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Alcohol in combination with malnutrition causes increased liver fibrosis in rats.

Rats were malnourished for 12 months with a highly inadequate fat-rich, calorie-sufficient but otherwise poly-deficient liquid diet composed of mashed potatoes with mayonnaise, comparable with the nutritional intake of many chronic alcoholics. When alcohol was incorporated into this diet, administered as whisky in drinking water available ad libitum, the livers of all eight rats showed increased fibrosis and cirrhosis as compared to the livers of the eight non-alcohol-treated, isocalorically fed, paired control rats. Alcohol-treated rats developed fibrosis and cirrhosis on a dietary fat content of 38% of total caloric intake and low blood alcohol levels, ranging from 50 to 126 mg/dl, due to gradual intake over the day and to low absolute intake (mean 11.9 +/- 0.6 g/kg per day). None of the rats died spontaneously. Malnutrition is likely to be an important factor in the development of the fibrosis of alcoholic liver disease, and this rat model may be used to study aspects of the pathogenesis.

Alanine Transaminase↗

The treatment of paediatric burns using topical papaya.

Due to the limited resources for the management of burns in most regions of Africa there is a significant role for many aspects of traditional African medicine. The active component of many traditional preparations is often of plant origin and more than 25 plants have been described as useful in relations to burns and wound healing. Carica papaya is currently used in The Gambia at the Royal Victoria Hospital, Banjul in the Paediatric Unit as the major component of burns dressings, where it is well tolerated by the children. Cheap and widely available, the pulp of the papaya fruit is mashed and applied daily to full thickness and infected burns. It appears to be effective in desloughing necrotic tissue, preventing burn wound infection, and providing a granulating wound suitable for the application of a split thickness skin graft. Possible mechanisms of action include the activity of proteolytic enzymes chymopapain and papain, as well as an antimicrobial activity, although further studies are required.

Administration, Topical↗

Effects of neonatal cocaine treatment and gender on opioid agonist-stimulated [(35)S]GTP gamma S binding in the striatum and nucleus accumbens.

Prenatal cocaine exposure increases mu-opioid receptor binding in dopaminergic terminal areas and enhances behavioral responsiveness to mu-opioid agonists. We investigated the influence of early postnatal cocaine treatment on in vitro mu- and delta-opioid receptor activation in male and female weanling rats. Pups received subcutaneous injections of either 20 mg/kg cocaine HCl or saline once daily on postnatal days 1 through 5. On postnatal day 25, animals were decapitated and their brains were removed and frozen for later sectioning. Opioid receptor activation was assessed in the striatum and the shell of the nucleus accumbens by autoradiographic analysis of agonist-stimulated [(35)S]GTP gamma S binding. Brain sections were incubated in the presence of [(35)S]GTP gamma S, GDP, and either the mu-opioid agonist [D-Ala(2)-N-MePhe(4)-Gly(5)-ol]enkephalin (DAMGO) or the delta-opioid agonist D-Pen(2)-D-Pen(5)-enkephalin (DPDPE). Baseline binding was assessed in the absence of agonist, and nonspecific binding was determined by the addition of unlabeled GTP gamma S. Film images were quantified using brain mash-calibrated [(14)C] standards. Neonatal cocaine treatment had no effect on either baseline or agonist-stimulated [(35)S]GTP gamma S binding. However, males exhibited significantly greater activation than females of delta-opioid receptors in both striatum and accumbens shell, regardless of neonatal treatment. These findings indicate a gender difference in delta-opioid receptor function that could mediate behavioral differences in response to opioid agonists.

Analgesics, Opioid↗

New use of broomcorn millets for production of granular cultures of aphid-pathogenic fungus Pandora neoaphidis for high sporulation potential and infectivity to Myzus persicae.

