Sperm Chromatin Dispersion Test Detects Sperm DNA Fragmentation Mainly Associated with Unviable Spermatozoa and Underestimates the Values with Respect to TUNEL Assay.

Several clinical laboratories assess sperm DNA fragmentation (sDF) in addition to semen analysis in male infertility diagnosis. Among tests evaluating sDF, TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) and SCD (Sperm Chromatin Dispersion) are widely used. Our lab developed a modified version of TUNEL (TUNEL/PI) able to distinguish two sperm populations (PI Brighter and PI Dimmer) differently associated with sperm viability and reproductive outcomes. The aim of this study was to compare sDF levels detected by SCD and TUNEL/PI in the semen samples from 71 male subjects attending our Andrology Laboratory. Our results demonstrate that SCD is less sensitive in determining sDF compared to TUNEL/PI. The statistically significant positive correlation found between sDF evaluated by SCD and PI Dimmer (consisting of all dead spermatozoa) suggests that SCD mainly detects sDF in unviable spermatozoa. We confirmed that most spermatozoa detected by SCD are unviable by performing SCD after incubation in hypo-osmotic medium to discriminate viable and unviable cells in 52 samples. Such results might explain the lower ability of this test in discriminating couples having successful ART outcomes demonstrated in published metanalyses. Overall, our results indicate that SCD is less sensitive in evaluating sDF for diagnostic purposes.

CDK9-55 guides the anaphase-promoting complex/cyclosome (APC/C) in choosing the DNA repair pathway choice.

DNA double-strand breaks (DSBs) contribute to genome instability, a key feature of cancer. DSBs are mainly repaired by homologous recombination (HR) and non-homologous end-joining (NHEJ). We investigated the role of an isoform of the multifunctional cyclin-dependent kinase 9, CDK9-55, in DNA repair, by generating CDK9-55-knockout HeLa clones (through CRISPR-Cas9), which showed potential HR dysfunction. A phosphoproteomic screening in these clones treated with camptothecin revealed that CDC23 (cell division cycle 23), a component of the E3-ubiquitin ligase APC/C (anaphase-promoting complex/cyclosome), is a new substrate of CDK9-55, with S588 being its putative phosphorylation site. Mutated non-phosphorylatable CDC23(S588A) affected the repair pathway choice by impairing HR and favouring error-prone NHEJ. This CDK9 role should be considered when designing CDK-inhibitor-based cancer therapies.

Effects of Benzo[a]pyrene on Human Sperm Functions: An In Vitro Study.

Benzo(a)pyrene (BaP) is considered one of the most dangerous air pollutants for adverse health effects, including reproductive toxicity. It is found both in male and female reproductive fluids likely affecting spermatozoa after the selection process through cervical mucus, a process mimicked in vitro with the swim-up procedure. In vitro effects of BaP (1, 5, 10 µM) were evaluated both in unselected and swim-up selected spermatozoa after 3 and 24 h of incubation. BaP reduced total, progressive and hyperactivated motility and migration in a viscous medium both in swim-up selected and unselected spermatozoa. Viability was not significantly affected in swim-up selected but was reduced in unselected spermatozoa. In swim-up selected spermatozoa, increases in the percentage of spontaneous acrosome reaction and DNA fragmentation were observed after 24 h of incubation, whereas no differences between the control and BaP-treated samples were observed in caspase-3 and -7 activity, indicating no effects on apoptotic pathways. ROS species, evaluated by staining with CellROX® Orange and Dihydroethidium, did not differ in viable spermatozoa after BaP treatment. Conversely, the percentage of unviable ROS-positive spermatozoa increased. Our study suggests that BaP present in male and female genital fluids may heavily affect reproductive functions of human spermatozoa.

Risk Influence of Some Environmental and Behavioral Factors on Air Contamination in the Operating Room: An Experimental Study.

Air contamination in operating rooms (ORs) depends on the conditions of the room and on activities therein performed. Methodologies of air quality assessment in ORs are often inadequately described in the scientific literature, and the time required for a change in status in air quality is never taken into account. The purpose of this study was to determine the influence of the state and the presence of human operators on air quality by implementing a precise measurement protocol that also took into account the time required for changes in the room to affect air pollution. As the main indicators of air pollution, bacterial load and concentration of airborne dust were measured. The results showed that: the use of surgical masks by operators in the OR did not significantly affect bacterial load within a distance of 2 m; keeping OR doors open did not induce a significant increase in bacterial load and of 5 µm particles while 10 µm particles concentration was positively affected; and air pollution measured with open doors was not significantly different from that due to the presence of two staff members, whether or not they were wearing masks. The results clarified the role of some factors on air pollution in ORs.

