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WS1279, a novel lipopeptide isolated from Streptomyces willmorei. Biological activities.

WS1279, a new lipopeptide isolated from the fermentation broth of Streptomyces willmorei No. 1279, stimulated the proliferation of mouse bone marrow cells in vitro and accelerated the recovery of granulocyte counts in bone marrow from leukopenia induced by mitomycin C (MMC) in mice. The glycerylcysteine moiety of WS1279 is necessary and the lipid peptide structure is required for manifestation of full stimulating activity in vitro. WS1279 was the most effective on the proliferation of bone marrow cells among the tested immunostimulants in vitro. However, the effect of WS1279 on restoration of reduced granulocyte counts in MMC-induced leukopenia in mice was less than that of FK-565, lipopolysaccharide, picibanil or forphenicinol. WS1279 augmented host resistance to infection with Staphylococcus aureus 47 in normal and immunosuppressed mice.

Adjuvants, Immunologic↗

Protective effect of fungal growth product (6MFA), assessed therapeutically against Ehrlich's ascites tumor in Swiss mice.

6MFA is a growth product of the fungus Aspergillus ochraceus (ATCC 28706) obtained by fermentation in stationary culture. It has both interferon inducing and antiviral properties, in vivo and in vitro, with a relatively high margin of safety (9, 17, 18). Ehrlich's ascites tumor bearing Swiss albino male mice were treated with 0.5 ml of acqueous preparation of 6MFA (0.75 mg total solids) i.p. in a therapeutic regimen schedule; the sham treated mice received only PBS. 6MFA treatment produced an increase in mean survival time over the untreated controls, restricted the body weight increase due to ascites and decreased the rate of mortality. As much as 100% of survival response was obtained in the group treated with 0.5 ml of 6MFA at the rate of one inoculation per week for 5 weeks. In general a dose-dependent response was seen in the antitumor effect of 6MFA against Ehrlich's ascites tumor in Swiss mice. Delay in administration of 6MFA to tumor bearing mice affected the survival rate.

Animals↗

Functional complementarity between the HMG1-like yeast mitochondrial histone HM and the bacterial histone-like protein HU.

The mitochondrial histone HM is the major DNA-binding protein in mitochondria and is necessary for maintenance of the mitochondrial genome in the yeast Saccharomyces cerevisiae during growth on fermentable sugars. HM and the Escherichia coli histone-like protein HU have similar activities in vitro, including DNA supercoiling, but share no sequence similarity. We show that HU can functionally complement the respiration deficiency associated with yeast strains lacking HM. Conversely, phenotypes of E. coli cells lacking HU protein, including nucleoid loss and a filamentous cell morphology, were alleviated by expression of HM in these cells. The HU protein of bacteria and the HM protein of mitochondria are therefore functionally complementary in vivo. Functional similarities among HM, HU, and the nuclear HMG1 proteins are implicated and discussed.

Bacterial Proteins↗

(+)-10 alpha-Hydroxy-4-muurolen-3-one, a new inhibitor of leukotriene biosynthesis from a Favolaschia species. Comparison with other sesquiterpenes.

A new inhibitor of leukotriene biosynthesis, (+)-10 alpha-hydroxy-4-muurolen-3-one (1), was isolated from fermentations of Favolaschia sp. 87129. Its structure was established by spectroscopic methods. The compound exhibited no antifungal or antibacterial activities. The effects of 1 on leukotriene biosynthesis were compared with (+)-T-cadinol, (-)-3-oxo-T-cadinol, and (+)-3 alpha-hydroxy-T-cadinol, three related sesquiterpenes.

Animals↗

Enzyme, bacterial inoculant, and formic acid effects on silage composition of orchardgrass and alfalfa.

