Lack of evidence for an immunosuppressive role for MUC1

The in vitro anti-proliferative properties of various supernatants from MUC1-expressing cell lines and of purified preparations of MUC1 were evaluated. We have observed that supernatants from the MUC1- and MUC3-positive cell line T47D, but not from the MUC1- and MUC4-positive cell line MCF7, were able to inhibit proliferation of cells from various haematopoietic cell lines. Although the activity of T47D supernatants could be abrogated by immunodepletion of MUC1, immunopurified MUC1 from T47D was unable to inhibit cell proliferation. Significantly, supernatants from mouse 3T3 cells transfected with a secreted form of MUC1 or from BHK-21 cells infected with a recombinant vaccinia virus coding for the secreted form of MUC1, as well as preparations of purified MUC1 from bile or urine, were likewise unable to inhibit T cell proliferation. Surprisingly, a crude mixture of bile mucins had a suppressive effect on T cell growth. Our results suggest that other molecules, such as amino sugars or other mucins, which can associate with MUC1, are likely to be responsible for the observed anti-proliferative effects of T47D cells. Received: 20 August 1998 / Accepted: 3 December 1998     

Lack of evidence for perfluorodecanoyl- or perfluorooctanoyl-coenzyme a formation in male and female rats

Perfluorodecanoic (PFDA) and perfluorooctanoic (PFOA) acids belong to the structurally diverse group of compounds known to cause peroxisomal proliferation. It has been hypothesized that the common mode of action of these compounds is that they act through an activated coenzyme A (CoA) thioester. Using rat liver microsomal and isolated rat hepatocyte incubation conditions that were effective in producing a COA conjugate of clofibric acid, no corresponding COA derivative could be found for either PFDA or PFOA.     

Genetic variants of eNOS gene may modify the susceptibility to idiopathic male infertility

Abstract

In testis, eNOS is responsible for synthesis of nitric oxide (NO) which is an essential gas message regulator in spermatogenesis, suggesting that eNOS gene plays a role in normal spermatogenesis and the genetic variants of eNOS gene may be potential genetic risk factors of spermatogenesis impairment. In this study, the polymorphic distributions of three common polymorphism loci including T-786C, 4A4B and G894T in eNOS gene were investigated in 355 Chinese infertile patients with azoospermia or oligozoospermia and 246 healthy fertile men and a meta-analysis was carried in order to explore the possible relationship between the three loci of eNOS gene and male infertility with spermatogenesis impairment. As a result, allele -786C of T-786C (11.4% versus 6.5%, p?=?0.004) and 4A of 4A4B (11.0% versus 6.3%, p?=?0.005) as well as genotype TC of T-786C (22.8% versus 13.0%, p?=?0.002) and AB of 4A4B (18% versus 11%, p?=?0.015) were significantly associated with idiopathic male infertility. The haplotypes T-4A-G (7.4% versus 4.1%, p?=?0.015) and C-4B-G (7.6% versus 4.4%, p?=?0.028) could increase the susceptibility to male infertility, whereas haplotype T-4B-G (67.0% versus 75.2%, p?=?0.002) might be a protective factor for male infertility. The results of meta-analysis revealed that the polymorphism of T-786C was associated with male infertility. These findings suggested that the variants of eNOS gene may modify the susceptibility to male infertility with impaired spermatogenesis.     

Synaptonemal complex analysis in human male infertility

The fine structural features of human spermatocytes from carriers of some of the most frequent chromosomal abnormalities are reviewed on the basis of original data and previous reports from the literature. Special emphasis is given to the Robert-sonian translocations t (13; 14), to one specific reciprocal translocation involving chromosome 21, and to Y disomy in spermatocytes from XYY men. Synaptonemal complex analysis shows that in many carriers of chromosomal aberrations that lead to pachytene configurations having terminal asynaptic segments in autosomes, there is a gradual association of these asynaptic segments with the XY body. This associations with the XY pair is assumed to trigger a process of germ cell deterioration, presumably through the spreading of the X-chromosome inactivation towards autosomal segments. Another different process of germ cell deterioration occurs when the X chromosome becomes an univalent, as in XYY men with persistence of two Y chromosomes in the germ line. The renewed interest in the examination of spermatocytes from human testicular biopsies is commented upon.     

