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Induction and characterization of an unusual alpha-D-galactosidase from Talaromyces flavus.

An extracellular alpha-d-galactosidase from Talaromyces flavus CCF 2686 with extremely broad and unusual acceptor specificity is produced exclusively in the presence of the specific inducer--6-deoxy-D-glucose (quinovose). The procedure for the preparation of this very expensive substance has been modified and optimized. Surprisingly, any of other common alpha-D-galactosidase inducers or substrates, e.g., D-galactose, melibiose and raffinose, did not stimulate its production. The crude alpha-D-galactosidase preparation was purified by anion-exchange chromatography and three isoenzymes with different substrate specificities were identified. The main isoenzyme (alphaGal1) was further purified by cation-exchange chromatography and fully characterized. When compared with other alpha-galactosidases and also with other isoenzymes produced by T. flavus, it showed a markedly different regioselectivity and also negligible hydrolytic activity towards melibiose. Moreover, it was active on polymeric substrates (locust bean gum, guar gum) and significantly inhibited by alpha-D-galactopyranosyl azide, D-galactose, D-xylose, melibiose, methyl alpha- and beta-D-galactopyranoside and lactose.

Deoxyglucose↗

The protein digestibility-corrected amino acid score method overestimates quality of proteins containing antinutritional factors and of poorly digestible proteins supplemented with limiting amino acids in rats.

The validity of the protein digestibility-corrected amino acid score (PDCAAS) method in predicting the quality of fourteen protein products was compared with the commonly used protein quality methods, protein efficiency ratio (RER) and net protein ratio (NPR). A rat growth and balance study was conducted to determine protein digestibility and quality of the animal and vegetable protein products by the PER and NPR methods. Amino acid compositions of the products were also determined, and PDCAAS were calculated using a rat and a human pattern of amino acid requirements. Compared to the biological methods, the scoring method overestimated protein quality of mustard flour [PDCAAS of 84-92% vs. relative PER (RPER) or relative NPR (RNPR) of 0], raw black beans (PDCAAS of 45-72% vs. RPER or RNPR of 0), alkaline-treated lactalbumin and soybean protein isolate (PDCAAS of 44-67% vs. RPER or RNPR of 0) and heated skim milk (PDCAAS of 29-31% vs. RPER and RNPR of 0-5%). The scoring method also overestimated the protein quality of zein (true protein digestibility of 63%) supplemented with Lys, Met, Thr and Trp (PDCAAS of 63-71% vs. RPER and RNPR of 3-44%). These data demonstrate that the PDCAAS method is inappropriate for predicting protein quality of those protein sources which may contain naturally occurring growth-depressing factors or antinutritional factors formed during alkaline and/or heat processing.

Amino Acids↗

Nut consumption, lipids, and risk of a coronary event.

In the past, many have avoided nuts because of their high fat content. The Dietary Approaches to Stop Hypertension (DASH) diet, however, recommends regular consumption of this food along with seeds and dried beans (4-5 servings per week) as part of a diet to control hypertension. Nuts are nutrient-dense and most of their fat is unsaturated. They are also perhaps the best natural source of vitamin E and are relatively concentrated repositories of dietary fiber, magnesium, potassium, and arginine, the dietary precursor of nitric oxide. Human feeding studies have demonstrated reductions of 8-12% in low-density lipoprotein (LDL) cholesterol when almonds and walnuts are substituted for more traditional fats. Other studies show that macadamias and hazelnuts appear at least as beneficial as fats in commonly recommended diets. Whether consuming modest quantities of nuts daily may promote weight gain is not known with certainty, but preliminary data suggest that this is unlikely. Four of the best and largest cohort studies in nutritional epidemiology have now reported that eating nuts frequently is associated with a decreased risk of coronary heart disease of the order of 30-50%. The findings are very consistent in subgroup analyses and unlikely to be due to confounding. Possible mechanisms include reduction in LDL cholesterol, the antioxidant actions of vitamin E, and the effects on the endothelium and platelet function of higher levels of nitric oxide. Although nuts may account for a relatively small percentage of dietary calories, the potential interacting effects of these factors on disease risk may be considerable.

Cholesterol, LDL↗

Exoglycosidase purity and linkage specificity: assessment using oligosaccharide substrates and high-pH anion-exchange chromatography with pulsed amperometric detection.

