Male infertility and mitochondrial DNA

The mitochondrial machinery plays a key role in the energy production and maintenance of spermatozoa motility. In this paper 200 idiopathic oligo-asthenozoospermic patients were classified on the basis of rapid progressive motility ("a") and sperm concentration. Mitochondrial enzymatic activity was studied and correlated to the viability of sperm cells. Mitochondrial DNA purified from both motile and non-motile sperm of the same individuals was amplificated using PCR. Results suggested that only motile sperm have organelles functional in oxygen consumption, unequivocally demonstrating that motility depends on the mitochondrial activity. Mitochondrial DNA of oligo-asthenozoospermic patients …

TheSCH9 protein kinase mRNA contains a long 5′ leader with a small open reading frame

The SCH9 yeast gene, that was previously identified as a suppressor of cdc25 and ras1- ras2-ts temperature-sensitive mutants, encodes a putative protein kinase that positively regulates the progression of yeast cells through the G1 phase of the cell cycle. We have determined the structure of the SCH9 transcription unit, using primer extension and S1 mapping techniques. The corresponding mRNA included an unusually long 5' region of more than 600 nucleotides preceding the major open reading frame (ORF). While the latter corresponded to a protein of 824 amino acids, an upstream open reading frame (uORF) within the 5' leader could potentially encode a 54 amino acid peptide. To investigate the r…

LA BIOLOGIA DELLA RIPRODUZIONE ANCHE IN AUSILIO PER IL PUBBLICO MINISTERO

LA BIOLOGIA DELLA RIPRODUZIONE ANCHE IN AUSILIO PER IL PUBBLICO MINISTERO E. CARRA Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche (STEBICEF), Università di Palermo; elena.carra@unipa.it Su incarico dell’Autorità Giudiziaria si è proceduto all’inventario del contenuto di fusti per la crioconservazione di gameti ed embrioni, ad analisi genetica su embrione umano degenerato ed alla ricostruzione del percorso dei gameti di pazienti sottoposti a procreazione medicalmente assistita (PMA) presso 4 centri di biologia della riproduzione (BDR) per evidenziare eventuali irregolarità significative ai sensi della normativa vigente. Trattasi delle prime indagini commissionate su…

DNA IN FORENSIC GENETICS

Forensic molecular genetics has been evolved from a rapidly developing field into accepted forensic science, leading DNA databases with DNA profiles from suspects and convicted offenders. DNA evidence has taken a central role by carrying a significant weight for convictions in the court of justice, as well as by excluding innocent suspects early on in a criminal investigation. At present, a robust system for DNA typing has been established for paternity testing and forensic stain analysis in criminal casework. In addition, forensic biologist should be aware about the ethical and legal consequences of their research on the society and the criminal justice system. New technologies will only b…

DNA TYPING OF BLOODSTAIN: A CASEWORK OF MORTAL CRASH

Since 1997 the analysis of DNA recovered from handled objects, or trace DNA has become routine and is frequently demanded from crime scene examination. However, this analysis often produces unpredictable results. The factor affecting the recovery of full profiles are numerous, and as demonstrated in casework and research, retrieving useable profiles from trace DNA exhibits is not guaranteed. There are numerous factors that affect the rate of recovery of full profiles, such as the ability of the individual to shed cells, the substrate surface, the time of contact and delay until analysis, and the environment. In addition to these external conditions, there are many varying methods used to co…

L'Embriologia della Procreazione Medicalmente Assistita (PMA) in ambito forense

L’EMBRIOLOGIA DELLA PROCREAZIONE MEDICALMENTE ASSISTITA (PMA) IN AMBITO FORENSE CARRA ELENA Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche,Università di Palermo Su incarico dell’Autorità Giudiziaria viene eseguita analisi genetica su embrione umano degenerato e ricostruzione del percorso dei gameti di pazienti sottoposti a PMA presso 4 centri di biologia della riproduzione per evidenziare eventuali irregolarità significative ai sensi della normativa vigente. Trattasi delle prime indagini commissionate sul territorio Nazionale che vedono impegnato il biologo forense in uno dei settori di frontiera della medicina della riproduzione. Con il pronunciamento della Consul…

DNA e realazioni di parentela

PMA E BIOLOGIA FORENSE IN AUSILIO DEL PUBBLICO MINISTERO

PMA E BIOLOGIA FORENSE IN AUSILIO DEL PUBBLICO MINISTERO Carra E., Di Natale M.V. Dipartimento di Scienze Tecnologie Molecolari Biomolecolari (STEMBIO)– Università di Palermo Edifio 16, viale delle Scienze.– elena.carra@unipa.it Nell’ambito di attività svolta su incarico dell’Autorità Giudiziaria si è proceduto all’inventario del contenuto di fusti per la crioconservazione di gameti ed embrioni sottoposti a sequestro e custoditi presso i locali di quattro centri di biologia della riproduzione per “verificarne la rispondenza alle registrazioni cartacee ed informatiche acquisite agli atti”. Si rendeva, altresì, necessaria analisi genetica su materiale biologico, costituito da embrione umano d…

