Preclinical study for treatment of hypospadias by advanced therapy medicinal products.

PURPOSE: This paper explores the feasibility of a new therapy for the treatment of hypospadias patients. Hypospadias is a very common congenital malformation of male genitals, with very high rate of recurrences after surgery. The field of regenerative medicine, which offers innovative solutions for many pathologies, still does not offer reliable solution for this pathology. Here, we propose quality, safety, and clinical feasibility assessment for an oral mucosa advanced therapy medicinal product (ATMP) grown on a biocompatible scaffold for a clinical study on urethral reconstruction of hypospadias patients. METHODS: Urethral and oral mucosal epithelia from donor biopsies were cultivated between two fibrin layers, under clinical-grade conditions for cell and tissue characterization and comparison, aimed at tissue engineering. In addition, single-clone analyses were performed to analyze gene expression profiles of the two epithelia by microarray technology. RESULTS: Oral mucosa appeared suitable for urethral reconstruction. The resulting ATMP was proven to maintain stem cells and regenerative potency. The preclinical safety studies were performed on human tissues to assess abnormalities and tumorigenicity, and confirmed the safety of the ATMP. Finally, the patient selection and the clinical protocol for the upcoming clinical trial were defined. CONCLUSIONS: Against this backdrop, in this paper, we are proposing a new reproducible and reliable ATMP for the treatment of hypospadias.

Effect of 6% hydroxyethyl starch 130/0.4 on kidney and haemostatic function in cardiac surgical patients: a randomised controlled trial.

Whether third-generation hydroxyethyl starch solutions provoke kidney injury or haemostatic abnormalities in patients having cardiac surgery remains unclear. We tested the hypotheses that intra-operative administration of a third-generation starch does not worsen postoperative kidney function or haemostasis in cardiac surgical patients compared with human albumin 5%. This triple-blind, non-inferiority, clinical trial randomly allocated patients aged 40-85 who underwent elective aortic valve replacement, with or without coronary artery bypass grafting, to plasma volume replacement with 6% starch 130/0.4 vs. 5% human albumin. Our primary outcome was postoperative urinary neutrophil gelatinase-associated lipocalin concentrations, a sensitive and early marker of postoperative kidney injury. Secondarily, we evaluated urinary interleukin-18; acute kidney injury using creatinine RIFLE criteria, coagulation measures, platelet count and function. Non-inferiority (delta 15%) was assessed with correction for multiple comparisons. We enrolled 141 patients (69 starch, 72 albumin) as planned. Results of the primary analysis demonstrated that postoperative urine neutrophil gelatinase-associated lipocalin (median (IQR [range])) was slightly lower with hydroxyethyl starch (5 (1-68 [0-996]) ng.ml-1 ) vs. albumin (5 (2-74 [0-1604]) ng.ml-1 ), although not non-inferior [ratio of geometric means (95%CI) 0.91 (0.57, 1.44); p = 0.15] due to higher than expected variability. Urine interleukin-18 concentrations were reduced, but interleukin-18 and kidney injury were again not non-inferior. Of 11 individual coagulation measures, platelet count and function, nine were non-inferior to albumin. Two remaining measures, thromboelastographic R value and arachidonic acid-induced platelet aggregation, were clinically similar but with wide confidence intervals. Starch administration during cardiac surgery produced similar observed effects on postoperative kidney function, coagulation, platelet count and platelet function compared with albumin, though greater than expected variability and wide confidence intervals precluded the conclusion of non-inferiority. Long-term mortality and kidney function appeared similar between starch and albumin.

Impact of erythrocyte species on assays for influenza serology.