Simplified HPLC protocols to determine the activity and linkage specificity and to detect the most commonly-encountered contaminants in available exoglycosidase preparations (Jacob and Scudder, Methods Enzymol., 230, 280-300, 1994) were developed. Monosaccharides and oligosaccharides were analyzed in a single chromatographic step using high-pH anion-exchange chromatography with pulsed amperometric detection. All analyses were performed with underivatized oligosaccharide substrates and by direct injection of unprocessed, diluted enzyme digests into the chromatograph. The sialidase from Newcastle disease virus was found to release both alpha (2-->3)- and alpha (2-->6)-linked Neu5Ac from a triantennary, lactosamine-type oligosaccharide. The activity of alpha-galactosidase from green coffee beans was assayed using Gal alpha(1-->3)[Fuc-alpha(1ar2)]Gal by detection of Gal and Fuc alpha(1-->3)Gal. The linkage specificities of beta-galactosidases from Streptococcus pneumoniae and bovine testis were assessed using Gal beta(1-->3 or 4)GlcNAc beta(1-->3)beta(1-->4)Glc as substrates. Contaminating beta-N-acetylhexosaminidase activity in the beta-galactosidase preparation was assayed using an agalactobiantennary oligosaccharide. The alpha(1-->3 or 4) linkage specificity of fucosidase III from almond meal was confirmed (Scudder et al., J. Biol. Chem. 265, 16472-16477, 1990) by its inactivity against a biantennary oligosaccharide with all Fuc residues linked alpha(1-->6). An alpha-fucosidase from chicken liver was found to cleave alpha(1-->2,3 or 6)-linked Fuc residues from oligosaccharides. The activity of jack bean (Canavalia ensiformis) alpha-mannosidase was assayed with a relatively resistant substrate, Man alpha(1-->3)- Man beta(1-->4)GlcNAc. A GlcNAc beta(1-->4)-terminated triantennary oligosaccharide was used to assay for contaminating beta-N-acetylhexosaminidase activity in alpha-mannosidase preparations and to determine the linkage and branch specificity of beta-N-acetylhexosaminidase at different enzyme concentrations.

Animals↗

Salmonellosis outbreak on transatlantic flights; foodborne illness on aircraft: 1947-1984.

In March 1984, 186 cases of gastroenteritis due to Salmonella enteritidis were reported after 29 flights to the United States on an international airline. An estimated 2,747 passengers on flights to the United States were affected. Illness was associated with flying supersonic or first class (odds ratio = 15, p less than 0.001). Eating food from the first-class menu was associated with illness (p = 0.09), and eating a tourist-class entree was protective (p less than 0.01). In 23 reported outbreaks of foodborne illness on aircraft, Salmonella has been the most common pathogen (seven outbreaks), followed by Staphylococcus (five outbreaks), and Vibrio species (five outbreaks). Outbreaks are most often the result of an improper temperature for preparation or for holding food in the flight kitchens. Serving the flight crew meals from one kitchen carries the risk that the entire crew will become ill.

Aircraft↗

Foodborne disease outbreaks in nursing homes, 1975 through 1987.

OBJECTIVE: --To describe the epidemiology of foodborne disease outbreaks in nursing homes and to identify where preventive efforts might be focused. DATA SOURCES: --Reports by state and local health departments of foodborne disease outbreaks occurring from January 1, 1975, through December 31, 1987. STUDY SELECTION: --Foodborne disease outbreaks reported to the Centers for Disease Control, Atlanta, Ga, on standard investigation forms. DATA EXTRACTION: --Each foodborne disease outbreak report was examined by an epidemiologist or statistician. Outbreaks were considered to have a known pathogen if confirmed by laboratory tests, and a known vehicle when an epidemiologic investigation implicated a specific food item. DATA SYNTHESIS: --From 1975 through 1987, 26 states reported 115 outbreaks of foodborne disease in nursing homes, causing illness in 4944 persons and death in 51. These outbreaks represented 2% of all reported foodborne disease outbreaks and 19% of outbreak-associated deaths in this period. Of 52 outbreaks with a known cause, Salmonella was the most frequently reported pathogen, accounting for 52% of outbreaks and 81% of deaths. Salmonella enteritidis outbreaks accounted for 56% of the Salmonella-associated deaths since 1981. The implicated food vehicles in S enteritidis outbreaks were made with eggs or prepared with equipment contaminated with eggs. Staphylococcal foodborne disease was the next most commonly identified cause, accounting for 23% of outbreaks. CONCLUSIONS: --Since the elderly are at high risk for serious morbidity from foodborne disease, nursing homes should practice careful food handling, preparation, and storage procedures; provide education for food handlers; and have active infection control programs to rapidly detect and control outbreaks of foodborne disease.

