Single nucleotide polymorphism C/T-13910, located upstream of the lactase gene, associated with adult-type hypolactasia: Validation for clinical practice

Objectives: To validate C/T-13910 polymorphism associated with primary hypolactasia for clinical practice. Design and methods: Lactose breath test and PCR-RFLP for the C/T-13910 polymorphism were performed. Results: Twenty-seven of 28 patients with genotype CC had positive breath tests, all twenty-two patients with genotypes CT or TT had negative breath tests. Agreement of tests was high (p<0.0001; Kappa Index 0.96). Conclusion: C/T-13910 polymorphism detection may be a new tool for primary hypolactasia diagnosis. (C) 2008 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Membrane transporter proteins are involved in Trichophyton rubrum pathogenesis

Trichophyton rubrum is a dermatophyte responsible for the majority of human superficial mycoses. The functional expression of proteins important for the initial step and the maintenance of the infection process were identified previously in T. rubrum by subtraction suppression hybridization after growth in the presence of keratin. In this study, sequences similar to genes encoding the multidrug-resistance ATP-binding cassette (ABC) transporter, copper ATPase, the major facilitator superfamily and a permease were isolated, and used in Northern blots to monitor the expression of the genes, which were upregulated in the presence of keratin. A sequence identical to the TruMDR2 gene, encoding an ABC transporter in T rubrum, was isolated in these experiments, and examination of a T rubrum Delta TruMDR2 mutant showed a reduction in infecting activity, characterized by low growth on human nails compared with the wild-type strain. The high expression levels of transporter genes by T. rubrum in mimetic infection and the reduction in virulence of the Delta TruMDR2 mutant in a disease model in vitro suggest that transporters are involved in T. rubrum pathogenicity.

Microbial and sensory changes throughout the ripening of Prato cheese made from milk with different levels of somatic cells

The objective of this research was to evaluate the effects of 2 levels of raw milk somatic cell count (SCC) on the composition of Prato cheese and on the microbiological and sensory changes of Prato cheese throughout ripening. Two groups of dairy cows were selected to obtain low-SCC (<200,000 cells/mL) and high-SCC (>700,000 cells/mL) milks, which were used to manufacture 2 vats of cheese. The pasteurized milk was evaluated according to the pH, total solids, fat, total protein, lactose, standard plate count, coliforms at 45 degrees C, and Salmonella spp. The cheese composition was evaluated 2 d after manufacture. Lactic acid bacteria, psychrotrophic bacteria, and yeast and mold counts were carried out after 3, 9, 16, 32, and 51 d of storage. Salmonella spp., Listeria monocytogenes, and coagulase-positive Staphylococcus counts were carried out after 3, 32, and 51 d of storage. A 2 x 5 factorial design with 4 replications was performed. Sensory evaluation of the cheeses from low- and high-SCC milks was carried out for overall acceptance by using a 9-point hedonic scale after 8, 22, 35, 50, and 63 d of storage. The somatic cell levels used did not affect the total protein and salt: moisture contents of the cheeses. The pH and moisture content were higher and the clotting time was longer for cheeses from high-SCC milk. Both cheeses presented the absence of Salmonella spp. and L. monocytogenes, and the coagulase-positive Staphylococcus count was below 1 x 10(2) cfu/g throughout the storage time. The lactic acid bacteria count decreased significantly during the storage time for the cheeses from both low- and high-SCC milks, but at a faster rate for the cheese from high-SCC milk. Cheeses from high-SCC milk presented lower psychrotrophic bacteria counts and higher yeast and mold counts than cheeses from low- SCC milk. Cheeses from low- SCC milk showed better overall acceptance by the consumers. The lower overall acceptance of the cheeses from high-SCC milk may be associated with texture and flavor defects, probably caused by the higher proteolysis of these cheeses.

