TUNEL assay—Standardized method for testing sperm DNA fragmentation

Sperm DNA integrity is important for normal functions such as fertilization, implantation, pregnancy and fetal development. Sperm DNA fragmentation (SDF) is more common in infertile men and may be responsible for poor reproductive function. Although there are a number of tests available to measure SDF, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling TUNEL) assay using flow cytometry is becoming more popular to measure the sperm DNA fragmentation. It is a direct test that measures both single- and double- DNA strand breaks. In this review, we describe the protocol, quality control and measurement of sperm DNA fragmentation using a benchtop flow cytometer

Rainfall thresholds for shallow landslide occurrence in Calabria, southern Italy

Abstract. In many areas, rainfall is the primary trigger of landslides. Determining the rainfall conditions responsible for landslide occurrence is important, and may contribute to saving lives and properties. In a long-term national project for the definition of rainfall thresholds for possible landslide occurrence in Italy, we compiled a catalogue of 186 rainfall events that resulted in 251 shallow landslides in Calabria, southern Italy, from January 1996 to September 2011. Landslides were located geographically using Google Earth®, and were given a mapping and a temporal accuracy. We used the landslide information, and sub-hourly rainfall measurements obtained from two complementary networks of rain gauges, to determine cumulated event vs. rainfall duration (ED) thresholds for Calabria. For this purpose, we adopted an existing method used to prepare rainfall thresholds and to estimate their associated uncertainties in central Italy. The regional thresholds for Calabria were found to be nearly identical to previous ED thresholds for Calabria obtained using a reduced set of landslide information, and slightly higher than the ED thresholds obtained for central Italy. We segmented the regional catalogue of rainfall events with landslides in Calabria into lithology, soil regions, rainfall zones, and seasonal periods. The number of events in each subdivision was insufficient to determine reliable thresholds, but allowed for preliminary conclusions about the role of the environmental factors in the rainfall conditions responsible for shallow landslides in Calabria. We further segmented the regional catalogue based on administrative subdivisions used for hydro-meteorological monitoring and operational flood forecasting, and we determined separate ED thresholds for the Tyrrhenian and the Ionian coasts of Calabria. We expect the ED rainfall thresholds for Calabria to be used in regional and national landslide warning systems. The thresholds can also be used for landslide hazard and risk assessments, and for erosion and landscape evolution studies, in the study area and in similar physiographic regions in the Mediterranean area

Geochemical and Sr-Nd isotopic features of the Zaro volcanic complex: insights on the magmatic processes triggering a small-scale prehistoric eruption at Ischia island (south Italy)

The prehistoric (< 7 ka) Zaro eruption at Ischia island (Southern Italy) produced a lava complex overlaying a pyroclastic deposit. Although being of low energy, the Zaro eruption might have caused casualties among the neolithic population that inhabited that area of Ischia, and damages to their settlements. A similar eruption at Ischia with its present-day population would turn into a disaster. Therefore, understanding the magmatic processes that triggered the Zaro eruption would be important for volcanic hazard assessment and risk mitigation, so as to improve a knowledge that can be applied to other active volcanic areas worldwide. The main Zaro lava body is trachyte and hosts abundant mafic and felsic enclaves. Here all juvenile facies have been fully characterized from petrographic, geochemical and isotopic viewpoints. The whole dataset (major and trace element contents; Sr-Nd isotopic composition) leads to rule out a genetic link by fractional crystallization among the variable facies. Thus, we suggest that the Zaro mafic enclaves could represent a deep-origin mafic magma that mingled/mixed with the main trachytic one residing in the Ischia shallow magmatic system. The intrusion of such a mafic magma into a shallow reservoir filled by partly crystallized, evolved magma could have destabilized the magmatic system presumably acting as a rapid eruption trigger. The resulting processes of convection, mixing and rejuvenation have possibly played an important role in pre- and syn-eruptive phases also in several eruptions of different sizes in the Neapolitan area and elsewhere in the world

Tracking research trends and hotspots in sperm DNA fragmentation testing for the evaluation of male infertility: a scientometric analysis.