Lack of a central role for calcium in the induction and release of human interleukin-1

This study examines the potential role of calcium in the activation of human monocytes by bacterial lipopolysaccharide (LPS) to produce interleukin-1 (IL-1). Monocytes cannot be triggered to produce IL-1 through addition of calcium ionophores. Triggering of IL-1 production by LPS cannot be blocked by depletion of extracellular calcium, blockade of calcium channels, or addition of agents which antagonize the effects of intracellular calcium. Finally, the addition of LPS does not induce the mobilization of intracellular free calcium as measured by quin-2 fluorescence.     

Screening for microdeletions in human Y chromosome--AZF candidate genes and male infertility

About 30% of couple infertilities are of male origin, some of them caused by genetic abnormalities of the Y chromosome. Deletions in AZF region can cause severe spermatogenic defects ranging from non-obstructive azoospermia to oligospermia. The intracytoplasmatic sperm injection technique (ICSI) is rapidly becoming a versatile procedure for human assisted reproduction in case of male infertility. The use of ICSI allows Y chromosome defects to be passed from father. The goal of our study is to evaluate the frequency of microdeletions in the long arm of Y chromosome, within the AZF regions, in these cases of infertilities, using molecular genetics techniques. Thirty infertile men with azoospermia or oligozoospermia, determined by spermogram, were studied after exclusion of patients with endocrine or obstructive causes of infertility. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the Y chromosome AZF zones. Each case was checked by multiplex PCR through coamplification with the SRY marker. Three men with microdeletions of the long arm of the Y chromosome were diagnosed among the 30 patients, corresponding to a proportion of 10%. The relatively high proportion of microdeletions found in our population suggest the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions. The molecular diagnostics was performed according to the current European Academy of Andrology laboratory guidelines for molecular diagnosis of Y chromosomal microdeletions.     

Morphological defects of sperm flagellum implicated in human male infertility

The assembly of sperm flagella involves specific components and processes that are still poorly defined. Several morphological defects of the different structures that compose the axoneme have been described and associated to human male infertility. These morphological defects can be classified in 15 main categories. Most of them have been associated to consanguinity and/or familial cases, suggesting their genetic origin. However, so far only few genes have been causally involved.     

Lack of evidence of mimivirus replication in human PBMCs

The Acanthamoeba polyphaga mimivirus (APMV) was first isolated during a pneumonia outbreak in Bradford, England, and since its discovery many research groups devoted efforts to understand whether this virus could be associated to human diseases, in particular clinical signs and symptoms of pneumonia. In 2013, we observed cytopathic effect in amoebas (rounding and lysis) inoculated with APMV inoculated PBMCs (peripheral blood mononuclear cell) extracts, and at that point we interpreted those results as mimivirus replication in human PBMCs. Based on these results we decided to further investigate APMV replication in human PBMCs, by transmission electron microscopy (TEM) and qPCR. No viral factory was observed in APMV inoculated PBMCs, at any analyzed time and M.O.I.s (multiplicity of infection), by checking 550 cells per condition tested. We also measured the variation of viral DNA by qPCR targeting helicase gene during the course of the TEM experiment in PBMCs, but the DNA levels stayed the same as the first time-point post infection. In summary, our newest qPCR and TEM results do not support previous statements (including ours) that mimivirus is able to replicate in humans PBMCs.     

The development of a qualitative assay for male infertility from a study of enzymes in human semen.

Various enzymes in the semen of men were examined to see if any could be related to measures of fertility. Fumarase activity was highly correlated with sperm number and percentage motility. Diamine oxidase activity was higher in samples with sperm counts of less than 20 X 10(6)/ml and aperm motility of less than 20%. Monoamine oxidase, adenine deaminase and prostaglandin dehydrogenase were undetectable in significant amounts in all samples, while peroxidase and adenosine deaminase were not correlated with sperm count and motility. It is suggested that the simple spectrophotometric assays for fumarase and diamine oxidase could form the basis of a routine assessment of human semen samples for estimation of male infertility.     

