Molecular study of the presence and transcriptional activity of HPV in semen.

PURPOSE: Human Papillomavirus (HPV) in semen represents a controversial topic. Recent evidence suggests a correlation with poor semen quality, but its detection is still unstandardized in this biological fluid. Thus, the aims of this study were to verify the ability of nested PCR to reveal HPV-DNA in semen; to evaluate association of seminal HPV with sperm parameters and risk factors for infection; to investigate the rate of HPV-DNA positivity in patients with and without risk factors; to assess HPV transcriptional activity. METHODS: We enrolled sexually active men and collected clinical and anamnestic data during andrological and sexually transmitted infections (STIs) evaluation. For each patient, we performed semen analysis and nested PCR to detect HPV-DNA in semen. In positive semen samples, we proceeded with genotyping and RNA quantification to detect HPV transcriptional activity. RESULTS: We enrolled 185 men (36.0 ± 8.3 years), of which 85 with (Group A) and 100 without HPV risk factors (Group B). Nested PCR was able to reveal HPV-DNA in semen, discovering a prevalence of 8.6% (11.8% in Group A and 6% in Group B, respectively). We observed no correlation between sperm quality and seminal HPV. Genital warts and previous anogenital infection were significantly associated with the risk of HPV positivity in semen. Moreover, no viral transcriptional activity was detected in positive semen samples. CONCLUSIONS: Our study suggests that searching for seminal HPV could be important in patients both with and without risk factors, especially in assisted reproduction where the risk of injecting sperm carrying HPV-DNA is possible.

Male reproductive health after 3 months from SARS-CoV-2 infection: a multicentric study.

PURPOSE: While SARS-CoV-2 infection appears not to be clinically evident in the testes, indirect inflammatory effects and fever may impair testicular function. To date, few long-term data of semen parameters impairment after recovery and comprehensive andrological evaluation of recovered patients has been published. The purpose of this study was to investigate whether SARS-CoV-2 infection affect male reproductive health. METHODS: Eighty patients were recruited three months after COVID-19 recovery. They performed physical examination, testicular ultrasound, semen analysis, sperm DNA integrity evaluation (TUNEL), anti-sperm antibodies (ASA) testing, sex hormone profile evaluation (Total testosterone, LH, FSH). In addition, all patients were administered International Index of Erectile Function questionnaire (IIEF-15). Sperm parameters were compared with two age-matched healthy pre-COVID-19 control groups of normozoospermic (CTR1) and primary infertile (CTR2) subjects. RESULTS: Median values of semen parameters from recovered SARS-CoV-2 subjects were within WHO 2010 fifth percentile. Mean percentage of sperm DNA fragmentation (%SDF) was 14.1 ± 7.0%. Gelatin Agglutination Test (GAT) was positive in 3.9% of blood serum samples, but no positive semen plasma sample was found. Only five subjects (6.2%) had total testosterone levels below the laboratory reference range. Mean bilateral testicular volume was 31.5 ± 9.6 ml. Erectile dysfunction was detected in 30% of subjects. CONCLUSION: Our data remark that COVID-19 does not seem to cause direct damage to the testicular function, while indirect damage appears to be transient. It is possible to counsel infertile couples to postpone the research of parenthood or ART procedures around three months after recovery from the infection.

Nasal cytology in children: scraping or swabbing?

Nasal cytology has become a valuable tool in the assessment of a multitude of nasal pathologies in children. Collection methods differ significantly and even though the use of the nasal curette is regarded as the most reliable in adults, most practitioners use the nasal swab in children. However, no studies have validated the reliability and supposed better tolerability of the latter. We have compared these two sampling methods regarding their tolerability and analysed the diagnostic accuracy of the cotton nasal swab (NSW) to identify nasal cytotypes and rhinitis phenotypes, using nasal scraping (NSC) for comparison. In a multicentric prospective study we recruited 208 children and performed nasal cytology by means of NSW and NSC. Microscopic evaluating of the nasal cytotypes was performed and tolerability of NSW and NSC was tested. Our data revealed a significantly inferior diagnostic accuracy of NSW compared to NSC regarding reliability and cell counts. Our study is the first to shed light on the role of the sampling tools for pediatric nasal cytology. We documented a poor diagnostic accuracy of NSW, thus suggesting using only the nasal curette in clinical practice. Furthermore, tolerability did not differ between the two, refuting the common thesis that swabs are to be preferred when doing nasal cytology in children.