Glutinous broomcorn millets from the crop Panicum miliaceum were first used as substrate to produce granular cultures of Pandora neoaphidis, an obligate fungal pathogen specific to aphids. Carrying a water content of 36.5% after being steamed in a regular autoclaving procedure, millet grains of each 15 g (dry weight) in a 100-ml flask were mixed with 3 ml modified Sabouraud dextrose broth containing half a mashed colony of P. neoaphidis grown on egg yolk milk agar and then incubated at 20 degrees C and a light/dark cycle of 12 h/12 h for 21 days. Based on individually monitoring conidial production potential of 20 millet grains sampled from an arbitrarily taken flask at 3-day intervals, the millet cultures incubated for 6-15 days were capable of producing 16.8-23.4 x 10(4) conidia per millet grain with conidial ejection lasting for up to 6 days. The cultured millet grains individually produced significantly more conidia than apterous adults of Myzus persicae killed by P. neoaphidis (8.4 x 10(4) conidia per cadaver) and sporulated twice longer. The modeling of time-dose-mortality data from bioassays on M. persicae apterae exposed to conidial showers from the cultured millet grains and the mycelial mats produced in liquid culture resulted in similar estimates of LC(50) (millets: 21.4, 7.3, and 4.9 conidia mm(-2) on days 5-7 after exposure; mycelial mats: 22.1, 10.6, and 7.7 conidia mm(-2)) although the LT(50) estimated at a given conidial concentration was slightly smaller for the millet cultures than for the mycelial mats. This indicates that the millet grains cultured with P. neoaphidis produced conidia as infective as or slightly more infective to M. persicae than those from the mycelial mats. Based on the sporulation potential, infectivity, and ease and cost of the millet cultures, the method developed in this study highly improved in vitro cultures of P. neoaphidis and may adapt to culturing other entomophthoralean fungi for microbial control of insect pests.

Animals↗

Distribution of Salmonella contamination in ten animal feedmills.

Detailed sampling of spillage and dust from milling equipment was carried out in nine animal feedmills, three of which were sampled twice. The salmonella isolation rate ranged from 1.1% to 41.7% of the samples and the most contaminated mills were those where the inside of the cooling systems for pellet or mash had been colonised by salmonella. A wide range of salmonella serotypes were isolated which included Salmonella typhimurium and S. enteritidis. Limited sampling every two weeks for an 18-month period in another animal feedmill showed marked variation in the contamination rate of samples and range of salmonella serotypes found. Contamination of ingredient intake pits and outloading gantries for finished products by wild bird droppings containing salmonella was also found in four mills.

Animal Feed↗

Monitoring and survival of Lactobacillus paracasei LTH 2579 in food and the human intestinal tract.

A PCR-based detection system specific for Lactobacillus paracasei LTH 2579 was developed and applied to follow the fate of the strain in complex ecosystems. This strain was isolated from fruit mash and was characterised as being highly resistant to low pH and bile at concentrations as they occur in the human digestive tract. The application of the subtraction hybridisation technique permitted to identify a 235 bp chromosomal DNA fragment of strain LTH 2579. Based on this target sequence a specific PCR system was developed and combined with the species-specific PCR system for L. paracasei. This combination of PCR based detection systems was successfully applied to monitor L. paracasei LTH 2579 in fermented sausages which were inoculated with this strain (2.0 x 10(7) CFU/g) together with the strongly competitive L. sakei LTH 681 (1.0 x 10(6) CFU/g). At the time of consumption of the sausages the respective counts were 1.8 x 10(7) and 1.4 x 10(8) CFU/g. After consumption of the sausages by three volunteers L. paracasei LTH 2579 was recovered from fecal samples. The counts determined for the strain ranged between 1.2 x 10(7) and 1.5 x 10(8) CFU/g of feces. The fortuitous lactobacilli constituted a share of 5-12% of the lactobacilli in the fecal flora.

DNA Primers↗

Feeding the disabled child.

Feeding difficulty and malnutrition are common in disabled children. Intake may be reduced because of anorexia, chewing and swallowing difficulties, or vomiting. Feeding is often time consuming, unpleasant, and may result in aspiration. Malnutrition may result in impaired growth and neurodevelopment, and impaired cardiorespiratory, gastrointestinal, and immune functions. Multidisciplinary assessment is recommended and should include a feeding history, oral-motor examination, and nutritional assessment. The energy requirements of most disabled children are less than those for a normal child of the same age but may be increased by spasticity, athetosis, convulsions, and recurrent infections. Micronutrient deficiencies may occur even in children receiving nutritionally complete feeds if the volume is reduced because of low energy requirements. Oral intake may be improved by a change of posture, special seating, feeding equipment, oral desensitization, mashing or pureeing of lumpy food, thickening of liquids, use of calorie supplements, and treatment of reflux/esophagitis. Non-oral feeding should be considered when oral feeding is unsafe, not enjoyable, inadequate, or very time consuming. Long-term support requires a gastrostomy. This is less obtrusive than a nasogastric tube, less likely to become displaced, less traumatic, and is associated with improved quality of life, but is also associated with significant morbidity. If there is symptomatic reflux a fundoplication may be required, but this is associated with significant mortality and substantial morbidity.

