Marianna Roccio

Infant sex, obstetric risk factors, and 2-year neurodevelopmental outcome among preterm infants

Aim  To evaluate the effect of the interaction between fetal sex and obstetric variables on the risk of neurodevelopmental impairment among preterm infants.

Cervical infections by multiple human papillomavirus (HPV) genotypes: Prevalence and impact on the risk of precancerous epithelial lesions

A large proportion of human papillomavirus (HPV) infections is sustained by multiple genotypes. The effect of multiple infections on the risk of cervical intraepithelial neoplasia (CIN) and the potential efficacy of vaccine on these infections are controversial. We performed viral typing by SFP10‐LIPA on a consecutive series of 1,323 women undergoing colposcopy, 69% of whom had cervical biopsy, and correlated CIN severity with the type and number of HPVs. Overall prevalence of HPV‐DNA was 68.9%, 97.3% in CIN1, and 98.1% in CIN≥2. HPV positivity correlated with younger age (35.9 vs. 37.3 years, P = 0.026) and history of CIN (P < 0.001). Multiple types were detected in 44.2% of cases, including 63.1% CIN1 and 80.8% CIN≥2. Twenty‐three different types were detected, HPV‐16, 31 and 52 being the most frequent. Infections by HPV‐6, 11, 16, or 18 occurred in 59.4% of CIN1 and 71.3% of CIN≥2. Number of viral types and class of oncogenic risk were linearly correlated with CIN severity (P < 0.0001) by univariate and multivariate analyses controlling for age and history of CIN. The effect of the number of HPV types was maintained after exclusion from the model of infections by HPV‐6, 11, 16, and 18. Frequency, distribution, and clinical correlates of multiple HPV infections highlight the importance of assessing individual types in the management and the prediction of outcome of women with abnormal baseline cytology and point to potential limitations in current vaccine strategies. J. Med. Virol. 81:703–712, 2009

Multiple human papillomavirus infection and high grade cervical intraepithelial neoplasia among women with cytological diagnosis of atypical squamous cells of undetermined significance or low grade squamous intraepithelial lesions

OBJECTIVE To evaluate the effect of multiple human papillomavirus (HPV) infection on the prevalence of cervical intraepithelial neoplasia (CIN) among women undergoing colposcopy following a cytological diagnosis of atypical squamous cells of undetermined significance (ASCUS) or low grade squamous intraepithelial lesions (LSIL). METHODS HPV type-specific sequences of 15 high-risk and 10 low risk types were detected by the line probe, INNO-LiPA HPV genotyping assay before colposcopic examination and targeted biopsies. Multinomial logistic regression was used to evaluate the effect of multiple infection on pathologic outcome adjusting for confounders. RESULTS The prevalence of HPV infection in the 1218 women enrolled was 69.9% (851/1218). HPV 16 (37.4%), 31 (26.1%), 51 (17.4%), 52 (15.7%) and 18 (14%) were the commonest viral types identified. Overall, the rates of multiple infection were 22.5% (153/680) among subjects with negative colposcopy/biopsy, 63.6% (218/343) and 79.5% (155/195) among those with CIN 1 and CIN>or=2, respectively (p for trend <.001). The corresponding rates among subjects uninfected by HPV 16 or 18 were 13.5% (77/572), 57.4% (112/195) and 62% (48/77), respectively (p for trend <.001). In multinomial logistic regression, the odds ratio of CIN>or=2 in multiple high risk as compared to single high risk HPV infection was 4.33 (95% confidence intervals=2.32-7.14) in the overall population and 2.76 (95% confidence intervals=1.36-5.59) among women uninfected by HPV 16 or 18. CONCLUSIONS Multiple HPV infection is a significant risk factor for CIN>or=2 among women undergoing colposcopy because of ASCUS/LSIL.

Conditioned Medium From Human Amniotic Mesenchymal Stromal Cells Limits Infarct Size and Enhances Angiogenesis

The paracrine properties of human amniotic membrane‐derived mesenchymal stromal cells (hAMCs) have not been fully elucidated. The goal of the present study was to elucidate whether hAMCs can exert beneficial paracrine effects on infarcted rat hearts, in particular through cardioprotection and angiogenesis. Moreover, we aimed to identify the putative active paracrine mediators. hAMCs were isolated, expanded, and characterized. In vitro, conditioned medium from hAMC (hAMC‐CM) exhibited cytoprotective and proangiogenic properties. In vivo, injection of hAMC‐CM into infarcted rat hearts limited the infarct size, reduced cardiomyocyte apoptosis and ventricular remodeling, and strongly promoted capillary formation at the infarct border zone. Gene array analysis led to the identification of 32 genes encoding for the secreted factors overexpressed by hAMCs. Among these, midkine and secreted protein acidic and rich in cysteine were also upregulated at the protein level. Furthermore, high amounts of several proangiogenic factors were detected in hAMC‐CM by cytokine array. Our results strongly support the concept that the administration of hAMC‐CM favors the repair process after acute myocardial infarction.

