Glicólise do poli(3-hidroxibutirato) por via enzimática

Poly(3-hydroxybutyrate), PHB, is a polymer with broad potential applications because of its biodegradability and biocompatibility. However, its high crystallinity is a limiting factor for many applications. To overcome this drawback, one strategy currently employed involves the reduction of the molecular weight of PHB with the concomitant formation of end-functionalized chains, such as those obtained via glycolysis. The glycolysis of PHB can be catalyzed by acid, base, or organometallic compounds. However, to our knowledge, there are no reports regarding PHB glycolysis catalyzed enzymatically. Among the major types of enzymes used in biocatalysis, the lipases stand out because they have the ability to catalyze reactions in both aqueous and organic media. Thus, in this study, we performed the enzymatic glycolysis of PHB using the lipase Amano PS (Pseudomonas cepacia) with ethane-1,2-diol (ethylene glycol) as the functionalizing agent. The results indicated that the glycolysis was successful and afforded hydroxyl-terminated oligomeric PHB polyols. Nuclear magnetic resonance spectra of the products showed characteristic signals for the terminal hydroxyl groups of the polyols, while thermogravimetric and differential scanning calorimetry analyses confirmed an increase in the thermal stability and a decrease in the crystallinity of the polyols compared with the starting PHB polymer, which were both attributed to the reduction in the molecular weight due to glycolysis.

Bionanocompósitos preparados por incorporação de nanocristais de celulose em polímeros biodegradáveis por meio de evaporação de solvente, automontagem ou eletrofiação

This review reports the preparation and characterization of bionanocomposites based on biodegradable polymers reinforced with cellulose nanocrystals (CNC) described in the literature. The outstanding potential of cellulose nanocrystals as reinforcement fillers of biodegradable polymers is presented with an emphasis on the solution casting process, which is an appropriate method to investigate the physico-chemical effects of the incorporation of CNC into the polymeric matrices. Besides solution casting, other small scale methods such as electrospinning and layer-by-layer are also covered.

HYDROLYTIC DEGRADATION BEHAVIOR OF PLLA NANOCOMPOSITES REINFORCED WITH MODIFIED CELLULOSE NANOCRYSTALS

Bionanocomposites derived from poly(L-Lactide) (PLLA) were reinforced with chemically modified cellulose nanocrystals (m-CNCs). The effects of these modified cellulose nanoparticles on the mechanical and hydrolytic degradation behavior of polylactide were studied. The m-CNCs were prepared by a method in which hydrolysis of cellulose chains is performed simultaneously with the esterification of hydroxyl groups to produce modified nanocrystals with ester groups. FTIR, elemental analysis, TEM, XRD and contact angle measurements were used to confirm and characterize the chemical modifications of the m-CNCs. These bionanocomposites gave considerably better mechanical properties than neat PLLA based on an approximately 100% increase in tensile strength. Due to the hydrophobic properties of the esterified nanocrystals incorporated into a polymer matrix, it was also demonstrated that a small amount of m-CNCs could lead to a remarkable decrease in the hydrolytic degradation rate of the biopolymer. In addition, the m-CNCs considerably delay the degradation of the nanocomposite by providing a physical barrier that prevents the permeation of water, which thus hinders the overall absorption of water into the matrix. The results obtained in this study show the nanocrystals can be used to reinforce polylactides and fine-tune their degradation rates in moist or physiological environments.

Infecção Assintomática do Líquido Amniótico

Objetivo: determinar a presença de infecção assintomática do líquido amniótico em gestantes, identificar os agentes bacterianos envolvidos na infecção e determinar o perfil de suscetibilidade a antimicrobianos in vitro. Métodos: foram obtidas 81 amostras de líquido amniótico, colhidas por amniocentese, em gestantes sem sinais de trabalho de parto e sem suspeita clínica de infecção, atendidas na Maternidade Escola Assis Chateaubriand, entre agosto/97 e janeiro/99. Pesquisou-se a presença de bactérias aeróbias, anaeróbias estritas/facultativas e micoplasmas genitais. As bactérias anaeróbias foram identificadas pelo sistema ATBÒ (Bio-Mérieux) e os micoplasmas pelo kit Micoplasmas ISTÒ (Bio-Mérieux). Resultados: entre as amostras obtidas, oito (9,8%) apresentaram culturas positivas, sendo que em duas foram identificadas duas espécies bacterianas. Os patógenos isolados foram: Ureaplasma urealyticum (7 casos, 8,6%), Mycoplasma hominis (1 caso, 1,2%) e Peptostreptococcus sp (2 casos 2,4%). O padrão de resistência aos antimicrobianos caracterizou-se pela maior resistência dos micoplasmas à eritromicina (37,5%) e nenhuma resistência às ciclinas. Conclusões: o percentual de infecções assintomáticas foi muito elevado, havendo necessidade de serem realizadas novas pesquisas para avaliar as conseqüências da infecção subclínica nas grávidas e em seus conceptos, que envolvam métodos que identifiquem micoplasmas genitais, já que foram as bactérias mais freqüentemente isoladas.