Study on endophytic strains of genus Acremonium in symptomless grapevines

Substance use, medication adherence and outcome one year following a first episode of psychosis

Both substance use and poor medication adherence are associated with poor outcome in psychosis. To clarify the contributions of substance use and poor medication adherence to poor outcome in the year following a first episode of psychosis, 205 patients were evaluated for use of tobacco, alcohol, cannabis and stimulants at their psychosis onset, and in a 1-year follow-up. Data on medication adherence and symptom remission were also collected. Patients had high rates of overall substance use before (37-65%) and after psychosis onset (45-66%). 44% showed poor medication adherence and 55% did not reach remission from psychosis. Nicotine dependence and cannabis use after psychosis onset signific…

DNA typing from vaginal smears slides and stained sperm-positive evidence in suspected rape cases.

Rape is a largely un-witnessed crime, and may be difficult to obtained statements from the victim, since most of the victims suffer from emotional damage as well as physical injury. The physical examination by forensic specialist is extremely important.

Mitochondrial DNA deletion and male infertility

Abstract The term “male infertility” does not constitute a defined clinical syndrome, but rather, a collection of different conditions exhibiting a variety of aetiologies and varying diagnosis. In men, oligozoospermia, asthenozoospermia, teratozoospermia and azoospermia are the main causes of infertility. The present research is aimed to investigate if mtDNA deletions can cause sperm defects in idiopathic astenozoospermic patients with different sperm motility and sperm concentration. The incidence of deletions in mtDNA (mtDNA) was co-related with diagnostic categorization of male infertility when 6.6kb out of the 16.6kb of the mitochondrial genome was amplified from whole sperm samples, f…

Study of the variability of an endophytic Acremonium population in symptomless grapevines.

Signature of recent historical events in the European Y-chromosomal STR haplotype distribution

Previous studies of human Y-chromosomal single-nucleotide polymorphisms (Y-SNPs) established a link between the extant Y-SNP haplogroup distribution and the prehistoric demography of Europe. By contrast, our analysis of seven rapidly evolving Y-chromosomal short tandem repeat loci (Y-STRs) in over 12,700 samples from 91 different locations in Europe reveals a signature of more recent historic events, not previously detected by other genetic markers. Cluster analysis based upon molecular variance yields two clearly identifiable sub-clusters of Western and Eastern European Y-STR haplotypes, and a diverse transition zone in central Europe, where haplotype spectra change more rapidly with longi…

STUDY ON AN ENDOPHYTIC POPULATION OF ACREMONIUM SP. IN SYMPTOMLESS GRAPEVINE

Determinazione del tempo di permanenza in acqua in base allo sviluppo di mitili impiantatisi su cadavere scheletrizzato e saponificato.

C10) DETERMINAZIONE DEL TEMPO DI PERMANENZA IN ACQUA IN BASE ALLO SVILUPPO DI MITILI IMPIANTATISI SU CADAVERE SCHELETRIZZATO E SAPONIFICATO L. Milone (*) – S. Procaccianti (*) – M. Grillo (*) – B. Salerno (**) - E. Carra (**) (*) Dipartimento Di Biopatologia E Biotecnologie Mediche E Forensi – Sezione Di Medicina Legale – Liviomilone@Virgilio.It - (**) Dipartimento Di Biologia Cellulare E Dello Sviluppo – Universita’ Degli Studi Di Palermo Introduzione: La valutazione tanatocronologica di cadaveri in avanzato stato di putrefazione, dovuto alla lunga permanenza in acqua salata, pone non pochi problemi al patologo forense che deve risalire all’epoca del decesso, che spesso viene indicata solt…

Comunicazione “DNA E RELAZIONI DI PARENTELA

Astenozoospermy and mitochondrial DNA

Male infertility, inability to fecundate the oocyte, is due to alterations of sperm characteristics (low number, low motility, altered morphology), to the quantity of semen or to the presence of alterations in the male genital line. 75% of infertile men deal with untreatable sub-fertility. Semen of 200 idiopathic astenozoospermic patients and individuals with normal seminal parameters as control, were analyzed. Results showed that mtDNA and Y chromosome regions from the motile fractions of patients and controls gave amplifications, while, the non-motile fractions of the same seminal samples did not give amplifications of ND5, ND6 and regions inside the D-loop (Carra E. et al.2002, 2004). We…

Localization of mitochondrial Hsp56 chaperonin during sea urchin development.

