Exposure levels for carbon monoxide in nuclear submarine atmosphere.

This work proposes exposure limits for carbon monoxide in the nuclear submarine environment. Linear and non-linear forms of the Coburn-Foster-Kane equation were used to evaluate carbon monoxide exposure for an environment with low oxygen content, different exposure times and crew physical activity levels. We evaluated the 90-day Continuous Exposure Guidance Level, 24-h and 1-h Emergency Exposure Guidance Levels and 10-day and 24-h Submarine Escape Action Levels. The results showed that the concentration of carbon monoxide in the environment must not exceed 9 ppm for the 90-day Continuous Exposure Guidance Level, 35 ppm for the 24-h Emergency Exposure Guidance Level, 90 ppm 1-h Emergency Exposure Guidance Level, 60 ppm for the 10-day Submarine Escape Action Level and 80 ppm for the 24-h Submarine Escape Action Level. Comparing these values with those established by the National Research Council for the United States Navy, the limits proposed by this work are verified to be lower, which may indicate a risk to the health of the crew. They also show the impact of the crew's level of physical activity on the formation of carboxyhemoglobin.

Brazil's Regulatory Context for Using New Approach Methodologies (NAMs) on the Registration of Products.

New Approach Methodologies (NAMs) are any non-animal-based approaches that can provide information in the context of chemical hazard and safety assessment. The goal is to develop information with equivalent or better scientific quality and relevance than that provided by traditional animal models. Starting with ethical issues, these approaches are gaining regulatory relevance in different global agencies. Since 2008, with the enactment of the Arouca Law-the first Brazilian legislation dedicated to laboratory animals, NAMs are gathering pace in Brazil's regulations. Specific regulations from different sectors include the acceptance of these new methods. However, some regulation is controversial about what is needed to address specific toxicological endpoints. The resulting regulatory uncertainty induces companies to keep on adopting the traditional methods, slowing NAM's development in the country. This work brings a perspective on the regulatory acceptance of NAMs in Brazilian Legislation for the registration of pharmaceuticals, medical devices, food/supplements, and agrochemical products. This text discusses the main issues of NAM adoption for each specific regulation. Therefore, legal acceptance of NAMs results in Brazil is still a process in progress. A collective effort including regulators, industry, contract research organizations (CROs), and the academic environment is needed to build regulatory confidence in the use of NAMs.

Safety assessment of the process Resinas del Ecuador, based on the Starlinger iV+ technology, used to recycle post-consumer PET into food contact materials.

The EFSA Panel on Food Contact Materials, Enzymes and Processing Aids (CEP) assessed the safety of the recycling process Resinas del Ecuador (EU register number RECYC230), which uses the Starlinger iV+ technology. The input is hot caustic washed and dried poly(ethylene terephthalate) (PET) flakes mainly originating from collected post-consumer PET containers, with no more than 5% PET from non-food consumer applications. The flakes are dried and crystallised in a first reactor, then extruded into pellets. These pellets are crystallised, preheated and treated in a solid-state polycondensation (SSP) reactor. Having examined the challenge test provided, the Panel concluded that the drying and crystallisation (step 2), extrusion and crystallisation (step 3) and SSP (step 4) are critical in determining the decontamination efficiency of the process. The operating parameters to control the performance of these critical steps are temperature, air flow and residence time for the drying and crystallisation step, and temperature, pressure and residence time for the extrusion and crystallisation step as well as the SSP step. It was demonstrated that this recycling process is able to ensure that the level of migration of potential unknown contaminants into food is below the conservatively modelled migration of 0.1 µg/kg food. Therefore, the Panel concluded that the recycled PET obtained from this process is not of safety concern when used at up to 100% for the manufacture of materials and articles for contact with all types of foodstuffs for long-term storage at room temperature, with or without hotfill. The final articles made of this recycled PET are not intended to be used in microwave and conventional ovens and such uses are not covered by this evaluation.

Safety assessment for temporary hospitals during the COVID-19 pandemic: A simulation approach.

