Post-transcriptional block in oxytocin gene expression within the seminiferous tubules of the bovine testis

Immunodeficiency is frequently invoked as an ethiopathogenetic factor for many somatic diseases. On the other hand, stress, depression, and psychotic disturbances are associated with severe immunological disorders. Taking into account that the benzodiazepines (BZ) are the psychoactive drugs more widely used than any other to treat psychological disturbances, it seems important to elucidate the immuno-enhancing or immunosuppressant potential of such drugs. Our goal was easily reached, since 69% of the outpatients visiting our Institute are chronic BZ consumers and because neurochemical, hormonal, immunological, and psychiatric investigations are routinely performed on all of our patients. In the present study, immune function was investigated on two occasions: while the patient was on active medication and 15 days after discontinuation. We concluded that chronic consumption of BZ provokes significant immunological disorders that should be further investigated. Said disorders could not be linked to a pre-existing affective disease or psychosis, since we only selected those BZ users in whom psychiatric investigations ruled out a past or present history of major psychiatric disease.     

Dynamics of development of the interstitial tissue and seminiferous tubules in the testis of human fetuses

Morphological changes in the testis of human fetuses (period of embryogenesis--from 8 to 28 weeks) during their differentiation were studied by histological and morphological methods. Investigations carried out demonstrated that complete cycle of transformation in the intestitial and embryonic tissue of the fetal testis could be divided into two main periods: the first period was attended by a gradual increase in the count and differentiation of Heydig's cells in the interstitial tissue (from the 8th to the 12th week of embryogenesis); gradual involution of Leydig's cells in the interstitial tissue and development of seminiferous tubules prevailed during the second period (from the 15th to the 28th week of embryogenesis). In the course of prenatal development of the human fetus testis Leydig's cells passed through a number of successive stages, each one being characterized by a definite functional activity level of these cells. The detected changes in the testes were apparently connected both with the changes in the Leydig's cells count and their functional activity, and with progressive growth of the tubules.     

Evidence for production and functional activity of nitric oxide in seminiferous tubules and blood vessels of the human testis

Previous studies have demonstrated that nitric oxide (NO) influences Leydig cell function. Here we provide evidence for NO production and activity in seminiferous tubules and blood vessels of the human testis. By immunohistochemistry, the soluble guanylyl cyclase (sGC), the intracellular NO receptor, and the second messenger, cyclic guanosine monophosphate (cGMP), were detected in myofibroblasts of the peritubular lamina propria in Sertoli cells, as well as in endothelial and smooth muscle cells of testicular blood vessels. Performed with isolated tubules and blood vessels, the biological activity of sGC could be proved by cGMP generation in response to treatments with the NO donor, sodium nitroprusside. The endothelial and neuronal subtypes of NO synthase (NOS) were localized immunohistochemically to the same cell types that express sGC and cGMP. In isolated tubules and vessels, the presence of endothelial NOS and neuronal NOS was confirmed by immunoblotting, and NOS activity was demonstrated by decreased cGMP production upon incubation with the NOS inhibitor L-nitro arginine methylester. These findings show that peritubular cells, Sertoli cells, and testicular blood vessels may be sites of NO production and activity, possibly involved in relaxation of seminiferous tubules and blood vessels to modulate sperm transport and testicular blood flow, respectively.     

Retinyl palmitate, a major retinyl ester in rat testis seminiferous tubules. II. The Sertoli cell, site of retinol esterification