Lack of evidence for perfluorodecanoyl- or perfluorooctanoyl-coenzyme A formation in male and female rats.

Perfluorodecanoic (PFDA) and perfluorooctanoic (PFOA) acids belong to the structurally diverse group of compounds known to cause peroxisomal proliferation. It has been hypothesized that the common mode of action of these compounds is that they act through an activated coenzyme A (CoA) thioester. Using rat liver microsomal and isolated rat hepatocyte incubation conditions that were effective in producing a CoA conjugate of clofibric acid, no corresponding CoA derivative could be found for either PFDA or PFOA.     

Genetic variation in human disease and a new role for copy number variants

While complex diseases, such as inflammatory bowel disease, do not follow distinctive Mendelian inheritance patterns, there is now considerable evidence from twin and pedigree studies to show that there are significant genetic influences in the development of many such diseases. In times past, this type of information was considered to be interesting, and was used mainly to alert other members of the families that they may also be at increased risk of developing the disease. However, with the ability to evaluate the genetic basis of common disease, this information will have important consequences for the diagnosis, prevention and treatment of the disorder. The genetic basis for common disease is likely to be more complicated than we had previously anticipated, since we now recognise epigenetic causes of disease, and other subtle gene regulatory mechanisms. Copy number variants have been highlighted in this review, as being a phenomenon that we have known about for a long time, but that has not previously been clearly associated with human disease. As complex disease is related to changes in gene expression, any variation in the human genome that alters gene expression is now a candidate for being involved in the disease process.     

The role of the microbial factor in the pathogenesis of male infertility

Microorganisms can be isolated from most seminal fluid samples, but the significance of bacteriospermia is uncertain because many males lack symptoms associated with the bacterial infection of the reproductive tract. The data on the influence of urogenital tract infections of fertility are contradictory. In many cases opportunistic microorganisms cause such classical infections of the urogenital tract as epididymitis and prostatitis, as well as subclinical reproductive tract infections. Some possible pathophysiological mechanisms of the development of infertility linked with infection of the ejaculate are considered: its direct effect on the fertile properties of the seminal fluid due to a decrease in the number of spermatozoa, the suppression of their motility, changes in their morphology and fertilizing capacity, its indirect influence due to the inhibition of spermatogenesis resulting from testicular damage, autoimmune processes induced by inflammation, secretory dysfunction of the male accessory sex glands as a consequence of the infection of the reproductive tract organs, leukocytospermia with its secondary influence of the ejaculate parameters, etc. The importance of the pathogenic properties of microorganisms for the localization of the inflammatory process in the urogenital tract of males and their role in the pathogenesis of male infertility are discussed.     

Lack of evidence for a role of hydrophobins in conferring surface hydrophobicity to conidia and hyphae of Botrytis cinerea

Background  

Hydrophobins are small, cysteine rich, surface active proteins secreted by filamentous fungi, forming hydrophobic layers on the walls of aerial mycelia and spores. Hydrophobin mutants in a variety of fungi have been described to show 'easily wettable' phenotypes, indicating that hydrophobins play a general role in conferring surface hydrophobicity to aerial hyphae and spores.     

