Production of proteinase A by Saccharomyces cerevisiae in a cell-recycling fermentation system: experiments and computer simulations

Overproduction of proteinase A by recombinant Saccharomyces cerevisiae was investigated by cultivations in a cell-recycling bioreactor. Membrane filtration was used to separate cells from the broth. Recycling ratios and dilution rates were varied and the effect on enzyme production was studied both experimentally and by computer simulations. Experiments and simulations showed that cell mass and product concentration were enhanced by high ratios of recycling. Additional simulations showed that the proteinase A concentration decreased drastically at high dilution rates and the optimal volumetric productivities were at high dilution rates just below washout and at high ratios of recycling. Cell-recycling fermentation gave much higher volumetric productivities and stable product concentrations in contrast to simple continuous fermentation.     

Conscious and anaesthetised Göttingen mini-pigs as an in-vivo model for buccal absorption – pH-dependent absorption of metoprolol from bioadhesive tablets

The potential of buccal mucosa as a site for systemic absorption has attracted increased attention in recent years creating a need for new predictive in-vivo models. The aim of this study was to evaluate anaesthetised and conscious Göttingen mini-pigs as a model for buccal drug absorption by testing pH-dependent absorption of metoprolol from a solid dosage form. Buccal tablets buffered to pH 6.2 and pH 8.9, oral liquid and intravenous injection were tested in four conscious and anaesthetised Göttingen mini-pigs in a non-randomised cross-over study. Blood samples were collected and processed before analysis by ultra-performance liquid chromatography with tandem mass spectrometry detection. An ex-vivo flow retention model was applied to study release and retention of the bioadhesive buccal tablets. The T_max obtained from the two buccal conscious groups (55?±?5 and 35?±?5?min) were significantly different to the buccal anaesthetised groups (120?±?0 and 165?±?15?min) for buccal tablet pH 6.2 and pH 8.9, respectively. Also, the absolute bioavailability from the anaesthetised buccal tablet pH 8.9 (20.7?±?4.0%) had a significant increase compared to all other buccal tablet groups. In conclusion, this study showed a pH-dependent absolute bioavailability of metoprolol when administrated as bioadhesive buccal tablets to anaesthetised mini-pigs. The anaesthesia was found to delay the time to reach maximal plasma concentration of metoprolol as compared to the conscious pig model when administrated as buccal tablets.     

Mucoadhesive Electrospun Nanofiber-Based Hybrid System with Controlled and Unidirectional Release of Desmopressin

The sublingual mucosa is an attractive route for drug delivery, although challenged by a continuous flow of saliva that leads to a loss of drug by swallowing. It is of great benefit that drugs absorbed across the sublingual mucosa avoid exposure to the harsh environment of the gastro-intestinal lumen; this is especially beneficial for drugs of low physicochemical stability such as therapeutic peptides. In this study, a two-layered hybrid drug delivery system was developed for the sublingual delivery of the therapeutic peptide desmopressin. It consisted of peptide-loaded mucoadhesive electrospun chitosan/polyethylene oxide-based nanofibers (mean diameter of 183 ± 20 nm) and a saliva-repelling backing film to promote unidirectional release towards the mucosa. Desmopressin was released from the nanofiber-based hybrid system (approximately 80% of the loaded peptide was released within 45 min) in a unidirectional manner in vitro. Importantly, the nanofiber–film hybrid system protected the peptide from wash-out, as demonstrated in an ex vivo flow retention model with porcine sublingual mucosal tissue. Approximately 90% of the loaded desmopressin was retained at the surface of the ex vivo porcine sublingual mucosa after 15 min of exposure to flow rates representing salivary flow.     

