CO2 pneumoperitoneum effects on proliferation and apoptosis in two different neuroblastoma cell lines.

PURPOSE: The proto-oncogene MYCN is considered a transcription factor involved in the regulation of neuroblastoma (NB) cell biology. Since minimally invasive-surgery represents a debated treatment of NB, we investigated CO2 effects on proliferative activity and apoptotic pathway in two NB cell lines, SH-SY5Y (MYCN-non-amplified) and IMR-32 (MYCN-amplified). METHODS: SH-SY5Y and IMR-32 were exposed to CO2 (100%) at a pressure of 15 mmHg for 4 h and then moved to normal condition for 24 h. Cell proliferation, caspase 3 activity and transcript levels of BAX, BCL-2, cyclin B, cyclin D and MMP-2 were evaluated. RESULTS: CO2 exposure caused a decrease in cell proliferation associated to increases in BAX/BCL-2 ratio and caspase 3 activity in SH-SY5Y, while opposite effects have been found in IMR-32. CO2 exposure induced a decrease of cyclin B1 in SH-SY5Y, while an increase in cyclin B1 and D1 was observed in IMR-32. A slight up-regulation of MMP-2 expression in SH-SY5Y and a significant increase of 2.2 folds in IMR-32 was observed (p < 0.05). CONCLUSIONS: Our results suggest that CO2 exposure may cause different effects on various NB cell lines, likely due to MYCN amplification status. Further in vitro and in vivo studies are needed to highlight the role of laparoscopy on NB behaviour.

Methylene blue dye for identification of processus vaginalis during hydrocele repair: experience in a teaching hospital.

BACKGROUND: The aim of this study is to investigate the use of methylene blue in perioperative identification of the patent processus vaginalis in a group of boys presenting with congenital or recurrent hydrocele where surgery was performed by junior surgeons in training. METHODS: We retrospectively reviewed the notes of 22 boys with hydrocele, of which two recurrences, who were operated via a standard inguinal approach, by trainees. Methylene blue 0.3-0.5 mL was injected into the hydrocele fluid through the scrotal wall. A processus vaginalis was identified as a blue line. RESULTS: Methylene blue injection clearly identified a patent processus vaginalis in 91% of patients. In 9% (N.=2), of which one recurrence, methylene blue injection demonstrated a hydrocele with an obliterated processus vaginalis. There were no intraoperative complications. No testicular atrophy was recorded. CONCLUSIONS: Injection of methylene blue into the hydrocele sac may be considered a useful aid for a clearer identification of a difficult patent processus vaginalis. In the present series, there were no complications, and thus we believe that this technique might be suitable and especially helpful, in cases of recurrent hydrocele, and for junior surgeons in training.

Stable Ozonides with Vitamin E Acetate versus Corticosteroid in the Treatment of Lichen Sclerosus in Foreskin: Evaluation of Effects on Inflammation.

BACKGROUND: Lichen sclerosus (LS) is a disease of the skin of unclear etiology that can occur in the foreskin. Topical therapy with corticosteroids is recommended, but they can have side effects. OBJECTIVES: We aimed to compare the effects of ozonides with vitamin E acetate (OZOILE) versus topical corticosteroid in children undergoing circumcision. METHOD: Twenty children undergoing circumcision were treated before surgery: 10 children with OZOILE cream and 10 with 0.1% mometasone furoate once a day for 7 days. Ten age-matched patients with LS of the foreskin without any treatment were recruited as controls. Transcript levels of proinflammatory and anti-inflammatory cytokines and e-cadherin were evaluated in removed foreskins by qRT-PCR. RESULTS: OZOILE and steroid topical treatment produced a similar reduction of TNF-α and IL-1ß mRNA levels in foreskins from patients with LS when compared to untreated patients (p < 0.001). OZOILE and steroid treatment caused an increase in the transcript levels of IL-13 and e-cadherin in the foreskin of patients affected by LS in comparison to untreated foreskin (p < 0.001). CONCLUSIONS: On the basis of our biochemical data, a randomized clinical trial might be useful to verify the actual clinical effect of OZOILE as alternative treatment to corticosteroids in children affected by LS of the foreskin.

Utility of uroflowmetry during the follow-up of children affected by balanitis xerotica obliterans (BXO).