Dalbavancin for the treatment of paediatric infectious diseases.

To review the topics of interest related to the use of dalbavancin in paediatric patients. PubMed was used to search for all of the studies published over the last 15 years using the key word "dalbavancin". A total of 36 manuscripts were selected, and due to the limited pediatric experience a further research was performed in order to identify clinical trials ongoing. Three studies that concerned children were found in clinicaltrials.gov. This review considers also the manuscripts published on the adult population in order to highlight the gaps requiring further research at pediatric age. Dalbavancin has emerged as a promising agent against resistant Gram-positive invasive infections. It is approved in the United States and Europe for the treatment of adult patients with acute bacterial skin and skin structure infections (SSTIs). Compared to other available antibiotics that are active against multi-resistant bacteria, the advantages of dalbavancin include a lower potential for drug interactions and the possibility of fewer required doses due to a longer half-life. Pharmacokinetic characteristics of dalbavacin are attractive for its clinical impact, especially for children who may avoid prolonged hospitalization and central venous access. However, further studies are needed to establish its appropriate paediatric dosage before it can be licensed for use in newborns and children. For younger patients, at a time when infections due to multidrug-resistant Gram-positive pathogens are increasing, dosage, efficacy and safety data for dalbavancin are needed to ensure the highest antimicrobial efficacy while also minimizing the risk of adverse events.

Pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections: an overview.

The acronym PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections) has been used to describe a syndrome characterized by various obsessions, compulsions, tics, hyperactivity, motor stereotypies, and paroxysmal movement disorders that are correlated with prior infection by group A beta-hemolytic Streptococcus pyogenes (GABHS) infections. Five clinical criteria can be used to diagnose PANDAS: (1) the presence of obsessive-compulsive disorder (OCD) and/or any other tic disorders; (2) prepuberal onset (between 3 years of age and the start of puberty); (3) abrupt onset and relapsing-remitting symptom course; (4) a distinct association with GABHS infection; and (5) association with neurological abnormalities during exacerbations (adventitious movements or motoric hyperactivity). The exact pathogenesis of PANDAS remains unclear, and several theories that focus on multiple etiologic or contributive factors have emerged. PANDAS appears to be a neurobiological disorder that potentially complicates GABHS infections in genetically susceptible individuals. The current standard of care for PANDAS patients remains symptomatic, and cognitive behavioral therapy, such as exposure and response prevention, combined with family counseling and psychoeducation, should be the first approach for treating PANDAS. This review examines current theories of PANDAS pathogenesis, identifies possible treatments for managing this complex condition, and highlights areas for future research. Moving forward, developing more standardized diagnostic criteria and identifying specific laboratory markers to facilitate PANDAS diagnoses are crucial.

Testosterone responses to standardized short-term sub-maximal and maximal endurance exercises: issues on the dynamic adaptive role of the hypothalamic-pituitary-testicular axis.

BACKGROUND: Few and conflicting data on the acute adaptive role of the hypothalamic-pituitary-testicular (HPT) axis to sub-maximal endurance exercise exist. AIMS: To investigate the acute HPT axis responses to standardized endurance exercises in a laboratory setting and the correlations between testosterone and classic adaptive hormones variations. SUBJECTS AND METHODS: 12 healthy male volunteers were recruited for this experimental study. Serum PRL, GH, ACTH, LH, cortisol, DHEAS, testosterone [total (TT), calculated free (cFT) and bioavailable (cBioT)], SHBG, and respective ratios, were evaluated before and after a 30-min sub-maximal exercise on cycle ergometer at individual anaerobic threshold (IAT) and a maximal exercise until exhaustion. Blood samples were collected before exercise (30, 15 min and immediately before), immediately after and at different time points during recovery (+15, +30 and +60 min) for hormones assays. Oxygen consumption and lactate concentration were evaluated. RESULTS: Testosterone (TT, cFT and cBioT) acutely increased in all volunteers after both exercises. Testosterone increased in parallel to GH after both exercises and to cortisol only after maximal exercise. Differently from other increased hormones, testosterone increases were not correlated to exercise-intensity-related variables. The anabolic/catabolic steroids ratios were higher after sub-maximal exercise, compared to maximal. CONCLUSIONS: A 30-min sub-maximal endurance exercise acutely increased serum testosterone similarly to maximal exercise, but without cortisol increases. Exercise-related testosterone peaks should be considered adaptive phenomena, but few data on their short- and long-term effects exist. Investigations on the mechanisms of adaptation to exercise in active individuals with physiological or pathological hypo-testosteronemia are warranted.

