Genetic characterisation of Measles virus variants identified during a large epidemic in Milan, Italy, March-December 2017.

In 2017, Italy experienced a large measles epidemic with 5408 cases and four deaths. As Subnational Reference Laboratory of the Measles and Rubella surveillance NETwork (MoRoNET), the EpiSoMI (Epidemiology and Molecular Surveillance of Infections) Laboratory (University of Milan) set up rapid and active surveillance for the complete characterisation of the Measles virus (Mv) responsible for the large measles outbreak in Milan and surrounding areas (Lombardy, Northern Italy). The aims of this study were to describe the genetic profile of circulating viruses and to track the pathway of measles transmission. Molecular analysis was performed by sequencing the highly variable 450 nucleotides region of the N gene (N-450) of Mv genome. Two-hundred and ninety-nine strains of Mv were analysed. The phylogenetic analysis showed five different variants, two not previously described in the studied area, belonging to D8 and B3 genotypes. Three events of continuous transmission of autochthonous variants (D8-Osaka, D8-London and B3-Milan variants) and two events of continuous transmission of imported variants (B3-Dublin and D8-Hulu Langat) tracked five different transmission pathways. These pathways outlined two epidemic peaks: the first in April and the second in July 2017. The correlation between Mv variant and the epidemiological data may enable us to identify the sources of virus importation and recognise long-lasting virus transmission pathways.

Acupuncture in clinical and experimental reproductive medicine: a review.

BACKGROUND: Acupuncture has been used as treatment for infertility for hundreds of years, and recently it has been studied in male and female infertility and in assisted reproductive technologies, although its role in reproductive medicine is still debated. AIM: To review studies on acupuncture in reproductive medicine, in experimental and clinical settings. METHODS: Papers were retrieved on PubMed and Google Scholar and were included in the review if at least the abstract was in English. RESULTS: There is evidence of benefit mainly when acupuncture is performed on the day of embryo transfer (ET) in the live birth rate. Benefit is also evident when acupuncture is performed for female infertility due to polycystic ovary syndrome (PCOS). There is some evidence of sperm quality improvement when acupuncture is performed on males affected by idiopathic infertility. Experimental studies suggest that acupuncture effects are mediated by changes in activity of the autonomic nervous system and stimulation of neuropeptides/neurotransmitters which may be involved in the pathogenesis of infertility. CONCLUSIONS: Acupuncture seems to have beneficial effects on live birth rate when performed on the day of ET, and to be useful also in PCOS as well as in male idiopathic infertility, with very low incidence of side effects. However, further studies are necessary to confirm the clinical results and to expand our knowledge of the mechanisms involved.

The impact of a diagnosis of couple subfertility on male sexual function.

AIM: The aim was to study: a) the prevalence of sexual dysfunction in male partners of subfertile couples; b) the possible correlation between sexual dysfunction and seminal profile. SUBJECTS AND METHODS: Male partners (no.=171) of subfertile couples were studied, aged between 25 and 40 yr, attending the Andrology Unit of Sant'Andrea Hospital. All the subjects responded to a modified International Index of Erectile Function (IIEF) questionnaire, made up of 5 questions: 2 regarding erectile function, 1 concerning orgasmic function, 1 question on sexual desire, and 1 on satisfaction with intercourse. The questionnaire investigated both spontaneous sex for pleasure and sex intended to lead to pregnancy. The subjects underwent standard semen analysis, according to World Health Organization guidelines. RESULTS: The data showed a disorder in sexual intercourse for reproductive purposes in 37 subjects (23.7%). Of these, only 14 (8.9%) also had problems with sex for pleasure. On the other hand, there was no significant variation in the prevalence of sexual dysfunction related to seminal profile. CONCLUSIONS: A diagnosis of subfertility represents a stressful situation which can reduce the pleasure of sex, especially in intercourse intended for reproductive purposes; this disorder in sexual activity does not seem to be directly correlated with awareness of the severity of the semen alterations.

The role of ejaculatory dysfunction on male infertility.