INTRODUCTION: To evaluate the outcome of circumcised patients with balanitis xerotica obliterans (BXO) using uroflowmetry (UF). METHODS: Between 2011 and 2013, 180 children underwent a circumcision for phimosis. The foreskin was examined on microscopy. Patients with an histological diagnosis of BXO were included in the study. Patients with BXO underwent UF two weeks after surgery and treatment with clobetasol propionate ointment. Patients were re-evaluated at 6, 12, 18 and 24 months postoperatively clinically and using UF. RESULTS: 75 of 180 circumcised patients (41.6%) were included. At two weeks, Thirtytwo of 75 patients (42.7%) displayed a pathological UF. At six months, 15 patients (20%) had pathological UF and a new cycle of clobetasol was prescribed. At one year, 10 patients (13.3%) displayed patholgocial UF and underwent progressive urethral dilatation or meatoplasty. At 18 months, 71 patients (94.7%) displayed regular UF, 3 underwent a meatoplasty and one a staged urethroplasty for a severe urethral stenosis. At two years, UF was normal in 74 out of 75 (98.7%). CONCLUSIONS: We recommend to send for hystological examination all foreskins excised after circumcision. We believe that a clinical and uroflowmetric follow-up of pediatric patients with BXO is mandatory for a prompt identification of post-voiding dysfunction.

Anti-Inflammatory and Tissue Regenerative Effects of Topical Treatment with Ozonated Olive Oil/Vitamin E Acetate in Balanitis Xerotica Obliterans.

Balanitis xerotica obliterans (BXO) is a chronic inflammatory skin disorder, considered the male genital variant of lichen sclerosus. Anti-inflammatory drugs are commonly used in BXO. We evaluated the effects of an innovative formulation of ozonated olive oil with vitamin E acetate (OZOILE®) on the inflammatory status and tissue remodeling in male children with BXO. The mRNA transcripts of proteins involved either in inflammation or in dynamics of tissue regeneration were analyzed by quantitative real-time PCR, in foreskins affected by BXO removed from patients untreated or treated with OZOILE® cream for 7 days before circumcision. We found a significant reduction in mRNA levels of IL-1ß, TNF-α, INF-γ, transglutaminase 2 and NOS2 in foreskins treated with OZOILE® in comparison to untreated ones (p < 0.001). No significant differences were observed in NF-κB activation in the specimens obtained from treated and untreated patients. Hence, OZOILE® treatment up-regulated hypoxia-inducible factor (HIF)-1alpha, vascular endothelial growth factor (VEGF) and E-cadherin gene expression (p < 0.001). The treatment with OZOILE® showed effective results in children affected by BXO by reducing the inflammatory process and stimulating mechanisms for tissue regeneration of the foreskin. A randomized clinical trial on a large number of children affected by BXO might be useful to verify the efficacy of topical treatment with OZOILE®.

Is inguinal orchidopexy still a current procedure in the treatment of intraabdominal testis in the era of laparoscopic surgery?

PURPOSE: To report our experience in surgical management of nonpalpable intraabdominal testis (NPIT) by inguinal orchidopexy without division of the spermatic vessels. METHODS: We reviewed the records of NPIT patients who underwent orchidopexy between 2012 and 2015. All patients were evaluated ultrasonographically. When the testis was not detected ultrasonographically, a laparoscopic exploration was performed. If the testis was found on laparoscopy, surgery was resumed through an inguinal incision. A follow-up was performed at 1week, 1, 3 and 6months. RESULTS: Twenty-one NPIT patients were treated, mean age 21.0±11.7months. Ultrasound identified 15 cases of NPIT (71%); diagnostic laparoscopy was performed in 6 (29%). All patients underwent an inguinal orchidopexy. At 1week, four testes were in a high scrotal position. At 6months follow-up, one testis was in a high scrotal position and one retracted up to the external inguinal ring. No atrophy was recorded. CONCLUSIONS: Despite several attempts to find a surgical technique without any significant complications, all described procedures failed to meet the target. In our experience, inguinal orchidopexy is a safe, reliable and successful surgical procedure for the management of NPIT. It should be preferred to a technique requiring vascular division, burdened with a higher incidence of atrophy. TYPE OF STUDY: Treatment study. LEVELS OF EVIDENCE: Case series with no comparison group.

Operative cystoscopy in the neonatal period.

Aim of the study is to report the experience in the last three years about endourological treatment of newborns affected by genito- and urinary malformation, implying critical urinary tract dilatation and bladder mass requiring a neonatal operative management. Seven - four males and three female - patients were treated cystoscopically during the neonatal period. Three newborns with a posterior urethral valves (PUV), underwent a successful ablation of PUV. In a male with infected persistent large mullerian duct and subsequent acute urinary retention, a cystoscopically-assisted catheterization was performed. Two females, one with cloaca and the other with high persistent urogenital sinus and massive associated hydrometrocolpos underwent a cystoscopical drainage of the vagina and a positioning of a balloon catheter into the bladder. Another female with a bladder mass had a endoscopical biopsy. Thanks to modern neonatal operative urethrocystoscope, today is possible to treat early many pathological genito-urinary conditions in newborns.

Neonatal bowel strangulation: Rare presentation of congenital diaphragmatic hernia.

