High-resolution array-CGH analysis on 46,XX patients affected by early onset primary ovarian insufficiency discloses new genes involved in ovarian function.

STUDY QUESTION: Can high resolution array-CGH analysis on a cohort of women showing a primary ovarian insufficiency (POI) phenotype in young age identify copy number variants (CNVs) with a deleterious effect on ovarian function? SUMMARY ANSWER: This approach has proved effective to clarify the role of CNVs in POI pathogenesis and to better unveil both novel candidate genes and pathogenic mechanisms. WHAT IS KNOWN ALREADY: POI describes the progression toward the cessation of ovarian function before the age of 40 years. Genetic causes are highly heterogeneous and despite several genes being associated with ovarian failure, most of genetic basis of POI still needs to be elucidated. STUDY DESIGN, SIZE, DURATION: The current study included 67 46,XX patients with early onset POI (<19 years) and 134 control females recruited between 2012 and 2016 at the Medical Cytogenetics and Molecular Genetics Lab, IRCCS Istituto Auxologico Italiano. PARTICIPANTS/MATERIALS, SETTING, METHODS: High resolution array-CGH analysis was carried out on POI patients' DNA. Results of patients and female controls were analyzed to search for rare CNVs. All variants were validated and subjected to a gene content analysis and disease gene prioritization based on the present literature to find out new ovary candidate genes. Case-control study with statistical analysis was carried out to validate our approach and evaluate any ovary CNVs/gene enrichment. Characterization of particular CNVs with molecular and functional studies was performed to assess their pathogenic involvement in POI. MAIN RESULTS AND THE ROLE OF CHANCE: We identified 37 ovary-related CNVs involving 44 genes with a role in ovary in 32 patients. All except one of the selected CNVs were not observed in the control group. Possible involvement of the CNVs in POI pathogenesis was further corroborated by a case-control analysis that showed a significant enrichment of ovary-related CNVs/genes in patients (P = 0.0132; P = 0.0126). Disease gene prioritization identified both previously reported POI genes (e.g. BMP15, DIAPH2, CPEB1, BNC1) and new candidates supported by transcript and functional studies, such as TP63 with a role in oocyte genomic integrity and VLDLR which is involved in steroidogenesis. LARGE SCALE DATA: ClinVar database (http://www.ncbi.nlm.nih.gov/clinvar/); accession numbers SCV000787656 to SCV000787743. LIMITATIONS, REASONS FOR CAUTION: This is a descriptive analysis for almost all of the CNVs identified. Inheritance studies of CNVs in some non-familial sporadic cases was not performed as the parents' DNA samples were not available. Addionally, RT-qPCR analyses were carried out in few cases as RNA samples were not always available and the genes were not expressed in blood. WIDER IMPLICATIONS OF THE FINDINGS: Our array-CGH screening turned out to be efficient in identifying different CNVs possibly implicated in disease onset, thus supporting the extremely wide genetic heterogeneity of POI. Since almost 50% of cases are negative rare ovary-related CNVs, array-CGH together with next generation sequencing might represent the most suitable approach to obtain a comprehensive genetic characterization of POI patients. STUDY FUNDING/COMPETING INTEREST(S): Supported by Italian Ministry of Health grants 'Ricerca Corrente' (08C203_2012) and 'Ricerca Finalizzata' (GR-2011-02351636, BIOEFFECT) to IRCCS Istituto Auxologico Italiano.

Crura augmentation with Bio-A® mesh for laparoscopic repair of hiatal hernia: single-institution experience with 100 consecutive patients.

BACKGROUND: The potential utility of both non-absorbable and absorbable meshes to reinforce the esophageal hiatus and prevent recurrent hernia has been investigated in observational studies and a few randomized clinical trials. Use of absorbable mesh has been associated with lesser side-effects, but the long-term safety and effectiveness are still debated. This rather scanty clinical evidence is due to heterogeneity and bias regarding the type of mesh and operation used, the modalities of follow-up, and the reporting of objective results. OBJECTIVES: The aim of the study was to assess safety, quality of life, and recurrence-free probability after laparoscopic repair of hiatal hernia reinforced with a synthetic absorbable mesh. METHODS: Observational, retrospective, single-center cohort study. All patients with hiatal hernia who underwent laparoscopic crura repair using a biosynthetic mesh (Gore Bio A® tissue reinforcement, Flagstaff, AZ) were included. Pre- and post-operative symptoms were assessed with the GERD-HRQL questionnaire. Objective follow-up consisted of upper gastrointestinal endoscopy and barium swallow study. RESULTS: From September 2011 to March 2016, a total of 100 patients underwent hiatal hernia repair using a Bio-A® mesh. All surgical procedures were completed laparoscopically. Postoperative morbidity rate was 10%. All patients had a minimum follow-up of 6 months, and the median follow-up was 30 (IQR = 22) months. No mesh-related complications occurred. The incidence of recurrent hernia ≥2 cm was 9%, and eight of the nine patients had a preoperative type III hernia. The median GERD-HRQL score was significantly reduced after operation (p < 0.001). The recurrence-free probability at 1 and 5 years was, respectively, 0.99 (CI 0.97-1.00) and 0.84 (CI 0.74-0.97), and no reoperation was required. No association was found between age, BMI, hernia size, previously failed surgical repairs and hernia recurrence. CONCLUSIONS: The use of a synthetic absorbable mesh to reinforce the esophageal hiatus is safe and appears to be effective and durable over a medium-term follow-up.