O uso de adoçantes na gravidez: uma análise dos produtos disponíveis no Brasil

Os adoçantes são freqüentemente utilizados por mulheres em idade reprodutiva. Esta é uma revisão narrativa da literatura a respeito dos adoçantes atualmente comercializados no mercado brasileiro. Existem poucas informações sobre o uso da sacarina e ciclamato na gestação, e seus efeitos sobre o feto. Devido às limitadas informações disponíveis e ao seu potencial carcinogênico em animais, a sacarina e o ciclamato devem ser evitados durante a gestação (risco C). O aspartame tem sido extensivamente estudado em animais, sendo considerado seguro para uso na gestação (risco B), exceto para mulheres homozigóticas para fenilcetonúria (risco C). A sucralose e o acessulfame-K não são tóxicos, carcinogênico ou mutagênicos em animais, mas não existem estudos controlados em humanos. Porém, como esses dois adoçantes não são metabolizados, parece improvável que seu uso durante a gestação possa ser prejudicial (risco B). A estévia, substância derivada de uma planta nativa brasileira, não produz efeitos adversos sobre a gestação em animais, porém não existem estudos em humanos (risco B). Os agentes de corpo usados na formulação dos adoçantes (manitol, sorbitol, xilitol, eritrol, lactilol, isomalte, maltilol, lactose, frutose, maltodextrina, dextrina e açúcar invertido) são substâncias consideradas seguras para o consumo humano. Concluindo, segundo as evidências atualmente disponíveis, o aspartame, a sucralose, o acessulfame e a estévia podem ser utilizados com segurança durante a gestação.

The effects of 2,3-diphosphoglycerate, adenosine triphosphate, and glycosylated hemoglobin on the hemoglobin-oxygen affinity of diabetic patients

The position of the oxygen dissociation curve (ODC) is modulated by 2,3-diphosphoglycerate (2,3-DPG). Decreases in 2,3-DPG concentration within the red cell shift the curve to the left, whereas increases in concentration cause a shift to the right of the ODC. Some earlier studies on diabetic patients have reported that insulin treatment may reduce the red cell concentrations of 2,3-DPG, causing a shift of the ODC to the left, but the reports are contradictory. Three groups were compared in the present study: 1) nondiabetic control individuals (N = 19); 2) insulin-dependent diabetes mellitus (IDDM) patients (on insulin treatment) (N = 19); 3) non-insulin-dependent diabetes mellitus (NIDDM) patients using oral hypoglycemic agents and no insulin treatment (N = 22). The overall position of the ODC was the same for the three groups despite an increase of the glycosylated hemoglobin fraction that was expected to shift the ODC to the left in both groups of diabetic patients (HbA1c: control, 4.6%; IDDM, 10.5%; NIDDM, 9.0%). In IDDM patients, the effect of the glycosylated hemoglobin fraction on the position of the ODC appeared to be counterbalanced by small though statistically significant increases in 2,3-DPG concentration from 2.05 (control) to 2.45 µmol/ml blood (IDDM). Though not statistically significant, an increase of 2,3-DPG also occurred in NIDDM patients, while red cell ATP levels were the same for all groups. The positions of the ODC were the same for control subjects, IDDM and NIDDM patients. Thus, the PO2 at 50% hemoglobin-oxygen saturation was 26.8, 28.2 and 28.5 mmHg for control, IDDM and NIDDM, respectively. In conclusion, our data question the idea of adverse side effects of insulin treatment on oxygen transport. In other words, the shift to the left reported by others to be caused by insulin treatment was not detected.