No evidence for the effectiveness of systemic corticosteroids in acute pharyngitis, community-acquired pneumonia and acute otitis media.

Corticosteroids have been used to treat infectious diseases for more than 50 years but, although it has been shown that they are highly effective in improving the clinical course of some diseases, their effects have not been clearly defined in others. Nevertheless, they are still used by a considerable number of physicians. This review analyses the role of systemic corticosteroids in the treatment of acute pharyngitis (AP), community-acquired pneumonia (CAP) and acute otitis media (AOM). A number of trials involving patients with AP have been carried out, but most are marred by methodological flaws that do not allow any firm conclusions to be drawn. The number of trials involving CAP patients is even higher, and the data suggest that corticosteroids may reduce the risk of death only in patients with severe disease. There are very few data concerning AOM, and there is currently no reason for prescribing corticosteroids to treat it. Overall, the data showed that there is, currently, no indication for the universal use of systemic corticosteroids in any of the reviewed diseases and, further, high-quality studies of all of these respiratory tract infections are needed in order to identify the patients for whom the prescription of corticosteroids is rationally acceptable.

A retrospective evaluation of microbiology of acute otitis media complicated by spontaneous otorrhea in children living in Milan, Italy.

PURPOSE: To evaluate the microbiology of acute otitis media (AOM) with otorrhea due to spontaneous tympanic membrane perforation (STMP) in children living in Milan, Italy. METHODS: We evaluated middle ear fluid (MEF) specimens taken from children affected by AOM associated with STMP and otorrhea between January 2001 and December 2011. The fluid was collected by means of direct swab sampling, sent for culture, and processed within 4 h. RESULTS: A total of 705 specimens were obtained from 458 children (233 boys; mean age ± SD 28.3 ± 19.9 months), and were positive for bacteria in 487 cases (69.1 %). The most frequently cultured infectious agent in single-pathogen cultures was Haemophilus. influenzae (51.0 %), followed by Streptococcus pneumoniae (19.4 %), Streptococcus pyogenes (17.4 %), and Staphylococcus aureus (10.7 %). After adjusting for the sub-period of data collection, age, gender, and previous full heptavalent pneumococcal conjugate vaccine (PCV-7) schedule, it was found that the prevalence of H. influenzae slightly increased in 2008-2010, and the prevalence of S. pneumoniae significantly decreased over time (p = 0.02). CONCLUSIONS: AOM with STMP is a particular form of AOM in which S. pyogenes plays a significant causative role although, as in uncomplicated cases, H. influenzae and S. pneumoniae retain their etiological importance. The frequency of the detection of S. aureus in MEF deserves further study because this pathogen can give rise to severe clinical problems. Finally, although the use of PCV-7 was relatively efficacious, the benefit of pneumococcal vaccination would be increased by vaccines including a larger number of serotypes.

Role of vitamin D in children with respiratory tract infection.

It has recently been shown that vitamin D (VitD) plays an important role in host defences, inflammation and immunity. We reviewed PubMed and selected all of the studies published over the last 15 years concerning VitD deficiency and VitD supplementation in children with respiratory tract infections. Our analysis showed that VitD seems to be very important because of its part in the complexity of the immune system. However, there are few pediatric studies and most have various limitations. First of all, the literature mainly refers to studies concerning the prevalence of VitD insufficiency and deficiency in specific pathologies. Secondly, it is extremely difficult to identify a common specific range of normal, insufficient and deficient VitD levels. Thirdly, the available studies of VitD supplementation often combined VitD with the use of other micronutrients, thus obscuring the role of VitD itself. Finally, different doses have been used for VitD supplementation. These observations clearly highlight the fact that further studies are needed to evaluate the impact of VitD deficiency and insufficiency in terms of the epidemiology and outcomes of pediatric respiratory tract infection, and whether VitD supplementation favours a positive outcome.