Aged↗

Clinical evaluation of a '3-in-1' intravenous nutrient solution.

Eighty-five consecutive general hospital patients requiring total parenteral nutrition (TPN) were prospectively studied in order to evaluate the safety and efficacy of a '3-in-1' nutrient mixture. All formulas were individualized to estimated requirements (average composition nitrogen 14 g, glucose 350 g, fat 50 g), mixed in the hospital pharmacy, contained within 3-litre EVA plastic bags, and given to the patients as a continuous 24-hour infusion. The average duration of TPN was 19 days per patient (range 8 - 84 days). Judging by nitrogen balance and plasma protein concentrations, the system was effective in maintaining or improving nutritional status in patients in a relatively stable condition but not in those who were critically ill (e.g. those in an intensive care unit). Development of magnesium and iron deficiencies was common during the period of TPN (25% of patients developing magnesium deficiency and 40% developing iron deficiency) despite daily supplementation with commercial trace element mixtures, but these states were easily corrected by high-dose administration. 'Creaming' of less than 5 mm on the surface of the emulsion was common, whereas that of more than 10 mm was rare (12 bags) and invariably associated with excessive addition of polyvalent cation or glucose. Deposition of lipid on the internal surface of the catheter was a common problem after 2 weeks' continuous administration. Temporary problems with faulty bag connections resulted in excessive catheter sepsis (14%) due to Staphylococcus epidermidis. Mild reversible disturbances in liver function occurred in one-third of the patients. The system appears safe and effective for the management of most patients requiring long-term TPN.(ABSTRACT TRUNCATED AT 250 WORDS)

Catheterization↗

Inhibition of Helicobacter pylori urease by phenyl phosphorodiamidates: mechanism of action.

Helicobacter pylori urease is a nickel-containing enzyme that hydrolyzes urea to bicarbonate and ammonia. Andrews et al. (J. Am. Chem. Soc. 1986, 108, 7124) have shown that amides and esters of phosphoric acid are slow, tight-binding inhibitors of urease isolated from jack bean. We show that 4-substituted phenyl phosphorodiamidates (4-R-PhOP(=O)(NH2)2) are slow-binding inhibitors of H. pylori urease with no evidence of kinetic saturation. Their second-order rates of inhibition ki are strongly correlated with phenol pKa (e.g. R = NO2, ki = 2.5 x 10(5) M-1s-1; R = OMe, ki = 1.2 x 10(4) M-1s-1). The Bronsted beta for inhibition is 0.4, similar to that of model system SN2(P) reactions. Based on these observations, we suggest that urease inhibition is covalent but reversible, involving a common phosphoacyl enzyme intermediate.

Helicobacter pylori↗

Analysis of Asn-linked glycans from vegetable foodstuffs: widespread occurrence of Lewis a, core alpha1,3-linked fucose and xylose substitutions.

The N-glycans from 27 "plant" foodstuffs, including one from a gymnospermic plant and one from a fungus, were prepared by a new procedure and examined by means of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). For several samples, glycan structures were additionally investigated by size-fractionation and reverse-phase high-performance liquid chromatography in conjunction with exoglycosidase digests and finally also (1)H-nuclear magnetic resonance spectroscopy. The glycans found ranged from the typical vacuolar "horseradish peroxidase" type and oligomannose to complex Le(a)-carrying structures. Though the common mushroom exclusively contained N-glycans of the oligomannosidic type, all plant foods contained mixtures of the above-mentioned types. Apple, asparagus, avocado, banana, carrot, celery, hazelnut, kiwi, onion, orange, pear, pignoli, strawberry, and walnut were particularly rich in Le(a)-carrying N-glycans. Although traces of Le(a)-containing structures were also present in almond, pistachio, potato, and tomato, no such glycans could be found in cauliflower. Coconut exhibited almost exclusively N-glycans containing only xylose but no fucose. Oligomannosidic N-glycans dominated in buckwheat and especially in the legume seeds mung bean, pea, peanut, and soybean. Papaya presented a unique set of hybrid type structures partially containing the Le(a) determinant. These results are not only compatible with the hypothesis that the carbohydrate structures are another potential source of immunological cross-reaction between different plant allergens, but they also demonstrate that the Le(a)-type structure is very widespread among plants.