Clustering patterns of human papillomavirus genotypes in multiple infections.

Many human papillomavirus (HPV) infections are sustained by multiple viral genotypes whose effect on the risk of cervical intraepithelial neoplasia (CIN) is unknown. The study investigated whether specific HPV types or species may affect the likelihood of multiple infections and have a clustered distribution in a consecutive series of 681 women with a histological diagnosis of CIN. HPV typing was performed by the SPF(10)-LIPA assay; associations were evaluated by loglinear analysis of multiple contingency tables after stratification by age and CIN grade. HPV prevalence was 99.4% with a 72.1% rate of coinfection. The risk of coinfection was higher for types 6, 11, 16, 18, 31, 33, 51, 52, 56. Significant interactions were found for species A7-A9-A10, A6-A9 and A7-A10. Coinfection by types 31-35-56, 16-51-52, 16-18 and 51-52 was more frequent than expected. Interactions between viral species and HPV 16-18 were maintained among CIN1, whereas interactions of 16-51-52 and 31-51-56 were significant only in CIN> or =2. Interactions between species and types were lost among women younger than 32 years. Significant clustering of HPV types and species occurs among women with CIN. This has implications for the assessment of the oncogenic potential and the prevention of HPV infections.

Prognostic significance of the interaction between abnormal umbilical and middle cerebral artery Doppler velocimetry in pregnancies complicated by fetal growth restriction

Objective. To evaluate the prognostic significance of the interaction between umbilical artery (UA) and middle cerebral artery (MCA) Doppler measurements in pregnancies complicated by fetal growth restriction (FGR). Design. Cohort study. Setting. Third‐level Perinatology Center in Northern Italy. Population. A study of 184 singleton pregnancies at 24–35 weeks’ gestational age complicated by FGR and abnormal UA Doppler measurements. Methods. FGR was diagnosed by serial ultrasonograms. Neonatal brain damage was defined as the presence of cystic leukomalacia or grade III‐IV intraventricular hemorrhage. Main outcome measures. Perinatal death and neonatal brain damage. Results. The prevalence of fetal/neonatal death or brain damage was 18.2% (16/88) in pregnancies with UA absent/reversed diastolic flow and 4.2% (4/96) in those with increased UA Doppler pulsatility. Stepwise logistic regression identified decreasing gestational age (OR = 1.75, 95% confidence interval, CI = 1.35–2.22) and absent/reversed UA blood flow (OR = 3.34, 95% CI = 1.1–10.9) as predictors of fetal/neonatal death or brain damage. A MCA pulsatility index below the 10th percentile was a risk factor for fetal/neonatal death or brain damage among women with absent/reversed UA diastolic flow (14/53 as compared to 2/35; OR = 5.9, CI =1.4–40.3) but not in pregnancies with forward velocity (1/33 as compared to 3/63; OR = 0.63, 95% CI = 0.02–6.13, Synergy index = 27.7, p = 0.007). Conclusions. In pregnancies complicated by FGR and absent/reversed UA end diastolic flow, vasodilatation of the MCA is a risk factor for neonatal death or brain damage.

Low-dose combined oral contraceptive and cervicovaginal shedding of human immunodeficiency virus

BACKGROUND The role of low-dose oral contraceptives on HIV cervicovaginal shedding among HIV-positive women is controversial. STUDY DESIGN We evaluated the effect of low-dose oral contraceptives on HIV cervicovaginal shedding in a cohort of 285 HIV-seropositive women followed for a median of 20 months. A sensitive, competitive polymerase chain reaction (cPCR) and a reverse transcription PCR (cRT-PCR) were applied for the quantification of HIV-associated and cell-free RNA and proviral DNA in cervicovaginal cells, as well as HIV-RNA in plasma. RESULTS In multivariable logistic generalized estimating equations, plasma viral load >100 copies/mL (OR=1.81, 95% CI=1.3-2.53) and bacterial vaginosis (OR=1.49, 95% CI=1.1-2.02) were associated with an increased risk of HIV-1 DNA shedding, whereas current use of oral contraceptive was associated with a reduced risk (OR=0.55, 95% CI=0.33-0.92). Oral contraceptives were also associated with a reduction of risk (OR=0.38, 95% CI=0.21-0.69) of cell-associated but not cell-free HIV-1 RNA. CONCLUSIONS In HIV-positive women with low levels of HIV viremia, low-dose oral contraceptives were associated with a modest but significant reduction of HIV-1 DNA and cell-associated HIV-1 RNA genital shedding.