Hypertonic saline is more effective than normal saline in seasonal allergic rhinitis in children.

Allergic rhinitis (AR) is a very common childhood disease that is associated with a significant reduction in the patients' quality of life. Its treatment combines educating the patients and their parents, immunotherapy and drug administration. However, even the best approach does not relieve the symptoms of a number of patients. Alternative therapies are particularly needed for children because the fear of adverse events frequently reduces parental compliance to the prescribed drugs, and immunotherapy is less easy to administer than in adults. In this prospective investigator-blinded study we evaluated whether children, with a documented history of seasonal grass pollen-related AR, benefit from nasal irrigation by assessing the effects on nasal signs and symptoms, on middle ear effusion and on adenoidal hypertrophy. We randomized children aged 5 to 9 years (median age 82 months) to normal saline or hypertonic saline (a 2.7% sodium chloride solution), administered twice-daily using a disposable 20 ml syringe, or no treatment. Nasal symptoms (rhinorrhea, itching, sneezing, nasal obstruction), swelling of turbinates, adenoid hypertrophy or middle ear effusion were assessed at baseline and after 4 weeks of treatment. Two hundred and twenty children (normal saline: 80; hypertonic saline: 80; no treatment: 60) completed the study. After four weeks, all the considered items were significantly reduced in the group receiving hypertonic saline (P < 0.0001), whereas in the group receiving normal saline only rhinorrhea (P = 0.0002) and sneezing (P = 0.002) were significantly reduced. There was no significant change in any of the items in the control group. The duration of oral antihistamines was significantly lower in the children receiving hypertonic saline than in those treated with normal saline or in controls. No adverse events were reported and parental satisfaction and compliance with the procedure were globally very good, regardless of the solution used. Using our procedure, hypertonic saline is effective, inexpensive, safe, well tolerated and easily accepted by children with seasonal grass pollen-related AR and their parents. Our data suggest that nasal irrigation with hypertonic saline might be included in the wide spectrum of therapies recommended for grass-pollen AR.

Use of complementary and alternative medicine in children with recurrent acute otitis media in Italy.

Controlling environmental factors, chemoprophylaxis, immunoprophylaxis and surgery are considered possible means of preventing recurrent acute otitis media (RAOM), but there are no available data concerning the paediatric use of complementary and alternative medicine (CAM). We evaluated the uses of CAM (homeopathy and/or herbal medicine) as means of preventing AOM in children with a history of RAOM. Eight hundred and forty Italian children with RAOM (≥3 episodes in six months) aged 1-7 years were surveyed in 2009 using a face-to-face questionnaire, filled by parents or caregivers, that explored the prevalence, determinants, reasons, cost, and perceived safety and efficacy of CAM. About one-half (46%) of the children used CAM, significantly more than the number who used immunoprophylaxis (influenza vaccine 15%; p<0.05), PCV-7 34%; p<0.05) or chemoprophylaxis (2%; p<0.001). Use of CAM in the family was the only important factor positively associated with the use of CAM in children (adjusted OR 7.94; 95% CI: 5.26-11.99). The main reasons for using CAM were a fear of the adverse effects of conventional medicine (40%) and to increase host defences (20%). CAM was widely seen as safe (95%) and highly effective (68%). CAM prescribers were paediatricians in 50.7% of cases; self-initiation was reported by 23% of respondents. CAM expenditure was between Euro 25 and Euro 50/month in 27.6% of cases and ≥ Euro 50/month in 16%. Children with RAOM should be considered among the categories of subjects likely to be using CAM. Together with the fact that paediatricians are the main prescribers, this is worrying because of the current lack of evidence regarding the efficacy, safety and cost-effectiveness of CAM in the prevention of RAOM.

Insights into infectious otitis media.