Amid the devastating effects caused by the pandemic of the new Coronavirus (COVID-19), health leaders around the world are adding efforts to search efficient and effective responses in the fight against the disease. Conventional health centers, such as hospitals and emergency departments have been registering an increase in demand and atypical patterns due to the high transmissibility of the virus. In this context, the adoption of Temporary Hospitals (THs) is effective in trying to relieve conventional hospitals and direct efforts in the treatment of suspected and positive patients for COVID-19. However, some requirements should be considered regarding the processes performed by THs to maintain the health and safety of patients and staff. Based on the literature, we evaluated aspects related to patient safety in THs, especially linked to biosafety of medical facilities, and patient transport and visit. We highlight the analysis of flows and layouts, hospital cleaning and patient care. We described two case studies to demonstrate the proposed approach. As result, simulation tests improved safety metrics, such as waiting time for procedures, movement intensity in each area, length of stay and TH capacity. We conclude that the approach allows us to provide better THs that prevent cross-contamination, provide suitable care, and meet the demand.

Safety Evaluation of Lactobacillus delbrueckii subsp. lactis CIDCA 133: a Health-Promoting Bacteria.

Lactobacillus delbrueckii subsp. lactis CIDCA is a new potential probiotic strain whose molecular basis attributed to the host's benefit has been reported. This study investigated the safety aspects of Lactobacillus delbrueckii subsp. lactis CIDCA 133 based on whole-genome sequence and phenotypic analysis to avoid future questions about the harmful effects of this strain consumption. Genomic analysis showed that L. delbrueckii subsp. lactis CIDCA 133 harbors virulence, harmful metabolites, and antimicrobial resistance-associated genes. However, none of these genetic elements is flanked or located within prophage regions and plasmid sequence. At a phenotypic level, it was observed L. delbrueckii subsp. lactis CIDCA 133 antimicrobial resistance to aminoglycosides streptomycin and gentamicin antibiotics, but no hemolytic and mucin degradation activity was exhibited by strain. Furthermore, no adverse effects were observed regarding mice clinical and histopathological analysis after the strain consumption (5 × 107 CFU/mL). Overall, these findings reveal the safety of Lactobacillus delbrueckii subsp. lactis CIDCA 133 for consumption and future probiotic applications.

A review of alternative methods to the use of animals in safety evaluation of cosmetics

ABSTRACT Alternative methods to the use of animals in research have been a global trend, mainly after the publication of the 3R's principle (Replacement, Reduction, and Refinement), proposed by Russel and Burch. In the cosmetic sector, safety and efficacy assessments using animals have generated controversial debates. For this reason, in vitro research techniques are widely used to assess acute toxicity; corrosivity and irritation; skin sensitization; dermal and percutaneous absorption; repeated dose toxicity; reproductive toxicity; mutagenicity and genotoxicity; carcinogenicity; toxicokinetic studies; photo-induced toxicity; and human data. Although there are many methodologies described, validated, and widely used in the cosmetic area, the evaluation of the safety of cosmetic ingredients and products is still an expanding field. It needs global collaboration among regulatory agencies, universities, and industry, to meet several unmet needs in the fields of sensitization, carcinogenicity, systemic action, among other issues involving safety of users of cosmetic products. This review article will cover the currently most relevant in vitro models regarding irritation, corrosion, sensitization, mutagenicity, genotoxicity, and phototoxicity, to help to choose the most appropriate test for evaluating the safety and toxicity of cosmetic ingredients and products.

Application of remote sensing in environmental impact assessment: a case study of dam rupture in Brumadinho, Minas Gerais, Brazil.

The collapse of mining tailing dams in Brumadinho, Minas Gerais, Brazil, that occurred in 2019 was one of the worst environmental and social disasters witnessed in the country. In this sense, monitoring any impacted areas both before and after the disaster is crucial to understand the actual scenario and problems of disaster management and environmental impact assessment. In order to find answers to that problem, the aim of this study was to identify and analyze the spatiality of the impacted area by rupture of the tailing dam of the Córrego do Feijão mine in Brumadinho, Minas Gerais, by using orbital remote sensing. Land use and land occupation, phytoplankton chlorophyll-a, water turbidity, total suspended solids on water, and carbon sequestration efficiency by vegetation (CO2Flux) were estimated by orbital imagery from the Landsat-8/OLI and MSI/Sentinel-2 sensors in order to assess the environmental impacts generated by the disaster. Data were extracted from spectral models in which the variables that best demonstrated the land use variation over the years were sought. Mean comparison by t-test was performed to compare the time series analyzed, that is, before and after the disaster. Through the analysis of water quality, it was observed that the environmental impact was calamitous to natural resources, especially water from Córrego do Feijão.

