Secreted products and extracellular matrix from testicular peritubular myoid cells influence androgen-binding protein secretion by Sertoli cells in culture

Metabolic cooperation mediated by secreted factors between Sertoli cells and peritubular myoid cells has been well documented. We have confirmed that factors secreted by peritubular myoid cells modulate androgen-binding protein (ABP) secretion by Sertoli cells and shown further that this can also be achieved with peritubular myoid cell extracellular matrix (ECM). While peritubular myoid cell ECM potentiated the stimulatory effect of dibutyryl cyclic AMP on Sertoli cell ABP secretion, secreted factors did not, suggesting that the two components influence Sertoli cells through distinct mechanisms. We also tested other factors and other cell lines for effects on ABP production by Sertoli cells. The addition of human plasma fibronectin or conditioned medium from the basement membrane-producing Englebreth-Holm- Swarm sarcoma also stimulated ABP secretion by Sertoli cells. Cocultures of epithelial Sertoli cells with the cells of mesenchymal origin, such as testicular peritubular myoid cells, embryonic skin fibroblasts, and bladder smooth muscle cells, significantly stimulated ABP secretion by Sertoli cells, but co-culture with the epithelial-derived Martin-Darby canine kidney cell line had no effect on Sertoli cell-secreted ABP levels. Our data further define the epithelial-mesenchymal cell interaction that exists between Sertoli cells and peritubular myoid cells in the mammalian testis.

Collagen biosynthesis in cultured rat testicular Sertoli and peritubular myoid cells

The incorporation of 3H-proline into protein was regarded as a measure of total protein synthesis and the incorporation into hydroxyproline as indicative of collagen synthesis. Relative collagen synthesis (expressed as percent of total protein synthesized) by Sertoli and peritubular myoid cells cultured from 20-22 day old rat testis was estimated. In both secreted and cellular pools, relative collagen synthesis by Sertoli cells was significantly greater than by peritubular myoid cells. Coculture of Sertoli and myoid cells resulted in a significant increase in relative collagen synthesis when compared to monocultures of each cell type. Addition of serum to peritubular myoid cells resulted in a stronger stimulation of relative collagen production. Sertoli cell extracellular matrix inhibited relative collagen synthesis by peritubular myoid cells in the presence or absence of serum. Radioactivity into hydroxyproline as corrected per cellular DNA also showed similar results. Immunolocalization studies confirmed that both cell types synthesize type I and type IV collagens. These results indicate that stimulation of collagen synthesis observed in Sertoli-myoid cell cocultures is due to humoral interactions, rather than extracellular matrix, and Sertoli cell extracellular matrix regulates serum-induced increase in collagen synthesis by peritubular myoid cells.

On extension and torsion of a compressible elastic circular cylinder

In this paper we examine the combined extension and torsion of a compressible isotropic elastic cylinder of finite extent. The equilibrium equations are formulated in terms of the principal stretches and then applied to the special case of pure torsion superimposed on a uniform extension (an isochoric deformation). Explicit necessary and sufficient conditions on the strain-energy function for the material to support this deformation with vanishing traction on the lateral surfaces of the cylinder are obtained. Some strain-energy functions satisfying these conditions are considered, existing results are recovered as special cases and new results are obtained. We also point out how the strain-energy functions generated from the considered isochoric deformation considered (of a compressible material) can be used to generate energy functions and corresponding solutions for the incompressible theory.

Behaviour and strength of hollow flange channel sections under torsion and bending

Hollow flange channel section is a cold-formed high-strength and thin-walled steel section with a unique shape including two rectangular hollow flanges and a slender web. Due to its mono-symmetric characteristics, it will also be subjected to torsion when subjected to transverse loads in practical applications. Past research on steel beams subject to torsion has concentrated on open sections while very few steel design standards give suitable design rules for torsion design. Since the hollow flange channel section is different from conventional open sections, its torsional behaviour remains unknown to researchers. Therefore the elastic behaviour of hollow flange channel sections subject to uniform and non-uniform torsion, and combined torsion and bending was investigated using the solutions of appropriate differential equilibrium equations. The section torsion shear flow, warping normal stress distribution, and section constants including torsion constant and warping constant were obtained. The results were compared with those from finite element analyses that verified the accuracy of analytical solutions. Parametric studies were undertaken for simply supported beams subject to a uniformly distributed torque and a uniformly distributed transverse load applied away from the shear centre. This paper presents the details of this research into the elastic behaviour and strength of hollow flange channel sections subject to torsion and bending and the results.