BACKGROUND: The aim of this study was to evaluate: 1) the prevalence of male infertility due to ejaculatory dysfunction (premature ejaculation-PE, intravaginal ejaculatory dysfunction-IVEjD, anejaculation-AE, and retrograde ejaculation-RE); and 2) the hormonal profile and semen characteristics of such subjects. METHODS: N.3280 subjects who were referred to our andrology unit for infertility were studied. Exclusion criteria: the presence of known causes of male infertility and erectile dysfunction. In all subjects were performed: medical history and andrological physical examination; hormonal profile; semen analysis or centrifugation/resuspension of post-orgasmic urine; IIEF-5 questionnaire for the diagnosis of ED; PEDT questionnaire for the diagnosis of EP. RESULTS: the prevalence of ejaculatory dysfunctions in infertile males was 1.8% (59/3280). The causes were: a) absence of ejaculation in 37/3280 subjects (1.1%); among them, 23/3280 (0.7%) subjects showed a condition of RE and 14/3280 (0.4%) of AE; b) PE in 16/3280 subjects (0.5%); and c) IVEjD in 6/3280 subjects (0.2%). Hormonal values and seminal parameters (when semen analysis was possible) were within the normal ranges in all the cases. In subjects with RE, sperm recovery was possible in 69.9% (16/23) subjects after centrifugation and resuspension of post-orgasmic urine. CONCLUSIONS: The prevalence of male infertility due to ejaculatory dysfunctions is overall just under 2%. The main cause is retrograde ejaculation; psychogenic origins could also have an important role. It is important to identify the cause of ejaculatory dysfunction in order to decide upon correct management (PE treatment, centrifugation and resuspension of post-orgasmic urine, penile vibratory stimulation, and psychological counselling).

Prevalence of diabetes mellitus in male partners of infertile couples.

AIM: The aim of this study was to investigate the prevalence of diabetes mellitus (DM) in a population of male partners of infertile couples, to evaluate their seminal and biochemical parameters and to plan therapeutic strategies to achieve pregnancy. METHODS: Standard semen analysis was carried out in 510 male partners of infertile couples, aged 25 to 51 years. RESULTS: The prevalence of DM in the subjects studied was 1.18%, 4 cases with non insulin dependent diabetes mellitus (NIDDM) and 2 cases with insulin dependent diabetes mellitus (IDDM). Three subjects with NIDDM and 1 with IDDM had normal ejaculation. Semen analysis of these subjects showed qualitative alteration; the most important effects concerned the kinetic properties, especially progressive motility. Sperm morphology was also significantly compromised. On the other hand, sperm concentration did not show significant alterations. The therapeutic iter in these subjects is similar to that for other dyspermic male partners of infertile couples. The 2 remaining subjects (one with NIDDM and one with IDDM) had retrograde ejaculation (sperm recovery from post-orgasmic urine) and were included in intrauterine insemination or in vitro fertilization programs. In addition, in the subjects with DM, the biochemical seminal profile showed significantly higher levels of fructose compared with the nondiabetic subjects. CONCLUSION: DM has a negative impact both in terms of sperm quality and of the ejaculation mechanism. The low prevalence of DM among this study population of 510 male partners of infertile couples is due to the low prevalence of DM in this age range.

New software for the automatic and semiautomatic assessment of sperm morphology.

AIM: An attempt was made to improve the standardization of sperm morphology assessment. METHODS: A software for computerized automatic/semiautomatic sperm morphology assessment was developed and a comparative study between this computerized system and the subjective micrometric method was carried out. RESULTS: The software has shown to be very reliable and reproducible. The subjective micrometric method significantly underestimated the percentage of atypical forms compared with the objective method. CONCLUSION: Until the development of new fully-automatic techniques, this system is certainly an effective first step in the objective, reproducible standardization of sperm morphology assessment.

An attempt to improve standardization of sperm motility class assessment using the superimposed image analysis system (SIAS) software.