The study was designed to 1) isolate and quantitate retinyl esters normally present in the testis seminiferous tubules (ST) and 2) determine whether the Sertoli (SE) cells in the ST have the enzyme system necessary to esterify retinol. In the first study testes from normal adult male rats were removed, the ST were isolated mechanically and retinol was extracted and separated into free and retinyl esters in a prestandardized alumina column. The fraction containing retinyl esters was hydrolyzed, and the fatty acid composition was determined gas chromatographically. The results showed that retinyl palmitate was the major retinyl ester in the testis ST constituting almost 55% of the total esters. In the second study adult male rats were given Busulfan (1,4, dimethylsulfonoxybutane) intraperitoneally twice six weeks apart. At the end of six weeks following injection, the testes from a few animals were examined histologically, and the results showed that Busulfan treatment destroyed the germinal epithelium completely but had no effect on the structural integrity of the SE cells. Labeled retinol (free) mixed in rat albumin solution was injected intratesticularly (in vivo study) or incubated (in vitro study) with ST which had been isolated from the testes of rats treated with Busulfan. The results of the studies indicate that labeled retinol (free) was converted to retinyl esters in the ST. Since the ratio of conversion of free retinol (labeled) to retinyl esters remained constant in the treated ST (which contained only Sertoli cells, the germinal epithelium having been destroyed) and in the control ST (which contained both Sertoli cells and germinal epithelium), we conclude that the Sertoli cells are the site of retinol esterification in the testis ST.     

Relaxin-like factor expression as a marker of differentiation in the mouse testis and ovary

Expression of the relaxin-like factor (RLF) was studied at the messenger RNA (mRNA) and protein levels in the testes and ovaries of the mouse, as well as through testicular development and differentiation in the mouse testis. In situ hybridization or RT-PCR, and immunohistochemistry using a polyclonal antibody raised against a recombinant protein, provided mutually confirmatory results for a high expression of RLF in the Leydig cells of the adult testis and at a much lower level of expression in the luteal cells of the ovary through the cycle, pregnancy, and in lactation. Analysis of protein and mRNA expression, through postnatal testicular development, indicated moderate RLF expression also in the fetal population of Leydig cells, even in the hpg mutant mouse, lacking an active pituitary-gonadal axis. Prepubertal Leydig cells, however, exhibit only very low-level RLF gene expression, this phenotype persisting in the adult hpg mouse. In summary, fetal Leydig cells express RLF in an LH/human CG-independent fashion, whereas LH/human CG is essential to induce RLF expression in the adult-type Leydig cell. In cultured adult Leydig cells or in the mouse tumor MA-10 cell line, RLF mRNA is expressed in a constitutive fashion. RLF thus seems to be a useful marker of Leydig cell differentiation status.     

Integration versus differentiation of fluid/crytallized intelligence in old age.

Used an intra-age simulation approach to evaluate several models of psychometric intelligence differing primarily along the developmental continuum of differentiation–integration for older adults. 109 adults (aged 60–89 yrs) were administered a battery of 17 ability tests selected to mark R. B. Catell's (1971) and J. L. Horn's (1970, 1978) fluid (Gf) and crystallized intelligence (Gc) and speed dimensions. Using the Gf/Gc model of intelligence and intermediate outcomes as guidelines, structural models with between 7 and 1 factors were successively evaluated by confirmatory factor analysis. Three main findings emerged. First, models with fewer factors provided better fits to the data. Second, a model with a general factor and 3 group factors was especially acceptable based on empirical and theoretical criteria. Third, it was not possible to obtain an acceptable solution for the older Ss that would directly parallel the Gf/Gc structural pattern of 1st-stratum factors reported for younger age groups. In concert, the findings support a neointegration, or dedifferentiation, view of psychometric intelligence in old age. (40 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

转录因子Ets1和Ets2在小鼠睾丸组织中的表达及其意义

目的:检测Ets家族转录因子Ets1和Ets2在小鼠睾丸组织中的表达,探讨其对小鼠睾丸发育的调控及其对精原干细胞(SSCs)增殖、分化的可能影响.方法:取生后第1、5、10、15、20、25、30、35、40、50和70天的小鼠睾丸组织,对成年小鼠进行白消安10 mg·kg-1腹腔注射,分别在注射后第0、3、5、8、10和18天取睾丸组织,用半定量RT-PCR方法对比内参β-actin在对应组织内的表达水平,分析Ets1、Ets2 mRNA在睾丸组织中的相对表达量.结果:Ets1的表达量在生后第1～30天显著高于出生后第35天(P<0.05或P<0.01),之后明显降低并保持稳定;Ets2在生后第1～25天表达量显著高于第35天(P<0.05或P<0.01),之后明显下降并保持稳定.白消安处理后,Ets1的表达量于第5天降至最低,随后逐渐恢复,第9天后基本达到处理前水平并保持相对稳定,其中第5、8天表达量均显著低于处理后第0天(P<0.05或P<0.01);Ets2的表达量在白消安处理后前期变化不明显,第10天时明显降低,与第0和18天比较差异有统计学意义(P<0.05),之后缓慢回升,第18天左右恢复至处理前水平.结论:转录因子Ets1和Ets2可能对睾丸早期发育、成年精子发生的维持及精原细胞的增殖、分化具有调节作用.     