Significant histamine formation in tuna (Thunnus albacares) at 2 degrees C--effect of vacuum- and modified atmosphere-packaging on psychrotolerant bacteria

Occurrence and importance of psychrotolerant histamine producing bacteria in chilled fresh tuna were demonstrated in the present study. The objective was to evaluate microbial formation of histamine and biogenic amines in chilled fresh tuna from the Indian Ocean and stored either vacuum-packed (VP) or modified atmosphere-packed (MAP). Firstly, biogenic amines and the dominating microbiota were determined in VP tuna involved in an outbreak of histamine fish poisoning in Denmark. Secondly, the microbiota of fresh MAP tuna was evaluated at the time of processing in Sri Lanka and chemical, microbial and sensory changes were evaluated during storage at 1-3 degrees C. To explain the results obtained with naturally contaminated tuna the effect of VP and MAP on biogenic amine formation by psychrotolerant bacteria was evaluated in challenge tests at 2 degrees C and 10 degrees C. The VP tuna that caused histamine fish poisoning had a histamine concentration of >7000 mg/kg and this high concentration was most likely produced by psychrotolerant Morganella morganii-like bacteria or by Photobacterium phosphoreum. Similar psychrotolerant M. morganii-like bacteria dominated the spoilage microbiota of fresh MAP tuna with 60% CO2/40% N2 and formed >5000 mg/kg of histamine after 24 days at 1.7 degrees C. These psychrotolerant bacteria were biochemically similar to M. morganii subsp. morganii and their 16S rDNA (1495 bp) showed >98% sequence similarity to the type strain of this species. Toxic concentrations of histamine were produced at 2.1 degrees C in inoculated VP tuna by both the psychrotolerant M. morganii-like bacteria (7400+/-1050 mg/kg) and P. phosphoreum (4250+/-2050 mg/kg). Interestingly, MAP with 40% CO2/60% O2, in challenge tests, had a strong inhibitory effect on growth and histamine formation by both the psychrotolerant M. morganii-like bacteria and P. phosphoreum. In agreement with this, no formation of histamine was found in naturally contaminated fresh MAP tuna with 40% CO2/60% O2 during 28 days of storage at 1.0 degrees C. To reduce current problems with histamine fish poisoning due to VP tuna it is suggested, for lean tuna loins, to replace vacuum packaging with MAP containing approximately 40% CO2 and approximately 60% O2.     

A home-based exercise program for nondisabled older adults

OBJECTIVES: This paper describes a videotaped, home-based, strength training program, titled Strong-for-Life and reports on its effectiveness in improving muscle strength, psychological well-being, and health status in a sample of older persons. DESIGN AND SETTING: We enrolled 102 nondisabled, community-dwelling older people aged 66 to 87, identified from the Medicare beneficiary list, into a randomized, controlled trial. MEASUREMENTS: Effectiveness was based on change in isokinetic upper and lower extremity muscle strength, psychologic well-being, and health status. RESULTS: Results revealed several statistically significant short-term benefits after 12 to 15 weeks of exercise, especially for men. Younger older adults demonstrated a 10% improvement in knee extensor strength relative to control subjects. Older male exercisers achieved significant differences relative to controls in perceived anger, tension, and overall social functioning. Male exercisers, in general, achieved significant improvement in perceived vigor. Women did not report psychological benefits following participation in the program. CONCLUSION: Study results reveal that the Strong for Life program, designed to be widely disseminated to the nondisabled older population, has many short-term positive benefits.     

Health status assessment in the occupational health setting

This article discusses the conceptual roots of measuring health- related outcomes based on a biopsychosocial model of health. The selection and applicability of instruments for measurement of health status are discussed and suggestions are made concerning how health-status instruments may be used in the occupational- health setting. The potential value of measuring health status in the occupation-health setting is explored with a brief review of current literature and an example from one investigation in which measurements of health status were used to examine outcomes of common musculoskeletal impairments.     