The hydrophilic extract from a new tomato genotype (named DHO) kills cancer cell lines through the modulation of the DNA damage response induced by Campthotecin treatment.

Introduction: DNA double-strand breaks are the most toxic lesions repaired through the non-homologous and joining (NHEJ) or the homologous recombination (HR), which is dependent on the generation of single-strand tails, by the DNA end resection mechanism. The resolution of the HR intermediates leads to error-free repair (Gene Conversion) or the mutagenic pathways (Single Strand Annealing and Alternative End-Joining); the regulation of processes leading to the resolution of the HR intermediates is not fully understood. Methods: Here, we used a hydrophilic extract of a new tomato genotype (named DHO) in order to modulate the Camptothecin (CPT) DNA damage response. Results: We demonstrated increased phosphorylation of Replication Protein A 32 Serine 4/8 (RPA32 S4/8) protein in HeLa cells treated with the CPT in combination with DHO extract with respect to CPT alone. Moreover, we pointed out a change in HR intermediates resolution from Gene Conversion to Single Strand Annealing through the modified DNA repair protein RAD52 homolog (RAD52), DNA excision repair protein ERCC-1 (ERCC1) chromatin loading in response to DHO extract, and CPT co-treatment, with respect to the vehicle. Finally, we showed an increased sensitivity of HeLa cell lines to DHO extract and CPT co-treatment suggesting a possible mechanism for increasing the efficiency of cancer therapy. Discussion: We described the potential role of DHO extract in the modulation of DNA repair, in response to Camptothecin treatment (CPT), favoring an increased sensitivity of HeLa cell lines to topoisomerase inhibitor therapy.

Effects of Meteo-Climatic Factors on Hospital Admissions for Cardiovascular Diseases in the City of Bari, Southern Italy.

The objective of this study was to determine the relationship between weather conditions and hospital admissions for cardiovascular diseases (CVD). The analysed data of CVD hospital admissions were part of the database of the Policlinico Giovanni XXIII of Bari (southern Italy) within a reference period of 4 years (2013-2016). CVD hospital admissions have been aggregated with daily meteorological recordings for the reference time interval. The decomposition of the time series allowed us to filter trend components; consequently, the non-linear exposure-response relationship between hospitalizations and meteo-climatic parameters was modelled with the application of a Distributed Lag Non-linear model (DLNM) without smoothing functions. The relevance of each meteorological variable in the simulation process was determined by means of machine learning feature importance technique. The study employed a Random Forest algorithm to identify the most representative features and their respective importance in predicting the phenomenon. As a result of the process, the mean temperature, maximum temperature, apparent temperature, and relative humidity have been determined to be the most suitable meteorological variables as the best variables for the process simulation. The study examined daily admissions to emergency rooms for cardiovascular diseases. Using a predictive analysis of the time series, an increase in the relative risk associated with colder temperatures was found between 8.3 °C and 10.3 °C. This increase occurred instantly and significantly 0-1 days after the event. The increase in hospitalizations for CVD has been shown to be correlated to high temperatures above 28.6 °C for lag day 5.

Soil heavy metal contamination in an industrial area: analysis of the data collected during a decade.

Soil contamination by heavy metals has become a serious problem mainly because, above certain concentrations, all metals have adverse effects on human health. In particular, the accumulation of heavy metals in agricultural soils leads to elevated uptake by crops and affects food quality and safety. In this paper, we present the results of a study carried out over a decade for evaluating the impact of a new industrial settlement in an area geared to agriculture and livestock and far from urban sites. We focus our study on the bioavailable fraction of Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn in soil samples. Heavy metal concentrations in soil are analysed with both univariate and multivariate statistical procedures. The main goal of this paper is the development of a statistical procedure, based on a mix of multivariate analysis, able to compare field surveys carried out during different years and to characterize spatial and temporal changes in soil heavy metals concentrations.

A method for the integration of satellite vegetation activities observations and magnetic susceptibility measurements for monitoring heavy metals in soil.