We evaluated the effects of cellulase (from Trichoderma longibrachiatum) application rates on neutral detergent fiber (NDF) concentration and fermentation products of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) silages harvested with decreasing dry matter (DM) digestibility. Additionally, the impacts of inoculant (Lactobacillus plantarum and Pediococcus cerevisiae), pectinase (from Aspergillus niger), or formic acid on silage composition were studied. Forages wilted to a DM content of about 320 g/kg were ensiled in laboratory silos for 60 d. Cellulase, combined with inoculant, was applied at 2, 10, and 20 ml/kg of herbage (at least 2500 IU/ml). Cellulase at 10 ml/kg was also applied alone or in combination with pectinase and inoculant or formic acid. The NDF concentration of orchardgrass silage decreased with increasing cellulase up to 20 ml/kg, at which NDF content was decreased by 30%. The NDF concentration of alfalfa silage decreased with increasing cellulase application up to 10 ml/kg, at which NDF content was decreased by 13%. Immature plants were more responsive to cellulase treatment than mature plants. Cellulase at 2 ml/kg combined with inoculant improved fermentation characteristics of the silages but generally, there was no effect on silage fermentation by higher cellulase applications, resulting in an accumulation of sugar. The improved fermentation of orchardgrass treated with cellulase and inoculant was mostly related to the effect of inoculant, whereas cellulase alone improved fermentation characteristics of alfalfa silage and this effect was enhanced by addition of inoculant. Decreased NDF and increased sugar concentrations did not improve the in vitro DM digestibility of cellulase-treated silages.

Aspergillus niger↗

Comparative in vitro activity of eight antibiotics against Pseudomonas aeruginosa and infrequently isolated gram-negative bacilli.

Seventy-eight Gram-negative non-fermentative bacilli have been tested against eight antimicrobial agents (wide spectrum penicillins, newer generation cephalosporins and aminoglycosides). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of each compound were determined by microdilution technique. Ceftazidime showed higher antibacterial activity against all bacterial strains.

Anti-Bacterial Agents↗

A review: acute and chronic effects of ethanol and alcoholic beverages on the pancreatic exocrine secretion in vivo and in vitro.

Whereas oral or intraduodenal ethanol causes a moderate stimulation of pancreatic bicarbonate and enzyme output, intravenous ethanol inhibits basal and hormonally stimulated pancreatic exocrine secretion in humans, dogs, cats, pigs, rabbits, and rats. This inhibition could be mediated by inhibitory cholinergic mechanisms or be the result of a direct cellular effect of ethanol. In vitro investigations have specified several signaling molecules that may be involved in the action of ethanol on stimulus-secretion coupling in the exocrine pancreas, including cyclic adenosine monophosphate, intracellular calcium, and cholecystokinin and somatostatin receptors. In difference to pure ethanol solutions and distilled spirits, beer strongly stimulates pancreatic enzyme output, probably by nonalcoholic fermentation products. During chronic alcoholism, the ethanol-induced inhibition is replaced by an enhanced enzyme output that causes intraductal protein precipitation. In vitro investigations suggest that this increase is reversible after alcohol withdrawal. The occurrence of protein precipitates is considered to be a crucial step in the development of chronic alcoholic pancreatitis in humans. Other ethanol-induced secretory alterations that may contribute to the development of alcoholic pancreatitis are a decreased secretion of trypsin inhibitor, an increased cholinergic tone, and changes in the concentration of lithostathine.

Alcoholic Beverages↗

Epithienamycins-novel beta-lactams related to thienamycin. I. Production and antibacterial activity.

The epithienamycins are cell wall active antibiotics structurally related to N-acetylthienamycin. We have found forty-three isolated of Streptomyces flavogriseus which are capable of producing members of the epithienamycin family. Six major epithienamycin components, and xanthomycin, have been isolated from fermentation broth. Fermentation conditions can be varied to enrich for certain members of the epithienamycin family. All six components show activity in vitro versus a broad spectrum of bacterial species. The weight potencies vary 27 fold from the most active to least active.

Animals↗

Highly potent anti-HIV-1 activity isolated from fermented Polygonum tinctorium Aiton.