The use of lectins identified with specific antibodies in lectin histochemistry of NZB/W F1 mouse kidney

The affinity of Helix pomatia, peanut, Pisum sativum, soy bean, and wheat germ agglutinins to various nephron parts of NZB/W F1 mice was different and is assumed to be age dependent. The affinity of Pisum sativum agglutinin to basal membranes of small renal vessels increased with the age of NZB/W F1 mice. The wheat germ agglutinin bound to structures with alkaline phosphatase activity.     

A unique variant of a homeobox gene related to Drosophila cut is expressed in mouse testis

The wild-type gene encoding granule-bound starch synthase (GBSS) is capable of both complementing the amylose-free (amf) potato mutant and inhibiting the endogenous GBSS gene expression in wild-type potato. Co-suppression of the endogenous GBSS gene, easily visualised by staining the starch with iodine, occurred when the full-size GBSS sequence (genomic), GBSS cDNA or even the mutant amf allele were introduced into the wild-type potato. Conversely, introduction of the GBSS promoter sequence alone, did not result in co-suppression in the 80 analysed transformants. Neither the orientation of the GBSS gene with respect to kanamycin resistance nor the presence of an enhancer influenced the frequency of plants showing a co-suppression phenotype. After crossing a partially complemented amf mutant with a homozygous wild-type plant, the F1 offspring segregated into plant phenotypes with normal and decreased expression of the GBSS gene. This decreased expression correlated with the presence of a linked block of five T-DNA inserts which was previously shown to be correlated with partial complementation of the amf mutant. This crossing experiment indicates that co-suppression can cause inhibition of gene expression of both inserted and endogenous wild-type GBSS genes. The frequency of partially complemented amf plants was equal to the frequency of co-suppressed wild types when a construct, with an enhancer in front of the GBSS promoter, was used (pWAM 101E). This might suggest that partial complementation of the amf genotype caused by unstable expression of the transgene can be overcome by inserting an enhancer in front of the GBSS promoter.     

Cloning of two isoforms of mouse DNA helicase Q1/RecQL cDNA; alpha form is expressed ubiquitously and beta form specifically in the testis

We cloned cDNAs encoding mouse homologues for the human DNA helicase Q1/RecQL (human helicase Q1) which has homology with the Escherichia coli RecQ protein and found that they encode two isoforms. The two isoforms are identical over the entire sequence except for the carboxyl terminal sequence spanning less than 30 amino acids. One of the two isoforms, alpha, contains a sequence, KKRK, in the carboxyl terminus, which is also contained in human helicase Q1 and was confirmed to function as the nuclear localization signal. The other form, beta, does not contain such a sequence. Expression of mouse helicase Q1 mRNA is extremely and relatively high in the testis and the thymus, respectively. RT-PCR analysis revealed that helicase Q1alpha was expressed in all tissues tested and the beta form was expressed only in the testis. A survey of expression of Q1alpha and Q1beta mRNA in the testis after birth revealed that Q1alpha mRNA is expressed in all testes of mice aged from 7 days to 8 weeks, and the expression of Q1beta mRNA begins 14 days after birth, corresponding to the appearance of cells in the pachytene stage.     