BACKGROUND: This article describes the research trends in sperm DNA fragmentation (SDF) over the past 20 years (1999-2018) using a scientometric approach. METHODS: A stepwise approach was adopted to retrieve scientometric data (articles per year, authors, affiliations, journals, countries) from Scopus and analyze the publication pattern of SDF with reference to key areas of research in the field of Andrology. RESULTS: A total of 2121 articles were retrieved related to SDF. Our data revealed an increasing research trend in SDF (n = 33 to n = 173) over the past 20 years (R2 = 0.894). Most productive country in publications was the USA (n = 450), while Agarwal A. (n = 129) being the most productive author. Most of the articles in SDF were primarily focused on lifestyle (n = 157), asthenozoospermia (n = 135) and varicocele (130). Mechanistic studies on SDF were published twice as much as prognostic/diagnostic studies, with significant emphasis on oxidative stress. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was the most widely used technique to evaluate SDF. Publications on SDF related to assisted reproductive techniques also showed a linear increasing trend (R2 = 0.933). CONCLUSIONS: Our analysis revealed an increasing trend in SDF publications predominantly investigating lifestyle, asthenozoospermia and varicocele conditions with TUNEL being the most widely used technique. A substantial increase in research is warranted to establish SDF as prognostic/diagnostic parameter to evaluate clinical scenarios and ART outcomes

Adding hydroxyurea in combination with ruxolitinib improves clinical responses in hyperproliferative forms of myelofibrosis

Ruxolitinib, an orally bioavailable and selective inhibitor of Janus kinase 1 (JAK1) and JAK2, significantly reduces splenomegaly and disease-related symptoms in patients with myelofibrosis (MF). However, no clear survival benefit has been demonstrated, which may in part reflect suboptimal drug exposure related to lower dosages needed to minimize hematological toxicity, specifically cytopenias. Furthermore, the optimal management of specific conditions such as leukocytosis or thrombocytosis in patients under ruxolitinib therapy is still undefined. In these cases, combining ruxolitinib with a cytoreductive agent like hydroxyurea might improve hematological response. This observational multi-center study enrolled 20 adult patients with intermediate- or high-risk primary MF, post- polycythemia vera MF, or postessential thrombocythemia MF with hyperproliferative manifestations of the disease and WBC and/or platelet counts not controlled by ruxolitinib therapy. The patients received treatment with a combination of ruxolitinib and hydroxyurea. A clinical response of any type was obtained in 8 patients (40%) during ruxolitinib monotherapy and in 17 patients (85%) during ruxolitinib-hydroxyurea combination (P = 0.003). After a median duration of 12.4 months of combination therapy, 16/20 patients had a hematological response; 14/17 patients who had started combination therapy to control WBC count and 2/3 who started in order to reduce platelets count. The number of patients requiring ruxolitinib dosage reduction or discontinuations was lower during combination therapy and, at the end of follow-up the median ruxolitinib dose was increased in 50% of patients. In conclusion, the combination of hydroxyurea with ruxolitinib yielded a high clinical response rate and increased ruxolitinib exposure in patients with hyperproliferative forms of MF

Developing and piloting a communication assessment tool assessing patient perspectives on communication with pharmacists (CAT-Pharm)

Background: Effective communication strategies in health care help to enhance patient empowerment and improve clinical outcomes. Objective: Adapt the original Communication Assessment (CAT) instrument for the pharmacist profession (CAT-Pharm) and to test its validity and reliability in two different settings. Setting: Five hospital pharmacies in Italy and five community pharmacies in Malta. Method: Pilot study involving a standardized multi-step process adhering to internationally accepted and recommended guidelines. Corrections and adjustments to the translation addressed linguistic factors and cultural components. CAT-Pharm, compared to the original CAT, maintained 10 out of the 14 items: one was slightly modified; three were changed to better fit the pharmacist role; one was added. Main outcome measure: CAT-Pharm development and testing its practicality to Assess patient perceptions of pharmacists’ interpersonal and communication skills. Results CAT-Pharm was tested on 97 patients in the Italian setting and 150 patients in the Maltese setting to assess the practicality of the tool and its usefulness in investigating gaps and priorities for improving pharmacist-patient communication. Results: Show reliability and internal validity of the CAT-Pharm tool. The analysis of patient perceptions of communication with the pharmacist in Italy indicated differences from that in Malta. The different settings provided insight into the utility of CAT-Pharm. Conclusion: This study provided a valid and reliable tool that could be applied to assess patient perception of the pharmacist's communication abilities

Molecular and clinical characterization of albinism in a large cohort of Italian patients.