Bending and torsion of hollow flange channel beams

The LiteSteel beam (LSB) is a cold-formed high strength steel channel section made of two torsionally rigid closed flanges and a slender web. Due to its mono-symmetric characteristics, its centroid and shear centre do not coincide. The LSBs can be used in floor systems as joists or bearers and in these applications they are often subjected to transverse loads that are applied away from the shear centre. Hence they are often subjected to combined bending and torsion actions. Previous researches on LSBs have concentrated on their bending or shear behaviour and strengths, and only limited research has been undertaken on their combined bending and torsion behaviour. Therefore in this research a series of nine experiments was first conducted on LSBs subject to combined bending and torsion. Three LSB sections were tested to failure under eccentric loading at mid-span, and appropriate results were obtained from seven tests. A special test rig was used to simulate two different eccentricities and to provide accurate simple boundary conditions at the supports. Finite element models of tested LSBs were developed using ANSYS, and the ultimate strengths, failure modes, and load–displacement curves were obtained and compared with corresponding test results. Finite element analyses agreed well with test results and hence the developed models were used in a parametric study to investigate the effects of load locations, eccentricities, and spans on the combined bending and torsion behaviour of LSBs. The interaction between the ultimate bending and torsional moment capacities was studied and a simple design rule was proposed. This paper presents the details of the tests, finite element analyses, and parametric study of LSBs subject to combined bending and torsion, and the results.

Immunization of male bonnet monkeys (M-radiata) with a recombinant FSH receptor preparation affects testicular function and fertility

Immunization of proven fertile adult male monkeys (n = 3) with a recombinant FSH receptor protein preparation (oFSHR-P) (representing amino acids 1-134 of the extracellular domain of the receptor Mr similar to 15KDa) resulted in production of receptor blocking antibodies. The ability of the antibody to bind a particulate FSH receptor preparation and receptors in intact granulosa cells was markedly (by 30-80%) inhibited by FSH. Serum T levels and LH receptor function following immunization remained unchanged. The immunized monkeys showed a 50% reduction (p<0.001) in transformation of spermatogonia(2C) to primary spermatocytes (4C) as determined by flow cytometry and the 4C:2C ratio showed a correlative change (R 0.81, p<0.0007) with reduction in fertility index (sperm counts X motility score). Breeding studies indicated that monkeys became infertile between 242-368 days of immunization when the fertility index was in the range of 123+/-76 to 354+/-42 (compared to a value of 1602+/-384 on day 0). As the effects observed ate near identical to that seen following immunization with FSH it is suggestive that oFSHR-P can substitute for FSH in the development of a contraceptive vaccine.

Determination of stress-intensity factors for cracks in tubes under torsion

An experimental investigation by two-dimensional photoelastic technique is carried out to study the stress distribution and to determine the stress-intensity factors for arbitrarily oriented cracks in thin cylindrical shells subjected to torsion. A new method is employed to evaluate the pure and mixed-mode SIF's.