We report a case of congenital diaphragmatic hernia (CDH) with perinatal bowel strangulation requiring intestinal resection. Ten hours after birth, the newborn started to be lethargic and developed bilious emesis. X-ray documented distended loops of bowel with air fluid levels in the abdomen and a gasless, non-homogeneous opacity of the left hemithorax, a right mediastinal shift and loss of the sharp left hemidiaphram line. On gastrographin enema the left colon was above the adjacent left diaphragm. Emergency surgery was performed at 16 h of age. The entire small bowel appeared reddish and compromised. After 24 h, second-look laparotomy was performed and only 25 cm of small bowel were viable. The postoperative period was uneventful. Neonatal bowel strangulation in CDH should be taken into account when estimating postnatal morbidity and mortality and, even if CDH treatment is not an emergency procedure, if gastrointestinal symptoms prevail over respiratory symptoms, surgery should be carried out without delay.

Clinical Pregenetic Screening for Stroke Monogenic Diseases: Results From Lombardia GENS Registry.

BACKGROUND AND PURPOSE: Lombardia GENS is a multicentre prospective study aimed at diagnosing 5 single-gene disorders associated with stroke (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, Fabry disease, MELAS [mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes], hereditary cerebral amyloid angiopathy, and Marfan syndrome) by applying diagnostic algorithms specific for each clinically suspected disease METHODS: We enrolled a consecutive series of patients with ischemic or hemorrhagic stroke or transient ischemic attack admitted in stroke units in the Lombardia region participating in the project. Patients were defined as probable when presenting with stroke or transient ischemic attack of unknown etiopathogenic causes, or in the presence of <3 conventional vascular risk factors or young age at onset, or positive familial history or of specific clinical features. Patients fulfilling diagnostic algorithms specific for each monogenic disease (suspected) were referred for genetic analysis. RESULTS: In 209 patients (57.4±14.7 years), the application of the disease-specific algorithm identified 227 patients with possible monogenic disease. Genetic testing identified pathogenic mutations in 7% of these cases. Familial history of stroke was the only significant specific feature that distinguished mutated patients from nonmutated ones. The presence of cerebrovascular risk factors did not exclude a genetic disease. CONCLUSIONS: In patients prescreened using a clinical algorithm for monogenic disorders, we identified monogenic causes of events in 7% of patients in comparison to the 1% to 5% prevalence reported in previous series.

Our Experience in Transcrotal Orchidopexy in Children Affected by Palpable Undescended Testis.

INTRODUCTION: Classically, surgical approach for palpable undescended testis (pUDT) consists is an inguinal orchidopexy. In fact, a double incision allows an adequate mobilization of the spermatic cord and an easy dissection of a patent processus vaginalis and also to perform a subdartos pouch. For reduce potential mobility of the inguinal approach, in 1989 Bianchi and Squire proposed a transcrotal orchidopexy, using a high scrotal incision. We report our experiences and retrospectively evaluate the feasibility and postoperative success of the transcrotal approach for treatment of pUDT. PATIENTS AND METHODS: From January 2012 to June 2014, 217 patients, affected by pUDT were treated at our Institution, for a total of 231 orchidopexies (203 monolateral pUDT, 14 bilateral pUDT). Patients, in whom, under anesthesia, the testis could be moved to the neck of the scrotum, have been treated with a transcrotal approach using a high scrotal incision. All patients have been clinically followed up at 1 week and at 1, 2, 3, and 6 months, at 1 year, and then annually and using scrotal sonography with color-Doppler at 2, 3, and 6 months. RESULTS: A total of 205 pUDT (88.7%) were considered eligible for transcrotal. Eight (3.9%) pUDT, that were first approached transcrotally, have been converted to inguinal approach. At follow-up, two moderate scrotal hematomas and one inguinal hernia were noted. No recurrence or testicular atrophy was showed in transcrotal approached testes, while 2 recurrences out of 26 procedures (7.7%) were displayed after inguinal orchidopexy. CONCLUSIONS: In our experience, transcrotal approach is possible in almost 90% of pUDT. No major complication, such as recurrence or testicular atrophy, has been complained. Just 3 out of the 205 cases (1.5%) reported minor complications and 1 of which required a successful day-case procedure. Our data confirm that transcrotal orchidopexy can be considered effective, safe, and with a success rate being equivalent or better to classical inguinal approach.

Bladder Mass in Newborn: Case Report and Review of Literature.

Benign proliferative lesions of the bladder are exceptional in neonates. We describe a case of a 3-day-old neonate, presenting with bloody meconium and sonographic diagnosis of bladder mass. Cystoscopic biopsies were performed and a diagnosis of polypoid cystitis was made. The patient was treated conservatively and the lesion healed during follow-up. We review the literature of other cases of neonatal bladder masses.