TY - JOUR T1 - Light element distribution in fresh and frozen-thawed human ovarian tissues: a preliminary study. JF - Reprod Biomed Online Y1 - 2018 A1 - Pascolo, Lorella A1 - Venturin, Irene A1 - Gianoncelli, Alessandra A1 - Bortul, Roberta A1 - Zito, Gabriella A1 - Giolo, Elena A1 - Salomè, Murielle A1 - Bedolla, Diana E A1 - Altissimo, Matteo A1 - Zweyer, Marina A1 - Ricci, Giuseppe KW - Cryopreservation KW - Female KW - Humans KW - Microscopy, Electron, Transmission KW - Organ Preservation KW - Ovarian Follicle KW - Ovary AB -

RESEARCH QUESTION: Does synchrotron X-ray fluorescence (XRF) provide novel chemical information for the evaluation of human ovarian tissue cryopreservation protocols?

DESIGN: Tissues from five patients undergoing laparoscopic surgery for benign gynaecological conditions were fixed for microscopic analysis either immediately or after cryopreservation. After fixation, fresh and slowly frozen samples were selected by light microscopy and transmission electron microscopy, and subsequently analysed with synchrotron XRF microscopy at different incident energies.

RESULTS: The distributions of elements detected at 7.3 keV (S, P, K, Cl, Fe, and Os) and 1.5 keV (Na and Mg) were related to the changes revealed by light microscopy and transmission electron microscopy analyses. The light elements showed highly informative findings. The S distribution was found to be an indicator of extracellular component changes in the stromal tissues of the freeze-stored samples, further revealed by the transmission electron microscopy analyses. Low-quality follicles, frequent in the freeze-thawed tissues, showed a high Na level in the ooplasm. On the contrary, good-quality follicles were detected by a homogeneous Cl distribution. The occurrence of vacuolated follicles increased after cryopreservation, and the XRF analyses showed that the vacuolar structures contained mainly Cl and Na.

CONCLUSIONS: The study demonstrates that elemental imaging techniques, particularly revealing the distribution of light elements, could be useful in establishing new cryopreservation protocols.

VL - 37 IS - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/29802069?dopt=Abstract ER - TY - JOUR T1 - Pitfalls and promises in FTIR spectromicroscopy analyses to monitor iron-mediated DNA damage in sperm. JF - Reprod Toxicol Y1 - 2016 A1 - Pascolo, Lorella A1 - Bedolla, Diana E A1 - Vaccari, Lisa A1 - Venturin, Irene A1 - Cammisuli, Francesca A1 - Gianoncelli, Alessandra A1 - Mitri, Elisa A1 - Giolo, Elena A1 - Luppi, Stefania A1 - Martinelli, Monica A1 - Zweyer, Marina A1 - Ricci, Giuseppe AB -

Many drugs, chemicals, and environmental factors can impair sperm functionality by inducing DNA damage, one of the important causes of reduced fertility potential. The use of vibrational spectromicroscopy represents a promising approach for monitoring DNA integrity in sperm, although some limitations exist, depending from the experimental conditions. Here, we report that when using FTIR spectromicroscopy to reveal oxidative stress mediated by Fenton's reaction on hydrated sperm samples, DNA damage interpretation is partially compromised by unexpected cell surface precipitates. The precipitates give a broad band in the 1150-1000cm(-1) infrared region, which partially covers one of the signatures of DNA (phosphate stretching bands), and are detected as iron and oxygen containing material when using XRF spectroscopy. On the other hand, the analyses further support the potential of FTIR spectromicroscopy to reveal cellular oxidative damage events such as lipid peroxidation, protein misfolding and aggregations, as well as DNA strain breaks.

VL - 61 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26923261?dopt=Abstract ER - TY - JOUR T1 - Effect of seminal leukocytes on in vitro fertilization and intracytoplasmic sperm injection outcomes. JF - Fertil Steril Y1 - 2015 A1 - Ricci, Giuseppe A1 - Granzotto, Marilena A1 - Luppi, Stefania A1 - Giolo, Elena A1 - Martinelli, Monica A1 - Zito, Gabriella A1 - Borelli, Massimo KW - Adult KW - Female KW - Fertilization in Vitro KW - Flow Cytometry KW - Humans KW - Leukocytes KW - Male KW - Pregnancy KW - Pregnancy Rate KW - Prospective Studies KW - Semen KW - Sperm Injections, Intracytoplasmic AB -

OBJECTIVE: To investigate the influence of seminal leukocytes on conventional IVF and intracytoplasmic sperm injection (ICSI) outcomes, using a flow cytometry method.

DESIGN: Prospective study.

SETTING: Tertiary infertility center and research institute.

PATIENT(S): One hundred sixty-four couples undergoing conventional IVF or ICSI.

INTERVENTION(S): Seminal leukocytes were counted by flow cytometry.

MAIN OUTCOME MEASURE(S): Correlation between seminal leukocytes concentration and reproductive outcomes in IVF and ICSI cycles.