Androgen receptor-dependent and independent functions of ARIP4

Androgen receptor (AR) is necessary for normal male phenotype development and essential for spermatogenesis. AR is a classical steroid receptor mediating actions of male sex steroids testosterone and 5-alpha-dihydrotestosterone. Numerous coregulators interact with the receptor and regulate AR activity on target genes. This study deals with the characterization of androgen receptor-interacting protein 4 (ARIP4). ARIP4 binds DNA, interacts with AR in vitro and in cultured yeast and mammalian cells, and modulates AR-dependent transactivation. ARIP4 is an active DNA-dependent ATPase, and this enzymatic activity is essential for the ability of ARIP4 to modulate AR function. On the basis of sequence homology in its ATPase domain, ARIP4 belongs to the SNF2 family of proteins involved in chromatin remodeling, DNA repair, and homologous recombination. Similar to its closest homologs ATRX and Rad54, ARIP4 does not seem to be a classical chromatin remodeling protein in that it does not appear to form large protein complexes in vivo or remodel mononucleosomes in vitro. However, ARIP4 is able to generate superhelical torsion on linear DNA fragments. ARIP4 is covalently modified by SUMO-1, and mutation of six potential SUMO attachment sites abolishes the ability of ARIP4 to bind DNA, hydrolyze ATP, and activate AR function. ARIP4 expression starts in early embryonic development. In mouse embryo ARIP4 is present mainly in the neural tube and limb buds. In adult mouse tissues ARIP4 expression is virtually ubiquitous. In mouse testis ARIP4 is expressed in the nuclei of Sertoli cells in a stage-dependent manner. ARIP4 is also present in the nuclei of Leydig cells, spermatogonia, pachytene and diplotene spermatocytes. Testicular expression pattern of ARIP4 does not differ significantly in wild-type, FSHRKO, and LuRKO mice. In the testis of hpg mice, ARIP4 is found mainly in interstitial cells and has very low, if any, expression in Sertoli and germ cells. Heterozygous Arip4+/ mice are fertile and appear normal; however, they are haploinsufficient with regard to androgen action in Sertoli cells. In contrast, Arip4 / embryos are not viable. They have significantly reduced body size at E9.5 and die by E11.5. Compared to wild-type littermates, Arip4 / embryos possess a higher percentage of apoptotic cells at E9.5 and E10.5. Fibroblasts derived from Arip4 / embryos cease growing after 2-3 passages and exhibit a significantly increased apoptosis and decreased proliferation rate than cells from wild-type embryos. Our findings demonstrate that ARIP4 plays an essential role in mouse embryonic development. In addition, testicular expression and AR coregulatory activity of ARIP4 suggest a role of ARIP4-AR interaction in the somatic cells of the testis.

Influence of electroslag refining on the hot ductility of AISI 4340 steel in torsion

Electroslag refining is a useful remelting process by which clean steels can be produced for sophisticated applications. In this investigation, AISI 4340 steel has been electroslag refined and the improvement in its hot ductility has been assessed using hot torsion tests; electroslag refining has improved the hot ductility considerably. The temperature at which peak ductility is obtained has also increased — from 1473 K in the unrefined steel to 1573 K in ESR steel. Results indicate that it should be possible to subject the ESR ingot to much higher strains per unit operation during industrial hot working processes such as forging, which would result in a considerable saving of power. The improvement in hot ductility in ESR steel has been attributed primarily to the removal of non-metallic inclusions and the reduction in sulphur content. From the apparent activation energy estimated from the hot torsion data, the dynamic recrystallization process is identified as the mechanism controlling the rate of hot deformation.

Changes in testicular function following specific deprivation of LH in the adult male rabbit

Sexually mature male rabbits actively immunized against highly purified ovine LH (oLH) were used as a model system to study the effects of endogenous LH deprivation (and therefore testosterone) on spermatogenesis as well as pituitary FSH secretion. Immunization against oLH generated antibody titres capable of cross-reacting and neutralizing rabbit LH and this resulted in a significant reduction (P<0.01) in serum testosterone levels by 2-4 weeks of immunization. A significant increase in circulating FSH concentration (from a basal level of similar to 1 ng to 60-100 ng/ml; P<0.01) was observed within 4-6 weeks of immunization, perhaps a consequence of the negative feedback effect of the lack of testosterone. The effect of LH deprivation on spermatogenesis assessed by DNA flow cytometry and histological analyses of testicular biopsy tissue revealed that lack of testosterone primarily results in a rapid reduction and complete absence of round (1C) and elongated (HC) spermatids. The immediate effect of LH/testosterone deprivation thus appears to be at the step of meiotic transformation of primary spermatocytes (4C) to 1C. A significant reduction (>80%; P<0.01) in the 4C population and a relative accumulation (>90%; P<0.01) in spermatogonia (2C) was also observed, suggesting a need for testosterone during the transformation of 2C to 1C. In all but one of the rabbits, both qualitative and quantitative recovery in spermatogenesis occurred during the recovery phase, even at a time when only a marginal increase in serum testosterone (compared with the preimmunization) levels was observed as a result of a rapid decline in the cross-reactive antibody titres. These results clearly show that LH/testosterone deprivation in addition to primarily affecting the meiotic step also regulates the conversion of 2C to 4C during spermatogenesis.