Child↗

Differentiation of embryonic stem cell-derived dopaminergic neurons is enhanced by survival-promoting factors.

Here, we describe the generation of viable and dopamine-producing neurons derived from pluripotent mouse embryonic stem cells. Neurotrophic factors in combination with survival-promoting factors, such as interleukin-1beta, glial cell line-derived neurotrophic factor, neurturin, transforming growth factor-beta(3) and dibutyryl-cyclic AMP, significantly enhanced Nurr1 and tyrosine hydroxylase (TH) mRNA levels, whereas En-1, mash-1 and dopamine-2-receptor mRNA levels were not upregulated. In parallel, mRNA levels of the anti-apoptotic gene bcl-2 were found to be upregulated at terminal stages. Double immunofluorescence analysis revealed increased numbers of TH- and dopamine transporter-, but not gamma-aminobutyric acid- and serotonin-positive neurons in relation to synaptophysin-labeled cells by survival-promoting factors. Moreover, high-performance liquid chromatography analysis showed detectable levels of intracellular dopamine. We conclude that survival-promoting factors enhance differentiation, survival and maintenance of dopaminergic neurons derived from embryonic stem cells.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Region-specific expression of the helix-loop-helix gene BETA3 in developing and adult brains.

Members of the basic helix-loop-helix (bHLH) transcription factor family are crucial regulators of neuronal cell generation and cell fate. A number of bHLH genes are expressed in the developing cerebral cortex, including MASH-1, neurogenin2 and NeuroD implying the existence of a regulatory and possibly redundant network of family members. BETA3 is a novel member originally cloned from pancreatic cells but we report here highly restricted expression patterns in developing forebrain structures that are highly stage-specific. We show that BETA3 mRNA is found in both neocortex and archicortex, mainly in cells that have reached their migratory destinations but is largely absent from proliferative zones. These expression data would suggest that BETA3 function is linked to the establishment rather than the initiation of neuronal fates.

Animals↗

A two-compartment model for zinc in humans.

The oral absorption of zinc, from a test meal of minced beef, mashed potatoes and peas, have been measured in 19 healthy adults using the radiotracer 65Zn. The oral absorption, expressed as a percentage of the administered dose, was 20 +/- 5% (mean +/- 1 SD) in good agreement with previous results. In a subset of 9 subjects, tracer retention in whole body and whole blood was followed out to one year. The data were fitted to a simple two compartment model yielding total body zinc (TBZn), the zinc content in each of the 2 compartments and zinc turnover. The TBZn values ranged from 15.5 to 35.9 mmol while zinc turnover ranged from 0.043 to 0.073 mmol/d in keeping with results reported for significantly more complicated compartmental models applied to more comprehensive 65Zn tracer data sets. Additionally, TBZn correlated well with total body potassium, a measure of lean body mass, measured by whole body counting of the naturally-occurring potassium radioisotope, 40K. The zinc content of the more rapidly turning over compartment ranged from 3.2 to 5.6 mmol in reasonable agreement with exchangeable zinc pool estimations reported for short term studies using stable zinc isotopes. Therefore, the simple dataset and model employed in the present study yielded information on the short- and long-term behaviour of zinc compatible with both more complex radiotracer studies and analytically more demanding stable isotope studies.

Administration, Oral↗

Analysis of the in vivo nitrosation capacity of the larvae used in the wing somatic mutation and recombination test of Drosophila melanogaster.