AIM: An attempt was made to improve the standardization of sperm motility assessment. METHODS: A computerized system based on image superimposition producing final image with a motion effect was employed. Numerical definition of straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) related to these motility classes were made, at different frame/rates (20 frames/s, 30 frames/s and 60 frames/s). RESULTS: At least 4 motility class categories were found to be necessary (instead of 3, WHO) for a valid assessment. According to the cut-offs suggested by the ROC curves, motility classes were defined as follows: Class 1 (straight progressive motility): VSL = or > 23 microm/s; LIN = or > 0.58 at 20 frames/s; LIN = or > 0.50 at 30 frames/s; LIN = or > 0.44 at 60 frames/s. Class 2 (straight slow motility): VSL > 10 microm/s and < 23 microm/s; LIN = or > 0.58 at 20 frames/s; LIN = or > 0.51 at 30 frames/s; LIN = or > 0.43 at 60 frames/s. Class 3 (progressive non straight motility): VSL > 10 microm/s; LIN < 0.58 at 20 frames/s; LIN < 0.51 at 30 frames/s; LIN<0.43 at 60 frames/s. Class 4 (non progressive motility): VSL <10 microm/s. A frame rate of 20 frames/s was found to be sufficient to distinguish sperm motility classes in standard semen analysis. CONCLUSIONS: The numerical definition of sperm motility classes may contribute towards standardization in the objective evaluation of sperm kinematics.

Prevalence of Diabetes Mellitus (DM) in a population of men affected by Erectile Dysfunction (ED).

AIMS: A) to evaluate the prevalence of patients affected by Diabetes Mellitus (DM) in a population of men with Erectile Dysfunction (ED); B) to define the epidemiological, biochemical and therapeutic aspects. MATERIALS AND METHODS: N.934 subjects referred at our Andrology Unit for ED were studied. The diagnosis of ED was evaluated using the IIEF-5 questionnaire (Total score ≤21). RESULTS: The prevalence of subjects affected by DM in a population of men with ED was 19.5% (182/934). The age ranges were: ≥55 years (108/182; 59.3%); ≥40<55 years (70/182; 38.5%); <40 years (4/182; 2.2%). HbA1c mean value was 7.9% ± 0.8%. No significant differences were found in DM onset timing or in anti-diabetic treatment. In n.125/182 cases (68.7%) the ED onset followed the diagnosis of DM; in n.34/182 cases (18.7%) it appeared at the same time; and in n.23/182 cases (12.6%) appeared before DM diagnosis. ED TREATMENT: in n.18/182 subjects (9.9%) there was a concomitant hypotestosteronemia; these patients were treated only with testosterone replacement; this treatment was efficacious (IIEF-5 total score ≥22) in 8/18 subjects (44.4%). In n.146/182 subjects (80.2%) a treatment with PDE5-i was given. Of these 146 subjects, the therapy was given "on demand" to 108 subjects (efficacy in 50.9%; 55/108) and "once a day" to the remaining 38 subjects (efficacy 63.1%, 24/38) (p=0.428, n.s.). N.15/182 subjects (8.2%) were treated with intracavernous injections of Alprostadil (efficacy in 8/15, 53.3%). In n.3/182 subjects (1.6%) a penile prosthesis was implanted. CONCLUSIONS: DM is one of the most frequent organic causes of ED; there were many strategies to treat this symptom without interfering with the antidiabetic treatment. Finally, ED can be predictive of DM.

Simultaneous cycle sequencing assessment of (TG)m and Tn tract length in CFTR gene.

The lengths of the dinucleotide (TG)m and mononucleotide Tn repeats, both located at the intron 8/exon 9 splice acceptor site of the cystic fibrosis transmembrane conductance regulator (CFTR) gene whose mutations cause cysticfibrosis (CF), have been shown to influence the skipping of exon 9 in CFTR mRNA. This exon 9-skipped mRNA encodes a nonfunctional protein and is associated with various clinical manifestations in CF As a result of growing interest in these repeats, several assessment methods have been developed, most of which are, however, cumbersome, multi-step, and time consuming. Here, we describe a rapid methodfor the simultaneous assessment of the lengths of both (TG)m and Tn repeats, based on a nonradioactive cycle sequencing procedure that can be performed even without DNA extraction. This method determines the lengths of the (TG)m and Tn tracts of both alleles, which in our samples ranged from TG8 to TG12 in the presence of T5, T7, and T9 alleles, and also fully assesses the aplotypes. In addition, the repeats in the majority of these samples can be assessed by single-strand sequencing, with no need to sequence the other strand, thereby saving a considerable amount of time and effort.

