Diagnóstico e conduta pré-natal em malformação adenomatóide cística pulmonar fetal: apresentação de um caso

A malformação adenomatóide cística fetal consiste em anomalia do desenvolvimento pulmonar, proveniente de crescimento anormal dos bronquíolos respiratórios terminais. Por tratar-se de malformação rara, nem sempre é lembrada como possibilidade diagnóstica. Apresentamos o relato de um caso de malformação adenomatóide cística pulmonar fetal em adolescente primigesta, ressaltando a importância do seu diagnóstico precoce e possibilidades terapêuticas. Apresentamos também o aspecto evidenciado após a colocação de cateter para drenagem contínua.

Relação entre os níveis séricos do hormônio anti-Mulleriano, inibina B, estradiol e hormônio folículo estimulante no terceiro dia e o status folicular ovariano

OBJETIVO: examinar a hipótese de que o nível sérico do hormônio anti-Mülleriano (HAM) reflete o status folicular ovariano. MÉTODOS: Desenho: estudo prospectivo. Pacientes: foram incluídas 101 candidatas à FIV-TE submetidas à estimulação ovariana controlada com agonista de GnRH e FSH. Depois de atingir a supressão da hipófise e antes da administração de FSH (dia basal), os níveis séricos de HAM, inibina B e FSH foram avaliados. O número de folículos antrais foi determinado pela ultra-sonografia (dia basal) (folículo antral precoce; 3-10 mm). RESULTADOS: as médias do nível sérico de HAM, inhibina B, E2, P4 e FSH (dia basal) foram 3,4±0,14 ng/mL, 89±4,8 pg/mL, 34±2,7 pg/mL, 0,22±0,23 ng/mL e 6,6±0,1 mUI/mL, respectivamente, e a média do número de folículos antrais precoces foi 17±0,39. O nível sérico do HAM foi negativamente correlacionado com a idade (r= -0,19, p<0,04) e positivamente correlacionado com o número de folículos antrais precoces (r=0,65, p<0,0001), mas isto não se aplicou aos níveis séricos de inibina B, E2 e FSH. CONCLUSÕES: esse dado demonstra a associação do HAM com a quantidade de folículo antral, sendo aquele, portanto, um provável biomarcador do status folicular ovariano.

The seasonal and ovarian status effects on in vitro production of domestic cat embryos between Equator and Tropic of Capricorn

From the Tropic of Capricorn to Equator, the seasonality of domestic cat is known to be absent, i.e., these animals are considered non-seasonal breeders at these regions. We hypothesized that this particularity might have some influence on in vitro embryo production. The aim of this experiment was to determine the percentage of cleavage and morulae and blastocyst formation produced from oocytes recovered from queen ovaries of three distinct status - follicular, luteal or inactive - during two different reproductive seasons experienced by cats in southeast of Brazil (22°53'09" S and 48°26'42" W) - non breeding season (NBS), comprehending January to March; and breeding season (BS), August to October. Thirty queens were neutered. Ovaries were classified according to their status and were sliced in PBS for cumulus oocyte complex (COC) releasing. Grade I COC were washed three times in H-MEM supplemented with BSA, glutamine, sodium pyruvate, cysteine, streptomycin and penicillin. Oocytes were incubated in groups of 20-30 in 400µL of DMEM supplemented with FSH, LH, estradiol, IGF-I and basic fibroblast growth factor under mineral oil for 30 or 36 hours at 38°C in humidified environment of 5% de O2, 5% CO2 and 90% N2. COC were fertilized in Ham's F-10 medium supplemented with BSA, cysteine, pyruvate and streptomycin/penicillin (culture medium) with fresh semen selected through swim up technique. Eighteen hours later, the presumptive zygotes were denuded, the percentage of cleavage was determined and every 10 zygotes were transferred to 100mL drops of culture medium for culture during three days. After 72 hours of culture the percentage of morulae formation was evaluated and these structures were transferred to drops of the same culture medium. At the eighth day of culture blastocyst formation was analyzed. During NBS, from a total of 272 (inactive), 162 (luteal) and 134 (follicular) fertilized oocytes, the percentage of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 24.63, 16.54 and 8.09 respectively; for those derived from luteal ovaries, the percentage was 21.6, 12.96 and 8.64, and for those from follicular ovaries, they were 24.62, 16.41 and 8.21. Considering BS, from a total of 102 (inactive), 198 (luteal) and 86 (follicular) fertilized oocytes, the relative frequency (%) of cleaved zygotes, morulae and blastocysts derived from inactive ovaries were 64.7, 41.17 and 23.53 respectively; for those derived from luteal ovaries, the percentage was 64.14, 40.41 and 23.73, and for those from follicular ovaries, they were 63.95, 39.54 and 24.41. The results of this experiment demonstrate that no statistically significant difference (P<0.05) was verified in the frequency of cleaved embryos and morulae and blastocyst formation when comparing the three ovarian conditions in the same season. However the breeding season presented better results considering cleavage and morulae and blastocyst formation.