Chemical safety assessment of transformation products of landfill leachate formed during the Fenton process.

Organic chemicals identified in raw landfill leachate (LL) and their transformation products (TPs), formed during Fenton treatment, were analyzed for chemical safety following REACH guidelines. The raw LL was located in the metropolitan region of Campina Grande, in northeast Brazil. We elucidated 197 unique chemical structures, including 154 compounds that were present in raw LL and 82 compounds that were detected in the treated LL, totaling 39 persistent compounds and 43 TPs. In silico models were developed to identify and prioritize the potential level of hazard/risk these compounds pose to the environment and society. The models revealed that the Fenton process improved the biodegradability of TPs. Still, a slight increase in ecotoxicological effects was observed among the compounds in treated LL compared with those present in raw LL. No differences were observed for aryl hydrocarbon receptor (AhR) and antioxidant response element (ARE) mutagenicity. Similar behavior among both raw and treated LL samples was observed for biodegradability; Tetrahymena pyriformis, Daphnia magna, Pimephales promelas and ARE, AhR, and Ames mutagenicity. Overall, our results suggest that raw and treated LL samples have similar activity profiles for all endpoints other than biodegradability.

Single-Arm, Multicenter Phase I/II Clinical Trial for the Treatment of Envenomings by Massive Africanized Honey Bee Stings Using the Unique Apilic Antivenom.

We evaluated the safety, optimal dose, and preliminary effectiveness of a new-approach Africanized honeybee (Apis mellifera) Antivenom (AAV) in a phase I/II, multicenter, non-randomized, single-arm clinical trial involving 20 participants with multiple stings. Participants received 2 to 10 vials of AAV depending on the number of stings they suffered, or a predefined adjuvant, symptomatic, and complementary treatment. The primary safety endpoint was the occurrence of early adverse reactions within the first 24 h of treatment. Preliminary efficacy based on clinical evolution, including laboratory findings, was assessed at baseline and at various time points over the four following weeks. ELISA assays and mass spectrometry were used to estimate venom pharmacokinetics before, during, and after treatment. Twenty adult participants, i.e., 13 (65%) men and 7 (35%) women, with a median age of 44 years and a mean body surface area of 1.92 m2 (median = 1.93 m2) were recruited. The number of stings ranged from 7 to > 2,000, with a median of 52.5. Symptoms of envenoming were classified as mild, moderate, or severe in 80% (16), 15% (3), and 5% (1) of patients, respectively; patients with mild, moderate, or severe envenoming received 2, 6, and 10 vials of AAV as per the protocol. None of the patients had late reactions (serum sickness) within 30 d of treatment. There was no discontinuation of the protocol due to adverse events, and there were no serious adverse events. One patient had a moderate adverse event, transient itchy skin, and erythroderma. All participants completed the intravenous antivenom infusion within 2 h, and there was no loss to follow-up after discharge. ELISA assays showed venom (melittin and PLA2) concentrations varying between 0.25 and 1.479 ng/mL prior to treatment. Venom levels decreased in all patients during the hospitalization period. Surprisingly, in nine cases (45%), despite clinical recovery and the absence of symptoms, venom levels increased again during outpatient care 10 d after discharge. Mass spectrometry showed melittin in eight participants, 30 d after treatment. Considering the promising safety results for this investigational product in the treatment of massive Africanized honeybee attack, and its efficacy, reflected in the clinical improvements and corresponding immediate decrease in blood venom levels, the AAV has shown to be safe for human use. Clinical Trial Registration: UTN: U1111-1160-7011, identifier [RBR-3fthf8].