The application of the differential pH method to the biochemical evaluation of seminal plasma.

OBJECTIVE: To apply the differential pH method to the biochemical evaluation of seminal plasma. DESIGN AND METHODS: Fructose, citric acid and free L-carnitine concentration in human seminal plasma were measured by the differential pH method. These are routinely taken as markers of seminal vesicle, prostate and epididymal function, respectively. The pH differential method was then compared with standard spectrophotometry. RESULTS: The linearity, recovery and imprecision calculation for each assay were found to be reliable and the method correlated well with the reference spectrophotometric technique. In addition, the pH differential method showed certain advantages compared to spectrophotometry: a) speed of procedure (40 to 120 sec/assay); b) ability to measure the analytes even in small volumes of seminal plasma (10 microL to 50 microL) without deproteinization or dilution, thus eliminating manual operations. CONCLUSIONS: The differential pH method is an innovative approach to the biochemical assessment of seminal plasma.

Superimposed Image Analysis System (SIAS) software: a new approach to sperm motility assessment.

OBJECTIVE: To apply Superimposed Image Analysis System (SIAS) software (Delta Sistemi, Rome, Italy) to the objective analysis of sperm motility. DESIGN: To test a new procedure for sperm motility assessment based on image superimposition. RESULTS: The system can superimpose six sequential frames onto a monitor producing a final image with a motion effect similar to the multiple exposure photography method, but bypassing the film processing stage. Viewing the final image of a complete series of six superimposed frames allows the evaluation of the percentage of motile spermatozoa and their kinetic characteristics. Compared with multiple exposure photography, SIAS has the following main advantages: [1] the procedure is quick; [2] projecting the first frame alone allows the easy evaluation of sperm concentration; and [3] the system allows quick and easy analysis and measurement of each sperm trajectory frame by frame, thus avoiding misleading interpretations because of crossing sperm trajectories, collisions, and overlapping of motile spermatoza with round cells or immotile spermatozoa. CONCLUSIONS: Superimposed image analysis is a new approach to sperm motility assessment. It seems to be useful particularly in selected situations, such as oligozoospermia or in cases of increased round cell concentration. In these situations, SIAS can even serve as a support or an alternative to the computer-aided sperm analysis system widely used for objective sperm analysis.

Response of adnexal glands to testosterone stimulation in the normal adult male.

Normal male volunteers were studied in order to set up a functional exploration test based on administration of testosterone propionate for use on the adnexal glands of the male genital tract. Seminal acid phosphatase and citric acid levels showed no significant variations compared with spontaneous physiologic oscillations. Concentration and total quantity of fructose were considerably reduced. Alkaline phosphatase showed an intermediate pattern with constant values of concentration and decrease of total quantity. The hypothesis is advanced that adnexal glands possess a secretory metabolism with a maximal basal response to androgen stimulation, but with a different mechanism of action: partly adaptable at the level of the prostate and of the "all or nothing" type at the level of the vesicles.

Superoxide anion in human semen related to seminal parameters and clinical aspects.

OBJECTIVE: To study the possible presence of extracellular superoxide anion (O2-) in untreated semen of normal and pathological subjects and to determine the possible relationship between superoxide anion levels, number of seminal leukocytes (WBCs) and seminal parameters. PATIENTS: Semen samples were analyzed from fertile normospermic subjects (n = 20) and patients (n = 132) attending the Outpatient Department of the 5th Medical Clinic. MAIN OUTCOME MEASURE: We evaluated superoxide anion levels by monitoring the reduction of cytochrome c. RESULTS: O2- was detected in 55.0% of fertile normospermic subjects. Both the percentage of subjects in whom O2- was detected and the mean values +/- SD of O2- were significantly higher in the male partners of infertile couples, in subjects with varicocele, and in subjects with positive sperm culture than normospermic subjects. No variations were seen in vasectomized subjects or in patients with past cryptorchidism. A high correlation was observed between O2- levels and WBCs (rs = 0.58), nonrapid, immotile sperm (rs = 0.64), percentage decrease of motility within a specified time frame (2 to 6 hours) (rs = 0.73), and sperm abnormalities (rs = 0.49). CONCLUSIONS: Superoxide anion in untreated semen is produced by WBCs and atypical and nonrapid, immotile sperm. A significant increase of superoxide anion was detected in the male partners of infertile couples and in some selected andrological conditions. The toxic effect of superoxide anion on sperm cells is dependent on the time of exposure and superoxide anion concentration.