Importance of considering food waste in the development of sustainable food packaging systems

Meeting the needs of both present and future generations forms the foundation of sustainable development. Concern about food demand is increasing alongside the continuously growing population. In the pursuit of food security preventing food waste is one solution avoiding the negative environmental impacts that result from producing food unnecessarily. Packages offer one answer to preventing food waste, as they 1) preserve and protect food, 2) introduce the user to the correct way to handle and use the food and package and 3) allow the user to consume the food in its entirety. This thesis aims to enhance the sustainability of food packages by giving special emphasis to preventing food waste. The focus of this thesis is to assist the packaging designer in being able to take into account the requirements for the sustainability of food packages and to be able to integrate these requirements into the product development process. In addition, life cycle methods that can be used as a tool in the packaging design process or in assessing the sustainability of finished food-packaging combinations are evaluated. The methods of life cycle costing (LCC) and life cycle working environment (LCWE) are briefly discussed. The method of life cycle assessment (LCA) is examined more thoroughly through the lens of the literature review of food-package LCA case studies published in the 21st century in three relevant journals. Based on this review and on experiences learned from conducting LCAs, recommendations are given as to how the LCA practitioner should conduct a food packaging study to make most of the results. Two case studies are presented in this thesis. The first case study relates the results of a life cycle assessment conducted for three food items (cold cut (ham), sliced dark bread (rye) and Soygurt drink) and the alternative packaging options of each. Results of this study show that the packaging constitutes only 1–12 % of the total environmental impacts of the food-packaging combination. The greatest effect is derived from the food itself and the wasted food. Even just a small percentage of wasted food causes more environmental impacts than does the packaging. The second case study presents the results of LCC and LCWE analysis done for fruit and vegetable transport packages. In this thesis, the specific results of the study itself are not the focus, but rather the study methods and scope are analysed based on how these complement the sustainability assessment of food packages. This thesis presents reasons why prevention of food waste should be more thoroughly taken into account in food packaging design. In addition, the task of the packaging designer is facilitated by the requirements of sustainable food packaging, by the methods and step-by-step guidance on how to integrate sustainability issues into the design process, and by the recommendations on how to assess the sustainability of food packages. The intention of this thesis is to express the issues that are important in the field of the food packaging industry. Having recognised and implemented these issues, businesses can better manage the risks that could follow from neglecting these sustainability aspects.

The pathogenesis of tropical spastic paraparesis/human T-cell leukemia type I-associated myelopathy

Tropical spastic paraparesis/human T-cell leukemia type I-associated myelopathy (TSP/HAM) is caused by a human T-cell leukemia virus type I (HTLV-I) after a long incubation period. TSP/HAM is characterized by a chronic progressive paraparesis with sphincter disturbances, no/mild sensory loss, the absence of spinal cord compression and seropositivity for HTLV-I antibodies. The pathogenesis of this entity is not completely known and involves a multivariable phenomenon of immune system activation against the presence of HTLV-I antigens, leading to an inflammatory process and demyelination, mainly in the thoracic spinal cord. The current hypothesis about the pathogenesis of TSP/HAM is: 1) presence of HTLV-I antigens in the lumbar spinal cord, noted by an increased DNA HTLV-I load; 2) CTL either with their lytic functions or release/production of soluble factors, such as CC-chemokines, cytokines, and adhesion molecules; 3) the presence of Tax gene expression that activates T-cell proliferation or induces an inflammatory process in the spinal cord; 4) the presence of B cells with neutralizing antibody production, or complement activation by an immune complex phenomenon, and 5) lower IL-2 and IFN-gamma production and increased IL-10, indicating drive to a cytokine type 2 pattern in the TSP/HAM subjects and the existence of a genetic background such as some HLA haplotypes. All of these factors should be implicated in TSP/HAM and further studies are necessary to investigate their role in the development of TSP/HAM.