The influenza viruses have the ability to agglutinate erythrocytes by binding to sialic acid receptors on the host cell. Human influenza viruses preferentially bind to sialic acid linked to galactose by α 2.6 linkage, while avian influenza viruses preferentially bind to sialic acid linked to Gal by α 2.3 linkage. There is a close correlation between the ability of influenza A viruses to agglutinate erythrocytes from different animal species and their receptor specificity. The haemagglutination and haemagglutination inhibition assays are influenced by the species of erythrocytes. To provide an overview of the expression of sialic acid receptors on different erythrocytes, avian (turkey, chicken, pigeon) and mammalian (sheep, horse, human) species have been analysed by flow cytometry. Chicken, turkey and human erythrocytes display both types of linkages. Horse and sheep erythrocytes show almost exclusively α 2.3 Gal linkages, while pigeon erythrocytes express almost exclusively α 2.6 Gal linkages. The erythrocytes from the same avian and mammalian species have been evaluated by haemagglutination and haemagglutination inhibition assays with seasonal and avian strains. Chicken and turkey erythrocytes seem to be the most appropriate for both assays with seasonal influenza strains, in addition to pigeon erythrocytes, particularly for the B strains. In the case of the avian strain, chicken erythrocytes are suitable for haemagglutination assay and horse erythrocytes for haemagglutination inhibition assay. The choice of erythrocytes has a significant impact on the titres measured by both assays.

"OKKIO alla salute 2014" results from the Tuscan sample.

BACKGROUND: The prevalence of overweight and obesity among children is rapidly increasing in many countries. For this reason we decided to investigate the eating habits of 8-9 years old Tuscany children, by paying attention to the frequency of their meals per day and to their food choices, also in relation to children's Body Mass Index (BMI) classes. In addition, we considered some environmental factors that could affect children eating behaviors, such as parents' BMI. METHODS: Data from the 2014 Tuscany nutritional surveillance system (part of the national periodical survey) and collected from children, parents and teachers, have been linked to study of children's eating habits. Logistic regression analyses were used to investigate the association between incorrect dietary habits and the potential predictors of overweight and obesity. RESULTS: More than half of the 2,163 children (68.64% overall, 67.61% females, 69.56% males) reported to have eaten an appropriate breakfast in the morning, while 5.9% skipped breakfast and only 33.5% of children ate an appropriate mid-morning snack. Logistic regression showed that having breakfast and especially an adequate breakfast are protective factors against obesity (OR 0.57, p<0.01; OR 0.84; p=0.03 respectively). CONCLUSION: In Tuscany Region, unhealthy dietary habits are common among children. The identified deficiencies may be a harbinger of future public health problems.

Epidemiology of herpes simplex virus type 1 and 2 in Italy: a seroprevalence study from 2000 to 2014.

Herpes simplex viruses (HSV) are among the most widespread causative agents of human viral infections. HSV-2 is one of the commonest causes of genital disease, while HSV-1 is associated primarily with orolabial ulceration; however, recent changes in HSV epidemiology showed an increase in genital and neonatal herpes particularly caused by HSV-1. The main purpose of this study was to assess the seroprevalence of HSV-1 and HSV-2 in a random population in Siena (central Italy) in 2000, 2005 and 2013-2014 and in Bari (southern Italy) in 2005. Moreover, a preliminary study was conducted to investigate the spread of HSV infection in a population of pregnant women and infants in Bari in 2003, 2004 and 2005. Human serum samples were tested for the presence of specific anti-HSV-1 and anti-HSV-2 IgG antibodies using a commercially available ELISA test. For the primary purpose, seroprevalence rates observed in Siena were compared over the years sampled and with the seroprevalence rate found in Bari. Results of seroprevalence in Siena show a decreased trend for both viruses, especially in adolescents and young adults; moreover, HSV-2 seroprevalence rates found in the two cities suggest geographical differences. For the secondary purpose, prevalence rates among pregnant women were compared with the seroprevalence found in women of the general population. No significant difference in prevalence rates were found among pregnant women, while results indicate both viruses are a source of infection in infants.

Water and air ozone treatment as an alternative sanitizing technology.

