de fornecer informações para o desenvolvimento de medidas de precaução que considerem a capacidade de suporte do meio ambiente e que visem à minimização dos efeitos prejudiciais da poluição atmosférica.

Cytotoxicity assays to evaluate tannery effluents treated by photoelectrooxidation

The advanced oxidation process (AOP) is used to increase the treatment efficiency of effluents however, it is necessary to compare the toxicity of treated and untreated effluents to evaluate if the decontamination process does not cause any biological harm. Cultured cells have been previously used to assess the genotoxic and cytotoxic potential of various compounds. Hence, the aim of this work was to assess the applicability of cytotoxicity assays to evaluate the toxicity related to the AOP treatment. Samples of an industrial effluent were collected after their treatment by a conventional method. Cytotoxicity of standard and AOP treated effluents was assessed in CRIB and HEp-2 cell line using the MTT and neutral red assays. We observed decrease at cell viability in the both assays (50% MTT and 13% NRU) when cells were exposed to the AOP treatment in the highest concentration. Thus, cytotoxic assays in cultured cells can be explored as an useful method to evaluate toxicity as well as to optimize effluents treatment process.

Resumo O processo de oxidação avançada (POA) tem sido usado para aumentar a eficiência do tratamento de efluentes; no entanto, é necessário comparar a toxicidade de efluentes tratados e não tratados para avaliar se o processo de descontaminação não é capaz de causar algum risco biológico. Cultivos celulares têm sido utilizados para avaliar o potencial genotóxico e citotóxico de vários compostos. Assim, o objetivo deste trabalho foi avaliar a aplicabilidade de ensaios de citotoxicidade para avaliar a toxicidade relacionada ao tratamento com POA. As amostras de um efluente industrial foram recolhidas após o tratamento por um método convencional. A citotoxicidade dos efluentes padrão e tratado com POA foi avaliada nas linhagens celulares CRIB e HEp-2 usando os ensaios do MTT e do vermelho neutro. Observou-se diminuição da viabilidade celular em ambos os ensaios (50% MTT e 13% VN) quando as células foram expostas à concentração mais elevada do efluente tratado com POA. Assim, os ensaios de citotoxicidade em cultivos celulares podem ser explorados como um método útil para avaliar a toxicidade, bem como para otimizar os processos de tratamento de efluentes.