RESULT(S): The median number of oocytes retrieved, the fertilization and cleavage rate, the median number and grade of embryos transferred, the median number of good-quality embryos transferred, and the median percentage of good-quality embryos from total embryos transferred, in leukocytospermic and non-leukocytospermic patients were not statistically different after either IVF or ICSI. Similarly, there were no significant differences between the two groups for implantation rate and clinical pregnancy rate. Multivariate logistic regression analysis showed that the reproductive outcomes were not influenced by adjustment for female age, infertility diagnosis, number of previous attempts, treatment protocol (GnRH agonist or antagonist), assisted reproduction procedure (IVF or ICSI), and leukocytospermia. By profiling the proper Poisson regression models, no leukocytospermia cut-off value was able to identify the subjects at risk for oocyte fertilization or embryo cleavage failure.

CONCLUSION(S): Using a flow cytometry method, we demonstrated that leukocytospermia does not significantly influence IVF or ICSI outcomes. The same results were obtained by using lower or higher cut-off values for leukocytospermia (from 0.2 to 2 × 10(6)/mL).

VL - 104 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25936234?dopt=Abstract ER - TY - JOUR T1 - Medical treatments for endometriosis-associated pelvic pain. JF - Biomed Res Int Y1 - 2014 A1 - Zito, Gabriella A1 - Luppi, Stefania A1 - Giolo, Elena A1 - Martinelli, Monica A1 - Venturin, Irene A1 - Di Lorenzo, Giovanni A1 - Ricci, Giuseppe KW - Endometriosis KW - Female KW - Gonadotropin-Releasing Hormone KW - Histone Deacetylase Inhibitors KW - Hormone Antagonists KW - Humans KW - Pain Management KW - Pelvic Pain KW - Progestins AB -

The main sequelae of endometriosis are represented by infertility and chronic pelvic pain. Chronic pelvic pain causes disability and distress with a very high economic impact. In the last decades, an impressive amount of pharmacological agents have been tested for the treatment of endometriosis-associated pelvic pain. However, only a few of these have been introduced into clinical practice. Following the results of the controlled studies available, to date, the first-line treatment for endometriosis associated pain is still represented by oral contraceptives used continuously. Progestins represent an acceptable alternative. In women with rectovaginal lesions or colorectal endometriosis, norethisterone acetate at low dosage should be preferred. GnRH analogues may be used as second-line treatment, but significant side effects should be taken into account. Nonsteroidal anti-inflammatory drugs are widely used, but there is inconclusive evidence for their efficacy in relieving endometriosis-associated pelvic pain. Other agents such as GnRH antagonist, aromatase inhibitors, immunomodulators, selective progesterone receptor modulators, and histone deacetylase inhibitors seem to be very promising, but there is not enough evidence to support their introduction into routine clinical practice. Some other agents, such as peroxisome proliferator activated receptors-γ ligands, antiangiogenic agents, and melatonin have been proven to be efficacious in animal studies, but they have not yet been tested in clinical studies.

VL - 2014 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25165691?dopt=Abstract ER - TY - JOUR T1 - Association between the JC polyomavirus infection and male infertility. JF - PLoS One Y1 - 2012 A1 - Comar, Manola A1 - Zanotta, Nunzia A1 - Croci, Eleonora A1 - Murru, Immacolata A1 - Marci, Roberto A1 - Pancaldi, Cecilia A1 - Dolcet, Ornella A1 - Luppi, Stefania A1 - Martinelli, Monica A1 - Giolo, Elena A1 - Ricci, Giuseppe A1 - Tognon, Mauro KW - Adult KW - Amino Acid Sequence KW - Amino Acid Substitution KW - Base Sequence KW - BK Virus KW - Capsid Proteins KW - DNA, Viral KW - Humans KW - Infertility, Male KW - JC Virus KW - Male KW - Molecular Sequence Data KW - Polyomavirus Infections KW - Semen KW - Sequence Analysis, DNA KW - Tumor Virus Infections AB -

In recent years the incidence of male infertility has increased. Many risk factors have been taken into consideration, including viral infections. Investigations into viral agents and male infertility have mainly been focused on human papillomaviruses, while no reports have been published on polyomaviruses and male infertility. The aim of this study was to verify whether JC virus and BK virus are associated with male infertility. Matched semen and urine samples from 106 infertile males and 100 fertile males, as controls, were analyzed. Specific PCR analyses were carried out to detect and quantify large T (Tag) coding sequences of JCV and BKV. DNA sequencing, carried out in Tag JCV-positive samples, was addressed to viral protein 1 (VP1) coding sequences. The prevalence of JCV Tag sequences in semen and urine samples from infertile males was 34% (72/212), whereas the BKV prevalence was 0.94% (2/212). Specifically, JCV Tag sequences were detected in 24.5% (26/106) of semen and 43.4% (46/106) of urine samples from infertile men. In semen and urine samples from controls the prevalence was 11% and 28%, respectively. A statistically significant difference (p<0.05) in JCV prevalence was disclosed in semen and urine samples of cases vs. controls. A higher JC viral DNA load was detected in samples from infertile males than in controls. In samples from infertile males the JC virus type 2 strain, subtype 2b, was more prevalent than ubiquitous type 1. JCV type 2 strain infection has been found to be associated with male infertility. These data suggest that the JC virus should be taken into consideration as an infectious agent which is responsible for male infertility.

VL - 7 IS - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22912758?dopt=Abstract ER -