Effect of beta-lactoglobulin polymorphism and seasonality on bovine milk composition

The objective was to evaluate the effect of beta-lactoglobulin (beta-lg) polymorphism and seasonality on milk composition (fat, lactose, total solids, milk urea nitrogen, total protein, true protein, casein and somatic cell counts) of Holstein and Girolando cows. Milk and blood samples from 278 Holsteins cows and 156 Girolando cows were taken during two dry seasons and two rainy seasons, for milk composition analysis and to determine beta-lg genotypes, respectively. BB genotype was the most frequent for both breeds, followed by AA genotype for Holstein (BB>AA>AB) and by AB for Girolando cows (BB>AB>AA). No differences were found in milk compositional characteristics among genetic variants of beta-lg (AA, AB and BB) either between Holstein or Girolando cows. No association between milk composition and beta-lg genetic polymorphism was observed. During the dry season, independently of the breed considered, higher contents of lactose, true protein, casein and casein :true protein ratio were found.

Sequence variation of the alpha-lactalbumin gene in Holstein and Nellore cows

The alpha-lactalbumin is a subunit of lactose-synthase, an enzyme responsible for lactose production, a disaccharide that influences milk production. Sequence variations of bovine alpha-lactalbumin have been associated with differences in milk yield. This study aimed to analyze allelic frequency differences at position-1689 (g. AG) and+15 (g. AG) of the alpha-lactalbumin gene in Holstein (Bos taurus) and Nellore (Bos indicus) cows. Blood samples were analyzed from 34 Holstein, 104 Nellore, and 99 Dairy Nellore cows using PCR-RFLP. The different RFLP patterns were sequenced and a novel sequence variation on nucleotide-46 was identified. An adenine at this position was designated as the A allele and a guanine was designated B allele. The frequencies of alleles A-1689, A-46, and A+15 differed between Holstein and both Nellore breeds. The results show that differences in alpha-lactalbumin allelic variants in the 5`-flanking and the 5`-UTR region might be associated with differences in milk production between Holstein cows and cows from Nellore breeds. However, the lack of difference between Nellore and Dairy Nellore suggests that other sequence variantions that regulate milk production might be responsible for the selection of Dairy Nellore cows with superior milk production.

Liquid distribution in wet granulation: dimensionless spray flux

This study investigates binder distribution in wet granulation and focuses on the nucleation zone, which is the area where the liquid binder and powder surface come into contact and form the initial nuclei. An equipment independent parameter, dimensionless spray flux Psi (a), is defined to characterise the most important process parameters in the nucleation process: solution flowrate, powder flux, and binder drop size. Ex-granulator experiments are used to study the relationship between dimensionless spray flux, process variables and the coverage of binder fluid on the powder surface. Lactose monohydrate powder on a variable speed riffler passed under a flat spray once only. Water and 7% HPC solution at two spray pressures were used as binders. Experiments with red dye and image analysis demonstrate that changes in dimensionless spray flux correlate with a measurable difference in powder surface coverage. Nucleation experiments show that spray flux controls the size and shape of the nuclei size distribution. At low Psi (a), the system operates in the drop controlled regime, where one drop forms one nucleus and the nuclei size distribution is narrow. At higher Psi (a), the powder surface cakes creating a broader size distribution. For controlled nucleation with the narrowest possible size distribution, it is recommended that the dimensionless spray flux be less than 0.1 to be in the drop-controlled regime. (C) 2001 Elsevier Science S.A. All rights reserved.

Virulence of Streptococcus pneumoniae: PsaA mutants are hypersensitive to oxidative stress

psaA encodes a 37-kDa pneumococcal lipoprotein which is part of an ABC Mn(II) transport complex. Streptococcus pneumoniae D39 psaA mutants have previously been shown to be significantly less virulent than wild-type D39, but the mechanism underlying the attenuation has not been resolved. In this study, we have shown that psaA and psaD mutants are highly sensitive to oxidative stress, i.e., to superoxide and hydrogen peroxide, which might explain why they are less virulent than the wild-type strain. Our investigations revealed altered expression of the key oxidative-stress response enzymes superoxide dismutase and NADH oxidase in psaA and psaD mutants, suggesting that PsaA and PsaD may play important roles in the regulation of expression of oxidative-stress response enzymes and intracellular redox homeostasis.