Single-arm, multicenter phase I/II clinical trial for the treatment of envenomings by massive africanized honey bee stings using the unique apilic antivenom

We evaluated the safety, optimal dose, and preliminary effectiveness of a new-approach Africanized honeybee (Apis mellifera) Antivenom (AAV) in a phase I/II, multicenter, non-randomized, single-arm clinical trial involving 20 participants with multiple stings. Participants received 2 to 10 vials of AAV depending on the number of stings they suffered, or a predefined adjuvant, symptomatic, and complementary treatment. The primary safety endpoint was the occurrence of early adverse reactions within the first 24 h of treatment. Preliminary efficacy based on clinical evolution, including laboratory findings, was assessed at baseline and at various time points over the four following weeks. ELISA assays and mass spectrometry were used to estimate venom pharmacokinetics before, during, and after treatment. Twenty adult participants, i.e., 13 (65%) men and 7 (35%) women, with a median age of 44 years and a mean body surface area of 1.92 m2 (median = 1.93 m2) were recruited. The number of stings ranged from 7 to > 2,000, with a median of 52.5. Symptoms of envenoming were classified as mild, moderate, or severe in 80% (16), 15% (3), and 5% (1) of patients, respectively; patients with mild, moderate, or severe envenoming received 2, 6, and 10 vials of AAV as per the protocol. None of the patients had late reactions (serum sickness) within 30 d of treatment. There was no discontinuation of the protocol due to adverse events, and there were no serious adverse events. One patient had a moderate adverse event, transient itchy skin, and erythroderma. All participants completed the intravenous antivenom infusion within 2 h, and there was no loss to follow-up after discharge. ELISA assays showed venom (melittin and PLA2) concentrations varying between 0.25 and 1.479 ng/mL prior to treatment. Venom levels decreased in all patients during the hospitalization period. Surprisingly, in nine cases (45%), despite clinical recovery and the absence of symptoms, venom levels increased again during outpatient care 10 d after discharge. Mass spectrometry showed melittin in eight participants, 30 d after treatment. Considering the promising safety results for this investigational product in the treatment of massive Africanized honeybee attack, and its efficacy, reflected in the clinical improvements and corresponding immediate decrease in blood venom levels, the AAV has shown to be safe for human use. Clinical Trial Registration: UTN: U1111-1160-7011, identifier [RBR-3fthf8].

Risk assessment of the antifungal and insecticidal peptide Jaburetox and its parental protein the Jack bean (Canavalia ensiformis) urease.

Jaburetox (JBTX) is an insecticidal and antifungal peptide derived from jack bean (Canavalia ensiformis) urease that has been considered a candidate for developing genetically modified crops. This study aimed to perform the risk assessment of the peptide JBTX following the general recommendations of the two-tiered, weight-of-evidence approach proposed by International Life Sciences Institute. The urease of C. ensiformis (JBU) and its isoform JBURE IIb (the JBTX parental protein) were assessed. The history of safe use revealed no hazard reports for the studied proteins. The available information shows that JBTX possesses selective activity against insects and fungi. JBTX and JBU primary amino acids sequences showed no relevant similarity to toxic, antinutritional or allergenic proteins. Additionally, JBTX and JBU were susceptible to in vitro digestibility, and JBU was also susceptible to heat treatment. The results did not identify potential risks of adverse effects and reactions associated to JBTX. However, further allergen (e.g. serum IgE binding test) and toxicity (e.g. rodent toxicity tests) experimentation can be done to gather additional safety information on JBTX, and to meet regulatory inquiries for commercial approval of transgenic cultivars expressing this peptide.

Transgene-product expression levels in genetically engineered breeding stacks are equivalent to those of the single events.

Transgene product expression levels are measured in genetically engineered (GE) crops containing single transformation events and the measured expression levels are then utilized in food, feed, and environmental safety assessments as part of the requirements for de-regulation of the event. Many countries also require measurement of expression levels and safety assessments for GE breeding stacks, even though the breeding stacks are composed of single events that have been previously assessed. Transgene product expression levels were measured in tissues of maize, soybean, and cotton breeding stacks and each of their component single events. Expression levels in the breeding stacks were plotted against expression levels in the single events to quantify the ability of the single events to predict transgene product expression levels in the breeding stacks. These results indicate that transgene product expression levels in single events are a reliable indicator of expression levels in breeding stacks. Based on these results it is concluded that safety assessments for breeding stacks can be conducted using transgene product expression levels from single events.

Elemental impurities in lipsticks: Results from a survey of the Portuguese and Brazilian markets.

