Maternal obesity and overnutrition increase oxidative stress in male rat offspring reproductive system and decrease fertility.

PURPOSE: Increasing evidence exists that maternal obesity (MO) and overnutrition during pregnancy and lactation have long-lasting consequences for progeny metabolism, cardiovascular and endocrine function. Data on effects of MO on offspring reproduction are limited. We hypothesized that MO during pregnancy and lactation in founder F(0) rat mothers would increase testicular and sperm oxidative stress (OS) and adversely impact male fertility in their F(1) offspring. METHODS: We induced pre-pregnancy MO by feeding F(0) females a high-fat diet from weaning through pregnancy and lactation. After weaning, all F(1) rats ate control (C) diet. We determined serum testosterone, malondialdehyde (MDA), reactive oxygen species (ROS) and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in F(1) testes and sperm at postnatal days (PNDs) 110, 450 and 650. RESULTS: At PNDs 450 and 650, MO offspring had lower luteinizing hormone while testosterone levels were lower at all ages. Testicular MDA and ROS concentrations and SOD and GPx activity were higher in MO F(1) at all ages. Nitrotyrosine immunostaining was higher at all ages in MO F(1) testes than C F(1). At PNDs 450 and 650, MO F(1) spermatozoa showed higher MDA concentrations and lower SOD and GPx activity with reduced sperm concentration, viability and motility, and more sperm abnormalities. Fertility rate was not affected at PND 110 but was lower in MO F(1) at PNDs 450 and 650. CONCLUSIONS: We conclude that MO during pregnancy and lactation increases F(1) testicular and sperm OS leading to premature aging of reproductive capacity.

Midregional pro-atrial natriuretic peptide and procalcitonin improve survival prediction in VAP.

Ventilator-associated pneumonia (VAP) affects mortality, morbidity and cost of critical care. Reliable risk estimation might improve end-of-life decisions, resource allocation and outcome. Several scoring systems for survival prediction have been established and optimised over the last decades. Recently, new biomarkers have gained interest in the prognostic field. We assessed whether midregional pro-atrial natriuretic peptide (MR-proANP) and procalcitonin (PCT) improve the predictive value of the Simplified Acute Physiologic Score (SAPS) II and Sequential Related Organ Failure Assessment (SOFA) in VAP. Specified end-points of a prospective multinational trial including 101 patients with VAP were analysed. Death <28 days after VAP onset was the primary end-point. MR-proANP and PCT were elevated at the onset of VAP in nonsurvivors compared with survivors (p = 0.003 and p = 0.017, respectively) and their slope of decline differed significantly (p = 0.018 and p = 0.039, respectively). Patients with the highest MR-proANP quartile at VAP onset were at increased risk for death (log rank p = 0.013). In a logistic regression model, MR-proANP was identified as the best predictor of survival. Adding MR-proANP and PCT to SAPS II and SOFA improved their predictive properties (area under the curve 0.895 and 0.880). We conclude that the combination of two biomarkers, MR-proANP and PCT, improve survival prediction of clinical severity scores in VAP.

Absorption of oral ethinylestradiol is delayed by its eutectic mixture with cholesterol.

A solid dispersion of ethinylestradiol-cholesterol (EE & CHOL; eutectic 1:4 W/W) was prepared by melting and rapid cooling. The fused material was then mixed with lactose as vehicle. Soft gelatin capsules were filled with 50 mg of the final mixture to give 0.050 mg of ethinylestradiol. Six female volunteers received, one capsule of the eutectic combination of EE:CHOL or one 50 micrograms tablet of ethinylestradiol (Dianor, Syntex), in a cross-over study and in fasting state. Venous blood samples were drawn at 0, 10, 20, 30, 40, 50, 60, 90, 120, 240, 360, 480, 720, 1440 minutes after dosing. Immunoreactive EE was measured by radioimmunoassay to assess the serum concentration-time course. All subjects exhibited a significant increase in EE levels after oral administration. Mean peak EE levels, 1350 pg/ml vs 91 pg/ml (p less than 0.001), were achieved 360 minutes and 90 minutes (p less than 0.01), after administration of the eutectic and reference formulation, respectively. Eutectic mixture showed a greater area under the serum concentration-time curve, longer mean residence time of the drug in the body, and four times the value of the elimination half-life of the reference formulation. It is concluded that the combination of ethinylestradiol with cholesterol forming an eutectic mixture, when administered orally to normal women, modulates the absorption and the bioavailability of the EE. This approach may be suitable for long-acting oral treatment with sex steroids.