We have previously demonstrated that Paracentrotus lividus nuclear genome encodes for the heat shock inducible chaperonin homolog Hsp 56 (1) and that the mature protein is localized in the mitochondrial matrix (2). In this paper we report that constitutive Hsp56 is maternally inherited, in fact it is present in the in unfertilized eggs, and that it has a perinuclear specific localization during cleavage. In the later stages both the constitutive and the heat shock inducible chaperonin has a specific territorial distribution. Moreover following heat shock, the Hsp56 appears in the cytoplasm and in the postmitochondrial supernatant beside the mitochondrial fraction.

The European Pine Marten Martes martes (Linnaeus, 1758) Is Autochthonous in Sicily and Constitutes a Well-Characterised Major Phylogroup within the Species (Carnivora, Mustelidae)

No molecular data are currently available for the Sicilian populations of the European pine marten Martes martes, thus preventing any sound inference about its native or non-native status on the island, as well as the local phylogeography of the species. In order to investigate these issues, we sequenced two mtDNA markers in road-killed specimens collected in Sicily. Both markers consistently demonstrated the existence of a well-characterised Sicilian clade of the species, which is endemic to the island and constitutes the sister group of a clade including the Mediterranean and Central–North European major phylogroups of the European pine marten. Such evidence supports the autochthony of Ma…

GENOMA MITOCONDRIALE ED INFERITLITA' MASCHILE

DNA Typing of Blood Stain: a casework of mortal crash

Ricerche sperimentali su colle in un caso giudiziario di effrazione di busta per gara d'appalto: aspetti tecnici ed identificativi

DNA extraction from phytoseiid specimens preserved on a glass slide and molecular diagnostic

This study focuses on the diagnostic of seven species of mite belonging to the family of Phytoseiidae; very important biocontrol agents of phytophagous mites. However, they are morphologically very similar and specific diagnostic is very difficult in fact using the traditional identification methods it is uncertain to identify a species especially when Authors use different characters, very often not stable in the time. In the 60’s M.me Athias-Henriot took into consideration the insemination apparatus that is considered a stable character in the time. According to her, the shape of this apparatus could be considered a major criterion for genus distinction. The aim of the present study is to…

ASTENOZOOSPERMY AND MITOCHONDRIAL DNA

A METHOD FOR STUDYING GENETICS OF THE PREDATORY MITE PHYTOSEIULUS PERSIMILIS

Two DNA extraction and purification procedures for single mite analysis, organic (phenol-chloroform) method and the Chelex® -100 DNA extraction procedure, have been compared concerning extraction efficiency, DNA purity and DNA suitability for amplification. Phenol-chloroform extraction takes a relatively long period of time (5 days) and involves the use of toxic and hazardous chemical reagents but it produces consistent results. Chelex is a chelating resin that has a high affinity for polyvalent metal ions, Singer-Sam et al. postulated that boiling a sample in the presence of chelex prevents the degradation of DNA. Both method were evaluated to extract DNA from single frozen mite or individ…

GENOMA MITOCONDRIALE ED INFERTILITÀ MASCHILE

Study of variability of an endophytic acremonium population in symptomless grapevine

The endophytism of genus Acremonium in symptomless grapevines of several cultivars, grown both in pots or in the field, is reported. The no-tillage management of tested grapevines and the lack of chemical treatments facilitated the investigation of the ecological plant-fungus interaction. The isolates were differentiated at the strain level by vegetative compatibility assay and randomly amplified polymorphic DNA (RAPD) analyses, using a set of four oligo-nucleotide primers. The endophytic Acremonium population in the healthy grapevine object of the survey seemed to show a low variability; all the techniques agreed in grouping the Acremonium strains in three different clusters and no relatio…

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Mitochondrial DNA deletion and male infertility

MITOCHONDRIAL DNA DELETIONS AND MALE INFERTILITY Salerno Barbara, Gerardi Eliana and Elena Carra Dipartimento di Biologia Cellulare e dello Sviluppo,Università di Palermo Viale delle Scienze, Palermo, Italy; In men, oligozoospermia, asthenozoospermia, teratozoospermia and azoospermia are the main causes of infertility. The present research is aimed to investigate if mtDNA deletions can cause sperm defects in idiopathic astenozoospermic patients with different sperm motility and sperm concentration. The aim of this investigation was to test the hypothesis that whole sperm samples with lower levels of motility would have a higher incidence of spermatozoa with deletions in mitochondrial genome…

DNA extraction from phytoseiid specimens preserved on glass slide

Mites belonging to the family of Phytoseiidae are important biocontrol agents of phytophagous mites (McMurtry and Croft, 1997). But the species used for controlling harmful mites have to be correctly identified. Sometimes using the traditional identification methods it is difficult or uncertain to identify a species especially as various Authors use different characters, very often not stable in the time. In the 60’s M.me Athias-Henriot (Athias-Henriot 1968; 1969) took into consideration the insemination apparatus that is considered a stable character in the time. According to her, the shape of this apparatus could be considered a major criterion for genus distinction; but other scientists …