Scale-up of mixer granulators for effective liquid distribution

There is considerable anecdotal evidence from industry that poor wetting and liquid distribution can lead to broad granule size distributions in mixer granulators. Current scale-up scenarios lead to poor liquid distribution and a wider product size distribution. There are two issues to consider when scaling up: the size and nature of the spray zone and the powder flow patterns as a function of granulator scale. Short, nucleation-only experiments in a 25L PMA Fielder mixer using lactose powder with water and HPC solutions demonstrated the existence of different nucleation regimes depending on the spray flux Psi(a)-from drop-controlled nucleation to caking. In the drop-controlled regime at low Psi(a) values. each drop forms a single nucleus and the nuclei distribution is controlled by the spray droplet size distribution. As Psi(a) increases, the distribution broadens rapidly as the droplets overlap and coalesce in the spray zone. The results are in excellent agreement with previous experiments and confirm that for drop-controlled nucleation. Psi(a) should be less than 0.1. Granulator flow studies showed that there are two powder flow regimes-bumping and roping. The powder flow goes through a transition from bumping to roping as impeller speed is increased. The roping regime gives good bed turn over and stable flow patterns. This regime is recommended for good liquid distribution and nucleation. Powder surface velocities as a function of impeller speed were measured using high-speed video equipment and MetaMorph image analysis software, Powder surface velocities were 0.2 to 1 ms(-1)-an order of magnitude lower than the impeller tip speed. Assuming geometrically similar granulators, impeller speed should be set to maintain constant Froude number during scale-up rather than constant tip speed to ensure operation in the roping regime. (C) 2002 Published by Elsevier Science B.V.

Drop penetration into porous powder beds

The kinetics of drop penetration were studied by filming single drops of several different fluids (water, PEG200, PEG600, and HPC solutions) as they penetrated into loosely packed beds of glass ballotini, lactose, zinc oxide, and titanium dioxide powders. Measured times ranged from 0.45 to 126 s and depended on the powder particle size,viscosity, surface tensions, and contact angle. The experimental drop penetration times were compared to existing theoretical predictions by M. Denesuk et al. (J. Colloid Interface Sci. 158, 114, 1993) and S. Middleman (Modeling Axisymmetric Flows: Dynamics of Films, Jets, and Drops, Academic Press, San Diego, 1995) but did not agree. Loosely packed powder beds tend to have a heterogeneous bed structure containing large macrovoids which do not participate in liquid flow but are included implicitly in the existing approach to estimating powder pore size. A new two-phase model was proposed where the total volume of the macrovoids was assumed to be the difference between the bed porosity and the tap porosity. A new parameter, the effective porosity (epsilon)eff, was defined as the tap porosity multiplied by the fraction of pores that terminate at a macrovoid and are effectively blocked pores. The improved drop penetration model was much more successful at estimating the drop penetration time on all powders and the predicted times were generally within an order of magnitude of the experimental results. (C) 2002 Elsevier Science (USA).

Ultra-high-temperature (UHT) treatment of milk: comparison of direct and indirect modes of heating

UHT processing of milk and its subsequent storage causes several changes which affect the shelf-life of UHT milk although it remains 'commercially sterile'. These changes include whey protein denaturation, protein-protein interaction, lactose-protein interaction, isomerisation of lactose, Maillard browning, sulphydryl compound formation, formation of a range of carbonyl and other flavoursome compounds, and formation of insoluble substances. They ultimately reduce the quality and limit the shelf life of UHT milk through development of off-flavours, fat separation, age gelation and sedimentation. The extent of these changes depends on many factors, a major one being the type of UHT heating. This review compares the effect heating milk by direct and indirect modes on various aspects of processing and quality of UHT milk.

Dietary influences on survival after ovarian cancer

We evaluated the effects of various food groups and micronutrients in the diet on survival among women who originally participated in a population-based case-control study of ovarian cancer conducted across 3 Australian states between 1990 and 1993. This analysis included 609 women with invasive epithelial ovarian cancer, primarily because there was negligible mortality in women with borderline tumors. The women's usual diet was assessed using a validated food frequency questionnaire. Deaths in the cohort were identified using state-based cancer registries and the Australian National Death Index (NDI). Crude 5-year survival probabilities were estimated using the Kaplan-Meier technique, and adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were obtained from Cox regression models. After adjusting for important confounding factors, a survival advantage was observed for those who reported higher intake of vegetables in general (HR = 0.75, 95% CI = 0.57-0.99, p-value trend 0.01 for the highest third, compared to the lowest third), and cruciferous vegetables in particular (HR = 0.75, 95% CI = 0.57-0.98, p-value trend 0.03), and among women in the upper third of intake of vitamin E (HR = 0.76, 95% CI = 0.58-1.01, p-value trend 0.04). Inverse associations were also seen with protein (p-value trend 0.09), red meat (p-value trend 0.06) and white meat (p-value trend 0.07), and modest positive trends (maximum 30% excess) with lactose (p-value trend 0.04), calcium and dairy products. Although much remains to be learned about the influence of nutritional factors after a diagnosis of ovarian cancer, our study suggests the possibility that a diet high in vegetable intake may help improve survival. (C) 2003 Wiley-Liss, Inc.

