A-factor and streptomycin biosynthesis inStreptomyces griseus

Accumulating data have shown that the metabolites with a -butyrolactone ring functions as an autoregulatory factor or a microbial hormone for the expression of various phenotypes not only in a variety ofStreptomyces spp. but also in the distantly related bacteria. A-factor, as a representative of this type of autoregulators, triggers streptomycin biosynthesis and cellular differentiation inStreptomyces griseus. A model for the A-factor regulatory cascade on the basis of recent work is as follows. At an early step in the A-factor regulatory relay, the positive A-factor signal is first received by an A-factor receptor protein that is comparable in every aspect to eukaryotic hormone receptors, and then, via one or more regulatory steps, transmitted to an A-factor-responsive protein that binds to the upstream activation sequence of thestrR gene, a regulatory gene in the streptomycin biosynthetic gene cluster. The StrR protein thus induced appears to activate the other streptomycin biosynthetic genes. This review summarizes the characteristics of A-factor as a microbial hormone and the A-factor regulatory relay leading to streptomycin production.     

Further studies on the metabolism of phospholipids inStreptomyces griseus: Turnover of phospholipid moieties

Phospholipid turnover inStreptomyces griseus was studied by pulse-chase techniques using 1-[¹⁴C]sodium acetate and [U-¹⁴C]glucose. Different phospholipids and their individual moieties were found to have different turnover rates. The moieties of inositol-containing phospholipids exhibited the fastest turnover rates among the major phospholipids, while only fatty acyl moieties of phosphatidylethanolamine showed rapid turnover. Cardiolipin did not show any significant turnover with both precursors.     

Regulation of biosynthesis of pesticidal metabolic complexes inStreptomyces griseus

The complex of pesticidal metabolites produced byStreptomyces griseus LKS-1 consists of a peptide antibiotic (A), nonactic acids (B), macrotetrolides (C), pyrrolizines (D), and of cycloheximide. The latter unwanted phytotoxic compound was eliminated by treatment with mutagens. Combined approaches, including both genetic and physiological manipulations, resulted in the following alterations in the biosynthetic capacity: (1) A more than 80-fold increase in the production of C under a substantial decrease in the yields ofA, B andD, the ratio of the components ofC being steered toward the required more active ones; (2) a more than 300-fold increase in the production ofB under suppression of the formation ofA andC: (3) a 10-fold increase in the yields ofD under suppression ofA andC; (4 a significant increase in the yields ofA with eliminatingB, C andD. The level of inorganic phosphate in fermentation media and the sensitivity of the organism to carbon catabolite repression were important factors participating in the regulation of the above biosynthetic processes.     

Silence of streptomycin 6-phosphotransferase gene derived by incubation at a high temperature inStreptomyces griseus

Summary The bald mutants from streptomycin (SM)-producingStreptomyces griseus 2247 obtained by incubation at high temperature (36° C), designated as HT strains, lost resistance to their own antibiotic and scarcely produced the antibiotic. Although SM susceptibility in the mutant was due to loss of SM 6-phosphotransferase activity produced in the cell, the gene coding for the enzyme cloned from an HT strain was surely expressed inS. lividans 1326 as a host. Northern blot analysis showed that the corresponding RNA is not detected in the mutant, indicating that though the gene encoding SM 6-phosphotransferase, at least, the structural gene is not deleted in the cell, the expression is silent.     

Nonlinear tensile properties of bovine articular cartilage and their variation with age and depth

Tensile stiffness of articular cartilage is much greater than its compressive stiffness and plays an essential role even in compressive properties by increasing transient fluid pressures during physiological loading. Recent studies of nonlinear properties of articular cartilage in compression revealed several physiologically pertinent nonlinear behaviors, all of which required that cartilage tensile stiffness increase significantly with stretch. We therefore performed sequences of uniaxial tension tests on fresh bovine articular cartilage slices using a protocol that allowed several hours to attain equilibrium and measured longitudinal and transverse tissue strain. By testing bovine cartilage from different ages (6 months to 6 years) we found that equilibrium and transient tensile modulus increased significantly with maturation and age, from 0 to 15 MPa at equilibrium and from 10 to 28 MPa transiently. Our results indicate that cartilage stiffens with age in a manner similar to other highly hydrated connective tissues, possibly due to age-dependent content of enzymatic and nonenzymatic collagen cross links. The long relaxation period used in our tests (5-10 hours) was necessary in order to attain equilibrium and avoid a very significant overestimation of equilibrium modulus that occurs when much shorter times are used (15-30 minutes). We also found that equilibrium and transient tensile modulus increased nonlinearly when cartilage is stretched from 0 to 10% strain without any previous tare load. Although our results estimate a nonlinear increase in tensile stiffness with stretch that is an order of magnitude lower than that required to predict nonlinear properties in compression, they are in agreement with previous results from other uniaxial tension tests of collagenous materials. We therefore speculate that biaxial tensile moduli may be much higher and thereby more compatible with observed nonlinear compressive properties.     

