Subject(s)
Antibodies, Monoclonal/immunology , Desmoglein 1/metabolism , Horse Diseases/pathology , Immunohistochemistry/veterinary , Pemphigus/veterinary , Animals , Biopsy/veterinary , Blotting, Western/veterinary , Desmoglein 1/immunology , Desmoglein 1/isolation & purification , Horse Diseases/diagnosis , Horse Diseases/immunology , Horses , Immunohistochemistry/methods , Pemphigus/diagnosis , Pemphigus/immunology , Pemphigus/pathology , Skin/immunology , Skin/pathology , Species SpecificityABSTRACT
In this study, we describe a pentaplex PCR to determine the parental origin of the X chromosome and the presence of mosaicism, via amplification of four polymorphic markers located along the X chromosome (DXS10011, DXS6807, HUMARA, DXS101) and the X-Y amelogenin marker, in 41 families having a daughter with Turner Syndrome. Our results confirmed the cytogenetic findings and we found that the parental origin of the single X chromosome to be maternal in 84% of cases.
Subject(s)
Chromosomes, Human, X , Chromosomes, Human, Y , Polymerase Chain Reaction/methods , Turner Syndrome/genetics , Adult , Amelogenin , Child , Child, Preschool , Dental Enamel Proteins/genetics , Family , Female , Genetic Markers , Humans , Mosaicism , Polymorphism, Genetic , Sex Chromosome AberrationsABSTRACT
Mitochondrial DNA (mtDNA) sequence variations at hypervariable regions HVI, HVII and HVIII were analysed in 100 unrelated Italians from Bologna. The values of the statistical parameters are in agreement with the range of European populations. We suggest that the less informative HVIII region may be useful to distinguish HVI-HVII identical sequences in forensic analysis especially when nuclear DNA cannot be investigated.
Subject(s)
Complementarity Determining Regions/genetics , DNA, Mitochondrial/genetics , Genetics, Population , Humans , Italy , Polymorphism, GeneticABSTRACT
The expression of class I human leucocyte antigen (HLA) has been investigated by reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry on human purified ejaculated spermatozoa from an infertile population. Similar investigation in a control fertile population has been performed. Among 22 fertile and 20 infertile men, 17 were selected for the study because they showed no contamination with non-sperm cells. HLA I molecules were expressed in four of nine fertile subjects (44.4%) and three of eight infertile subjects (37.5%), with no significant difference between groups. These data demonstrate for the first time the presence of class I HLA antigens on spermatozoa of infertile subjects. In addition, considering that similar results have been obtained in both fertile and infertile populations, the hypothesis that the fertilizing capacity of the spermatozoon is independent of class I HLA-antigen expression on their surface may be advanced.
Subject(s)
Histocompatibility Antigens Class I/biosynthesis , Infertility, Male/immunology , Spermatozoa/immunology , Humans , Male , Polymerase Chain ReactionABSTRACT
Human leukocyte antigens (HLA) class II transcripts in mature spermatozoa of healthy volunteers have recently been demonstrated using reverse transcription polymerase chain reaction (RT-PCR). HLA II expression was investigated on ejaculated sperm cells in fertile and infertile men by RT-PCR and flow cytometry. Among 22 fertile and 20 infertile men, 18 were selected for the study because they showed no contamination with non-sperm cells. HLA II mRNA transcripts were expressed in all but 1 of 8 infertile subjects and in only 2 of 10 fertile ones. The cytofluorometric analysis on three RT-PCR positive samples confirmed the presence of class II antigens on cell surfaces. These data clearly confirm the presence of both HLA II mRNA and surface molecules on human sperm cells. In addition, an interesting nonrandom distribution of positivity among fertile and infertile samples regarding HLA II expression (p < .025) suggests a possible correlation with infertility.
Subject(s)
Fertility/immunology , Histocompatibility Antigens Class II/genetics , Infertility, Male/immunology , Spermatozoa/immunology , Transcription, Genetic , Adult , Antigens, CD/analysis , Fertility/physiology , Flow Cytometry , Histocompatibility Antigens Class II/analysis , Humans , Infertility, Male/genetics , Infertility, Male/physiopathology , Male , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sperm Count , Sperm MotilityABSTRACT
The myotonic dystrophy (DM) CTG repeat polymorphism has been studied in an Italian population sample. Polymerase chain reaction (PCR) amplification, manual polyacrylamide gel electrophoresis (PAGE), and silver staining were employed. Alleles were typed by comparison with a sequenced allelic ladder. A total of 25 different alleles, spanning the range from 5 to 31 CTG triplets, was observed. The heterozygosity was 79%, and no significant deviation from Hardy-Weinberg equilibrium was found. Eighty-one meioses from parentage testing were also analyzed, and a Mendelian pattern of inheritance was observed in all cases. In addition, we could successfully type the DM locus in 20 laboratory-prepared bloodstains, with 1 ng of DNA allowing clear definition of alleles. We conclude that the CTG repeats at the DM locus may be useful for forensic applications.
