ABSTRACT
Chronic alcohol intake is associated with male reproductive function impairment. However, no longitudinal studies have been carried out to determine the recovery of alcohol-related spermatogenetic failure subsequent to moderate periods of abstinence. The present report describes the achievement of a pregnancy 3 months after withdrawal from alcohol consumption in the partner of a patient with azoospermia secondary to heavy alcoholic intake (mean daily alcohol consumption: 90 g). Alcoholism was the putative cause of the infertile condition of this patient because, during alcohol consumption, he first had teratozoospermia characterized by a never reported high percentage of spermatozoa with large heads (associated with a nonmegaloblastic macrocytic anaemia in the blood smear), and subsequently azoospermia.
Subject(s)
Alcoholism/complications , Oligospermia/etiology , Oligospermia/physiopathology , Pregnancy , Adult , Ethanol/administration & dosage , Female , Humans , MaleABSTRACT
Activator of cAMP-responsive element modulator (CREM) in testis (ACT) has recently been found in the mouse testis where it activates CREM, a transcription factor essential for the differentiation of spermatids into mature spermatozoa. The importance of CREM in human spermatogenesis prompted us to examine whether ACT was also present in the human testis. Western blot analysis, performed with an anti-mouse ACT serum, showed the presence of a single immunoreactive band of a size similar to murine ACT. A library screening resulted in the isolation and characterization of the complete cDNA which showed 88% homology with the mouse counterpart. The human ACT gene is composed of five coding exons, being the first untranslated, and the mRNA spans 835 nucleotides coding for a 284 amino acid protein. Expression studies by RT-PCR confirmed that ACT is present in normal human testis. The human ACT gene is localized on the chromosome 6.
Subject(s)
Repressor Proteins , Testis/metabolism , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Cyclic AMP Response Element Modulator , DNA Primers/genetics , DNA, Complementary/genetics , DNA-Binding Proteins/metabolism , Gene Expression , Humans , LIM Domain Proteins , Male , Mice , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In this work we have developed reverse transcription polymerase chain reaction (RT-PCR) methods for detecting specific mRNA from enterococci, particularly vanA and vanB genes, responsible for glycopeptide resistance in this genus. mRNA from the two genes was detected immediately after RNA extraction of a midlog phase culture, determined by growth rate analysis. Because of the short half-life associated with many bacterial RNA species (1.5-2 min), time is an important factor in obtaining RNA of good yield and high purity. Our results showed that: (i) the transcription of mRNA related to vanA ligase in enterococci showing Van A phenotype happens only after induction with both vancomycin and teicoplanin; (ii) the transcription of mRNA related to vanB ligase happens only in the presence of vancomycin and (iii) there was no transcription of mRNA in the two strains positive to vanA gene after PCR experiments. RT-PCR methodology can have numerous applications in microbiology for studying gene expression in isolated bacteria and also in nonculturable cells in environmental samples, for studies of mechanisms and/or as an indicator of viability in bacterial communities.