Acyl lipid composition variation related to culture age and nitrogen concentration in continuous culture of the microalga Phaeodactylum tricornutum

The influence of culture age and nitrogen concentration on the distribution of fatty acids among the different acyl lipid classes has been studied in continuous cultures of the microalga Phaeodactylum tricornutum. The culture age was tested in the range of 1.15-7 days, controlled by adjusting the dilution rate of fresh medium supplied. The effect of nitrogen concentration was tested from saturating conditions to starvation by modifying nitrate concentration in the fresh medium. Culture age had almost no influence on the fatty acid content; 16:0, 16:3 and 20:5 increased moderately wherein the level of 16:1 decreased when the culture age decreased. Culture age had no effect on the total fatty acid content that remained around 11% of dry weight. Conversely, culture age had a greater impact on lipid classes, producing changes in amounts of triacylglycerols (TAG) which ranged between 43% and 69%, and galactolipids (GLs) that oscillated between 20% and 40%. In general, the content of polar lipids of the biomass decreased with culture age. The other factor assayed, nitrogen content, affected the fatty acid profile. Saturated and monounsaturated fatty acids accumulated when the nitrogen concentration was decreased. The experiments regarding the effect of nitrogen concentration on lipid species were carried out with cells of an average age of 3.5 days. A decrease of the nitrogen concentration caused the GL fraction to decrease from 21 to 12%. Conversely, both neutral lipids (NLs) and phospolipids (PLs) increased from about 73 to 79% and from 6 to 8%, respectively. In these experiments, TAG was the lipid class with the highest increase, from 69 to 75%.     

Leukaemomycin-blocked mutants of Streptomyces griseus and their pigments

In a continued search for leukaemomycin-blocked mutants of three leukaemomycin-producing strains IMET JA 3933, IMET JA 5142 and IMET JA 5570 of Streptomyces griseus, 32 mutants producing aerial mycelium and spores were detected. Furthermore, in all mutants cosynthetic capability has been observed. This report describes characterization of leukaemomycin-blocked mutants obtained by mutagenic treatment experiments using NTG and combined UV-/X-rays. According to the biosynthetic capability for anthracyclinones or other pigments the mutants could be divided into six classes. The first class contains 14 leukaemomycin-blocked mutants unable to synthesize anthracyclinones. Besides two classes of mutants (12)synthesizing well-known anthracyclinones as epsilon-rhodomycinone, 7-deoxy-epsilon-rhodomycinone, 11-deoxy-derivatives of daunomycinone, three new classes of mutants (6) synthesizing reddish-brown, brown and blue-violet pigments on solid media with structures not elucidated as yet, will be described.     

Ventilatory threshold: measurement and variation with age

The purpose of this study is to present measurement of ventilatory threshold (VeT) and maximal oxygen uptake (VO2max) in a large group of predominantly older subjects using a bicycle ergometer and an automated measuring system. One hundred and twenty-seven healthy elderly subjects (mean age: 68) and 44 young and middle-aged subjects (mean age: 39) underwent a maximal exercise test with breath-by-breath measurement of ventilation and gas exchange variables. Ventilatory threshold was determined by visual inspection of the breakpoints in the VE/VO2 and PETO2 data curves. Additional measures were made in a subset of subjects to determine the reproducibility and interobserver variability of VeT and the relationship between VeT and the venous lactate threshold (LaT). Day-to-day reproducibility of VeT was good with a mean difference in VO2 at VeT on two occasions of 40.23 +/- 125 ml/min. Interobserver variability was low (intraclass correlation coefficient of r = 0.941) and VeT was found to correlate to LaT (r = 0.79, P less than 0.05) with LaT occurring a mean 2.3 min after VeT. VeT declined significantly with age in both males and females but less rapidly than VO2max. Both VO2max and VeT were found to vary with age, sex, height, and weight in a stepwise multiple-linear regression analysis. Age-associated changes in skeletal muscle composition may be in part responsible for the less precipitous decline in VeT with age compared with VO2max.     