AIMS: We investigated the effectiveness of ozone (aqueous and gaseous) treatment as an alternative sanitizing technology to common conventional disinfectants in reducing the microbial contamination of both water and air. METHODS: Ozone was added for 20 minutes to a well-defined volume of water and air by the system named "Ozonomatic®". The effectiveness of ozonation was determined by counting CFU/ m3 or ml of bacteria present in samples of air or water collected before (T0) and after (T1) the addition of ozone and comparing the microbial load of different bacteria present in ozonized and nonozonized samples. RESULTS: When the ozonisation equipment was located at 30 cm from the surface of the water in the bath tub in which the bacteria investigated were inoculated, the treatment was able to reduce the total microbial load present in the aerosol by 70.4% at a temperature of 36°C for 48 hours. Conversely, at 22°C for 5 days, only a modest decrease (9.1%) was observed. Escherichia coli and Pseudomonas aeruginosa were completely eliminated. A 93.9% reduction was observed for Staphylococcus aureus, followed by Streptococcus faecalis (25.9%). The addition of ozone to water was able to almost eliminate Staphylococcus aureus (98.9% reduction) and also to exert a strong impact on Legionella pneumophila (87.5% reduction). Streptococcus faecalis and Pseudomonas aeruginosa showed a decrease of 64.2% and 57.4%, respectively. Conversely, only a 26.4% reduction was observed for the bacterium Escherichia coli. This study showed that the addition of ozone in the air exerted a modest reduction on microbial load at 36°C, whereas no effect was observed at 22°C. CONCLUSIONS: Aqueous and gaseous ozone treatments were effective against microbial contaminants, reducing the CFU of the microorganisms studied. These results confirm the efficacy of the ozone disinfection treatment of both water and air; particularly, it constitutes an extremely promising alternative, allowing the possibility to reuse contaminated water.

Cross-sex hormone therapy for gender dysphoria.

PURPOSE: Gender identity is the sense one has of being male or female. Gender dysphoria (GD) refers to the distress caused by the incongruence between gender identity and biological sex in gender-nonconforming individuals. Cross-sex hormone therapy (CHT) aims at easing GD, improving well-being, and quality of life of gender-nonconforming individuals. This can be achieved by inducing and maintaining the desired-sex characteristics in accordance with the specific aspirations and expectations of each individual. Nevertheless, CHT can be associated with potentially serious long-term complications. METHODS: Here, we review when, how, and how long to prescribe CHT to adult transsexuals as well as what to expect and monitor once it has been initiated. RESULTS: In recent years, transsexualism has become more and more recognized and depathologized. To manage GD, National and International Standards of Care have been established. Nevertheless, the needs of transgender patients can still be ignored or dismissed. Moreover, some questions remain unanswered because of the lack of specific retrospective or prospective studies on CHT. CONCLUSION: Education and culturally sensitive training must be supplied to healthcare professionals to overcome the existing issues on GD management and change the perspectives of transsexual people.

Sexual outcomes in men who have sex with men who underwent radical prostatectomy.

BACKGROUND: Sexual difficulties are a recognized consequence of prostate cancer (PCa) treatments. An estimated one in three men who have sex with men (MSM) receive PCa a diagnosis during their lifetime. MSM may experience all types of sexual dysfunction as reported in men who have sex with women (MSW), along with a number of more specific bothersome problems. This systematic literature review aims to evaluate sexual outcomes in MSM who have undergone radical prostatectomy (RP). METHODS: A systematic review was carried out following the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The searches were made using relevant keywords in the PubMed, Scopus, and Web of Science databases, thus including the whole literature from January 2000 to November 2023. Studies which did not allow to retrieve data on sexual outcomes on MSM treated with RP for PCa were excluded. Data on sexual outcomes and health-related quality of life (HRQoL) were retrieved, mostly including changes in libido, erectile function, ejaculatory disorders, orgasm, climacturia, changes in role-in-sex identity, changes in sexual partnerships, and the presence of painful receptive anal intercourses (AI). PROSPERO ID: CRD42024502592. RESULTS: Six articles met the inclusion criteria. In total, data of 260 patients were analyzed. Three main themes emerged: (a) MSM may experience specific sexual dysfunctions due to the different dynamics of their intimacy; (b) the lack of tool validated on gay and bisexual population to assess sexual outcomes (c) the need for a tailored approach that also takes into account sexual orientation throughout the oncological journey. CONCLUSIONS: MSM undergoing RP may experience similar sexual problems as MSW. Painful AI should be considered a potential post-operative adverse outcome in MSM. Future studies should prioritize validating a questionnaire that explores AI. Healthcare providers should adopt a tailored approach that takes into account sexual orientation throughout the cancer journey.