A54145, a new lipopeptide antibiotic complex: factor control through precursor directed biosynthesis.

A54145 is a complex of new lipopeptide antibiotics produced by Streptomyces fradiae. Eight factors, containing four similar peptide nuclei in combination with three different fatty acid acyl side chains, have been isolated from the natural fermentation and characterized. The nuclei differ only in valine/isoleucine and glutamate/3-CH3-glutamate substitutions at one or both of two locations on the peptide ring. Prior deacylation of all four nuclei with Actinoplanes utahensis had permitted chemical reacylation of each nucleus with new fatty acid acyl chains for structure-activity relationship studies. In an effort to induce the native biosynthesis of preferred factors or analogs by S. fradiae, the effect of fatty acid precursors on the fermentation was examined. Many fatty acids were extremely toxic to S. fradiae, which limited experiments to slow, continuous feeding of the lipids in stirred bioreactors that were equipped for on-line respiration analysis by mass spectrometry. These studies determined that precursing with aliphatic fatty acids of various chain lengths did enhance the biosynthesis of factors containing specific fatty acid acyl side chains. Caprate, for example, increased the n-decanoyl-containing factors from the natural level of approximately 14% to approximately 80%. The percentage of factors containing branched-chain fatty acid acyl substituents was also increased, in shaken-flask studies, by enriching the medium with valine or isoleucine. These amino acids additionally enhanced the percentage of nuclei containing either valine or isoleucine.

A47934, a novel glycopeptide-aglycone antibiotic produced by a strain of Streptomyces toyocaensis taxonomy and fermentation studies.

A47934, a novel glycopeptide-aglycone antibiotic, is produced by a strain of Streptomyces toyocaensis, NRRL 15009. A47934 is unique among reported glycopeptides in that it contains a sulfate ester. Like several other glycopeptides, the majority of the A47934 produced remained associated with the producing biomass, from which it could be released into aqueous media by alkalization. Antibiotic biosynthesis was depressed when initial levels of phosphate phosphorus in the medium exceeded the normal level of 35 micrograms/ml. Enrichment of the fermentation medium with tyrosine depressed A47934 yields while enrichment with p-hydroxyphenylglycine or p-hydroxyphenylglyoxylic acid stimulated antibiotic biosynthesis.

Deacylation of A21978C, an acidic lipopeptide antibiotic complex, by Actinoplanes utahensis.

A21978C, produced by Streptomyces roseosporus NRRL 11379, is an acidic lipopeptide antibiotic complex that inhibits Gram-positive bacteria. Individual factors of the complex possess an identical peptide core or "nucleus", and are differentiated by the distinctive fatty acid acyl group attached to the N-terminus of the nucleus. Certain members of the family Actinoplanaceae deacylated A21978C to yield the unaltered nucleus, which was then reacylated to form new analogs. Actinoplanes utahensis NRRL 12052 was the most efficient of these cultures, producing up to 500 micrograms of nucleus per ml of culture broth per hour. Eacylation was also accomplished with semi-pure and tert-butoxycarbonyl (tert-BOC)-A21978C. In the latter, the ornithine amino group was blocked to prevent formation of diacyl analogs during reacylation. The acylase was an endoenzyme present in submerged cultures of A. utahensis from less than 18 to greater than 168 hours of incubation. Whole cells suspended in phosphate buffer or entrapped in polyacrylamide gel also deacylated A21978C efficiently.

A41030, a complex of novel glycopeptide antibiotics produced by a strain of Streptomyces virginiae. Taxonomy and fermentation studies.

A41030 is a complex of novel glycopeptide antibiotics produced by a culture isolated from a soil. Taxonomic studies have identified the microorganism, NRRL 15156, as a strain of Streptomyces virginiae. The major factor, A41030A, and three of the six minor factors are unique among glycopeptides in that they are naturally occurring aglycones, containing no neutral or amino sugars. The A41030 that was not spontaneously released into the fermentation broth could be released from the biomass into aqueous media at pH 10.5. In contrast to the vancomycin and N-demethylvancomycin fermentations, A41030 biosynthesis was stimulated by enriching the medium with K2HPO4 at a level of 1 mg/ml. Enrichment with putative precursors of the aglycone, however, did not increase the biosynthesis of A41030.

Deacylation of echinocandin B by Actinoplanes utahensis.