For safety reasons, European regulations prohibit the use of a long list of metal(loid)s as ingredients of cosmetic products. However, their presence as impurities in finished products is virtually unavoidable, even under GMP conditions. This study aimed at determining the elemental profile of lipsticks available in the Portuguese and Brazilian markets. A total of 96 lipsticks were purchased in Brazil (n = 53; 9 brands) and Portugal (n = 43; 7 brands) and the content of 44 elements was determined. Results ranged from <1 µg/g to several tens of µg/g (e.g., Sn, Mn, Zn). Significant differences were found between Portuguese and Brazilian products for several elements, particularly for Pb. For the elements of major toxicological concern (Pb, Cd, As, Sb, Hg), mean values were always below the current limits set by the German competent authority. However, a significant percentage of exceedances were observed for Pb (24%) and Cd (21%). A safety assessment was carried out for the toxicologically relevant elements. Results showed that, except for Pb, the systemic exposure resulting from lipstick use represents less than 0.2% (ca. 3% for Pb) of the respective permitted daily exposure even in the worst-case scenario (i.e., ingestion of the total amount of product applied).

Natural Variability of Allergen Levels in Conventional Soybeans: Assessing Variation across North and South America from Five Production Years.

Soybean (Glycine max L. Merrill) is one of eight major allergenic foods with endogenous proteins identified as allergens. To better understand the natural variability of five soybean allergens (Gly m 4, Gly m 5, Gly m 6, Gly m Bd 28k, and Gly m Bd 30k), validated enzyme-linked immunosorbent assays (ELISAs) were developed. These ELISAs measured allergens in 604 soybean samples collected from locations in North and South America over five growing seasons (2009-2013/2014) and including 37 conventional varieties. Levels of these five allergens varied 5-19-fold. Multivariate statistical analyses and pairwise comparisons show that environmental factors have a larger effect on allergen levels than genetic factors. Therefore, from year to year, consumers are exposed to highly variable levels of allergens in soy-based foods, bringing into question whether quantitative comparison of endogenous allergen levels of new genetically modified soybean adds meaningful information to their overall safety risk assessment.

Safety assessment of a traditionally used extract from leaves of Boldoa purpurascens.

ETHNOPHARMACOLOGICAL RELEVANCE: Boldoa purpurascens Cav. (Nyctaginaceae) is a plant species used in traditional medicine in Cuba as a diuretic. AIM OF THE STUDY: The aim of the present investigation was to evaluate the safety profile of a hydroalcoholic extract from leaves of Boldoa purpurascens. MATERIALS AND METHODS: First, an experimental study to assess the oral acute toxicity at a dose of 2000mg/kg body weight of the extract was carried out. Potential genotoxicity of the extract was evaluated using the Ames test and the micronucleus induction assay in mouse bone marrow. In the Ames test a concentration range of 50, 100, 150, 300 and 500µg/plate was tested. In the micronucleus induction assay, doses of 500, 1000 and 2000mg/kg of body weight were tested. For completeness, since the extract contains saponins, the evaluation of the hemolytic activity, ocular and skin irritation were included. RESULTS: No signs or symptoms of toxicity were observed in the oral acute toxicity test (body weight at baseline, seven days and end of the experiment of 236.41±20.07, 256.81±30.44 and 240.02±26.16 respectively for the treated group). The hydroalcoholic extract from the leaves was not mutagenic in the Ames test, and no genotoxicity was observed in the micronucleus assay. A hemolysis test at concentration of 1mg/mL confirmed hemolytic activity, which is not a safety concern since saponins are not absorbed after oral administration. In order to evaluate the percentage of protein denaturation, the ocular irritability index was calculated. The extract was found to be irritating. Finally, skin irritability was evaluated and the irritation index was equal to zero. CONCLUSIONS: Based on the toxicological evaluation of a traditionally used hydroalcoholic extract from the leaves of Boldoa purpurascens we can confirm the safety of its oral use.