Possible etiologies for tropical spastic paraparesis and human T lymphotropic virus I-associated myelopathy

The epidemiology of tropical spastic paraparesis/human T lymphotropic virus I (HTLV-I)-associated myelopathy (TSP/HAM) is frequently inconsistent and suggests environmental factors in the etiology of these syndromes. The neuropathology corresponds to a toxometabolic or autoimmune process and possibly not to a viral disease. Some logical hypotheses about the etiology and physiopathology of TSP and HAM are proposed. Glutamate-mediated excitotoxicity, central distal axonopathies, cassava, lathyrism and cycad toxicity may explain most cases of TSP. The damage caused to astrocytes and to the blood-brain barrier by HTLV-I plus xenobiotics may explain most cases of HAM. Analysis of the HTLV-I/xenobiotic ratio clarifies most of the paradoxical epidemiology of TSP and HAM. Modern neurotoxicology, neuroimmunology and molecular biology may explain the neuropathology of TSP and HAM. It is quite possible that there are other xenobiotics implicated in the etiology of some TSP/HAMs. The prevention of these syndromes appears to be possible today.

Skin morphology of the mutant hairless USP mouse

The morphology of the skin of the mutant hairless USP mouse was studied by histological, histochemical and immunohistochemical methods and compared to the skin of BALB/c mice. Representative sections of the dorsal skin from mice of both strains aged 18 days, and 1, 3, 6, and 8 months were studied. Sections stained with hematoxylin and eosin showed cystic formations called utricles and dermal cysts in the dermis that increased in size and number during growth. Skin thickness increased significantly at 8 months. Sections stained with picrosirius and examined with polarized light, displayed different colors, suggesting different thicknesses of dermal collagen fibers (probably types I and III). Weigert, Verhoeff and resorcin-fuchsin stains revealed fibers of the elastic system. The PAS and Alcian blue methods revealed neutral and acid glycosaminoglycans in the skin ground substance of both mouse strains. Immunohistochemical staining for fibronectin and laminin did not show differences between the mutant and BALB/c mice. Mast cells stained by the Gomori method and macrophages positive for HAM 56 antibodies were observed in both mouse strains. Except for the presence of enlarged cysts in the hairless strain, no qualitative differences were found during development of the skin of BALB/c and the mutant hairless mice.

Autoimmunity and molecular mimicry in tropical spastic paraparesis/human T-lymphotropic virus-associated myelopathy

Viruses share antigenic sites with normal host cell components, a phenomenon known as molecular mimicry. It has long been suggested that viral infections might trigger an autoimmune response by several mechanisms including molecular mimicry. More than 600 antiviral monoclonal antibodies generated against 11 different viruses have been reported to react with 3.5% of cells specific for uninfected mouse organs. The main pathological feature of tropical spastic paraparesis/human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (TSP/HAM) is a chronic inflammation of the spinal cord characterized by perivascular cuffing of mononuclear cells accompanied by parenchymal lymphocytic infiltration. We detected the presence of autoantibodies against a 98- to 100-kDa protein of in vitro cultured human astrocytes and a 33- to 35-kDa protein from normal human brain in the serum of HTLV-I-seropositive individuals. The two cell proteins exhibited molecular mimicry with HTLV-I gag and tax proteins in TSP/HAM patients, respectively. Furthermore, the location of 33- to 35-kDa protein cross-reaction correlated with the anatomical spinal cord areas (in the rat model) in which axonal damage has been reported in several cases of TSP/HAM patients. Our experimental evidence strongly suggests that the demyelinating process occurring in TSP/HAM may be mediated by molecular mimicry between domains of some viral proteins and normal cellular targets of the spinal cord sections involved in the neurodegeneration.

Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co-hydroxyvalerate) as surfaces for Vero cell culture

Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37°C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces.