Effects of Influenza Vaccine on the Immune Responses to SARS-CoV-2 Vaccination.

A number of studies have suggested that influenza vaccination can provide protection against COVID-19, but the underlying mechanisms that could explain this association are still unclear. In this study, the effect of the 2021/2022 seasonal influenza vaccination on the immune response to the booster dose of anti-SARS-CoV-2 vaccination was evaluated in a cohort of healthy individuals. A total of 113 participants were enrolled, 74 of whom had no prior COVID-19 diagnosis or significant comorbidities were considered for the analysis. Participants received the anti-influenza tetravalent vaccine and the booster dose of the anti-SARS-CoV-2 vaccine or the anti-SARS-CoV-2 vaccine alone. Blood was collected before and 4 weeks after each vaccination and 12 weeks after SARS-CoV-2 vaccination and analyzed for anti-flu and anti-spike-specific antibody titers and for in vitro influenza and SARS-CoV-2 neutralization capacity. Results indicated an increased reactivity in subjects who received both influenza and SARS-CoV-2 vaccinations compared to those who received only the SARS-CoV-2 vaccine, with sustained anti-spike antibody titers up to 12 weeks post-vaccination. Immune response to the influenza vaccine was evaluated, and individuals were stratified as high or low responders. High responders showed increased antibody titers against the SARS-CoV-2 vaccine both after 4 and 12 weeks post-vaccination. Conversely, individuals classified as low responders were less responsive to the SARS-CoV-2 vaccine. These data indicate that both external stimuli, such as influenza vaccination, and the host's intrinsic ability to respond to stimuli play a role in the response to the vaccine.

Role of US in acute scrotal pain.

BACKGROUND: The acute scrotum is a common emergency department (ED) presentation and can be defined as any condition of the scrotum or intrascrotal contents requiring emergent medical or surgical intervention. Although rarely fatal, acute scrotal pathology can result in testicle infarction and necrosis, testicular atrophy, infertility, and significant morbidity. METHODS: Scrotal US is best performed with a linear 7.5- to 12-MHz transducer. In addition to imaging in the longitudinal and transverse planes, it is helpful to obtain simultaneous images of both testes for comparison. Color Doppler is used to evaluate for abnormalities of flow and to differentiate vascular from nonvascular lesions. Attention to appropriate color Doppler settings to optimize detection of slow flow is critical. RESULTS: The evaluation of acute scrotal pain can be challenging for the clinician initially examining and triaging the patient. Acute scrotal conditions due to traumatic, infectious, vascular, or neoplastic etiologies can all present with pain as the initial complaint. Additionally, the laboratory and physical examination findings in such conditions may overlap; this, coupled with potential patient guarding and lack of collaboration, may result in a limited, non-specific physical examination. Therefore, scrotal ultrasound has emerged to play a central role in the evaluation of the patient presenting with acute scrotal pain. CONCLUSIONS: In conclusion, we are firmly convinced that a scrotal ultrasound should always be performed in the presence of acute scrotal pain. Moreover, urologist should be able to perform a scrotal ultrasound but, if imaging does not supply a clear diagnosis, surgical exploration is still mandatory.

Retrospective Study Evaluating Seroprevalence of Hepatitis E Virus in Blood Donors and in Swine Veterinarians in Italy (2004).

Hepatitis E is an emerging viral disease in developed countries, with sporadic cases occasionally linked to the consumption of raw or undercooked pork, wild boar or deer meat. Cases due to transfusion or transplantation have also been reported. In developed countries, hepatitis E is considered a zoonosis and pig is the main reservoir. In the last few years, several studies conducted in Europe reported variable seroprevalence rates among the general population, ranging between 0.26% and 52.5%. A higher seroprevalence was described among workers who come in contact with pigs. The aim of this retrospective study was to evaluate the seroprevalence of anti-HEV IgG and IgM antibodies in blood donors (170) and in pig veterinarians (83). Archival sera were collected in Italy in 2004. The observed seroprevalence was 9.64% and 8.82% in veterinarians and blood donors, respectively. Overall, only three sera from blood donors were positive for IgM, but no HEV-RNA was detected.