The in vivo nitrosation capacity of third-instar larvae of Drosophila melanogaster was assessed using the wing somatic mutation and recombination test (SMART). Larvate derived from two different crosses, the standard cross (ST) and the high bioactivation cross (HB) both involving the recessive wing cell markers multiple wing hairs (mwh) and flare (flr3), were used. The HB cross is characterised by an increased cytochrome P450-dependent bioactivation capacity for promutagens and procarcinogens. The larvae were treated either with methyl urea, sodium nitrite or its combination. N-Nitrosomethylurea was used as a positive control. The wings of the resulting flies were analysed for the occurrence of mutant spots produced by various types of mutational events or by mitotic recombination. Methyl urea is negative in the ST and the HB cross, whereas sodium nitrite is weakly genotoxic in both crosses. However, the combination of both compounds produces highly increased frequencies of mutations and recombinations predominantly in the HB cross. The genotoxic effects produced by the combined treatments were considerably increased when mashed potatoes or an agar-yeast medium were used for the treatment instead of the standard instant medium. Treatment of larvae with the mixture resulting from the in vitro reaction of nitrosation precursors also resulted in high frequencies of induced spots comparable to those recorded with the potent genotoxin N-nitrosomethylurea. Further experiments showed that the genotoxic effect resulting from the in vivo exposure to nitrosation precursors can be reduced by co-treatment with catechin, a known nitrosation inhibitor. The present study demonstrates that the wing spot test is well suited for the determination of genotoxicity produced by in vivo nitrosation processes and for the study of their modulation by individual compounds or dietary complex mixtures.

Animals↗

Fuzzy control of bioprocess in Japan.

Process control of bioprocess has been carried out by the judgment of the experts, who are the skilled operators and have lots of experiences for the control of the process. In almost all cases, those experiences are described linguistic IF-THEN rules. Fussy inference is one of the powerful tools to incorporate the linguistic rules to the computer for process control. Fuzzy control are divided into two types; one is the direct fuzzy control of process variables such as sugar feed rate in fed-batch culture and fermentation temperature in batch operation. The other is the indirect control of bioprocess, in which at first the phase recognition is carried out by fuzzy inference and the control strategies constructed in each phase are used for the control of process variables. In Japan, the fuzzy control has already been applied to practical industrial productions, such as pravastatin precursor, vitamin B2, and Japanese sake mashing process. In this review, these industrial approaches of fuzzy control are introduced.

Alcoholic Beverages↗

Production and some properties of salt-tolerant beta-xylosidases from a shoyu koji mold, Aspergillus oryzae in solid and liquid cultures.

Beta-xylosidase production from a shoyu (soy sauce) koji mold, Aspergillus oryzae HL15, cultured in solid and liquid media was examined and some properties of the enzymes were studied. Three beta-xylosidases (Xy11, Xy12 and Xy13) were easily extracted with 0.5% NaCl from a solid medium and purified homogeneously on SDS-PAGE by chromatography. On the other hand, in a liquid medium, A. oryzae HL15 produced mainly cell-wall-bound beta-xylosidases which could not be extracted with 0.5% NaCl or any detergent. Cell-wall-bound beta-xylosidases, Xy11-CB and Xy12-CB, were liberated by digestion of mycelia with Yatalase and purified to a homogeneous state on SDS-PAGE by HPLC column chromatography. Four beta-xylosidases (Xy11, Xy12, Xy11-CB and Xy12-CB) exhibited not only high activity at high NaCl concentrations, but also similar properties; on the other hand, Xy13 differed in terms of thermostability and halophilic properties. The salt tolerance of beta-xylosidases in A. oryzae suggests that these enzymes are highly active and involved in releasing xylose in shoyu moromi mash.

Journal Article↗

Fuzzy control of bioprocess.

Bioprocesses have been operated according to the judgment of experts who are skilled operators and have long experience. In almost all cases, these experiences are described by linguistic IF-THEN rules. Fuzzy inference is one of the powerful tools to incorporate linguistic rules into computational algorithms for application to process control. Fuzzy control is categorized into two types: the direct fuzzy control of process variables such as sugar feed rate in fed-batch culture and broth temperature in batch operation, and the indirect control of bioprocesses in which the phase recognition is first done by fuzzy inference using process variables such as DO, sugar concentration, pH and so on and then the control strategies having been constructed in each phase are used for the process operation. Fuzzy control has already been applied to practical industrial processes, such as production of a pravastatin precursor and vitamin B2, and the Japanese sake mashing process. In this review, these industrial practices of fuzzy control are introduced.

Journal Article↗

Molecular breeding of Aspergillus kawachii overproducing cellulase and its application to brewing barley shochu.