Human T-cell lymphotropic virus type I (HTLV-I) proviral DNA viral load among asymptomatic patients and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis

To evaluate the human T-cell lymphotropic virus type I (HTLV-I) proviral DNA load among asymptomatic HTLV-I-infected carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), real time PCR using TaqMan probes for the pol gene was performed in two million peripheral blood mononuclear cells (PBMC). The albumin gene was the internal genomic control and MT2 cells were used as positive control. The results are reported as copies/10,000 PBMC, and the detection limit was 10 copies. A total of 89 subjects (44 HAM/TSP and 45 healthy HTLV-I-infected carriers) followed up at the Institute of Infectious Diseases "Emilio Ribas" and in the Neurology Division of Hospital of Clínicas were studied. The asymptomatic HTLV-I-infected carriers had a median number of 271 copies (ranging from 5 to 4756 copies), whereas the HAM/TSP cases presented a median of 679 copies (5-5360 copies) in 10,000 PBMC. Thus, HAM/TSP patients presented a significantly higher HTLV-I proviral DNA load than healthy HTLV-I carriers (P = 0.005, one-way Mann-Whitney test). As observed in other persistent infections, proviral DNA load quantification may be an important tool for monotoring HTLV-I-infected subjects. However, long-term follow-up is necessary to validate this assay in the clinical setting.

High HTLV-1 proviral load, a marker for HTLV-1 associated myelopathy/tropical spastic paraparesis, is also detected in patients with infective dermatitis associated with HTLV-1

Salvador (BA, Brazil) is an endemic area for human T-cell lymphotrophic virus type 1 (HTLV-1). The overall prevalence of HTLV-1 infection in the general population has been estimated to be 1.76%. HTLV-1 carriers may develop a variety of diseases such as adult T-cell leukemia/lymphoma, HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and infective dermatitis associated with HTLV-1 (IDH). IDH is a chronic and severe form of childhood exudative and infective dermatitis involving mainly the scalp, neck and ears. It has recently been observed that 30% of patients with IDH develop juvenile HAM/TSP. The replication of HTLV-1 has been reported to be greater in adult HAM/TSP patients than in asymptomatic HTLV-1 carriers. In the current study, the proviral load of 28 children and adolescents with IDH not associated with HAM/TSP was determined and the results were compared to those obtained in 28 HTLV-1 adult carriers and 28 adult patients with HAM/TSP. The proviral load in IDH patients was similar to that of patients with HAM/TSP and much higher than that found in HTLV-1 carriers. The high levels of proviral load in IDH patients were not associated with age, duration of illness, duration of breast-feeding, or activity status of the skin disease. Since proviral load is associated with neurological disability, these data support the view that IDH patients are at high risk of developing HAM/TSP.

In vivo fluctuation of Tax, Foxp3, CTLA-4, and GITR mRNA expression in CD4+CD25+ T cells of patients with human T-lymphotropic virus type 1-associated myelopathy

HTLV-1 Tax expression exerts an inhibitory effect on the Foxp3 transcription factor in CD4+CD25+ T-regulatory cells (Treg). For a better understanding of the role of Tax mRNA in the gene expression of cellular markers we measured Tax, Foxp3, CTLA-4, GITR, TGF-β, and IL-10 mRNA in Treg cells of 50 patients with human T-lymphotropic virus type 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP; 27 women and 23 men; mean age: 56.7 years). The control group consisted of 23 non-infected subjects (12 women and 11 men) with a mean age of 51.3 years. Real-time PCR was used to measure mRNA of Tax proteins and several cellular markers of Treg function. Determinations revealed a high level of Tax mRNA in HAM/TSP (124.35 copies/100 CD4+CD25+ T cells). Foxp3, GITR, and CTLA-4 mRNA levels were lower in HAM/TSP patients (mean ± SD, 22.07 ± 0.78, 9.63 ± 0.36, and 4.54 ± 0.39, respectively) than in non-infected controls (47.15 ± 12.94, 22.14 ± 1.91, and 21.07 ± 2.31). Both groups had similar levels of TGF-β and IL-10. An inverse relationship was found between Tax levels and Foxp3, CTLA-4, and GITR levels. Conversely, there was a direct correlation between levels of Foxp3, GITR, and CTLA-4. Disease severity and evolution time did not correlate with Tax or Foxp3 levels. The present results suggest that Tax and Foxp3 mRNA vary with the same degree of disease severity in HAM/TSP patients. Tax fluctuations may affect CTLA-4 and GITR expression via the Foxp3 pathway, causing virus-induced dysfunction of CD4+CD25+ T cells in HAM/TSP patients.