The protein kinase Cdr2, related to Nim1/Cdr1 mitotic inducer, regulates the onset of mitosis in fission yeast

Cdc2-Cyclin B, the protein kinase that catalyzes the onset of mitosis, is subject to multiple forms of regulation. In the fission yeast Schizosaccharomyces pombe and most other species, a key mode of Cdc2-Cyclin B regulation is the inhibitory phosphorylation of Cdc2 on tyrosine-15. This phosphorylation is catalyzed by the protein kinases Wee1 and Mik1 and removed by the phosphatase Cdc25. These proteins are also regulated, a notable example being the inhibition of Wee1 by the protein kinase Nim1/Cdr1. The temperature-sensitive mutation cdc25-22 is synthetic lethal with nim1/cdr1 mutations, suggesting that a synthetic lethal genetic screen could be used to identify novel mitotic regulators. Here we describe that such a screen has identified cdr2(+), a gene that has an important role in the mitotic control. Cdr2 is a 775 amino acid protein kinase that is closely related to Nim1 and mitotic control proteins in budding yeast. Deletion of cdr2 causes a G2-M delay that is more severe than that caused by nim1/cdr1 mutations. Genetic studies are consistent with a model in which Cdr2 negatively regulates Wee1. This model is supported by experiments showing that Cdr2 associates with the N-terminal regulatory domain of Wee1 in cell lysates and phosphorylates Wee1 in vitro. Thus, Cdr2 is a novel mitotic control protein that appears to regulate Wee1.     

Sustained proliferation, multi-lineage differentiation and maintenance of primitive human haemopoietic cells in NOD/SCID mice transplanted with human cord blood

Time course studies of sublethally irradiated non-obese mice with severe combined immunodeficiency (NOD/ SCID mice) transplanted intravenously with 10(7) human cord blood cells showed a rapid and parallel regeneration of human erythroid, granulopoietic, megakaryopoietic and B-lymphoid progenitors, as well as more primitive subpopulations of CD34+ cells (defined by their multi-lineage in vitro colony-forming ability, coexpression of Thy-1, or functional activity in long-term culture-initiating cell [LTC-IC] assays), in the marrow, spleen and blood. Maximum numbers of human cells were reached within 6 weeks and were then sustained for another 18-20 weeks. 3H-thymidine suicide studies showed all types of in vitro clonogenic human progenitors tested and the human LTC-IC to be proliferating in vitro throughout this period. A 2-week course of injections of human Steel factor, interleukin-3, granulocyte-macrophage colony-stimulating factor and erythropoietin given just prior to assessment of the mice had no effect on any of these human engraftment parameters. 4-6 weeks post-transplant, the marrow of primary NOD/SCID recipients contained human cells that were able to regenerate lymphopoiesis and/or myelopoiesis in secondary irradiated NOD/SCID mice. These findings establish a baseline for the kinetics of engraftment, multi-lineage differentiation and self-renewal of human cord blood stem cells in this xenogeneic transplant model and thus set the stage for future studies of their regulation in vivo.     

Mouse beta-globin DNA-binding protein B1 is identical to a proto-oncogene, the transcription factor Spi-1/PU.1, and is restricted in expression to hematopoietic cells and the testis

The hematopoietic-specific DNA-binding protein B1 binds to the DNA consensus sequence AAAGRGGAARYG located twice in intervening sequence 2 of both of the mouse beta-globin genes (D. L. Galson and D.E. Housman, Mol. Cell. Biol. 8:381-392, 1988). B1 was cloned by expression of a murine erythroleukemia (MEL) cell cDNA library in transfected COS cells and screening by electrophoretic mobility shift analysis. B1 is identical to the proto-oncogene Spi-1/PU.1 (Spi-1), an ets family member. Protein-DNA contacts are shown to resemble those of the helix-turn-helix homeodomain proteins. By Northern (RNA) analysis, we found that Spi-1 mRNA is present at low levels during murine CFU-E maturation and is at least 20-fold higher in uninduced MEL, a transformed proerythroblast-like cell line which contains an activating/transforming insertion of spleen focus-forming virus at the Spi-1 locus. Dimethyl sulfoxide-induced MEL cell differentiation decreases Spi-1 mRNA to approximately 20% of the uninduced level before commitment occurs. In addition to erythroid cells, Spi-1 mRNA is present in B cells, myelomonocytes, and mast cells but not in T cells and nonhematopoietic cell types. In situ hybridization demonstrated Spi-1 mRNA expression in bone marrow, spleen, interstitial nonhepatocytes of the liver, and interstitial nontubular cells of the testis. The Spi-1 locus was mapped on human chromosome 11 to the same interval as ACP2 (lysosomal acid phosphatase), between the anonymous DNA markers D11S33 and D11S14. This region has not yet been found to be associated with a human malignancy.     