Ionic charge on phospholipids and their interaction with the mitochondrial adenosine triphosphatase

The activity of the lipid-depleted, oligomycin-sensitive mitochondrial ATPase has been measured in the presence of liposomes prepared from mixtures of phosphatidylglycerol and phosphatidylglycerol lysine. Enzyme activity increased linearly with an increase in the negative charge of liposomes prepared from the phosphatidylglycerol-phosphatidylglycerol lysine mixtures. The electrophoretic mobility and activating capacity of liposomes of several other phospholipids were determined. A linear relationship between electrophoretic mobility of the liposomes and oligomycin-sensitive activity was again apparent. These observations demonstrate that the activity of the ATPase is directly proportional to the ionic charge on phospholipid activators if the acyl chain composition of the phosphoglycerides is relatively constant.     

Discrimination of different generations of Zeugodacus scutellata using age grading technique and their local genetic variation

The striped fruit fly, Zeugodacus scutellata, exhibits generation overlap due to its relatively long lifespan. This study clarified the fly generations by age-grading technique and traced migration of local populations using genetic markers. Male adults were monitored using Cue-lure in two different places (Andong and Guri) of mainland Korea during the entire host cultivating period. They showed two occurrence peaks (from late July to early August and from mid-August to mid-October). A developmental model of Z. scutellata using rate summation method predicted four generations during this period. Pterin amounts in fly heads increased with adult age and indicated at least 3 cycles of age variation during the occurrence period with an additional adult generation after the third cycle. The additional peak was supported by field monitoring using protein diet supplemented with terpinyl acetate, which showed an additional occurrence peak during October. These seasonal populations were monitored through their migration by gene flow analysis using randomly amplified polymorphic DNA (RAPD) markers. In each locality, three seasonal populations at July, August, and September were discrete. Although local populations were genetically discrete at July, they were not separate at late seasons except for Jeju populations. These results indicate the usefulness of the head pterin quantification in age-grading of adult populations and suggest that Z. scutellata occurs at least four generations a year. In addition, gene flow analysis using RAPD suggests frequent movements among local populations along with season progression.     

Synthesis of lipid A type carboxymethyl derivatives with ether chains instead of ester chains and their LPS-antagonistic activities

Synthesis of lipid A type carboxymethyl derivatives having ether chains at both the C-3 and C-3' positions and their LPS-antagonistic activities toward human U937 cells are described.     

Growth patterns,and age and size at maturity in femaleCottus hangiongensis,with special reference to their life-history variation

Growth patterns of the 1982 year-class, individual growth patterns, age and size at sexual maturity and longevity in females of the river-sculpin,Cottus hangiongensis (Cottidae), were examined along the course of the Daitobetsu River of southern Hokkaido, Japan. Growth of females slightly varied both along the river course and among individual fishes: slow growth occurs in females from the lower reaches, while more rapid growth occurs in females from upstream areas. Body size and age at the first sexual maturity of females slightly increased towards the upstream, from 52 mm SL and 2 years in the most downstream area to 72 mm SL and 2–3 years in the uppermost site. Longevity was estimated to be 7 years in the downstream areas and 8–9 years in the upstream sites. These results suggest that female life history varies along the course of the river and thus allow us to consider the following alternative reproductive tactics: when females stay in the lower reaches, they attain sexual maturity at a smaller body size and younger age, and have a small clutch size, but when females migrate into the upper reaches, their maturity is delayed until they reach a larger body size and older age, and have a greater clutch size.     

Fatty chains of alkenylacyl, alkylacyl and diacyl phospholipids in sea urchin spermatozoa.

