Detection of oncogenic human papillomavirus genotypes on spermatozoa from male partners of infertile couples

Objective: To evaluate the prevalence of human papillomavirus (HPV) sperm infection and its correlation with sperm parameters in patients who attended a fertility clinic. Design: Cross-sectional clinical study. Setting: University-affiliated Reproductive Medicine Clinic. Patients: A total of 308 male partners of couples undergoing in vitro fertilization techniques. Interventions: Specimens of semen were collected from all patients. Main Outcome Measures: Sperm parameters were evaluated according to the World Health Organization manual. The presence of HPV-DNA was researched by the combined use of two HPV assays and a highly sensitive nested PCR assay, followed by HPV genotyping. To examine whether HPV was associated with the sperm, in situ hybridization (ISH) analysis was performed. Results: Results of HPV investigation were compared to sperm parameters and ISH analysis. Twenty-four out of 308 (7.8%) semen samples were HPV DNA positive but HPV infection does not seem to affect semen quality. Moreover, ISH revealed a clear HPV localization at the equatorial region of sperm head in infected samples. Conclusions: Oncogenic HPV genotypes were detected on spermatozoa from asymptomatic subjects but a role of the infection in male infertility was not demonstrated

Immune mediators of sea-cucumber Holothuria tubulosa (Echinodermata) as source of novel antimicrobial and anti-staphylococcal biofilm agents

The present study aims to investigate coelomocytes, immune mediators cells in the echinoderm Holothuria tubulosa, as an unusual source of antimicrobial and antibiofilm agents. The activity of the 5kDa peptide fraction of the cytosol from H. tubulosa coelomocytes (5-HCC) was tested against a reference group of Gram-negative and Gram-positive human pathogens. Minimal inhibitory concentrations (MICs) ranging from 125 to 500 mg/ml were determined against tested strains. The observed biological activity of 5-HCC could be due to two novel peptides, identified by capillary RP-HPLC/nESI-MS/MS, which present the common chemical-physical characteristics of antimicrobial peptides. Such peptides were chemically synthesized and their antimicrobial activity was tested. The synthetic peptides showed broad-spectrum activity at 12.5 mg/ml against the majority of the tested Gram-positive and Gram-negative strains, and they were also able to inhibit biofilm formation in a significant percentage at a concentration of 3.1 mg/ml against staphylococcal and Pseudomonas aeruginosa strains. The immune mediators in H. tubulosa are a source of novel antimicrobial peptides for the development of new agents against biofilm bacterial communities that are often intrinsically resistant to conventional antibiotics

New Bioactive Peptides from the Mediterranean Seagrass Posidonia oceanica (L.) Delile and Their Impact on Antimicrobial Activity and Apoptosis of Human Cancer Cells

The demand for new molecules to counter bacterial resistance to antibiotics and tumor cell resistance is increasingly pressing. The Mediterranean seagrass Posidonia oceanica is considered a promising source of new bioactive molecules. Polypeptide-enriched fractions of rhizomes and green leaves of the seagrass were tested against Gram-positive (e.g., Staphylococcus aureus, Enterococcus faecalis) and Gram-negative bacteria (e.g., Pseudomonas aeruginosa, Escherichia coli), as well as towards the yeast Candida albicans. The aforementioned extracts showed indicative MIC values, ranging from 1.61 μg/mL to 7.5 μg/mL, against the selected pathogens. Peptide fractions were further analyzed through a high-resolution mass spectrometry and database search, which identified nine novel peptides. Some discovered peptides and their derivatives were chemically synthesized and tested in vitro. The assays identified two synthetic peptides, derived from green leaves and rhizomes of P. oceanica, which revealed interesting antibiofilm activity towards S. aureus, E. coli, and P. aeruginosa (BIC50 equal to 17.7 μg/mL and 70.7 μg/mL). In addition, the natural and derivative peptides were also tested for potential cytotoxic and apoptosis-promoting effects on HepG2 cells, derived from human hepatocellular carcinomas. One natural and two synthetic peptides were proven to be effective against the “ in vitro” liver cancer cell model. These novel peptides could be considered a good chemical platform for developing potential therapeutics