Otitis media is a multifactorial disease. It represents a major public health burden, as it is the most common diagnosis in children and is characterized by both immediate and long-term morbidity. For years the knowledge about risk factors and microbiology of otitis media has been relatively stable. In recent years new data have been gathered regarding different microbiological aspects of otitis media. This new knowledge may contribute toward the development and implementation of therapeutic and preventive strategies.

Effectiveness of a propolis and zinc solution in preventing acute otitis media in children with a history of recurrent acute otitis media.

Recurrent acute otitis media (rAOM) is frequently encountered in infants and children and the lack of any definitive treatment has led parents and physicians to try complementary and alternative therapies. We evaluated the efficacy of a propolis and zinc suspension in preventing AOM in 122 children aged 1-5 years with a documented history of rAOM, who were prospectively, blindly, randomized 1:1 to receive the suspension plus elimination of environmental risk factors or elimination of environmental risk factors only. AOM- and respiratory-related morbidity were assessed at study entry and every four weeks. In the 3-month treatment period AOM was diagnosed in 31 (50.8%) children given the propolis and zinc suspension and in 43 (70.5%) controls (p=0.04). The mean number of episodes of AOM per child/month was 0.23+/-0.26 in the propolis and zinc group and 0.34+/-0.29 in controls (reduction 32.0%, p=0.03). The administration of a propolis and zinc suspension to children with a history of rAOM can significantly reduce the risk of new AOM episodes and AOM-related antibiotic courses, with no problem of safety or tolerability, and with a very good degree of parental satisfaction. No effect can be expected on respiratory infections other than AOM.

Urinary and serum hormones profiles after testosterone enanthate administration in male hypogonadism: concerns on the detection of doping with testosterone in treated hypogonadal athletes.

OBJECTIVE: To describe serum and urinary hormones, androgens metabolites and testosterone/epitestosterone ratio profiles after testosterone administration in male hypogonadal volunteers, and to evaluate their possible usefulness in detecting doping with testosterone in treated hypogonadal athletes. DESIGN: Controlled open label design vs placebo; pharmacokinetic study. PARTICIPANTS: Ten male volunteers affected by severe hypogonadism (serum testosterone <2.31 ng/ml). INTERVENTIONS AND MAIN OUTCOME MEASURES: Serum and urinary parameters were evaluated, by radioimmunoassay and gas chromatography-mass spectrometry, before and at different time points for 7/3 weeks after a single administration of testosterone enanthate (250 mg) or placebo, respectively. RESULTS: As partially known, testosterone administration increased, with great individual variability, urinary concentrations of glucuronide testosterone, androsterone, etiocholanolone, 5alpha-androstane- 3alpha,17beta-diol, 5beta-androstane-3alpha,17beta-diol, testosterone/ epitestosterone and testosterone/LH ratios; and decreased epitestosterone and 5alpha-androstane-3beta,17beta-diol/5beta-androstane- 3alpha,17beta-diol ratio. Serum testosterone and dihydrotestosterone increased in all volunteers, and concentrations higher than the upper reference limits were observed in many volunteers until 2 weeks after testosterone administration. CONCLUSION: Whereas the observed prolonged hyperandrogenism partially limited data interpretation, the report ed characteristics of variation of urinary parameters might be used to suspect testosterone misuse in hypogonadal athletes treated with testosterone enanthate. In this sense, while the actual threshold for tes tos terone/epites tos ter one ratio was confirmed to be of reduced usefulness, we suggest a contemporary evaluation of whole urinary androgen metabolites profile and serum androgens, at specific time points after testosterone enanthate administration. Moreover, an adequate tailoring of treatment, to avoid transitory hyperandrogenism, is highly advisable. Further studies on strategies for detecting doping with testosterone in hypogonadal athletes are warranted.

The plant ribosome inactivating protein saporin induces micronucleus formation in peripheral human lymphocytes in vitro.

Saporin belongs to the family of plant enzymes known as ribosome inactivating proteins (RIPs) for their property to depurinate the major rRNA, thus leading to inactivation of ribosomes. In this work we analyzed the genotoxic effects of saporin, purified from root cultures of Saponaria officinalis, by evaluating micronucleus formation and by the quantitative determination of cytosolic histone-associated DNA fragments. Saporin induces micronuclei formation in cultured human lymphocytes in a dose dependent manner; treated lymphocytes show a decrease in cell viability and a concomitant increase in the apoptotic response evidenced by the appearance of cytosolic oligonucleosomes. On the other hand saporin treatment failed to induce sister-chromatid exchange (SCE) at any of the doses tested.