HIV shedding in cervico-vaginal secretions in pregnant women

OBJECTIVE The purpose of this study was to evaluate the presence of HIV-1 in cervico-vaginal secretions of pregnant as compared to non-pregnant HIV-seropositive women. PATIENTS AND METHODS We compared 43 known HIV seropositive pregnant patients versus 241 age-matched (± 2 years) control non-pregnant HIV-seropositive subjects. In pregnant patients blood and cervico-vaginal samples were obtained during each trimester of pregnancy. In control subjects the same samples were obtained at enrolment. HIV-1 RNA was measured in plasma; proviral HIV-1 DNA, cell-associated and cell-free HIV-1 RNA in cervico-vaginal secretion by competitive polymerase chain reaction (cRT-PCR) and reverse transcriptase PCR. RESULTS The genital shedding of HIV-DNA (22/43 as compared to 79/241, p = 0.02), and cell-free HIV-RNA detection (26/43 as compared to 72/241, p < .001) was more common in first-trimester pregnant than in non pregnant women. Pregnancy correlated with a significant positive trend in the cervico-vaginal load of HIV-DNA (Spearman Rho= 0.149, p= 0.012), and cell-free HIV-RNA (Spearman Rho= 0.253, p < .001), but not of HIV-RNA transcripts (Spearman Rho = 0.06, p= 0.31). After correction for potential confounders, first trimester pregnant women had increased rates of genital HIV- DNA (odds ratio = 1.94, 95% confidence interval = 1.01 3.78) and cell-free HIV-RNA (odds ratio = 4.07, 95% confidence interval = 1.97 8.41) detection compared to nonpregnant controls. CONCLUSION The shedding of genital HIV was increased in pregnant compared to non pregnant subjects, even in patients with undetectable viremia. In this low-risk HIV-positive population the risks of vertical or horizontal transmissions should not be underestimated.

Untypable human papillomavirus infection and risk of cervical intraepithelial neoplasia among women with abnormal cervical cytology.

The risk of cervical intraepithelial neoplasia and/or invasive cervical cancer associated with untypable human papillomavirus (HPV) infections has been not investigated fully. HPV infection caused by 18 high‐risk and 7 low‐risk genotypes as detected by the INNO‐LIPA genotyping system, was investigated in 4,258 women with abnormal Pap smear referred to a colposcopic service. The prevalence of HPV infection was 76.1%. Rates of cervical intraepithelial neoplasia grade 3+ were 0.88% (9/1,017) in HPV‐negative subjects, 1.8% (7/380) in subjects with untypable HPV infection, 3.2% (11/343) in subjects with single/multiple low‐risk types, 28.3% (201/709) in subjects with multiple low and high‐risk types, 15.2% (162/1,069) in subjects with single high‐risk types, and 31.2% (229/733) in those with multiple high‐risk types. Compared to women without any HPV infection, the odds ratios of cervical intraepithelial neoplasia grade 2+ or grade 3+ in subjects with untypable or low‐risk HPV genotypes were 5.73 (95% CI = 2.79–11.78) and 12.4 (95% CI = 6.31–24.5, P = 0.014 compared to untypable) and 3.1 (95% CI = 1.11–8.16) and 7.1 (95% CI = 2.9–17.2, P = 0.07 compared to untypable), respectively. In the subgroup of subjects with cervical intraepithelial neoplasia grade 1 or negative colposcopy/biopsy, the progression to cervical intraepithelial neoplasia grade 2+ at follow‐up (median 25 months, range 6–70) was 2% (14/684), 3.4% (7/205), and 5.6% (11/195, P = 0.04 compared to negative) among negative, untypable, and low‐risk HPV infection, respectively. The risk of cervical intraepithelial neoplasia associated with untypable HPV infection was higher than that recorded among uninfected women, but lower than the risk associated with low‐ or high‐risk HPV genotypes. J. Med. Virol. 86:1145–1152, 2014.

Human 3D Cultures as Models for Evaluating Magnetic Nanoparticle CNS Cytotoxicity after Short- and Repeated Long-Term Exposure

Since nanoparticles (NPs) can translocate to the brain and impact the highly vulnerable central nervous system (CNS), novel in vitro tools for the assessment of NP-induced neurotoxicity are advocated. In this study, two types of CNS spheroids have been developed from human D384 astrocyte- and SH-SY5Y neuronal-like cells, and optimized in combination with standard assays (viability readout and cell morphology) to test neurotoxic effects caused by Fe3O4NPs, as NP-model, after short- (24–48 h; 1–100µg/ml) and long-term repeated exposure (30days; 0.1–25µg/ml). Short-term exposure of 3D-spheroids to Fe3O4NP induced cytotoxicity at 10 µg/mL in astrocytes and 25 µg/mL neurons. After long-term repeated dose regimen, spheroids showed concentration- and time-dependent cell mortality at 10 µg/mL for D384 and 0.5 µg/mL for SH-SY5Y, indicating a higher susceptibility of neurons than astrocytes. Both spheroid types displayed cell disaggregation after the first week of treatment at ≥0.1 µg/mL and becoming considerably evident at higher concentrations and over time. Recreating the 3D-spatial environment of the CNS allows cells to behave in vitro more closely to the in vivo situations, therefore providing a model that can be used as a stand-alone test or as a part of integrated testing strategies. These models could drive an improvement in the species-relevant predictivity of toxicity testing.