Genetic and pharmacological factors that influence reproductive aging in nematodes

Age-related degenerative changes in the reproductive system are an important aspect of aging, because reproductive success is the major determinant of evolutionary fitness. Caenorhabditis elegans is a prominent organism for studies of somatic aging, since many factors that extend adult lifespan have been identified. However, mechanisms that control reproductive aging in nematodes or other animals are not well characterized. To use C. elegans to measure reproductive aging, we analyzed mated hermaphrodites that do not become sperm depleted and monitored the duration and level of progeny production. Mated hermaphrodites display a decline of progeny production that culminates in reproductive cessation before the end of the lifespan, demonstrating that hermaphrodites undergo reproductive aging. To identify factors that influence reproductive aging, we analyzed genetic, environmental, and pharmacological factors that extend lifespan. Dietary restriction and reduced insulin/insulin-like growth factor signaling delayed reproductive aging, indicating that nutritional status and a signaling pathway that responds to environmental stress influence reproductive aging. Cold temperature delayed reproductive aging. The anticonvulsant medicine ethosuximide, which affects neural activity, delayed reproductive aging, indicating that neural activity can influence reproductive aging. Some of these factors decrease early progeny production, but there is no consistent relationship between early progeny production and reproductive aging in strains with an extended lifespan. To directly examine the effects of early progeny production on reproductive aging, we used sperm availability to modulate the level of early reproduction. Early progeny production neither accelerated nor delayed reproductive aging, indicating that reproductive aging is not controlled by use-dependent mechanisms. The implications of these findings for evolutionary theories of aging are discussed.     

Genetic and environmental factors that influence production and quality of milk of Alpine and Saanen goats

Data from 1279 lactations of 783 Alpine and Saanen goats of the herd of our university in Minas Gerais, Brazil, were used to study environmental effects on and to estimate genetic parameters for milk production until 270 days of lactation (MP270) and for production and percentages of fat (PFAT and %FAT), protein (PPROT and %PROT), lactose (PLACT and %LACT), and total dry extract (PEXTR and %EXTR). Environmental effects were estimated by a statistical model that included contemporary group effect, type of kidding, genetic grouping, and kidding order. A multi-trait animal model with animal and permanent environment random effects was used to estimate genetic parameters and the significant environmental effects (fixed). Contemporary group influenced all traits; genetic grouping did not influence %LACT; type of kidding did not influence PFAT, %PROT or %LACT, and kidding order did not influence %FAT or %EXTR. Heritability and repeatability estimates were, respectively, 0.19 and 0.37 (MP270); 0.10 and 0.20 (PFAT); 0.12 and 0.24 (PPROT); 0.15 and 0.27 (PLACT); 0.13 and 0.24 (PEXTR); 0.21 and 0.34 (%FAT); 0.39 and 0.44 (%PROT); 0.17 and 0.29 (%LACT); 0.31 and 0.47 (%EXTR). Estimates of genetic correlations among MP270 and production of milk constituents were positive and high, but correlations between MP270 and %FAT, MP270 and %PROT, MP270 and %ESTR were moderate and negative. These heritability estimates show that satisfactory genetic gains can be obtained by selection, especially for milk constituents.     

Genetic variables that influence phenotype

Characterization of genetically engineered mice requires consideration of the gene of interest and the genetic background on which the mutation is maintained. A fundamental prerequisite to deciphering the genetic factors that influence the phenotype of a mutant mouse is an understanding of genetic nomenclature. Mutations and transgenes are often maintained on segregating or mixed backgrounds of often-unspecified origin. Minimizing the importance of strain and substrain differences, especially among 129 strains, can lead to poor experimental design or faulty interpretations of data. Genetic factors that influence phenotype can be categorized as traits that are unique to the background strain, unique to the gene of interest, or an interaction of both the background strain and the gene of interest. The commonly used inbred strains are generally well characterized and understood; however, specific genetic alterations combined with genes unique to the background inbred strain may lead to unexpected results. Genetic background effects can be analyzed and controlled for by using specific targeting and breeding strategies. Selection of appropriate experimental controls is critical. Ideally, mutations or transgenes should be characterized on more than one genetic background and in hybrids of the two progenitor strains. This approach may lead to the identification of novel genetic modifiers of the "gene of interest." Conditional mutagenesis technologies increase the options for controlling genetic background effects in addition to permitting the study of developmental and temporal changes in gene and protein expression and thus phenotype.     