Treatment and follow-up of patients with infertility due to spermagglutinins.

A therapeutic trial based on pharmacologically induced azoospermia followed by the administration of corticosteroids was carried out in 48 patients with infertility due to spermagglutinins. Three types of responses were observed: type A, no modifications in the blood and spermagglutinating (SA) titer (19 cases); type B, disappearance or significant decrease in SA titer during treatment, with return to normal values upon resumption of spermatogenesis (7 cases); and type C, disappearance or significant decrease in SA titer for at least 1 year after stopping treatment (22 cases). Within the 1st year after stopping therapy only men who responded in type C fashion were able to impregnate partners, three pregnancies ended in abortion and nine in normal births.

Azoospermia: incidence, and biochemical evaluation of seminal plasma by the differential pH method.

BACKGROUND: Recent artificial reproductive technologies (ART) have enabled even azoospermic subjects to take part in "in vitro fertilization" programs. The aim of the present study was: a) to evaluate the incidence of azoospermic male partners of infertile couples; b) to determine whether biochemical evaluation of such azoospermic males can help to discriminate between obstructive and secretory azoospermia. METHODS: Semen samples (n. 28,339; period 1990-1997) were analyzed according to WHO guidelines. In addition, in 108 azoospermic subjects, fructose, citric acid and L-carnitine (taken respectively as markers of seminal vesicle, prostate and epididymal function) were assayed using a new, faster technique, the differential pH method. RESULTS: The incidence of azoospermic semen samples was 4.76%. The biochemical analyses were indicative diagnostically, especially with regard to obstructive azoospermia. However, in secretory azoospermia, biochemical analysis of seminal plasma did not offer particularly helpful indications, except in cases of hypogonadism. CONCLUSIONS: Azoospermia accounts for a relatively high number of cases and justifies detailed attention to the diagnostic and therapeutic approach. Biochemical analysis of the seminal markers is a valid support for differential diagnosis between secretory obstructive azoospermia. It can therefore help in the correct recruitment of such patients for ART programs.

Semen analysis and seminal plasma biochemistry in two brothers with familial reciprocal translocation 46,XY,t(8;9)(p10;p10) transmitted by the father.

Here we report familial balanced translocation (46,XY,t(8;9)(p10;p10) inherited by two brothers from the father. The first subject, 33 years old, was examined after 3 years of childless marriage. Semen analysis showed severe oligozoospermia, hypomotility and teratozoospermia. His brother, 29 years old, volunteered to join this study. He had not yet tried to start a family. Semen analysis showed dyspermia, but the condition was much less serious than his brother's. In both subjects, the secretory function of the epididymis and the genital tract accessory glands did not seem to be affected by this particular translocation. This case report highlights the importance of cytogenetic analysis in dealing with infertile patients suffering from severe dyspermia apparently sine causa. This is particularly the case when certain Assisted Reproductive Technology (ART) programs are recommended, where natural selection of the sperm no longer takes place (e.g. ICSI). The possible risk of an increase in fetal chromosomal abnormalities would suggest genetic counselling in all such situations.

[Trial of a new spermatocidal agent in suppository form with local disinfectant action]. / Indagine su un nuovo spermicida in candelette ad azione disinfettante locale.