An observational retrospective evaluation of 79 young men with long-term adverse effects after use of finasteride against androgenetic alopecia.

Concern regarding adverse effects of finasteride is increasing. We aimed to determine the type and frequency of symptoms in men having long-term sexual and non-sexual side effects after finasteride treatment (a condition recently called post-finasteride syndrome, PFS) against androgenetic alopecia (AGA). Subjects were recruited at the Urology Unit of the Trieste University-Hospital, and from a dedicated website. Out of 79 participants, 34% were white Italians, mean age was 33.4 ± 7.60 years, mean duration of finasteride use was 27.3 ± 33.21 months; mean time from finasteride discontinuation was 44.1 ± 34.20 months. Symptoms were investigated by an ad hoc 100 questions' questionnaire, and by validated Arizona Sexual Experience Scale (ASEX) and Aging Male Symptom Scale (AMS) questionnaires. By ASEX questionnaire, 40.5% of participants declared getting and keeping erection very difficult, and 3.8% never achieved; reaching orgasm was declared very difficult by 16.5%, and never achieved by 2.5%. By the ad hoc questionnaire, the most frequent sexual symptoms referred were loss of penis sensitivity (87.3%), decreased ejaculatory force (82.3%), and low penile temperature (78.5%). The most frequent non-sexual symptoms were reduced feeling of life pleasure or emotions (anhedonia) (75.9%); lack of mental concentration (72.2%), and loss of muscle tone/mass (51.9%). We contributed to inform about symptoms of PFS patients; unexpectedly loss of penis sensitivity was more frequent than severe erectile dysfunction and loss of muscle tone/mass was affecting half of the subjects. Further studies are necessary to investigate the pathophysiological and biochemical pathways leading to the post-finasteride syndrome.

Acute peritonitis due to introital stenosis and perforation of a bowel neovagina in a transsexual.

BACKGROUND: Stenosis of the neovagina is a late postoperative complication of male-to-female sex reassignment surgery when patients do not have frequent sexual intercourse or do not perform vaginal dilation. CASE: A 39-year-old male-to-female transsexual who had sex reassignment surgery, in which a segment of sigmoid colon was used for neovagina construction, developed total introital stenosis and subsequent peritonitis caused by bowel perforation of the colon conduit. CONCLUSION: To avoid stenosis of the neovagina, an inflatable silicon vaginal stent should be used all day for 30 days, then for 3 months overnight or until sexual function is regular.

Rare association of renal angiomyolipoma and oncocytoma.

The association of renal angiomyolipoma and renal oncocytoma in the same kidney is rare, only 7 cases having been previously reported. We report a 69-year-old woman affected by the simultaneous presence of renal angiomyolipoma and oncocytoma in the right side with a decrease of renal function on the left side due to a stenosis of the superior polar branch that was successfully treated preoperatively by angioplasty. An enucleation of renal masses on the right side was performed with renal function preservation after 2 years.

Long-term results with Nesbit's procedure as treatment of Peyronie's disease.

The objective was to assess sexual function at long-term follow-up after the Nesbit operation for Peyronie's disease. One hundred and fifty seven out of 213 patients treated between 1986 and 1998 using the Nesbit procedure were reassessed by means of IIEF-5 questionnaire together with two questions about residual deformity and treatment satisfaction. The results from this questionnaire together with the patient case records constitute the basis of this paper. After an average 72-month follow-up subjective patient determination of satisfaction indicated that 87.9% were satisfied with the results of surgery, 136 patients (86.7%) had good erectile function (IIEF-5 > 21). Shortening of the penis (from 1.5 to 3 cm) occurred in 22 patients (14%), but only in 2 (1.3%) was intercourse difficult because of excessive shortening. In conclusion, the Nesbit's operation results in the greatest amount of patient satisfaction about sexual function. When penile shortening occurs, it has not been a significant problem for patients who are properly counselled.

Monolateral tubulovasostomy. Report of four cases.