Asparagine↗

Experience of explantation of Trilucent breast implants.

On 6 June 2000, the Medical Devices Agency (MDA) recommended that all Trilucent (soya-bean-oil filled) breast implants should be removed. This accelerated a trend that had already begun in our unit. This study follows our previous report on a 3 year series of women with Trilucent breast implants. The aim was to present our operative findings at explantation, and to correlate them with the preoperative signs and symptoms, the postoperative complications and the details of the hospital stay. In total, 44 patients (82 implants) underwent explantation, of whom 34 were cosmetic cases and 10 were reconstructive. Five patients had their implants removed before the MDA announcement. Implant rippling was the most common problem reported (25%), followed by pain (18%), implant deflation (9.1%) and capsular contracture (4.5%). Free oil was seen around the implant in 15 cases; four of these presented with clinical deflation, and three with rippling. The remaining eight patients were asymptomatic. We conclude that these implants tend to bleed, as evidenced by the presence of free oil around the implant in 34% of patients. The absence of free oil in 73% of the patients who presented with rippling suggests that the leaking oil is often metabolised and absorbed. The findings of this study are significant for women in whom free oil was found around the implant during explantation, and for those who still have Trilucent implants in place, for whatever reason, in spite of the MDA recommendation.

Adult↗

Polypeptide composition and amino-terminal sequence of broad bean polyphenoloxidase.

Polyphenoloxidase was purified from broad bean (Vicia faba) leaves. The purified enzyme contained two immunocross-reactive proteins of approximately 60 to 65 and 43 to 45 kilodaltons. Further electrophoretic separation resolved these proteins into doublets with molecular mass of 61.5, 60, 44.5, and 43 kilodaltons, respectively. Each of the four polypeptides was transferred to Immobilon and subjected to microprotein sequencing. All the polypeptides showed the same amino acid sequence up to residue 9 but some variations occurred thereafter. The amino-terminal sequence contained a large number of proline and serine residues. These results suggest that the four polypeptides were derived from a common parent form and that a posttranslational modification(s) must have occurred to account for the difference in their apparent size.

Journal Article↗

Antioxidant phenolics and flavonoids in common Indian foods.

To determine antioxidant phenolics and flavonoids in commonly consumed Indian foods we chemically analysed 85 food-stuffs comprising of cereals, pulses, nuts, oilseeds, vegetables, fruits and beverages. Total phenolics were measured biochemically and flavonoids were measured as a sum of quercetin, kaempferol, luteolin and pelargonidin. High flavonoid content (> 100 mg/100 gm) was present in tea, coffee, apple, guava, terminalia bark, fenugreek seeds, mustard seeds, cinnamon, red chili powder, cloves and turmeric. Medium levels (50-100 mg) were found in Indian gooseberry, omum, cumin, cardamom, betel leaf and brandy. Small but significant amounts were also present in food-items of large consumption such as kidney beans, soyabeans, grapes, ginger, coriander powder, bajra and brinjal.

Antioxidants↗

The nucleolar RNA-binding protein B-36 is highly conserved among plants.