PIP: The in vivo spermicidal action of a pharmacological association consisting of 0.080 g of sodium tetraborate decahydrate, 0.075 g lactic acid, 0.020 g 8-hydroxyquinoline sulphate, 0.005 g sodiopropionate, and hydrosoluble excipient 1820 g polyethyl englycol in the form of vaginal containers, has been investigated. The study is part of a more general investigation of alternative systems to the use of estro-progestins. The experimental scheme applied in the present study provided for the use of this association in 28 volunteer couples who were definitely fertile and during the pre- and postovulatory periods. A normal postcoital test was carried out on each, with samples taken from the posterior fornix and cervical canal using a speculum. The results showed that the preparation possesses profound spermicidal activity and is therefore considered a useful alternative to oral contraceptives. To be absolutely safe, however, other contraceptive systems should be used simultaneously during ovulation (condom, diaphragm). (author's)^ieng

[Study of a drug combination for topical use, with contraceptive, antibacterial and antimycotic action and venereal disease preventive action. II. Optical microscopic and scanning microscopic studies of morphological changes of human spermatozoa in vitro after contact with the oxyquinoline sulfate and copper sulfate association]. / Studio su un'associazione farmacologica per uso topico ad azione anticoncezionale, antibatterica, antimicotica e profilattica per le malattie veneree. Nota 2. Alterazioni morfologiche al microscopio ottico e al microscopio a scansione degli spermatozoi umani in "vitro" dopo contatto con l'associazione ossichinolina solfato e solfato di rame.

PIP: A new spermicidal contraceptive agent consisting of an association of oxyquinoline sulfate and copper sulfate was tested in vitro to study its morphological effects on fresh sperm. Notable changes, such as a crumpling of the head, and variations in linear geometry of spermatozoa, were noted at the optic microscope. The scanning microscope discovered further erosions at the head and at the neck membranes level. It would be advisable to further study the association of these 2 drugs, to be eventually used as a strong topical spermicidal agent, and as a valid alternative to hormonal contraception.^ieng

[Intra-spermatic L-carnitine and survival of sperm motility]. / L-carnitina intraspermatica e survival della motilità nemaspermica.

BACKGROUND: The aim of this work was to evaluate the intrasperm carnitine (L-C) content related to sperm motility survival in bovine cervical mucus and in culture medium (Tyrode solution). METHODS: The following subjects were selected: 15 fertile normospermic subjects (according to WHO guidelines) and 31 male partners of infertile couples (semen profile: volume > 2.0 ml; concentration > 20 x 10(6)/ml, progressive motility > or = 25% (WHO categories "a" and "b") after fluidification, abnormal forms < 70%; round cells < 1.0 x 10(6)/ml). After standard semen analysis, the samples were subdivided into three aliquots in order to carry out: a) intrasperm L-C (free, total and acetylated) assay; b) sperm motility survival in bovine cervical mucus; c) sperm motility survival in culture medium. RESULTS: A strict correlation was found between L-C (total and acetylated) content and motility survival in cervical mucus. This is probably due to the fact that in cervical mucus lipids are an important energy source for sperm and to metabolize these lipids intrasperm L-C is essential. Therefore, L-C content can be considered as an indicator of sperm motility life-span in cervical mucus. A significant correlation, even if reduced compared with cervical mucus, was also observed between L-C (total and acetylated) content and sperm motility survival in the biological medium. This probably is because the L-C system modulates the reserves of free CoA, essential to the tricarboxylic acid cycle function. CONCLUSIONS: The intrasperm L-C deficit could be due to; a) alterations in the L-C uptake mechanisms in the epididimys due to inflammatory processes; b) lack of testosterone (L-C uptake is androgen-dependent). Therefore, the therapeutic implication of this finding is that where hypomotility is due to intrasperm L-C deficit, exogenous L-C administration or improvement of L-C epididymal testosterone-dependent uptake could promote the acquisition of sperm motility.

Improved human sperm recovery using superoxide dismutase and catalase supplementation in semen cryopreservation procedure.

The aim of this work was to evaluate the effects of ROS scavenger supplementation in human semen samples undergoing cryopreservation procedures.After screening out andrological pathologies, we selected 25 male partners of infertile couples with the following semen profile: volume >/= 2.0 ml, normal viscosity, sperm count >/=20 x 10(6)/ml, straight progressive motility (classes 1 and 2) >/= 40% (Mazzilli, Rossi, Delfino and Nofroni (1999) Andrologia 31: 187-194), atypical forms