Echinocandin B (ECB) is a lipopeptide antifungal agent produced by several species of Aspergillus. The lipid side chain of cyclic lipopeptides is known to be an important determinant of their antibiotic activity and toxicity. Deacylation of another lipopeptide antibiotic, A21978C, had formerly been accomplished with Actinoplanes utahensis. In spite of the structural dissimilarities between the peptide cores and acyl side chains of A21978C and ECB, A. utahensis also removed the linoleoyl acyl unit from the amino terminus of ECB to yield the bioinactive cyclic peptide core, or "nucleus". The ECB nucleus, which contained a new titratable group at the N-terminus, was subsequently employed for chemical reacylation with other side chains to yield a variety of novel ECB analogs. One of these, cilofungin (LY121019), containing an N-(4-n-octyloxybenzoyl)acyl unit, is currently undergoing clinical evaluation.

Biosynthesis of thiopeptide antibiotic A10255 in stirred reactors using a chemically defined medium supplemented with continuous nutrient feeds.

A10255 is a complex of new thiopeptide antibiotics produced by Streptomyces gardneri. When stirred reactors were operated in batch mode using a defined medium with a glucose feed, 250 micrograms/ml of A10255 were produced during a four-day fermentation cycle. The linear growth phase of S. gardneri was extended through seven days by supplementing the defined medium with continuous feeds of hydrolyzed casein and methyl caprate. With the supplementary feeds, antibiotic biosynthesis paralleled growth during the extended cycle and attained levels of 1,750 micrograms/ml. Increasing the standard glucose feed rate increased titers principally by increasing cell mass. Supplementing the standard glucose feed with lipids such as caprylate or caprate, and decyl alcohol, affected cell mass minimally but produced higher titers by increasing the specific biosynthesis of A10255 per unit of biomass.

16-Deethylindanomycin (A83094A), a novel pyrrole-ether antibiotic produced by a strain of Streptomyces setonii. Taxonomy, fermentation, isolation and characterization.

16-Deethylindanomycin (A83094A) is a novel pyrrole-ether antibiotic produced by a strain of Streptomyces setonii. The antibiotic, which is structurally similar to indanomycin (X-14547A), is active in vitro against Gram-positive bacteria as well as coccidia.

Biosynthesis of thiopeptide antibiotic A10255: incorporation of isotopically-labeled precursors.

The biosynthetic origin of antibiotic A10255 was investigated using 14C- and 13C-labeled amino acids. DL-[1-(13)C]Serine labeled 15 of the 17 amino acid residues present in A10255G. These included the oxazole, thiazole, dehydroalanine, masked glycine, masked alanine and pyridine moieties. The same 15 residues labeled by serine were labeled by [2-(13)C]glycine, apparently by conversion of the glycine to [2,3-(13)C]serine. Formation of the pyridine ring occurred via a C3 to C3 condensation of two serines. The results indicated origin of the masked alanine from alanine; the masked glycine from glycine; the thiazole residues from cysteine; and the threonine, masked dehydrobutyrine, masked dehydronorvaline and masked dehydroleucine residues from threonine. L(-)[CH3-(13)C]Methionine labeled the methyl carbon of the masked dehydronorvaline moiety in factor B and the two methyl carbons of the masked dehydroleucine moiety in factor E. The results demonstrate that A10255 originates exclusively from amino acids in a manner similar to the closely related thiopeptide antibiotics nosiheptide and thiostrepton.

A10255, a complex of novel growth-promoting thiopeptide antibiotics produced by a strain of Streptomyces gardneri. Taxonomy and fermentation studies.

A10255 is a complex of new thiopeptide antibiotics characterized structurally by a cyclic peptide core to which is attached a side chain composed of dehydroalanine moieties. The complex contained 80-85% factor B, 15-20% factor G, and trace amounts of factors C, D, E, F, H, and J. Taxonomic studies indicated the producing microorganism to be a strain of Streptomyces gardneri. The major portion of the antibiotic produced remained associated with the mycelial biomass, from which it was extracted with polar solvents such as aqueous methanol or aqueous acetone. Initial A10255 yields of < 2 micrograms/ml were increased to over 300 micrograms/ml in stirred reactors through strain selection, nutritional studies, and conversion of the batch fermentation to a fed-batch mode.

A30641, a new epidithiodiketopiperazine with antifungal activity.

A new antibiotic, designated A30641, having in vitro activity against Gram-positive bacteria and fungi has been isolated from a strain of Aspergillus tamarii. Chemical and physical characterization indicate that it is a member of the class of antibiotics containing the epidithiodiketopiperazine moiety.

A32390A, a new biologically active metabolite. I. Discovery and fermentation studies.