The nucleolar protein B-36 is an RNA-associated protein which has a number of properties in common with pre-mRNA-binding proteins (hnRNP proteins). Like the hnRNP proteins, B-36 appears to be evolutionarily conserved among various eukaryotes (protists and several animal species). The conservation of B-36 throughout the plant kingdom has been investigated using a panel of nine monoclonal antibodies previously shown to recognize a minimum of four different epitopes in Physarum B-36, the protein used to generate the monoclonal antibodies. Two of the epitopes (I and III) are widely conserved in 34 kDa proteins (presumably B-36 homologues) from the various species tested (Chlamydomonas, moss, fern, oat, onion, carrot, and bean). Using immunofluorescence localization in moss and carrot protoplasts, the cross-reacting proteins were shown to be restricted to the nucleolus, further confirming their probable homology to B-36. Epitopes I and III are also unique to the B-36 homologues as demonstrated by the failure of any other bands to cross-react. Another epitope (IV) was specifically recognized in the plant B-36 homologues but exhibited greatly reduced affinity for the monoclonal antibody relative to Physarum B-36. The remaining epitope (II), unlike the others, exhibited variable conservation in the plant B-36 homologues and, in addition, was present in several other seemingly unrelated proteins. Finally, several of the plant species exhibited two cross-reacting variants at roughly the 34 kDa position and in at least one of these cases a single monoclonal antibody was able to distinguish between the two variants, a result indicating that the variants do have bona fide structural differences.(ABSTRACT TRUNCATED AT 250 WORDS)

Carrier Proteins↗

Analysis of oxylipins by high-performance liquid chromatography with evaporative light-scattering detection and particle beam-mass spectrometry.

The metabolism of 13S-hydroperoxy-9Z,11E,15Z-octadecatrienoic acid was investigated in a crude enzyme extract from mung bean seedlings (Phaseolus radiatus L.). Hydroperoxide-metabolizing activity was mainly due to a hydroperoxide lyase and, to a lesser extent, to an allene oxide synthase and a peroxygenase. Oxylipins originating from hydrolysis and cyclization of the allene oxide synthase product 12,13-epoxy-9Z,11,15Z-octadecatrienoic acid and from peroxygenase catalysis were identified by high-performance liquid chromatography (HPLC) particle beam-mass spectrometry (PB-MS) and quantified by normal-phase HPLC with an evaporative light-scattering detector (ELSD). An advantage of this methodology was the possibility to avoid extensive derivatization procedures commonly used for the gas chromatographic analysis of oxylipins. Owing to a comparable sample inlet system, the ELSD served an important analytical pilot function for the PB-MS: Qualitatively identical chromatographic patterns were obtained with both detection systems. The HPLC system enabled the separation of methyl 12-oxo-phytodienoate, methyl 11-hydroxy-12-oxo-9Z,15Z-octadecadienoate, methyl 12-oxo-13-hydroxy-9Z,15Z-octadecadienoate, methyl 9-hydroxy-12-oxo-10E,15Z-octadecadienoate, methyl 13-hydroxy-9Z,11E,15Z-octadecatrienoate, methyl 15,16-epoxy-13-hydroxy-9Z,11E-octadecadienoate, and methyl 13-hydroperoxy-9Z,11E,15Z-octadecatrienoate on a Lichrospher DIOL column within 33 min. Compared with a diode array detector, the ELSD proved to be more sensitive, in the case of methyl 12-oxo-13-hydroxy-9Z, 15Z-octadecadienoate by a factor of about 15. In addition, volatile metabolites were analyzed by capillary gas chromatography. The yield of the hydroperoxide lyase product 2E-hexenal was 49%, whereas the sum of oxylipins reached about 15%.

Aldehyde-Lyases↗

Dietary choices of people with ostomies.

OBJECTIVE: The objective was to collect data related to individual dietary recommendations and restrictions for people with ostomies. DESIGN: A descriptive survey was used that asked people about food choices/avoidances that occurred because of their ostomy. SUBJECTS: Subjects were members of the United Ostomy Association who had an ostomy and were living at home. METHODS: A 14-item questionnaire was mailed to the United Ostomy Association membership (approximately 40,000 current members) in their quarterly newsletter. RESULTS: Six hundred and four questionnaires were returned. The most frequent reason for ostomy surgery among the respondents was colon cancer (71%; n = 424), and the most common type of ostomy was a colostomy. The majority of respondents (88.4%) stated that they were not following a special diet because of the ostomy; 11.8% (n = 69) stated that they had modified their diet. Of the people who had modified their diet, 55% stated that they were using the special diet indefinitely and were adhering to it very well. More than half of the participants had received dietary instructions, provided mainly by a WOC nurse, at the time of surgery. The foods avoided because of the ostomy included fresh fruits, nuts, coconut, and vegetables such as corn, popcorn, cabbage, beans, and onions. The major problems reported were blockage and increased gas. CONCLUSION: Many respondents reported problems with certain foods. Preoperative teaching and postoperative follow-up must include anticipatory guidance on food selection.