Desenvolvimento de epiderme humana reconstruída (RHE) como plataforma de testes in vitro para irritação, sensibilização, dermatite atópica e fotoimunossupressão / Development of a Reconstructed Human Epidermis (RHE) as platform to in vitro assays: irritation, sensitization, atopic dermatitis and photoimmunosuppression

O desenvolvimento de novos modelos de pele e novas metodologias in vitro segue uma tendência mundial na busca pela redução ou substituição de testes em animais. Nesse contexto, kits de epiderme humana reconstruída (RHE) apresentam-se como uma plataforma promissoras para essa proposta e, alguns modelos encontram-se validados para ensaios de irritação e corrosão cutânea in vitro. Entretanto, em países como o Brasil, enfrentam-se questões alfandegárias e perda do material por perecibilidade, dificultando e até impedindo, a importação desses kits para utilização por parte das indústrias e laboratórios nacionais. Em contrapartida, o desenvolvimento de um modelo de RHE apresenta-se como um avanço tecnológico e ganho de autonomia para esses países. Assim, no capítulo 1 explorou-se o desenvolvimento de um modelo nacional de RHE (USP-RHE) que atendesse às exigências internacionais descritas no guia OECD 439. O modelo desenvolvido apresentou uma epiderme bem diferenciada e atendeu aos parâmetros de qualidade (histologia, viabilidade e função barreira) bem como da funcionalidade, a qual é expressa na capacidade de distinção entre irritantes e não irritantes, apresentando 85,7% de especificidade, 100% sensibilidade e 92,3% de acurácia quando comparada com a classificação in vivo obtida pelo ensaio do linfonodo local (LLNA). No capítulo 2, células monocíticas THP-1 em monocamada foram capazes de distinguir entre agentes sensibilizantes e não sensibilizantes por meio da expressão de CD86, CD54 e liberação de IL-8. Após a obtenção de RHE e THP-1 funcionais, um cross-talking foi estabelecido gerando uma RHE imunocompetente. A RHEI distinguiu satisfatoriamente entre agentes sensibilizantes e não sensibilizantes por meio da expressão de CD86 e CD54 na membrana das células THP-1. A liberação de IL-8 também foi avaliada na RHEI, mas, não demonstrou ser um bom indicador para a avaliação de sensibilização, ao contrário de IL-1α, que distinguiu satisfatoriamente agentes sensibilizantes de não-sensibilizantes, mas não foi capaz de hierarquizá-los. No capítulo 3, avaliou-se o papel de interleucinas do tipo Th2 e da depleção de colesterol na membrana plasmática no desenvolvimento de características morfológicas e moleculares da dermatite atópica (DA) in vitro em um modelo de RHE. Os resultados demonstram que o uso de IL-4, IL-13 e IL-25 em combinação com a depleção de colesterol na membrana plasmática mimetiza in vitro, as principais características da DA. No capítulo 4, buscou-se avaliar os efeitos imunossupressores da radiação ultravioleta na RHEI. Os ensaios foram realizados em diferentes períodos de exposição, entretanto, não foi possível observar tais efeitos. Os resultados justificam-se pela ausência da liberação de IL-10 pelo RHE imunocompetente, por exemplo, e demonstram uma limitação do RHE imunocompetente para avaliações de inativação da reposta imune. Neste trabalho, concluímos que foi possível obter uma RHE competitiva, similar aos modelos internacionais validados e que pode ser utilizada como plataforma para ensaios de irritação e sensibilização cutânea, além de ser uma plataforma para estudos da dermatite atópica. No modelo é possível estudar a ativação do sistema imune, o que o torna promissor como uma plataforma para avaliação de resposta imunológica in vitro. Conclui-se, portanto, que os objetivos foram amplamente atendidos além de oferecermos um protocolo de livre acesso para reprodução por outros laboratórios e um modelo para validação futura