Bone morphogenetic proteins: from structure to clinical use

Bone morphogenetic proteins (BMPs) are multi-functional growth factors belonging to the transforming growth factor ß superfamily. Family members are expressed during limb development, endochondral ossification, early fracture, and cartilage repair. The activity of BMPs was first identified in the 1960s but the proteins responsible for bone induction were unknown until the purification and cloning of human BMPs in the 1980s. To date, about 15 BMP family members have been identified and characterized. The signal triggered by BMPs is transduced through serine/threonine kinase receptors, type I and II subtypes. Three type I receptors have been shown to bind BMP ligands, namely: type IA and IB BMP receptors and type IA activin receptors. BMPs seem to be involved in the regulation of cell proliferation, survival, differentiation and apoptosis, but their hallmark is their ability to induce bone, cartilage, ligament, and tendon formation at both heterotopic and orthotopic sites. This suggests that, in the future, they may play a major role in the treatment of bone diseases. Several animal studies have illustrated the potential of BMPs to enhance spinal fusion, repair critical-size defects, accelerate union, and heal articular cartilage lesions. Difficulties in producing and purifying BMPs from bone tissue have prompted the attempts made by several laboratories, including ours, to express these proteins in the recombinant form in heterologous systems. This review focuses on BMP structure, molecular mechanisms of action and significance and potential applications in medical, dental and veterinary practice for the treatment of cartilage and bone-related diseases.

Protein turnover, amino acid requirements and recommendations for athletes and active populations

Skeletal muscle is the major deposit of protein molecules. As for any cell or tissue, total muscle protein reflects a dynamic turnover between net protein synthesis and degradation. Noninvasive and invasive techniques have been applied to determine amino acid catabolism and muscle protein building at rest, during exercise and during the recovery period after a single experiment or training sessions. Stable isotopic tracers (13C-lysine, 15N-glycine, ²H5-phenylalanine) and arteriovenous differences have been used in studies of skeletal muscle and collagen tissues under resting and exercise conditions. There are different fractional synthesis rates in skeletal muscle and tendon tissues, but there is no major difference between collagen and myofibrillar protein synthesis. Strenuous exercise provokes increased proteolysis and decreased protein synthesis, the opposite occurring during the recovery period. Individuals who exercise respond differently when resistance and endurance types of contractions are compared. Endurance exercise induces a greater oxidative capacity (enzymes) compared to resistance exercise, which induces fiber hypertrophy (myofibrils). Nitrogen balance (difference between protein intake and protein degradation) for athletes is usually balanced when the intake of protein reaches 1.2 g·kg-1·day-1 compared to 0.8 g·kg-1·day-1 in resting individuals. Muscular activities promote a cascade of signals leading to the stimulation of eukaryotic initiation of myofibrillar protein synthesis. As suggested in several publications, a bolus of 15-20 g protein (from skimmed milk or whey proteins) and carbohydrate (± 30 g maltodextrine) drinks is needed immediately after stopping exercise to stimulate muscle protein and tendon collagen turnover within 1 h.

Metronomic chemotherapy in the neoadjuvant setting: results of two parallel feasibility trials (TraQme and TAME) in patients with HER2+ and HER2− locally advanced breast cancer

Neoadjuvant chemotherapy has practical and theoretical advantages over adjuvant chemotherapy strategy in breast cancer (BC) management. Moreover, metronomic delivery has a more favorable toxicity profile. The present study examined the feasibility of neoadjuvant metronomic chemotherapy in two cohorts [HER2+ (TraQme) and HER2− (TAME)] of locally advanced BC. Twenty patients were prospectively enrolled (TraQme, n=9; TAME, n=11). Both cohorts received weekly paclitaxel at 100 mg/m2 during 8 weeks followed by weekly doxorubicin at 24 mg/m2 for 9 weeks in combination with oral cyclophosphamide at 100 mg/day (fixed dose). The HER2+ cohort received weekly trastuzumab. The study was interrupted because of safety issues. Thirty-six percent of patients in the TAME cohort and all patients from the TraQme cohort had stage III BC. Of note, 33% from the TraQme cohort and 66% from the TAME cohort displayed hormone receptor positivity in tumor tissue. The pathological complete response rates were 55% and 18% among patients enrolled in the TraQme and TAME cohorts, respectively. Patients in the TraQme cohort had more advanced BC stages at diagnosis, higher-grade pathological classification, and more tumors lacking hormone receptor expression, compared to the TAME cohort. The toxicity profile was also different. Two patients in the TraQme cohort developed pneumonitis, and in the TAME cohort we observed more hematological toxicity and hand-foot syndrome. The neoadjuvant metronomic chemotherapy regimen evaluated in this trial was highly effective in achieving a tumor response, especially in the HER2+ cohort. Pneumonitis was a serious, unexpected adverse event observed in this group. Further larger and randomized trials are warranted to evaluate the association between metronomic chemotherapy and trastuzumab treatment.