Consumer Organizations↗

Hydrogenase genes from Rhizobium leguminosarum bv. viciae are controlled by the nitrogen fixation regulatory protein nifA.

Rhizobium leguminosarum bv. viciae expresses an uptake hydrogenase in symbiosis with peas (Pisum sativum) but, unlike all other characterized hydrogen-oxidizing bacteria, cannot express it in free-living conditions. The hydrogenase-specific transcriptional activator gene hoxA described in other species was shown to have been inactivated in R. leguminosarum by accumulation of frameshift and deletion mutations. Symbiotic transcription of hydrogenase structural genes hupSL originates from a -24/-12 type promoter (hupSp). A regulatory region located in the -173 to -88 region was essential for promoter activity in R. leguminosarum. Activation of hupSp was observed in Klebsiella pneumoniae and Escherichia coli cells expressing the K. pneumoniae nitrogen fixation regulator NifA, and in E. coli cells expressing R. meliloti NifA. This activation required direct interaction of NifA with the essential -173 to -88 regulatory region. However, no sequences resembling known NifA-binding sites were found in or around this region. NifA-dependent activation was also observed in R. etli bean bacteroids. NifA-dependent hupSp activity in heterologous hosts was also absolutely dependent on the RpoN sigma-factor and on integration host factor. Proteins immunologically related to integration host factor were identified in R. leguminosarum. The data suggest that hupSp is structurally and functionally similar to nitrogen fixation promoters. The requirement to coordinate nitrogenase-dependent H2 production and H2 oxidation in nodules might be the reason for the loss of HoxA in R. leguminosarum and the concomitant NifA control of hup gene expression. This evolutionary acquired control would ensure regulated synthesis of uptake hydrogenase in the most common H2-rich environment for rhizobia, the legume nodule.

Bacterial Proteins↗

Immediate early genes expressed in chlorovirus infections.

Twenty-three chlorovirus genes expressed in host cells as early as 5-10 min postinfection (p.i.), or immediate early, were isolated and characterized. Some showed significant homology with those for transcriptional factors and mRNA-processing proteins including TFIIB, helicases, mRNA capping enzyme, nucleolin, and bean transcription factor. Others code for (i) factors influencing translation such as aminoacyl tRNA synthetases and ribosomal protein, and (ii) unknown proteins. Enzymes involved in polysaccharide synthesis were also found. All transcripts of these genes had a poly(A) tail, which decreased in size after 20 min p.i., possibly caused by the shortening by an exonuclease. Often, due to readthrough either from an upstream ORF or into a downstream ORF, a few extra transcripts for each gene appeared after 40 min p.i., suggesting a change in promoter selection and termination accuracy at this point. A typical TATA-box and a common element 5'-ATGACAA were in the promoter region of almost all of the immediate early genes, which may be recognized by host RNA polymerase and transcription factors.

Animals↗

Dietary factors and stomach cancer: a case-control study in Korea.

BACKGROUND: Stomach cancer is the most common cancer among Koreans. There is wide agreement that dietary factors are important in gastric carcinogenesis, but the role of many Korean food items remains unknown. METHODS: A case-control investigation involving 213 incident cases of histologically confirmed stomach cancer and an equal number of controls, matched by age (within 2 years) and sex, was conducted from June 1990 to October 1991. RESULTS: An increased risk of stomach cancer was noted among those with high consumption of stewed foods such as soybean paste stew and hot pepper-soybean stew, broiled fish, and those who liked salty food. However, mung bean pancake, tofu (soybean curd), cabbage, spinach, and sesame oil decreased the risk of stomach cancer. Stratified analysis by salt in combined foods, such as stewed foods and pickled vegetables, disclosed salt as being an important risk factor. Analysis of cooking methods showed that broiling and salting increased the risk of stomach cancer, but that frying tended to decrease the risk. These results suggest that the cooking method might modify the gastric carcinogenicity of foods. As non-dietary factors, smoking and a family history of stomach cancer increased the risk, but use of a refrigerator decreased the risk of cancer. CONCLUSIONS: Heavy salt consumption and cooking methods like broiling and salting seem to play a major role in gastric carcinogenesis among Koreans.

Adult↗