Estudo da cinética e influência dos métodos de secagem sobre as propriedades físico-químicas de soro de leite

O soro de leite é um subproduto da fabricação do queijo, seja por acidificação ou por processo enzimático. Em condições ideais, a caseína do leite se agrega formando um gel, que posteriormente cortado, induz a separação e liberação do soro. É utilizado de diversas formas em toda a indústria alimentícia, possui rica composição em lactose, sais minerais e proteínas. A desidratação é um dos principais processos utilizados para beneficiamento e transformação do soro. Diante disto, o objetivo deste trabalho foi avaliar a influência dos métodos de secagem: liofilização, leito de espuma (nas temperaturas de 40, 50, 60, 70 e 80ºC) e spray-dryer (nas temperaturas de 55, 60, 65, 70 e 75ºC), sobre as características de umidade, proteína, cor e solubilidade do soro, bem como estudar o seu processo de secagem. O soro foi obtido e desidratado após concentração por osmose reversa, testando 11 tratamentos, em 3 repetições, utilizando um delineamento inteiramente casualizado. Os resultados demonstraram que o modelo matemático que melhor se ajustou foi o modelo de Page, apresentado um coeficiente de determinação ajustado acima de 0,98 e erro padrão da regressão em todas as temperaturas abaixo de 0,04 para o método por leito de espuma. Para o método de liofilização os respectivos valores foram 0,9975 e 0,01612. A partir disso, pode-se elaborar um modelo matemático generalizado, apresentando um coeficiente de determinação igual a 0,9888. No caso do leito de espuma, observou-se que à medida que se aumenta a temperatura do ar de secagem, o tempo de secagem diminui e os valores do coeficiente de difusão efetiva aumentam. Porém, a redução no tempo de secagem entre os intervalos de temperatura, diminui com o aumento da mesma. A energia de ativação para a difusão no processo de secagem do soro foi de 26,650 kJ/mol e para todas as avaliações físico-químicas e tecnológicas, a análise de variância apresentou um valor de F significativo (p<0,05), indicando que há pelo menos um contraste entre as médias dos tratamentos que é significativo.

Nôvo meio para isolamento de enterobacteriáceas e diferenciação simultânea das colônias de Proteus e Providência

Um nôvo meio é proposto para o isolamento de enterobacteriáceas. Baseia-se na propriedade de desanimar a fenil-alanina que possuem tôdas as bactérias dos grupos Proteus e Providencia. As colônias dêstes microrganismos apresentam no meio descrito coloração marrom característica. As colônias das outras enterobacteriáceas, fermentadoras rápidas ou lentas e não fermentadoras da lactose, apresentam aspecto diferente.