Recombinant protein production and streptomycetes

The biopharmaceutical market has come a long way since 1982, when the first biopharmaceutical product, recombinant human insulin, was launched. Just over 200 biopharma products have already gained approval. The global market for biopharmaceuticals which is currently valued at over US$99 billion has been growing at an impressive compound annual growth rate over the previous years. To produce these biopharmaceuticals and other industrially important heterologous proteins, different prokaryotic and eukaryotic expression systems are used. All expression systems have some advantages as well as some disadvantages that should be considered in selecting which one to use. Choosing the best one requires evaluating the options--from yield to glycosylation, to proper folding, to economics of scale-up. No host cell from which all the proteins can be universally expressed in large quantities has been found so far. Therefore, it is important to provide a variety of host-vector expression systems in order to increase the opportunities to screen for the most suitable expression conditions or host cell. In this overview, we focus on Streptomyces lividans, a Gram-positive bacterium with a proven excellence in secretion capacity, as host for heterologous protein production. We will discuss its advantages and disadvantages, and how with systems biology approaches strains can be developed to better producing cell factories.     

Aptamers that bind to the antibiotic moenomycin A

Nuclease-resistant moenomycin-binding aptamers with dissociation constants in the range of 300 to 400 nM have been selected. Competition experiments have demonstrated that these aptamers recognize a disaccharide analogue of moenomycin. The results offer the opportunity of setting up a selective and sensitive assay for identifying moenomycin biosynthetic precursors.     

Beta-lactam and aminoglycoside production from streptomycetes

Streptomyces cattleya, S. fradiae and S. griseus produced different amounts of growth when cultured sequentially through sporulation, vegetative and antibiotic production media. Only S. griseus grew well on all three types of medium. Streptomyces cattleya grew poorly on both sporulation and vegetative media. Growth was 1.6 and 8.0 mg/1/h respectively. For all three species, biomass yield in the final antibiotic production medium was dependent on amount of inoculum. Antibiotic yields were obtained only from production media. Under slow growth conditions L-cysteine and L-valine supplementation stimulated S. cattleya beta-lactam production, giving 1000 micrograms/ml beta-lactam equivalents compared with 45 micrograms/ml beta-lactam equivalents for no supplementation. For aminoglycosides the agar well diffusion bioassay was more sensitive towards the hydrochloride than the neutral salt. Paper chromatography confirmed the main antibiotic classes. RF values for replicate samples indicated aminoglycoside homogeneity and beta-lactam heterogeneity.     

Aptamers that recognize the lipid moiety of the antibiotic moenomycin A

Moenomycin A is an amphiphilic phosphoglycolipid antibiotic that interferes with the transglycosylation step in peptidoglycan biosynthesis. The antibiotic consists of a branched pentasaccharide moiety, connected to the moenocinol lipid via a glycerophosphate linker. We have previously described the selection of aptamers that require the lipid group and the disaccharide epitopes of the oligosaccharide moiety for moenomycin binding. Here we report that the enriched moenomycin-binding library contains sequences that evolved for specific recognition of the unpolar lipid group of the antibiotic. These results suggest that the evolution of hydrophobic binding pockets in RNA molecules may be much more common than previously assumed.     

Genetic determinants of active antibiotic-producing soil streptomycetes.

Plasmids or covalently closed circular (CCC)-DNA molecules are abundant in the genus Streptomyces, and have been suggested to be involved in the genetic control of the production of many antibiotics in these organisms. In this study, 21 active antibiotic-producing Streptomyces isolates were screened for their plasmid content by an alkaline lysis method which revealed the presence of a small plasmid DNA in the positive control Streptomyces lividans ATCC 35287, containing pIJ702 plasmid (5.65 kb in size). However, no low molecular weight plasmids were observed in the tested antibiotic-producing Streptomyces strains suggesting that antibiotic production in these strains is likely chromosomally encoded DNA. Treatment of 2 Streptomyces strains with 10 mM ethidium bromide (EB) resulted in the failure to produce aerial mycelia and antibiotic activity.     