The development of new in vitro skin models and new methodologies follows a global trend in search for reductions or replacement of animal testing. In this context, Reconstructed Human Epidermis kits (RHE) are presented as a promising platform in the search for alternative methods to animal use, and some models are validated for skin irritation and corrosion in vitro tests. However, in countries such as Brazil, who face customs issues and loss of material due to perishability, making it challenging and even compromising the importation of these kits for use by industries and laboratories. In contrast, the development of an RHE model is presented as a technological breakthrough and gain of autonomy for these countries. Thus, in Chapter 1 we explored the development of a national model of RHE (USP-RHE) that meet international requirements described in OECD TG 439. The developed model presented a well-differentiated epidermis and met the quality parameters, for instance, histology, viability, and barrier function as well as the functionality expressed in the capacity of screening between irritants and nonirritants, with 85.7 % of specificity, 100 % of sensitivity and 91.7% of accuracy in comparision to in vivo UN GHS classification from Local limph node assay (LLNA). In chapter 2, monocytic THP-1 cell line, as monolayers, were able to distinguish between sensitizers and non-sensitizers by expression of CD86, CD54, and IL-8 release. In this model, functional RHE and THP-1 were used in a cross-talking, and thus an immunocompetent RHE (RHEI) was generated. The RHEI has distinguished satisfactorily between sensitizers and non-sensitizers through CD86 and CD54 expression that was larger and more sensitive in this model. The release of IL-8 was also evaluated in RHEI, however, did not demonstrate to be a good parameter for this evaluation, unlike IL-1α, which satisfactorily distinguished sensitizers from non-sensitizers, but was not able to hierarchize them. In chapter 3, we evaluated the role of Th2-related cytokines and plasma membrane cholesterol depletion (CD) in the development of atopic dermatitis (AD) morphological and molecular characteristics in an in vitro model of RHE. The results showed that combination of IL-4, IL-13 and IL-25 in combination with CD can reproduce the major features of AD in vitro. In Chapter 4, we sought to evaluate the ultraviolet radiation-induced immunosuppressive effects in RHE. The tests were performed at different times. However, it was not possible to observe such effects. The results are justified by the absence of IL-10 release by RHEI, for example, and show a limitation of RHEI for rating inactivation of the immune response. In this work, we conclude that it was possible to obtain a competitive RHE similar to the validated international models that can be used as a platform for irritation and skin sensitization tests, besides being a platform for the study of atopic dermatitis. Using this model is possible to explore the activation of immune system, which makes it promising as a platform for the evaluation of immune response in vitro. We conclude, therefore, that the objectives have been met as well as it is offering an open source protocol for breeding by other laboratories, thus offering the RHE model developed here for future validation tests

Model-based tolerance intervals derived from cumulative historical composition data: application for substantial equivalence assessment of a genetically modified crop.

Compositional analysis is a requisite component of the substantial equivalence framework utilized to assess genetically modified (GM) crop safety. Statistical differences in composition data between GM and non-GM crops require a context in which to determine biological relevance. This context is provided by surveying the natural variation of key nutrient and antinutrient levels within the crop population with a history of safe use. Data accumulated from various genotypes with a history of safe use cultivated in relevant commercial crop-growing environments over multiple seasons are discussed as the appropriate data representative of this natural variation. A model-based parametric tolerance interval approach, which accounts for the correlated and unbalanced data structure of cumulative historical data collected from multisite field studies conducted over multiple seasons, is presented. This paper promotes the application of this tolerance interval approach to generate reference ranges for evaluation of the biological relevance of statistical differences identified during substantial equivalence assessment of a GM crop.

Análisis histológico de lesiones pódales de pollos comerciales en la Planta de Sacrificio de Pollo Olympico S. A / Histological analyses of toe lesions for commercial broiler at the slaughterhouse of Pollo Olympico S. A

Realizar el análisis histológico de lesiones pódales de pollos comerciales en la planta de sacrificio de Pollo Olympico S.A., Colombia con el fin de discutir aspectos sobre su retención sanitaria en plantas de sacrificio. Materiales y métodos: Se utilizó la población avícola de 10 granjas de la empresa Pollo Olympico S.A evaluando macroscópicamente 129.551 animales y microscópicamente 330 lesiones podales. El análisis histológico evaluó el tipo y la profundidad de las lesiones inflamatorias y la presencia de estructuras micóticas. Resultados: La evaluación macroscópica evidenció lesiones podales en el 64,7% de la población total de aves. La evaluación microscópica mostró un predominio de procesos inflamatorios con presentación de heterófilos (73,1%), el 19% fueron infiltrados mixtos y el 7.9% de predominio mononuclear. Las lesiones encontradas mostraron una localización principalmente subcorneal (77.9%) y la ausencia de estructuras micóticas, células gigantes o lesiones vasculares...