Effect of TNF-α production inhibitors on the production of pro-inflammatory cytokines by peripheral blood mononuclear cells from HTLV-1-infected individuals

Human T lymphotropic virus type 1 (HTLV-1) is the causal agent of myelopathy/tropical spastic paraparesis (HAM/TSP), a disease mediated by the immune response. HTLV-1 induces a spontaneous proliferation and production of pro-inflammatory cytokines by T cells, and increasing interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) levels are potentially involved in tissue damage in diseases related to HTLV-1. This exaggerated immune response is also due to an inability of the natural regulatory mechanisms to down-modulate the immune response in this group of patients. TNF-α inhibitors reduce inflammation and have been shown to improve chronic inflammatory diseases in clinical trials. The aim of this study was to evaluate the ability of pentoxifylline, forskolin, rolipram, and thalidomide to decrease in vitro production of TNF-α and IFN-γ in cells of HTLV-1-infected subjects. Participants of the study included 19 patients with HAM/TSP (mean age, 53 ± 11; male:female ratio, 1:1) and 18 HTLV-1 carriers (mean age, 47 ± 11; male:female ratio, 1:2.6). Cytokines were determined by ELISA in supernatants of mononuclear cell cultures. Pentoxifylline inhibited TNF-α and IFN-γ synthesis with the minimum dose used (50 µM). The results with forskolin were similar to those observed with pentoxifylline. The doses of rolipram used were 0.01-1 µM and the best inhibition of TNF-α production was achieved with 1 µM and for IFN-γ production it was 0.01 µM. The minimum dose of thalidomide used (1 µM) inhibited TNF-α production but thalidomide did not inhibit IFN-γ production even when the maximum dose (50 µM) was used. All drugs had an in vitro inhibitory effect on TNF-α production and, with the exception of thalidomide, all of them also decreased IFN-γ production.

Perfil sensorial e aceitação de presuntos crus produzidos por métodos tradicionais e acelerado

Existem muitos tipos de presuntos crus com perfis sensoriais particulares, em decorrência de diferentes matérias-primas e técnicas de processamento, que são apreciados por seus sabores e texturas característicos. Este trabalho objetivou caracterizar o perfil sensorial de presuntos crus através da Análise Descritiva Quantitativa e verificar a aceitação dos produtos pelo consumidor. Foram avaliados dois presuntos crus experimentais, produzidos por processo acelerado (denominados CTC 3,5 e 5,0% devido ao teor inicial de sal adicionado), e quatro produtos comercializados no Brasil, um Serrano espanhol, um Italiano e dois brasileiros (Tipo Serrano e Tipo Parma). Os produtos diferiram pelos seguintes atributos: CTC 3,5% - sabor mais ácido, menor intensidade de sabor de ranço e aroma de ranço, aparência: menor intensidade de cor vermelha e menor intensidade de suculência; CTC 5,0% - mais fibrosidade, menores intensidade e persistência de sabor e maciez; Serrano - maiores aroma de ranço, cor vermelha, intensidade e persistência de sabor e menor sabor salgado; Tipo Serrano - maior sabor de ranço e menor sabor doce; Italiano - maiores sabor salgado e maciez; Tipo Parma - sabor de carne, marmoreado e amarelo da gordura mais intensos. Todos os produtos obtiveram boa aceitação pelo consumidor. O presunto Tipo Serrano foi o mais aceito e o Serrano foi o menos aceito pelos consumidores brasileiros entrevistados. Os produtos CTC foram considerados de boa qualidade, apresentando características típicas de um presunto cru, apesar do curto período de maturação.

Partial substitution of nitrite by chitosan and the effect on the quality properties of pork sausages

The aim of the study was to evaluate the influence of partial nitrite replacement by chitosan on the quality of Ham Visking (a type of pork sausages). Five Ham Visking formulations were elaborated modifying the sodium nitrite (0.011; 0.016 or 0.0212%) and chitosan concentrations (0.25 or 0.5%) in the products. Sausages were stored at 4 ºC and physicochemical, microbiological, and sensorial evaluations were performed in order to estimate their shelf life. Chitosan can be used in pork sausages without affecting ensory attributes such as color although the panelists detected textural differences among the samples with chitosan, which suggests that there is some influence of deacetylation degree of chitosan on the textural behavior of sausages which still needed to be explained for a successful application of chitosan in meat products. The reduction of residual sodium nitrite did not affect the color and flavor of such products, but the use of chitosan increasedsignificantly the shelf life of sausages.