Genetic factors in broodstock management for seed production

Reviews in Fish Biology and Fisheries -     

Identification of molecular genetic factors that influence migraine

Migraine is a common neurological disorder with a strong genetic basis. However, the complex nature of the disorder has meant that few genes or susceptibility loci have been identified and replicated consistently to confirm their involvement in migraine. Approaches to genetic studies of the disorder have included analysis of the rare migraine subtype, familial hemiplegic migraine with several causal genes identified for this severe subtype. However, the exact genetic contributors to the more common migraine subtypes are still to be deciphered. Genome-wide studies such as genome-wide association studies and linkage analysis as well as candidate genes studies have been employed to investigate genes involved in common migraine. Neurological, hormonal and vascular genes are all considered key factors in the pathophysiology of migraine and are a focus of many of these studies. It is clear that the influence of individual genes on the expression of this disorder will vary. Furthermore, the disorder may be dependent on gene–gene and gene–environment interactions that have not yet been considered. In addition, identifying susceptibility genes may require phenotyping methods outside of the International Classification of Headache Disorders II criteria, such as trait component analysis and latent class analysis to better define the ambit of migraine expression.     

Standardization of factors that influence human urine metabolomics

In nutritional metabolomics a large inter- and intra-subject variability exists, and thus, it becomes important to limit the variance introduced by external factors. In a composite controlled study with full provision of all food for the standardized intervention, human urinary metabolite profiles were investigated for different factors, such as handling of urine collections, diet standardization, diet culture, cohabitation and gender. In study A, 8 healthy subjects (4 men; 4 women) collected 24-h urine, splitting each void into two specimens stored either at 4°C or at room temperature. In study B, 16 healthy subjects (7 men; 9 women) collected 24-h urine for three days while being on a standardized diet. Samples were analyzed by ¹H NMR and chemometrics. The NMR profiles indicated the presence of metabolites presumably originating from bacterial contamination in 3 out of 16 sample collections stored at room temperature. On the contrary, no changes in the NMR profiles due to contamination occurred in the 24-h urine samples stored at 4°C. The study also showed a trend towards a reduced inter- and intra-individual variation during 3 days of diet standardization. In study A, the urine metabolome showed a clear effect of diet culture and cohabitation, but these effects significantly attenuated after diet standardization (study B). Besides, gender-specific differences were found in both studies. Our results emphasize that best practice for any metabolomic study is a standardized, chilled sample collection procedure, and recommend that diet standardization is performed prior to dietary interventions in order to reduce intra- and inter-subject variability.     

β-Lactam and aminoglycoside production from streptomycetes

Streptomyces cattleya, S. fradiae and S. griseus produced different amounts of growth when cultured sequentially through sporulation, vegetative and antibiotic production media. Only S. griseus grew well on all three types of medium. Streptomyces cattleya grew poorly on both sporulation and vegetative media. Growth was 1·6 and 8·0 mg/1/h respectively. For all three species, biomass yield in the final antibiotic production medium was dependent on amount of inoculum. Antibiotic yields were obtained only from production media. Under slow growth conditions l -cysteine and l -valine supplementation stimulated S. cattleya β-lactam production, giving 1000 μg/ml β-lactam equivalents compared with 45 μg/ml β-lactam equivalents for no supplementation. For aminoglycosides the agar well diffusion bioassay was more sensitive towards the hydrochloride than the neutral salt. Paper chromatography confirmed the main antibiotic classes. R _F values for replicate samples indicated aminoglycoside homogeneity and β-lactam heterogeneity.     

Potential sites of geosmin production by streptomycetes in and around reservoirs

Water from several oligotrophic reservoirs of the North West Water Authority at Longridge, Lancashire, supported only sparse growth of streptomycetes isolated from the reservoirs. Growth was enhanced by nutrient-amendment of the water and one common isolate, identified as Streptomyces albidoflavus , produced geosmin in water which was supplemented with sufficient concentrations of available carbon, nitrogen and phosphorus. The minimum concentrations required for geosmin production were higher than those recorded in natural reservoir water. Synthesis of geosmin in water also required a minimum temperature of about 15dEC. Sterilized samples of reservoir sediment, sediment extract, plant debris from banks and surrounding soil also supported geosmin production by S. albidoflavus. Both bank debris and exposed sediment developed earthy odours without inoculation. The structure of reservoir banks influenced the quantity of plant growth and accumulation of litter, and therefore also the potential for contamination of reservoir water by geosmin. The potential of these various sites of geosmin production to contaminate reservoir water is assessed.