PURPOSE: The purpose of this study was to identify the molecular basis of albinism in a large cohort of Italian patients showing typical ocular landmarks of the disease and to provide a full characterization of the clinical ophthalmic manifestations. METHODS: DNA samples from 45 patients with ocular manifestations of albinism were analyzed by direct sequencing analysis of five genes responsible for albinism: TYR, P, TYRP1, SLC45A2 (MATP), and OA1. All patients studied showed a variable degree of skin and hair hypopigmentation. Eighteen patients with distinct mutations in each gene associated with OCA were evaluated by detailed ophthalmic analysis, optical coherence tomography (OCT), and fundus autofluorescence. RESULTS: Disease-causing mutations were identified in more than 95% of analyzed patients with OCA (28/45 [62.2%] cases with two or more mutations; 15/45 [33.3%] cases with one mutation). Thirty-five different mutant alleles were identified of which 15 were novel. Mutations in TYR were the most frequent (73.3%), whereas mutations in P occurred more rarely (13.3%) than previously reported. Novel mutations were also identified in rare loci such as TYRP1 and MATP. Mutations in the OA1 gene were not detected. Clinical assessment revealed that patients with iris and macular pigmentation had significantly higher visual acuity than did severe hypopigmented phenotypes. CONCLUSIONS: TYR gene mutations represent a relevant cause of oculocutaneous albinism in Italy, whereas mutations in P present a lower frequency than that found in other populations. Clinical analysis revealed that the severity of the ocular manifestations depends on the degree of retinal pigmentation

Highly cited articles in the field of male infertility and antioxidants: a scientometric analysis

Purpose The objective of this scientometric analysis was to recognize the top 100 cited articles on ‘Male infertility and Antioxidants’ and analyze its publication characteristics. Materials and Methods The Scopus database was used to retrieve related articles and the top 100 identified based on citation rate. Results The articles were published in 56 journals between 1995 and 2019 with a median (interquartile range) citation score of 17 (5–62). Among the top 100 articles, 69 were clinical studies, which included controlled and blinded (33.33%), prospective (27.54%), randomized-controlled trials (26.09%), uncontrolled (11.59%), and retrospective (1.45%) studies. In addition to conventional semen parameters, advanced sperm function tests such as oxidative stress (51%) and sperm DNA damage (23%) were reported. Pregnancy rate (33%) was found to be the most reported reproductive outcome. Antioxidant therapy was mostly investigated in male cohorts with sperm abnormalities such as asthenozoospermia (28%) and clinical conditions such as idiopathic male infertility (20%), varicocele/varicocelectomy (17%) and general male infertility (16%). Conclusions The most influential publications on antioxidants and male infertility were identified for the first time in the literature. This will serve as a reliable source of information for researchers and clinicians alike

Polyphenol intake and cardiovascular risk factors in a population with\ua0type 2 diabetes: The TOSCA.IT study.

Summary Background The role of polyphenol intake on cardiovascular risk factors is little explored, particularly in people with diabetes. Aim: To evaluate the association between the intake of total polyphenols and polyphenol classes with the major cardiovascular risk factors in a population with type 2 diabetes. Methods Dietary habits were investigated in 2573 males and females participants of the TOSCA.IT study. The European Prospective Investigation on Cancer and Nutrition (EPIC) questionnaire was used to assess dietary habits. In all participants, among others, we assessed anthropometry, plasma lipids, blood pressure, C-reactive protein and HbA1c following a standard protocol. The USDA and Phenol-Explorer databases were used to estimate the polyphenol content of the habitual diet. Results Average intake of polyphenols was 683.3 \ub1 5.8 mg/day. Flavonoids and phenolic acids were the predominant classes (47.5% and 47.4%, respectively). After adjusting for potential confounders, people with the highest intake of energy-adjusted polyphenols (upper tertile) had a more favorable cardiovascular risk factors profile as compared to people with the lowest intake (lower tertile) (BMI was 30.7 vs 29.9 kg/m2, HDL-cholesterol was 45.1 vs 46.9 mg/dl, LDL-cholesterol was 103.2 vs 102.1 mg/dl, triglycerides were 153.4 vs 148.0 mg/dl, systolic and diastolic blood pressure were respectively 135.3 vs 134.3 and 80.5 vs 79.6 mm/Hg, HbA1c was 7.70 vs 7.67%, and C-reactive Protein was 1.29 vs 1.25 mg/dl, p < .001 for all). The findings were very similar when the analysis was conducted separately for flavonoids or phenolic acids, the two main classes of polyphenols consumed in this population. Conclusions Polyphenol intake is associated with a more favorable cardiovascular risk factors profile, independent of major confounders. These findings support the consumption of foods and beverages rich in different classes of polyphenols particularly in people with diabetes. Clinical trial http://www.clinicaltrials.gov; Study ID number: NCT00700856