Dysfunctional D1A receptor-G-protein coupling in proximal tubules of spontaneously hypertensive rats is not due to abnormal G-proteins

Improving immunization coverage is vital to promoting child health and reducing childhood diseases and deaths. In spite of being actively promoted as a major public health intervention for national development since the late 1970s, immunization coverage in Ghana remains low. We investigated factors that influence attendance to immunization sessions in the Komenda-Edina-Eguafo-Abrem District of Ghana. The major factors hindering attendance were poor knowledge about immunization, lack of suitable venues and furniture at outreach clinics, financial difficulties, long waiting times, transport difficulties, poorly motivated service providers and weak intersectoral collaboration. The timing of immunization sessions, length of prior notice to the community, attitude of service providers and fear of side-effects generally did not deter attendance.     

A study of the fluctuating factors in the seminiferous epithelial cycle in mice with special attention to statistical studies on diurnal fluctuations and on variations due to testes and testis loci in the relative frequencies of the stages

The influence of the testis and loci within testes on diurnal variations and fluctuations in the seminiferous epithelial cycle was investigated in 30 50-60 day old mice. Histological examination of the seminiferous tubules revealed a typical cellular arrangement for each stage of spermatogenesis in virtually all of the specimens. No apparent diurnal fluctuations were observed in the relative frequency of the various stages between specimens obtained at 6 different times of sacrifice or among the uniform periods over a 24-hour period. However, a significant (p less than .05) difference in the frequencies of the stages was observed between testes among individuals and loci within testes.     

Underreporting of energy intake in 1 to 18 year old German children and adolescents

AIMS: Intraosseous jaw cysts with a solely orthokeratinized lining epithelium have been suggested to differ from the typical odontogenic keratocysts (OKC) by exhibiting a less aggressive behaviour. We report 15 cases of such cyst type under the term of 'orthokeratinized odontogenic cyst (OOC)' and compared their clinical, histological and immunocytochemical features with that of OKC. METHODS AND RESULTS: The cysts of the present series were all solitary lesions, occurred mostly in young male patients, and showed a predilection for the posterior mandible areas. Follow-up of 14 patients, nine of whom were treated by simple enucleation, revealed no recurrence over a period of 3.5-12 years after surgery. None of the patients had any association with the naevoid basal cell carcinoma syndrome. Furthermore, histological and immunocytochemical comparison between OOC and OKC revealed marked differences in their morphology and epithelial expression. The lining epithelium of OOC lacked the typical features of OKC and appeared to show a lower proliferative activity. CONCLUSION: These findings suggest that OOC is clinicopathologically separate from other types of odontogenic cysts and may thus constitute a distinct clinical entity.     

Response of stem cells in the mouse testis to fission neutrons of 1 MeV mean energy and 300 kV x rays. Methodology, dose-response studies, relative biological effectiveness

Microelectrode recording in the thalamus of pigeons subjected to tilt and sinusoidal rotational stimuli around the vertical, longitudinal and transversal axes revealed vestibularly driven units in two thalamic nuclei, the nucleus posteroventralis and the nucleus principalis precommissuralis. Many of these units responded in a complex manner suggesting that inputs from contralateral and ipsilateral cupulae and maculae converged on them. A few units received additional visual or proprioceptive information. The homology relationship with a mammalian vestibular thalamic nucleus is discussed briefly.