Testicular function in adolescent boys with Klinefelter syndrome

Klinefelter syndrome (KS) is the most frequent karyotype disorder of male reproductive function. Since its original clinical description in 1942 and the identification of its chromosomal basis 47,XXY in 1959, the typical KS phenotype has become well recognized, but the mechanisms behind the testicular degeneration process have remained unrevealed. This prospective study was undertaken to increase knowledge about testicular function in adolescent KS boys. It comprised a longitudinal follow-up of growth, pubertal development, and serum reproductive hormone levels in 14 prepubertal and pubertal KS boys. Each boy had a testicular biopsy that was analyzed with histomorphometric and immunohistochemical methods. The KS boys had sufficient testosterone levels to allow normal onset and progression of puberty. Their serum testosterone levels remained within the low-normal range throughout puberty, but from midpuberty onwards, findings like a leveling-off in testosterone and insulin-like factor 3 (INSL3) concentrations, high gonadotropin levels, and exaggerated responses to gonadotropin-releasing hormone stimulation suggest diminished testosterone secretion. We also showed that the Leydig cell differentiation marker INSL3 may serve as a novel marker for onset and normal progression of puberty in boys. In the KS boys the number of germ cells was already markedly lower at the onset of puberty. The pubertal activation of the pituitary-testicular axis accelerated germ cell depletion, and germ cell differentiation was at least partly blocked at the spermatogonium or early primary spermatocyte stages. The presence of germ cells correlated with serum reproductive hormone levels. The immature Sertoli cells were incapable of transforming to the adult type, and during puberty the degeneration of Sertoli cells increased markedly. The older KS boys displayed an evident Leydig cell hyperplasia, as well as fibrosis and hyalinization of the interstitium and peritubular connective tissue. Altered immunoexpression of the androgen receptor (AR) suggested that in KS boys during puberty a relative androgen deficiency develops at testicular level. The impact of genetic features of the supernumerary X chromosome on the KS phenotype was also studied. The present study suggests that parental origin of the supernumerary X chromosome and the length of the CAG repeat of the AR gene influence pubertal development and testicular degeneration. The current study characterized by several means the testicular degeneration process in the testes of adolescent KS boys and confirmed that this process accelerates at the onset of puberty. Although serum reproductive hormone levels indicated no hypogonadism during early puberty, the histological analyses showed an already markedly reduced fertility potential in prepubertal KS boys. Genetic features of the X chromosome affect the KS phenotype.

Mixed-mode stress intensity factors for arbitrarily oriented cracks in cylindrical shells under axial pull and torsion

Results of photoelastic investigations conducted on cylindrical tubes (made of Araldite material) containing cracks oriented at 0°, 30°, 45°, 60° and 90° to the axis of the tube and subjected to axial and torsional loads are reported. The stress-intensity factors (SIFs) were determined by analysing the crack-tip stress fields. Smith and Smith's method [Engng Fracture Mech.4, 357–366 (1972)] and a new method developed by the authors by modifying Rakesh et al.'s method [Proc. 26th Congress of ISTAM, India (1981)] were employed to evaluate the mixed-mode SIFs.

Bond length distortions associated with torsion potentials; ab-initio studies on methanedio

The 4-31G basis set is used to study the bond length variations as functions of dihedral angels in methanediol. This study is compared with O---C---O bond angle optimization studies by Gorenstein and Kar and the possible reason for bond length shorteing in the trans---trans configuration is analysed.