Aumento da vida útil e microbiologia da carne suína embalada em atmosfera modificada

Com o objetivo de avaliar o efeito da embalagem de carne suína em atmosfera modificada, obteve-se assepticamente amostras de lombo (Longissimus dorsi) de suíno de aproximadamente 3x3cm que depois de lavadas em solução salina (0,35%) estéril, foram introduzidas em bolsas de plástico (Cryovac BB4L) de baixa permeabilidade. Dividiram-se em 4 lotes e se embalaram com aproximadamente 1,0l de ar (100%), nitrogênio (100%), 20/80 e 40/60 CO2/O2 e se termo-selaram. Finalmente, cada lote se subdividiu em 2, armazenando-se a 1 e 7ºC. Durante o armazenamento tomaram-se amostras determinando-se a composição da atmosfera, o pH e a composição da microbiota (microbiota total, bactérias lácticas, Enterobacteriaceae, Brochothrix thermosphacta e contagens em ágar pseudomonas). Os parâmetros de crescimento (fase de latência e tempo de duplicação) dos microrganismos foram determinados mediante a equação de Gompertz. Os resultados demonstraram que a vida útil da carne suína é prolongada quando se armazena em atmosferas modificadas. As atmosferas enriquecidas com CO2 e a 1ºC mostraram uma maior eficácia. As atmosferas com CO2, mantiveram o pH das amostras constante durante todo o período de armazenamento. A 1 e 7ºC, as fases de latência e os tempos de duplicação da microbiota total foram progressivamente maiores na seguinte ordem: atmosfera de ar (100%), N2 (100%), 20/80 e 40/60 CO2/O2. Pode-se concluir que, tanto a 1 como a 7ºC, a utilização das atmosferas modificadas retardou o crescimento das bactérias alterantes da carne suína, favorecendo o prolongamento da vida útil, principalmente nas atmosferas com CO2.

Avaliação da utilização de embalagem em atmosfera modificada sobre a conservação de sardinhas (Sardinella brasiliensis)

A vida útil dos alimentos perecíveis conservados em atmosfera normal é limitada principalmente pelo efeito do oxigênio atmosférico e o crescimento de microorganismos aeróbios produtores de alterações, que promovem mudanças de odor, sabor, cor e textura, conduzindo à perda da qualidade. A modificação da atmosfera prolonga significativamente a vida útil dos alimentos, quando comparados à refrigeração, podendo chegar a um aumento de três a quatro vezes. O trabalho teve como objetivo avaliar o efeito da embalagem de sardinhas (Sardinella brasiliensis) em atmosfera modificada. As amostras foram divididas em 8 lotes, embaladas com aproximadamente 1,5 L de ar (100%), CO2 (100%), 50/50 CO2/O2 e vácuo e termo-seladas, sendo armazenadas por um período de 22 dias. A cada 2 dias as amostras foram avaliadas quanto ao pH, bases voláteis totais (BVT) e contagens totais de microrganismos heterotróficos aeróbios mesófilos. Os valores de pH aumentaram durante o tempo de estocagem, com exceção das amostras armazenadas em 100 e 50% CO2. A evolução de BVT foi compatível com outros parâmetros, como pH e contagem de bactérias totais, sendo sua evolução mais lenta nas amostras em que ocorreu o uso de atmosfera modificada. Os microrganismos mesófilos, nas atmosferas enriquecidas com CO2, apresentaram fases de latência e tempos de duplicação maiores, quando comparados com os das amostras em que não foi aplicado o método. Sob o ponto de vista microbiológico, as embalagens enriquecidas com CO2 demonstraram ser o melhor método de conservação, como também os resultados de BVT se encontraram dentro dos limites de 30 mg.100 g -1 de acordo com o estabelecido pela RIISPOA8. Recomenda-se o uso da atmosfera de 100% de CO2 como forma de conservação da sardinha por apresentar melhores parâmetros de vida útil.

Methods of discard ewes carcass suspension and the quality of meat

The objective of this study was to evaluate the carcass suspension method concerning quality of sheep meat. Ten discard ewes were used, with approximately 62 kg of body weight. After slaughtering, flaying, evisceration and removal of head and paws, carcasses were longitudinally divided into two parts. Alternated sides of half carcasses were hanged by the tendon of the gastrocnemius (Treatment 1 - T1) and by the pelvic bone (Treatment 2 - T2) in cold store for a 24-hour period. Subsequently, the Semimembranosus muscle was removed from all half carcasses for meat quality analyses. The Semimembranosus muscles from the carcasses hanged by the pelvis suspension method presented higher softness than the same muscles from the carcasses hanged by the tendon of the gastrocnemius, with values of 1.99 kgf.cm-2 and 3.15 kgf.cm-2, respectively. Treatment 2 presented lower meat cooking losses than Treatment 1, with average values of 32.14 and 33.44%, respectively. The remaining meat quality parameters evaluated were not influenced by the carcass suspension method. We concluded that the carcass suspension method influenced meat softness and losses by cooking, with better results for carcasses hanged by the pelvic bone.