A water-soluble extract of fermented Polygonum tinctorium Aiton (Polygonaceae) called Sukumo, exhibited a potent inhibitory activity against HIV type 1 in vitro. The extract potently suppressed acute HIV-1 (IIIB) infection in MT-4 cells with EC50 values of 0.5 microg/ml but exhibited low cytotoxicity to MT-4 cells even at a high concentration (CC50 > 1000 microg/ml). It also inhibited giant cell formation in co-cultures of HIV-infected cells and uninfected Molt-4 cells. Sukumo extract was found to interact with both the viral envelope glycoprotein and cellular receptors, thus blocking virus-cell binding and virus-induced syncytium formation. There was a good correlation between the extract's anti-HIV-1 activity and its inhibitory effects on HIV-1 binding. It also suppressed replication of herpes simplex virus type 1 in Vero cells with an EC 50 of 11.56 microg/ml. On the other hand, there was no appreciable activity against influenza A virus, poliovirus or SARS corona virus when tested at concentrations ranging from 3.2-400 microg/ml as shown by microscopic image analysis for cytopathic effect (CPE). Physico-chemical studies revealed that the anti-HIV activity in the extract was essentially maintained after boiling at 100 degrees C in 1N HCl or 1N NaOH, and after treatment with 100 mM NaIO4. The inhibitory activity of the extract was also not reduced after pronase digestion. The active factor in the extract is likely to be a novel compound(s) having a polyanionic substructure and a molecular weight of 10,000-50,000.

Anti-HIV Agents↗

Methylsulfomycin I, a new cyclic peptide antibiotic from a Streptomyces sp. HIL Y-9420704.

Methylsulfomycin I (1) is a new cyclic peptide antibiotic isolated from the fermentation broth of a Streptomyces sp. HIL Y-9420704. Its structure was elucidated by NMR and GC-MS. The in vitro activity (MIC) against a wide range of Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-, and teicoplanin-resistant strains, is described.

Animals↗

In vitro effects on polydextrose by colonic bacteria and caco-2 cell cyclooxygenase gene expression.

A 4-stage colon simulator and a cell culture-based human intestinal epithelial function model were combined to study the effects of a soluble fiber, polydextrose (PDX), on intestinal microbes and mucosal functions relevant to the risk of colon cancer. We observed sustained degradation of PDX throughout the different stages of the model. The fermentation was characterized by gradual degradation of PDX, production of short-chain fatty acids, and no increasing in putrefactive markers. We observed less marked effects in the microbial densities. When we applied colon fermentation metabolites obtained from the simulators with PDX to Caco-2 colon cancer cell line, a significant dose-dependent decreasing effect on cyclooxygenase-2 (COX-2) and an increasing effect on COX-3 expression levels were observed. PDX concentration appeared not to have effect on the expression levels of COX-1. Overexpression of COX-2 and decreased expression of COX-1 have been suggested to be characteristics of colon cancer. The exact physiological role of COX-3, an intron-retaining splice variant of COX-1, is not known, but it is suspected to play a role in transcriptional regulation of COX-1 and COX-2. In vitro modulation of COX expression by colon microbial fermentation products of polydextrose offers an interesting starting point for further studies on possible risk-decreasing effect of PDX on the development of colon cancer.

Bacteria↗

Evaluation of pumpkin seed products for bread fortification.

Pumpkin seed products (raw, roasted, autoclaved, germinated, fermented, pumpkin protein concentrate and pumpkin protein isolate) were incorporated into wheat flour to produce blends with protein levels of 15, 17, 19 and 21%. Dough properties were evaluated by a farinograph; loaves of breads were evaluated by a taste panel for crust color, crumb color, crumb texture, flavor, and overall quality. Results indicated that pumpkin seed products can be added to wheat flour up to a 17% protein level for raw, roasted and autoclaved pumpkin meal, 19% level for germinated, fermented and pumpkin protein concentrate and 21% level for pumpkin protein isolate without a detrimental effect on dough or loaf quality. On the other hand, the addition of pumpkin seed proteins resulted in increasing protein, lysine and mineral contents compared to the control. While lysine and tryptophan were the first and second limiting amino acids in the control bread, tryptophan and lysine were the first and second limiting amino acids for raw, roasted, autoclaved, germinated and fermented pumpkin meal; valine and lysine and valine and total sulfur amino acids were the first and second limiting amino acids for pumpkin protein concentrate and isolate, respectively. In vitro protein digestibility improved when the pumpkin seed proteins were added.

Amino Acids↗

3-amino-5-hydroxybenzoic acid in antibiotic biosynthesis. XI. Biological origins and semisynthesis of thionaphthomycins, and the structures of naphthomycins I and J.