We present a procedure for monitoring heavy metals in soil based on the integration of satellite and ground-based techniques, tested in an area affected by high anthropogenic pressure. High resolution multispectral satellite data were elaborated to obtain information on vegetation status. Magnetic susceptibility measurements of soils were collected as proxy variable for monitoring heavy metal presence. Chemical analyses of heavy metals were used for supporting and validating the integrated monitoring procedure. Magnetic and chemical measurements were organized in a GIS environment to be overlapped to satellite-based elaborations and to analyze the pattern distribution. Results show the presence of correlation between anomalies in vegetation activity and soil characteristics. The relationship between the distribution of normalized difference vegetation index anomalies and magnetic susceptibility values provides hints for adopting the integrated procedure as preliminary screening to minimize monitoring efforts and costs by supporting the planning activities of field campaigns.

Magnetic susceptibility measurements as proxy method to monitor soil pollution: the case study of S. Nicola di Melfi.

The development of in situ, cheep, noninvasive, and fast strategies for soil monitoring is a crucial task for environmental research. In this paper, we present the results of three field surveys carried out in an industrial area of Southern Italy: S. Nicola di Melfi. The monitoring procedure is based on soil magnetic susceptibility measurements carried out by means of experimental protocols that our research group developed during the last years. This field surveys is supported by both geological characterization of the area and analytical determinations of metal concentrations in soils. Magnetic studies were carried out not only in situ but also in laboratory. Results show that, taking into account the influence due to the geomorphologic difference, soil magnetic susceptibility is an optimal indicator of the anthropogenic impact. So, our monitoring strategy discloses that the combined use of magnetic susceptibility measurements and soil geomorphology information may be used as a useful tool for the temporal monitoring of pollution evolution and for a fast screening of polluted zones.

Magnetic susceptibility measurements as proxy method to monitor soil pollution: development of experimental protocols for field surveys.

In the framework of the development of new methods for measuring and monitoring soil pollution, this paper deals with the use of magnetic methodologies to monitor the heavy metals presence in soils. In particular it shows a procedure for collecting magnetic susceptibility measurements in order to interpret them as proxy variable for monitoring heavy metals in soils. Magnetic measurements are carried out using a magnetic susceptibility meter with two different probes for in situ field surveys. The experimental procedure is divided in two parts. In the first part we carry out laboratory tests aimed to evaluate, for both the probes, the effective investigation depth for soil, the measurement reproducibility under different conditions, and the influence of water content. We complete this part comparing in situ measurements obtained by means of two probes with different characteristics. In the second part we carry out tests to evaluate the relationships between heavy metal levels and magnetic susceptibility values of soil samples. We investigate the variability of the magnetic susceptibility measurements contaminating different soil samples with well known concentration of heavy metals. Moreover we study the correlation between magnetic susceptibility values and metal concentrations, determined by means of AAS, in soil samples collected during a field survey. Results suggest that a careful check of the experimental procedure play a crucial role for using magnetic susceptibility measurements for heavy metals in situ monitoring. This is very helpful both for improving the quality of data and for making simpler data interpretation.

Metal levels in fodder, milk, dairy products, and tissues sampled in ovine farms of Southern Italy.

We measured Cd, Cr, Hg, Mn, and Pb levels in samples of fodder, milk, dairy products, and tissues collected from 12 ovine farms in the regions of Campania and Calabria (Southern Italy). The areas in which the farms are located show different levels of anthropogenic pressure. The main purpose of this study is the identification and the analysis of relationships among metal concentrations observed in samples representative of different links in the food chain. Particularly, we apply univariate, bivariate, and multivariate statistical techniques to identify the correlation structure of our data set and to evaluate the influence of anthropogenic activity. We discuss the results, focusing the analysis on the spatial and the temporal patterns of metal concentrations.

Heavy metals in ryegrass species versus metal concentrations in atmospheric particulate measured in an industrial area of Southern Italy.

The aim of this paper is to evaluate the reliability of ryegrass species as active biomonitors by assessing atmospheric metal concentrations. We show a procedure for measuring atmospheric concentrations of heavy metals by means of biomonitors and present the data collected between July 1997 and October 2000 in the industrial area of Tito Scalo (Basilicata region, Southern Italy). In particular, we discuss the reproducibility of the biomonitoring measures, the influence of plant age and the correlation between metal concentrations in plants and in atmospheric particulate. Statistical analysis of measured data suggests us that in the investigated site, Cd, Cr and Ni are suitable to be monitored by means of ryegrass species. For the other metals, their emission patterns in atmosphere make it difficult to identify the correlation structure between plants and particulate, and as a result the interpretation of the biomonitoring data is complex. On the basis of the results, we believe that for correct application of active biomonitoring procedure, a careful preliminary analysis of the monitoring site and integration of the biomonitoring and chemical-physical observation is necessary.