1. Alkenylacyl, alkylacyl and diacyl phospholipids were analyzed in the spermatozoa of the sea urchin, Hemicentrotus pulcherrimus. 2. Choline phosphoglycerides (CPG) contained alkylacyl component (19%) in addition to the diacyl component (81%), and alkenylacyl analog was present in a trace amount. The ethanolamine phosphoglycerides (EPG) contained alkenylacyl (51%), alkylacyl (2%) and diacyl (47%) components and the serine phosphoglycerides (SPG), alkylacyl (9%) and diacyl (91%) derivatives. 3. Analysis by gas-liquid chromatography indicated that the fatty chain at the 1-position in alkenylacyl, alkylacyl and diacyl compounds of CPG, EPG and SPG was mainly composed of saturated and monoenoic types (16:0, 18:0, 18:1 and 20:1). In contrast, considerable amounts of polyunsaturated types (20:4 and 20:5) were noted at the 2-position.     

Isolation and characterization of glycocalyx inStreptomyces aureofaciens

The surface layer, considered to be glycocalyx according to electron-microscopic observations, was separated from a lowproduction strain ofStreptomyces aureofaciens by solubilization with urea and subsequent sonication. The isolation procedure was developed using various agents; neutral phosphatase served as a marker indicating the amount of the material released. The peripheral structure consisted predominantly of glycoprotein and differed from S-layers.     

Isolation of glycosaminoglycans and variation with age in the feline brain

M any laboratories have reported the presence of glycosaminoglycans (GAG) in the brains of a variety of species, including monkey, man, cat, pig, rabbit, sheep, rat and chicken (C handrasekaran and B achhawat , 1969; C lausen and H ansen , 1963; C lausen and R osenkast , 1962; C ustod and Y oung , 1968; M argolis , 1967; O nodera , H irona , H oriuchi and K ashimura , 1966; S aigo and E gami , 1970; S ingh and B achhawat , 1965 , 1968; S ingh , C handrasekaran , C herian and B achhawat , 1969). Because of the interest of our laboratory in the GAG of the cat brain (C ustod and Y oung , 1968; Y oung , 1963), we initiated the present studies to quantify these various brain polysaccharides and to investigate their changes as a function of postnatal age.     

Morphometric analysis of variation in the sternum with sex and age

Age and sex‐related variations in sternum morphology may affect the thoracic injury tolerance. Male and female sternum size and shape variation was characterized for ages 0–100 from landmarks collected from 330 computed tomography scans. Homologous landmarks were analyzed using Procrustes superimposition to produce age and sex‐specific functions of 3D‐sternum morphology representing the combined size and shape variation and the isolated shape variation. Significant changes in the combined size and shape variation and isolated shape variation of the sternum were found to occur with age in both sexes. Sternal size increased from birth through age 30 and retained a similar size for ages 30–100. The manubrium expanded laterally from birth through age 30, becoming wider in relation to the sternal body. In infancy, the manubrium was 1.1–1.2 times the width of the sternal body and this width ratio increased to 1.6–1.8 for adults. The manubrium transformed from a circular shape in infancy to an oval shape in early childhood. The distal sternal body became wider in relation to the proximal sternal body from birth through age 30 and retained this characteristic throughout adulthood. The most dramatic changes in sternum morphology occur in childhood and young adulthood when the sternum is undergoing ossification. The lesser degree of ossification in the pediatric sternum may be partly responsible for the prevalence of thoracic organ injuries as opposed to thoracic skeletal injuries in pediatrics. Sternum fractures make up a larger portion of thoracic injury patterns in adults with fully ossified sternums. The lack of substantial size or shape changes in the sternum from age 30–100 suggests that the increased incidence of sternal fracture seen in the elderly may be due to cortical thickness or bone mineral density changes in the sternum as opposed to morphological changes. J. Morphol. 275:1284–1299, 2014. © 2014 Wiley Periodicals, Inc.     

Role of ATP-glucokinase and polyphosphate glucokinase inStreptomyces aureofaciens

The activity of ATP-glucokinase and of polyphosphate glucokinase was examined during growth of the actinomyceteStreptomyces aureofaciens 8425 under conditions of intense chlortetracycline (CTC) synthesis. ATP-glucokinase was active in the strain only during the logarithmic phase of culture growth; the activity of polyphosphate glucokinase appears only at the end of the logarithmic phase of growth and rises in parallel with the rate of CTC biosynthesis in the stationary phase. During the rise of activity of polyphosphate glucokinase and of CTC biosynthesis the cells accumulate sugar phosphates, mainly glucose-6-phosphate. It appears that the biosynthesis of CTC inStreptomyces aureofaciens takes place at the expense of glycolysis, using up the high-energy phosphate of high-molecular polyphosphates.