Fermentations of Streptomyces sp. E/784 produce low levels of the novel C-30 alkylthio-substituted ansamycin antibiotics naphthomycins J (9) and I (10), in addition to the more abundant C-30 hydroxylated analogues actamycin (1) and naphthomycin D (2) and C-30 chlorinated analogues naphthomycins H (3) and A (4). The addition of N-acetyl-L-cysteine to the fermentation medium substantially increases the production of the thionaphthomycins J and I at the expense of their chloro analogues H and A. Other thiols and thiol progenitors are similarly utilised, including N-acetyl-L-cysteine methyl ester which affords the known naphthomycin F (8) and its novel 2-demethyl homologue (7). The formation of thioansamycins from chloroansamycins and thiols in vivo is probably non-enzymic since similar conversions can be effected in vitro.

Acetylcysteine↗

Evaluation of antioxidative and mutagenic properties of 50% ethanolic extract from red beans fermented by Aspergillus oryzae.

Various bean products fermented by microorganisms are commonly consumed in Asian diets; however, the safety or functional properties of fermented beans can vary with different microbial species and with different processes being applied to different beans. The objectives of this study were to evaluate the antioxidative and mutagenic properties of 50% ethanolic extracts from red beans fermented by Aspergillus oryzae. The extracts' antioxidative activities, including alpha,alpha;-diphenyl-beta-picryl-hydrazyl (DPPH) radical-scavenging effects, Fe(2+)-chelating ability, and reducing power, were studied in vitro. The antioxidative effects provided by the extracts depended strongly on their concentrations. In general, antioxidative activity increased with extract concentration to a certain point and then leveled off as the concentration further increased. The fermented red bean extracts showed less of a scavenging effect on the DPPH radical and less reducing power than the commercial antioxidants alpha-tocopherol and butylated hydroxytoluene, but better Fe(2+)-chelating ability. No mutagenicity or toxicity effect on any of the tested strains (Salmonella Typhimurium TA97, TA98, TA100, TA102, and TA1535) was found for the 50% ethanolic extracts of fermented red beans with the Ames mutagenicity assay. These results suggest that the 50% ethanolic extracts were not mutagenic.

Antioxidants↗

Halovirs A-E, new antiviral agents from a marine-derived fungus of the genus Scytalidium.

Marine micro-organisms represent an under explored resource for the discovery of novel antiviral agents. Here, we describe a series of peptides designated halovirs A-E (1-5) that are produced during the saline fermentation of a marine-derived fungus of the genus Scytalidium. These lipophilic, linear peptides are potent in vitro inhibitors of the herpes simplex viruses 1 and 2. Evidence is presented that the halovirs directly inactivate herpes viruses, a mechanism of action that could be applicable in the prevention of HSV transmission. The total structures of these new compounds were established by a combination of spectral and chemical techniques. Salient structural features of the halovir hexapeptides include a nitrogen terminus acylated by myristic (C14) or lauric (C12) acid, an unusual Aib-Hyp dipeptide segment, and a carboxyl terminus reduced to a primary alcohol. A qualitative analysis of the secondary structures of these molecules using variable temperature NMR experiments and NOE analyses is also reported.

Acylation↗

Effects of increased transaldolase activity on D-xylulose and D-glucose metabolism in Saccharomyces cerevisiae cell extracts.

In vitro metabolism of D-xylulose and D-glucose in extracts obtained from D-glucose- and D-xylulose-fermenting Saccharomyces cerevisiae cells was investigated with 10- and 100-fold-increased activity of the enzyme transaldolase (EC 2.2.1.2). The rate of sugar consumption was the same in most cases, whereas the rate of ethanol formation decreased with increased levels of transaldolase. The formation of glycerol, pentitols, and acetic acid was not dependent on added transaldolase but was dependent on the sugar used as the growth substrate and on the sugar used in the in vitro metabolism experiments. The carbon balance showed that the dissimilated carbon could not be accounted for in products when transaldolase was added. The concentration of D-fructose-1,6.-diphosphate in the extracts was not influenced by added transaldolase but was higher with D-xylulose than with D-glucose. Levels of pyruvate, comparable with the two substrates, decreased with increasing levels of transaldolase. Exogenously added transaldolase decreased D-sedoheptulose-7-phosphate levels when D-xylulose was the substrate. The results are discussed in relation to the dissimilation of carbon through the upper part of glycolysis and the pentose phosphate pathway.

Biological Transport↗