Inhibition of L-tyrosine-induced micronuclei production by phenylthiourea in human melanoma cells.

It was previously found that L-tyrosine oxidation product(s) are cytotoxic, genotoxic and increase the sister chromatid exchange (SCE) levels in human melanoma cells. In this work, the micronucleus assay has been performed on human melanotic and amelanotic melanoma cell lines (Carl-1 MEL and AMEL) in the presence of 1.0, 0.5 and 0.1 mM L-tyrosine concentrations to investigate if melanin synthesis intermediate(s) increase micronuclei production. L-Tyrosine oxidation product(s) increased the frequency of micronuclei in melanoma cells; 0.1 mM phenylthiourea (PTU), an inhibitor of L-tyrosine oxidation by tyrosinase, lowered the micronucleus production to the control levels. The culture of melanoma cells with high L-tyrosine in the culture medium resulted in a positive response to an ELISA-based apoptotic test. For comparison the effect of L-tyrosine on micronuclei production in human amelanotic melanoma cells was also investigated; the micronucleus production in the presence of 1 mM L-tyrosine in the culture medium was lower than that found with melanotic melanoma cells of the same cell line. The data suggest that melanin synthesis intermediates arising from L-tyrosine oxidation may cause micronuclei production in Carl-1 human melanoma cells; the addition of PTU in the presence of L-tyrosine decreased the frequency of micronuclei to about the control values thus the inhibition of melanogenesis may have some clinical implication in melanotic melanoma.

Biochemical diagnosis of gastroenteropancreatic endocrine tumors.

Functioning gastroenteropancreatic endocrine tumors produce and secrete different substances that can be detected in the plasma and cause hormone-related syndromes. Symptoms such as diarrhea associated either with typical skin rash or peptic ulcer disease may be suggestive of the presence of intestinal carcinoid or gastrinoma. Other clinical manifestations such as severe hypoglycemia, diabetes, necrolytic erythema and gallbladder disease may also indicate an endocrine tumor. Sometimes, patients present no, or just vague, symptoms such as dyspepsia or abdominal pain and nonfunctioning endocrine tumors in these patients can be found incidentally during diagnostic imaging procedures or at operation. Usually, the diagnosis is established by the measurement of the specific tumor marker in the plasma and, sometimes, in the urine. In some cases, normal basal hormone levels are observed even in the presence of typical symptoms. Therefore, stimulatory tests such as the secretin test for gastrinomas are required to establish the diagnosis. General markers for the diagnosis of gastroenteropancreatic endocrine tumors are also available. Among these, chromogranin A has proved to be of great value for diagnosing nonfunctioning tumors and is considered the most sensitive general marker. The availability of both specific and general markers as well as stimulatory tests may enable the clinician to diagnose functioning gastroenteropancreatic endocrine tumors at an early stage and to recognize nonfunctioning tumors.

Effect of simulated intraoperative heating and shaping on mechanical properties of a bioabsorbable fracture plate material.

Heating bioabsorbable fracture fixation plates to above their glass transition temperature renders them temporarily malleable, thus facilitating their adaptation to the underlying bone geometry, although the consequence of heating is not well understood. Poly (L-lactide-co-glycolide) copolymer specimens were heated under various conditions, and the effects on specimen mechanics were assessed. Heating temporarily increased toughness while slight reducing flexural modulus. No lasting effects on in vitro material degradation were seen.

Onychomycosis caused by Blastoschizomyces capitatus.

Blastoschizomyces capitatus was cultured from the nail of a healthy patient with onychomycosis. The identity of the isolate was initially established by standard methods and ultrastructural analysis and was verified by molecular probing. Strains ATCC 200929, ATCC 62963, and ATCC 62964 served as reference strains for these analyses. To our knowledge, this is the first case of nail infection secondary to paronychia caused by this organism reported in the English literature.