A32390A is an isonitrile-containing derivative of diacyl D-mannitol. The compound is produced in fermentation as the major component of a metabolic complex known as A32390. A32390A inhibits dopamine-beta-hydroxylase reduces heart and adrenal norepinephrine levels, lowers blood pressure in hypertensive rats, and possesses antibiotic activity vs. Gram-positive bacteria and fungi, including Candida albicans. A32390 is produced in submerged culture by a mold, a species of Pyrenochaeta, NRRL-5786. Glucose and sucrose are among the best carbon sources for the biosynthesis of A32390. Mannitol, although a substituent of the A32390A molecule, supports little or no biosynthesis of the compound when employed as the major carbon source for the fermentation. The addition of crotonic acid derivatives. ethanol, or L-histidine to the fermentation medium enhances the level of A32390 produced.

New azasteroidal antifungal antibotics from Geotrichum flavo-brunneum. I. Discovery and fermentation studies.

Although Geotrichum species occur ubiquitously, antibiotic production by members of this genus has not previously been reported. The antibiotic complex designated A25822, consisting of one major and six minor structurally-related components active primarily against Candida and Trichophyton, represents a new family of naturally-occurring compounds. Approximately 90% of the antibiotic activity synthesized remained associated with the fungal cell mass, from which it was recovered by multiple methanolic extractions for quantitation. Antibiotic production was enhanced by tryptophan, iron, zinc, and high levels of dextrin.

Angiotensin converting enzyme inhibitors produced by Streptomyces chromofuscus. Discovery, taxonomy and fermentation.

Culture A58365.1, NRRL 15098, identified as a new strain of Streptomyces chromofuscus, was found to produce two novel angiotensin converting enzyme (ACE) inhibitors, A58365A and A58365B. Fermentation medium studies afforded an increase in ACE inhibitor titers from less than 1 microgram/ml to greater than 20 micrograms/ml. Proline was the obligatory supplement for ACE inhibitor biosynthesis.

The discovery, fermentation, isolation, and structure of antibiotic A33853 and its tetraacetyl derivative.

The structure of antibiotic A33853, isolated from the culture broth of Streptomyces sp., NRRL 12068, is reported. The structure was deduced from an X-ray crystallographic study of its tetraacetyl derivative. Tetraacetyl A33853 is unique because it contains an anhydride moiety, an unexpected product from the reaction of A33853 with acetic anhydride and pyridine.

N-demethylvancomycin, a novel antibiotic produced by a strain of Nocardia orientalis. Taxonomy and fermentation.

A novel vancomycin analog, N-demethylvancomycin, is produced by a soil isolate collected in Yucatan, Mexico. Taxonomic studies indicated this microorganism, designated NRRL 15232, is a strain of Nocardia orientalis. Unlike some glycopeptide antibiotics, virtually none of the N-demethylvancomycin synthesized remained bound to the cells of the producing culture. Antibiotic production was markedly depressed by the addition of orthophosphate to the fermentation medium. Enrichment of the medium with tyrosine, p-hydroxyphenylglycine, p-hydroxyphenylglyoxylic acid, or leucine, all putative precursors of the aglycone, stimulated the biosynthesis of N-demethylvancomycin.

A83586C, a new depsipeptide antibiotic.

A culture identified as Streptomyces karnatakensis was found to produce a novel cyclic hexadepsipeptide antibiotic designated A83586C. The structure was elucidated by X-ray crystallography, and full 1H and 13C NMR assignments are reported. The absolute configuration was confirmed by the detection of D-threonine in the acid hydrolysate of A83586C. A83586C had potent Gram-positive activity in vitro but lacked in vivo efficacy in mice.

A54145, a new lipopeptide antibiotic complex: discovery, taxonomy, fermentation and HPLC.

A54145 is a complex of new lipopeptide antibiotics that inhibits Gram-positive bacteria and acts as a growth promotant for broiler chicks. Eight factors; A, B, C, D, E, F, A1 and B1; have been isolated and characterized. They contain four similar peptide nuclei, each of which is acylated with either an 2-decanoyl, n-decanoyl, or undecanoyl side chain. Taxonomic studies ascertained that the producing microorganism was a strain of Streptomyces fradiae. Fermentation studies determined that superior antibiotic yields were obtained in stirred bioreactors in a soybean flour-molasses medium employing a continuous glucose feed. These findings, interwoven with the selection of hyper-productive mutants, increased fermentation yields from less than 50 micrograms/ml to more than 1 mg/ml. An analytical HPLC system was developed for the identification and subsequent quantitation of each factor of the A54145 complex.