After conventional vasoepididymostony, the epididymal lumen may become occluded or obstructed, resulting in recurring azoospermia. Recently, two-layer direct anastomosis has been advocated on the basis of experimental models of tubulovasostomy performed in rats. Monolateral tubulovasostomy was performed in four cases under 16-40 times magnification. Nylon or 11/0 Vycril was used as suture material for the internal layer of the anastomosis, and 10/0 Ethilon was used for the external layer. After three and six months, semen samples were evaluated for sperm count and motility. The presence of spermatozoa in all the samples confirmed 100% patency of the duct. Three patients had a number of spermatozoa compatible with fertility, and one pregnancy occurred.

Tubulovasostomy in the rat. A new experimental model.

Tubular patency is the primary concern in the evaluation of procedures for epididymovasostomy. The epididymal lumen easily becomes occluded or obstructed, with the resulting recurrence of azoospermia. Recently, two-layer direct anastomosis has been advocated. The aim of the present study was to present an experimental model of tubulovasostomy in the rat. Microsurgery was performed in 18 rats (16-40 X magnification). Suture material used was 11/0 Vycryl or nylon for the internal layer of the anastomosis and 10/0 Ethilon for the external layer. After 6 and 12 weeks, semen samples were obtained by electroejaculation and evaluated for sperm count and motility.

Retinal and epiretinal glia--an immunohistochemical study.

Immunohistochemical techniques were used to examine the distribution of cells containing glial fibrillary acidic protein (GFAP) in normal and pathological human specimens, including 22 globes (13 of which contained epiretinal membranes 'in situ'), 16 surgically excised epiretinal membranes, and monolayers of cells obtained from five epiretinal membranes placed in tissue culture. The astrocytic cells of normal and pathological retinae stained with the glial-cell marker, but Müller cells were GFAP-negative in normal retinae at the antisera dilutions used. Müller cells did, however, stain in retinae from glaucomatous eyes and in eyes with prolonged retinal detachment. Electron microscopy did not reveal any obvious morphological difference between the intermediate filaments of normal (GFAP-negative) and GFAP-positive Müller cells. Ten of the 13 epiretinal membranes 'in situ', all 16 excised membranes, and three of the five monolayers contained glial cells. Purely glial membranes were not associated with retinal puckering or detachment, while all membranes causing tractional complications had a prominent fibrous, non-glial component. Our findings suggest that glial cells do not contribute significantly to the contractile forces generated by epiretinal membranes. They may, however, provide a scaffold on which other cells proliferate and contract and an anchorage by means of which tangential forces are transmitted into and through the retina.

Immunohistochemical demonstration of metallothionein in eyes with choroidal melanomas.

PURPOSE: The since immunohistochemically detectable metallothionein (MT) overexpression has been described in a variety of human tumours, including skin melanomas, in relation to different stages of tumour development and progression. MATERIALS AND METHODS: We used a monoclonal antibody to investigate the distribution of MT in 18 formalin-fixed, paraffin-embedded, surgically enucleated eyes with choroidal melanomas, from 18 patients (8 male, 10 female; age range 30-83 years, mean 58.7). Clinico-pathological details and follow-up data (2-124 months, mean 36.1) were also available. MT immunoreactivity was recorded and the percentage of stained cells was graded for semiquantitative purposes. Correlations between immunohistochemical data and morphological characteristics of melanomas were investigated using non-parametric methods; survival analysis was done by the Kaplan-Meier method and the survival curves were compared by the Mantel-Cox log-rank test. RESULTS: MT immunoexpression was found in 15/18 cases (83.3%) with staining scores from 1 to 3; MT staining varied in intensity and was mainly localized in the cytoplasm, although a combined nuclear/cytoplasmic reactive pattern was seen in neoplastic elements. No differences in MT immunostaining were seen in relation to age or sex, tumour size, histotype and amount of pigment; univariate analysis of survival data showed no prognostic significance regarding MT expression. CONCLUSIONS: The immunohistochemical evidence of MT in neoplastic elements could be related to the production of this scavenging protein in the tumour for cell defense mechanisms against hydroxyl free radicals, and to act as a Zn donor, since Zn is required for the synthesis of DNA and DNA-repair enzymes.