The "drought-inducible" histone H1s of tobacco play no role in male sterility linked to alterations in H1 variants

Tobacco ( Nicotiana tabacum L.) has two major H1 variants (H1A and H1B), which account for over 80% of chromatin linker histones, and four minor variants: H1C, H1D, H1E and H1F. We have shown previously [M. Prymakowska-Bosak et al. (1999) Plant Cell 11:2317-2329] that reversal of the natural proportion of major to minor H1 variants in transgenic tobacco plants results in a characteristic male-sterility phenotype identical to that occurring in many plant species subjected to water deficit at the time of male meiosis. It has been proposed by others that the drought-induced arrest of male gametophyte development is linked to decreased sugar delivery to reproductive tissues. Within the family of angiosperm H1s there is a well-defined class of minor H1 variants named "drought inducible" because some of its members have been shown to be induced by water deficit. We have identified and cloned the tobacco H1C gene, which, based on sequence similarity, represents a "drought-inducible" minor H1 variant. Analysis of the un-translated mRNA and promoter regions of H1C suggests a regulation by sucrose concentration. Antisense silencing of H1C and its close homologue H1D in plants that do not express H1A and H1B does not affect the characteristic H1A(-)/ H1B(-) male-sterility phenotype. Silencing of H1C and H1D also has no effect on growth and development of plants. Our findings demonstrate that H1C and H1D are dispensable for normal growth and development of tobacco, and that the compensatory up-regulation of "drought-inducible" H1s observed in H1A(-)/ H1B(-) plants is not the direct cause of male sterility linked to alterations in H1 variants.     

Lack of evidence for changing virulence of HIV-1 in North America

Background

Several long-term cohort studies and in-vitro fitness assays have resulted in inconsistent reports on changes in HIV-1 virulence, including reports of decreasing, stable, and increasing virulence over the course of the AIDS pandemic. We tested the hypothesis of changing HIV-1 virulence by examining trends in prognostic clinical markers of disease progression from 1984 to 2005 among nearly 400 antiretroviral-naïve participants in the United States Multicenter AIDS Cohort Study (MACS), a longitudinal study of HIV infection in men who have sex with men (MSM).

Methodology/Principal Findings

Because clinical AIDS endpoints could not be used (due to antiretroviral therapies and prophylaxis), three prognostic markers of disease progression were used as proxies for HIV-1 virulence: plasma viral RNA load and CD4+ T cell count at “set point” (between ∼9 and ∼15 months after seroconversion), and rate of CD4 cell decline within three years after seroconversion. We performed multivariate analyses of the association between these markers and seroconversion year, with covariates including MACS site, race/ethnic group, seroconversion age, and CCR5Δ32 status. No statistically significant association was found between year of seroconversion and “set point” plasma viral load (at ∼9 months after seroconversion: slope = −0.004 log₁₀ copies/mL/year, p = 0.76; at ∼15 months: slope = −0.005 log₁₀ copies/mL/year, p = 0.71), CD4 cell count after seroconversion (at ∼9 months: slope = −0.112 cells/µL/year, p = 0.22; at ∼15 months: slope = −0.047 cells/µL/year, p = 0.64), or rate of CD4 cell decline over the first three years after seroconversion (slope = −0.010 cells/ul/yr², p = 0.88).

Conclusions/Significance

The lack of significant trends from 1984 to 2005 in these prognostic markers of HIV disease progression suggests no major change in HIV-1 virulence over the AIDS pandemic in MSM in the US.     

PIWIL1在不同肿瘤细胞中的差异性表达

PIWI作为AGO蛋白家族的成员,在睾丸中特异表达,在精子形成过程中扮演着重要角色.已有文献报道,PIWIL2也广泛存在于多种肿瘤中,与肿瘤的发生相关.本文旨在确定PIWL1在不同肿瘤细胞中的表达差异性,进而初步探讨PIWIL1的表达差异与肿瘤发生发展的关系.半定量RT-PCR和Western 印迹检测多种肿瘤细胞中,PIWIL1在mRNA水平及蛋白水平的表达情况,进一步用免疫组化和免疫荧光检测PIWIL1在细胞中的表达及定位.PIWIL1在多种肿瘤细胞中的表达存在差异,其中一些肿瘤细胞中的表达水平较高,包括卵巢癌,前列腺癌,肝癌,胃癌等细胞.对于肿瘤细胞而言,PIWIL1定位于细胞浆中.因此,PIWIL1在多种肿瘤细胞中的表达差异性可能为肿瘤发生发展的研究提供新的线索.