Changes in liquid‐holding capacity,water distribution and microstructure during chill storage of smoked salmon

BACKGROUND: During recent years, increases in liquid loss and soft texture in cold smoked salmon have been reported, possibly due to increased lipid content and a change in muscle structure. These problems can lead to downgrading of the product. This study aimed to investigate the influence of raw material composition and chill storage on quality parameters of cold smoked salmon. RESULTS: Initial smoked fish had a higher liquid‐holding capacity (LHC) than samples stored for 20 days. Large fish lost more liquid than small fish. The difference in LHC was consistent with a change in water distribution, which could indicate denaturation of muscle protein. Studies of the microstructure showed the influence of both processing and chill storage. An indication of lipid released from the cells was seen after 20 days of chill storage, which could be related to the reduced LHC at that time. CONCLUSION: Both raw material composition and chill storage affected the quality parameters of smoked salmon. This study has improved knowledge about the relationships between muscle structure, liquid‐holding properties and water distribution in smoked salmon. Copyright © 2007 Society of Chemical Industry     

Freshwater fish as a dietary source of vitamin A in Cambodia

Vitamin A deficiency is a public health problem among children and women. Common Cambodian fish species were sampled and screened for vitamin A content. Contents of vitamin A-active compounds (all-trans retinol, all-trans dehydroretinol, 13-cis retinol, 13-cis dehydroretinol and β-carotene) were determined by high-performance liquid chromatography in samples of raw, whole fish from 29 fish species and in raw, edible parts from 24 species. Replicate samples were analysed in seven selected species. Two species, Parachela siamensis and Rasbora tornieri had very high vitamin A contents >1500 RAE/100 g raw, whole fish, and six species (Barbodes altus, Barbodes gonionatus, Dermogenys pusilla, Puntioplites proctozysron and Thynnichthys thynnoides) had high contents of 500–1500 RAE/100 g raw, whole fish. Two species, Puntioplites proctozysron and Thynnichthys thynnoides had high vitamin A contents in raw, edible parts, after employing traditional cleaning practices. (RAE: The amount of vitamin A active compounds in food is expressed as retinol activity equivalents (RAE), defined as the bioefficacy relative to all-trans-retinol [ West, C. E., & Eilander, A. (2002). Consequences of revised estimates of carotenoid bioefficacy for the control of vitamin A deficiency in developing countries. Journal of Nutrition, 132, 2920S–2926S]. Dehydroretinoids (vitamin A₂) are not converted to all-trans-retinol but have similar metabolic functions. In this paper, RAE refers to the functional bioefficacy as defined by Brouwer et al. [ Brouwer, I. A., Dusseldorp, M. V., West, C. E., & Steegers-Theunissen, R. P. M. (2001). Bioavailability and bioefficacy of folate and folic acid in man. Nutrition Research Review, 14, 267–293]).     

Concepts for measuring maintenance performance and methods for analysing competing failure modes

Measurement of maintenance performance is done on the basis of component history data in which service sojourns are distinguished according to whether they terminate in corrective or preventive maintenance. From the viewpoint of data analysis, corrective and preventive maintenance constitute competing failure nudes. This article examines ways to assess maintenance performance without introducing statistical assumptions, then introduces a plausible statistical model for describing the interaction of preventive and corrective maintenance, and finally illustrates these with examples from the Nordic TUD data system.     

Oxidative stress, genotoxicity, and vascular cell adhesion molecule expression in cells exposed to particulate matter from combustion of conventional diesel and methyl ester biodiesel blends

Our aim was to compare hazards of particles from combustion of biodiesel blends and conventional diesel (D(100)) in old and improved engines. We determined DNA damage in A549 cells, mRNA levels of CCL2 and IL8 in THP-1 cells, and expression of ICAM-1 and VCAM-1 in human umbilical cord endothelial cells (HUVECs). Viability and production of reactive oxygen species (ROS) were investigated in all cell types. We collected particles from combustion of D(100) and 20% (w/w) blends of animal fat or rapeseed oil methyl esters in light-duty vehicle engines complying with Euro2 or Euro4 standards. Particles emitted from the Euro4 engine were smaller in size and more potent than particles emitted from the Euro2 engine with respect to ROS production and DNA damage, but similarly potent concerning cytokine mRNA expression. Particles emitted from combustion of biodiesel blends were larger in size, and less or equally potent than particles emitted from combustion of D(100) concerning ROS production, DNA damage and mRNA of CCL2 and IL8. ICAM-1 and VCAM-1 expression in HUVECs was only increased by D(100) particles from the Euro4 engine. This suggests that particle emissions from biodiesel in equal mass concentration are less toxic than conventional diesel.