A Novel Peptide with Antifungal Activity from Red Swamp Crayfish Procambarus clarkii

The defense system of freshwater crayfish Procambarus clarkii as a diversified source of bioactive molecules with antimicrobial properties was studied. Antimicrobial activity of two polypeptideenriched extracts obtained from hemocytes and hemolymph of P. clarkii were assessed against Gram positive (Staphylococcus aureus, Enterococcus faecalis) and Gram negative (Pseudomonas aeruginosa, Escherichia coli) bacteria and toward the yeast Candida albicans. The two peptide fractions showed interesting MIC values (ranging from 11 to 700 g/mL) against all tested pathogens. Polypeptideenriched extracts were further investigated using a high-resolution mass spectrometry and database search and 14 novel peptides were identified. Some peptides and their derivatives were chemically synthesized and tested in vitro against the bacterial and yeast pathogens. The analysis identified a synthetic derivative peptide, which showed an interesting antifungal (MIC and MFC equal to 31.2 g/mL and 62.5 g/mL, respectively) and antibiofilm (BIC50 equal to 23.2 g/mL) activities against Candida albicans and a low toxicity in human cells

Procreazione Medicalmente Assistita: dalla qualità ovocitaria al transfer embrionario

One of the most difficult aspects of assisted reproductive technology (ART) is the determination of which embryos are most suitable for transfer into the uterus. Two factors requiring consideration include the choice of embryos with the best developmental competence and the risk of multiple pregnancy associated with the number of embryos transferred. The development of technological advances such as micromanipulation and a wealth of experience in embryo culturing techniques have resulted in an increase in embryo implantation potential. Numerous criteria have been suggested to optimize the selection process. Despite all the advances in determining embryo developmental competence, no consensus has been reached on how many embryos to transfer. Key words: IVF, oocyte selection, embryo culture, PG

“Outcome” della FIV dopo coltura e transfer di blastocisti ottenute con e senza l’ausilio della cocoltura

La coltura "in vitro" di blastocisti fu messa a punto agli inizi degli anni '80 in alcune specie di animali d'allevamento (Camous, 1984) e solo successivamente applicata agli embrioni umani. Scopo della tecnica era quello di incrementare le percentuali di impianto nei cicli FIV riducendo il numero di embrioni trasferiti. La coltura di blastocisti consentiva, infatti, di effettuare il trasferimento di embrioni con elevate potenzialità evolutive e in perfetto sincronismo con l'endometrio uterino. Attraverso il sistema della cocoltura fu possibile supportare lo sviluppo embrionario "in vitro" dallo stadio di singola cellula a quello di blastocisti con l'ausilio di monostrati cellulari costituiti da cellule del tratto genitale , per lo più dell'epitelio tubarico. Fu solo nei primi anni '90 che la tecnica di cocoltura si avvalse dell'utilizzo di cellule extra-genitali, quali le cellule epiteliali di rene di scimmmia, meglio note come cellule VERO. Le cellule VERO avevano in comune con le cellule del tratto genitale l'origine mesodermica e presentavano, inoltre, notevoli vantaggi: la facile distribuzione commerciale,la disponibilità illimitata e la sicurezza. Negli ultimi anni la cocoltura è stata soppiantata dall'utilizzo dei nuovi terreni sequenziali stadio-specifici, con una composizione simile a quella fisiologica delle secrezioni del tratto genitale e, senza dubbio, più pratici e sicuri. Lo scopo del nostro studio è quello di mettere a confronto i due sistemi di coltura embrionaria "in vitro" valutandone l'efficacia in termini di percentuali di formazione di blastocisti, impianto e gravidanza

La Sindrome da Iperstimolazione ovarica

ABSTRACT The ovarian hyperstimulation syndrome, is characterized by a broad spectrum of clinical and biochemical modifications which are caused by the administration of medications used ovulation induction, assisted reproduction techniques, and the induction of multiple follicular growth. Therefore, it represents the most serious medical complication associated with ovulation induction. Key words: ovarian hyperstimulation syndrome, ovulation induction, assisted reproduction technique