Diagnóstico ambiental de uma queijaria e proposta de melhorias

O presente trabalho tem como objectivo o diagnóstico ambiental da empresa Lacticinios do Paiva, S.A, a avaliação da água do processo e da ETARI e o estudo da fermentação do soro de queijo com o intuito de produção de bioetanol. No diagnóstico ambiental da empresa, observou-se que 18.227.731 litros de leite usados anualmente geram 5.031 ton/ano de queijo, 7.204 ton/ano de soro de queijo, 74.201 m3/ano de efluente liquido, 14 ton/ano de plástico e 20 ton/ano de cartão. Os principais problemas com necessidade de optimização são a recuperação de água das lavagens, avaliação da produção de biogás no digestor anaeróbio, recuperação do volume de leite que é desperdiçado na produção de queijo fresco de longa duração, avaliação da eficiência energética da empresa, valorização das natas e do soro de queijo. Decidiu-se neste trabalho avaliar a possibilidade de reciclagem das águas de lavagem, avaliar o funcionamento da ETARI face à legislação existente e estudar a possibilidade de valorização do soro de queijo. Na avaliação das águas de processo das lavagens para posterior reciclagem, verifica-se que relativamente ao pH e aos sólidos suspensos não existe problema, podendo encarar-se a hipótese de reciclagem directa. No entanto, no que respeita à carga orgânica das águas de lavagem do sistema de ultrafiltração do queijo fresco de longa duração, constata-se que esta não poderia ser utilizada novamente, uma vez que apresenta valores elevados de CQO. Para a sua reutilização, será necessário remover a CQO, hipótese que se estudou com resultados positivos. Verificou-se que, um tratamento por adsorção em carvão activado precedido de microfiltração, reduz a CQO de forma significativa permitindo admitir a hipótese de reciclagem da água, nomeadamente para as 1ª e 3ª águas de lavagem. As outras águas teriam necessidade de mais tempo de contacto com o carvão activado. No sentido de avaliar o funcionamento da ETARI, foram analisadas várias correntes da mesma, em particular a do efluente final, no que respeita a parâmetros como: pH, Sólidos Suspensos Totais, Carência Química de Oxigénio, Carência Bioquímica de Oxigénio, Turvação, Nitratos, Fósforo Total, Azoto Kjeldalh, Azoto Amoniacal e Cloretos. Observou-se que os valores para o efluente final da ETARI são os seguintes: pH compreendido entre [7,21 – 8,69], SST entre [65,3 – 3110] mg/L, CQO entre [92,5 – 711,5] mg/L, CBO5 entre [58 – 161] mg/L, NO3- entre [10,8 – 106,7] mg/L, fósforo total entre [8,3 – 64,3] mg/L, turvação entre [67,7 – 733,3] FTU e cloretos entre [459,9 – 619,81] mg/L; pode-se dizer que os parâmetros analisados se encontram quase sempre dentro da gama de valores impostos pela Câmara Municipal de Lamego pelo que o efluente pode ser lançado no Colector Municipal de Cambres. Relativamente à fermentação alcoólica do soro de queijo, verifica-se que a levedura Kluyveromyces Marxianus consegue degradar praticamente todo o açúcar presente no permeado produzindo assim uma quantidade razoável de etanol. Quando se utilizou a levedura Saccharomyces Cerevisiae, a produção de etanol foi muito reduzida, como esperado, dado que esta levedura apresenta dificuldades na metabolização da lactose. Constatou-se assim que a melhor levedura para a fermentação do permeado do soro de queijo é a Kluyveromyces Marxianus, estimando-se em 150 mg a produção de etanol por L de soro.

Man-tailored biomimetic sensor of molecularly imprinted materials for the potentiometric measurement of oxytetracycline

A novel biomimetic sensor for the potentiometric transduction of oxytetracycline is presented. The artificial host was imprinted in methacrylic acid and/or acrylamide based polymers. Different amounts of molecularly imprinted and non-imprinted polymers were dispersed in different plasticizing solvents and entrapped in a poly(vinyl chloride) matrix. Only molecularly imprinted based sensors allowed a potentiometric transduction, suggesting the existence of host–guest interactions. These sensors exhibited a near-Nernstian response in steady state evaluations; slopes and detection limits ranged 42–63 mV/decade and 2.5–31.3 µg/mL, respectively. Sensors were independent from the pH of test solutions within 2–5. Good selectivity was observed towards glycine, ciprofloxacin, creatinine, acid nalidixic, sulfadiazine, cysteine, hydroxylamine and lactose. In flowing media, the biomimetic sensors presented good reproducibility (RSD of ±0.7%), fast response, good sensitivity (65 mV/decade), wide linear range (5.0×10−5 to 1.0×10−2 mol/L), low detection limit (19.8 µg/mL), and a stable baseline for a 5×10−3M citrate buffer (pH 2.5) carrier. The sensors were successfully applied to the analysis of drugs and urine. This work confirms the possibility of using molecularly imprinted polymers as ionophores for organic ion recognition in potentiometric transduction.