To conduct histological analyses of toe lesions for commercial broiler at the slaughterhouse of Pollo Olympico S.A., Colombia, with the ultimate goal of proposing recommendations about sanitary containment. Materials and Methods: Poultry subjects were sampled in 10 farms of Pollo Olympico S.A. Macroscopic inspections were conducted on 129.551 animals while the microscopic ones were on 330 toe lesions. Histological analyses evaluated type and depth of inflammatory lesions as well as the presence of fungal infections Results: Macroscopic analyses revealed inflammatory process mediated by heterophiles (73,1%), mixed infiltrates (19%) and mononuclear (7.9%). Lesions were mainly localized at the sub-corneal level (77.9%) and showed no signs of fungal structures, giant cells or vascular presence...

Physicochemical and biochemical characterization of transgenic papaya modified for protection against Papaya ringspot virus.

BACKGROUND: Papaya, a nutritious tropical fruit, is consumed both in its fresh form and as a processed product worldwide. Major quality indices which include firmness, acidity, pH, colour and size, are cultivar dependent. Transgenic papayas engineered for resistance to Papaya ringspot virus were evaluated over the ripening period to address physicochemical quality attributes and food safety concerns. RESULTS: With the exception of one transgenic line, no significant differences (P > 0.05) were observed in firmness, acidity and pH. Lightness (L*) and redness (a*) of the pulps of non-transgenic and transgenic papaya were similar but varied over the ripening period (P < 0.05). Fruit mass, though non-uniform (P < 0.05) for some lines, was within the range reported for similar papaya cultivars, as were shape indices of female fruits. Transgene proteins, CP and NPTII, were not detected in fruit pulp at the table-ready stage. CONCLUSION: The findings suggest that transformation did not produce any major unintended alterations in the physicochemical attributes of the transgenic papayas. Transgene proteins in the edible fruit pulp were low or undetectable.

Safety assessment of cosmetic products, with emphasis on the ocular area: regulatory aspects and validation processes

Before marketing a cosmetic product, a series of biological assays, such as ocular irritation tests, must be conducted in order to prove that the product is safe. However, a few scientific articles mention the discussion and evolution of cosmetic products testing performed in the eyes area. The aim of this study was to review the available literature on the evolution of tests carried out with cosmetics, in the ocular area, as well as to describe the methodologies that have been used and that are currently accepted. In Brazil, tests performed on animals are still allowed. However, the international laws strongly recommend the use of alternative methods for evaluating the risk of cosmetic ingredients and products. Regulatory requirements involving the registration of these products also request safety support of them in human beings. To perform ocular tests in human beings, it is necessary to involve an ophthalmologist for conducting clinical protocols. These protocols signed by the expert physician are sent to the National Health Surveillance Agency in order to endorse the product manufacturer concerning its safety. The safety support of a cosmetic product is very important, taking into account that the consumer has free access to these products of widespread use in today's society.

Com o objetivo de comprovar que um produto cosmético é seguro, antes que este seja colocado no mercado, este deve passar por uma série de ensaios biológicos, que avaliem sua segurança, como, por exemplo, os testes de irritação ocular. Porém, poucos artigos científicos trazem a discussão e a evolução sobre os testes de produtos cosméticos realizados na área dos olhos. O objetivo desse trabalho foi realizar uma revisão bibliográfica sobre a evolução dos testes realizados com cosméticos, na região ocular, bem como descrever as metodologias que já foram utilizadas e as que são aceitas atualmente. No Brasil, são ainda permitidos testes em animais, entretanto, as legislações internacionais indicam fortemente a utilização de métodos alternativos para avaliação de risco de ingredientes e produtos cosméticos. As exigências regulatórias que envolvem o registro desses produtos solicitam também a comprovação de segurança destes produtos em serem humanos. Para a realização dos testes oculares em humanos, é necessário o envolvimento de um oftalmologista na condução de protocolos clínicos. Esses protocolos assinados pelo médico especialista são enviados à Agencia Nacional de Vigilância Sanitária, a fim de respaldar o fabricante do produto sobre a segurança do mesmo. A comprovação da segurança de um produto cosmético é bastante importante, considerando-se o livre acesso aos consumidores e o amplo uso desses produtos na sociedade atual.