In order to improve fermentation of barley without addition of commercial cellulase, a white koji mold, Aspergillus kawachii IFO4308, was transformed with the egl1 gene encoding endoglucanase I (EGI) of Trichoderma viride and the endogenous cekA gene encoding endoglucanase (CekA). Transformants with egl1 under the control of the strong glaA promoter produced EGI in both submerged and solid-state cultures. However, the EGI produced in solid-state culture was unstable due to the acidic condition of this culture. A transformant N10 with two additional copies of the cekA gene exhibited endoglucanase activities against carboxymethyl-cellulose, which are 21- and 1.8-fold higher than that of the wild-type (wt) strain when the cells were cultivated in submerged and solid-state cultures, respectively. Cultivation of strain N10 in steamed barley for preparing koji followed by fermentation with Saccharomyces cerevisiae resulted in improved fermentation assessed based on higher productions of ethanol, amino acids, and organic acids, the reduction of residual sugar, and the low viscosity of barley mash. The overall fermentation result for the transformant carrying cekA was comparable with that for the wt strain using commercial cellulase. These results demonstrate that acquisition of only two-fold CekA activity by A. kawachii in the solid-state culture allows us to improve the brewing of barley shochu.

Journal Article↗

Cloning and analysis of the AWA1 gene of a nonfoaming mutant of a sake yeast.

Almost all sake yeasts form a thick foam layer on sake mash during fermentation. To reduce the amount of foam, nonfoaming mutants were bred from foam-forming sake yeasts. To elucidate the mechanism of this foam formation, we have cloned a gene from a foam-forming sake yeast that confers foam-forming ability to a nonfoaming mutant. This gene, named AWA1, encodes a glycosylphosphatidylinositol (GPI) anchor protein that is localized to the cell wall and is required for cell surface hydrophobicity. In this paper, we describe the genomic analysis of the AWA1 gene in a nonfoaming mutant strain K701 derived from a foam-forming sake yeast strain K7. K701-AWA1 was cloned in a cosmid and its sequence was compared with that of K7-AWA1. Although the 5' half of K701-AWA1 was identical to that of K7-AWA1, the 3' half of K701-AWA1 was different from that of K7-AWA1, resulting in a loss of the C-terminal hydrophobic sequence of Awa1p. Since this sequence is considered to be required for the anchoring of Awa1p to the cell wall, K7-Awa1p could not confer both cell surface hydrophobicity and foam-forming ability to strain K701 cells. Since the change found in K701-AWA1 was not a point mutation but a larger scale event, we analyzed chromosome rearrangement by pulsed-field gel electrophoresis Southern blot analyses. The results suggest that the left subtelomeric region of chromosome IX in strain K7 was translocated to the AWA1 gene in chromosome XV by a nonreciprocal recombination.

Journal Article↗

Effect of Ganoderma lucidum on the quality and functionality of Korean traditional rice wine, yakju.

The goal of this study was to develop a high value Korean traditional rice wine possessing the pharmaceutical functionality of Ganoderma lucidum. The effects of the fruiting body of G. lucidum on the alcohol fermentation of Korean traditional rice wine, yakju, were investigated. Optimal fermentation conditions for the preparation of G. lucidum-yakju consisted of the koji added at 15% and a fermentation period of 15 d at 25 degrees C. The effects of the amount of G. lucidum added on the acceptability and functionality of G. lucidum-yakju were investigated. G. lucidum GL-1 yakju brewed by adding 0.1% G. lucidum into the mash showed the best acceptability and its angiotensin I-converting enzyme (ACE) inhibitory activity and SOD-like activity were 63% and 42%, respectively, both of which are higher than those of yakju. The high ACE inhibitory activity of G. lucidum GL-1 yakju was found to result from ganoderic acid K in G. lucidum on the basis of physical and spectral data. However, the fibrinolytic activity and antioxidant activity of G. lucidum GL-1 yakju were very low, while tyrosinase inhibitory activity was not determined. From these results, G. lucidum GL-1 yakju may become a new functional Korean traditional rice wine with antihypertensive properties.

Journal Article↗

Variations in the apparent nutrient content of foods: a study of sampling error.

1. Successive portions of boiled and mashed potatoes, roast pork, cooked, freeze-dried peas and ice-cream were taken in order to determine the total (sampling and experimental) error involved in the 'duplicate analysis' method of dietary survey. 2. These samples were analysed for water, nitrogen, fat, iron and energy. 3. The experimental error of the methods was also studied separately. 4. The sampling and experimental errors ranged from 0-3% for water in potatoes, to 23% for Fe in ice-cream. Sampling error was significantly greater than experimental error for all nutrients studied. 5. It is concluded that in reporting the results from dietary surveys, it is necessary to take account of the erros inherent even in the most precise methods.

Analysis of Variance↗