Advances in Poly(4-aminodiphenylaniline) Nanofibers Preparation by Electrospinning Technique.

Polyaniline (PANI) nanofibers are drawing a great deal of interest from academia and industry due to their multiple applications, especially in biomedical field. PANI nanofibers were successfully electrospun for the first time by MacDiarmid and co-workers at the beginning of the millennium and since then many efforts have been addressed to improve their quality. However, traditional PANI prepared from aniline monomer shows some drawbacks, such as presence of toxic (i.e., benzidine) and inorganic (salts and metals) co-products, that complicate polymer post-treatment, and low solubility in common organic solvents, making hard its processing by electrospinning technique. Some industrial sectors, such as medical and biomedical, need to employ materials free from toxic and polluting species. In this regard, the oxidative polymerization of N-(4-aminophenyl)aniline, aniline dimer, to produce poly(4-aminodiphenylaniline), P4ADA, a kind of PANI, represents an innovative alternative to the traditional synthesis because the obtained polymer results free from carcinogenic and/or polluting co-products, and, moreover, more soluble than traditional PANI. This latter feature can be exploited to obtain P4ADA nanofibers by electrospinning technique. In this paper we report the advances obtained in the P4ADA nanofibers electrospinnig. A comparison among polyethylene oxide (PEO), polymethyl methacrylate (PMMA) and polystyrene (PS), as the second polymer to facilitate the electrospinning process, is shown. In order to increase the conductivity of P4ADA nanofibers, two strategies were adopted and compared: selective insulating binder removal from electrospun nanofibers by a rinsing tratment, afterwards optimizing the minimum amount of binder necessary for the electrospinning process. Moreover, the effect of PEO/P4ADA weight ratio on the fibers morphology and conductivity was highlighted.

Multicystic peritoneal mesothelioma treated by surgical cytoreduction and hyperthermic intra-peritoneal chemotherapy (HIPEC).

BACKGROUND: Multicystic peritoneal mesothelioma (MPM) is an extremely uncommon lesion with uncertain malignant potential. Multiple recurrences after surgical interventions and transition to aggressive malignancies have been reported. Here, we review our experience with cytoreduction and hyperthermic intraperitoneal chemotherapy (HIPEC) in the management of MPM. PATIENTS AND METHODS: Five women with MPM underwent 6 procedures of cytoreduction and close-abdomen HIPEC with cisplatin and doxorubicin. Three patients had recurrent disease after 1, 2 and 4 previous debulkings, respectively. RESULTS: Optimal cytoreduction (residual tumor nodules < or =2.5 mm) was performed in all the procedures. One grade 4 postoperative complication (NCI/CTCAE v.3.0) and no operative mortality occurred. Median follow-up was 31 months (range 3-102). MPM recurred in two patients: one is presently disease-free after a second cytoreduction with HIPEC and the other is alive with minimal stable disease. CONCLUSION: Definitive eradication by means of cytoreduction and HIPEC seems a safe and effective therapeutic option for MPM.

High-speed gas chromatography with direct resistively-heated column (ultra fast module-GC)-separation measure (S) and other chromatographic parameters under different analysis conditions for samples of different complexities and volatilities.

The influence of GC speed on the separation capability of a chromatographic system is reported measuring a series of parameters including separation measure (S), peak capacity (n), peak width (w), analysis time, t(b) (determined on the last eluting compound) and separation measure/analysis time ratio (S/t(b)) determined by analyzing a bergamot essential oil sample and a standard mixture of pesticides. Conventional GC, fast GC (with 10 m (FGC10) and 5 m (FGC5) narrow-bore columns), and direct resistively-heated ultra fast module-GC (UFM-GC) were the GC speed approaches used. The influence of different heating rates with a constant flow for FGC5, FGC 10, and UFM-GC and with variable flows for UFM-GC on S, n, w, S/t(b), and t(b) was also studied. The results of this study show that: (a) separation capability of the chromatographic system (i.e. S and n) and analysis time depend on the GC approaches. Within each GC approach, S and n and analysis time depend on the heating rates, although to a different extent, and S and n decrease much less than the gain in analysis time, in particular when fast heating rates are applied; (b) in UFM-GC, the loss of separation capability with heating rate can also be partially compensated by the choice of an appropriate flow rate that, within each heating rate, may contribute to increase S while reducing t(b); (c) within a specific GC approach, the chromatographic system (column and stationary phase) and conditions (heating and flow rates) must be such to achieve a suitable S-value when two analytes must be separated with a given resolution in a minimum analysis time.

Conventional inner diameter short capillary columns: an approach to speeding up gas chromatographic analysis of medium complexity samples.

Short capillary columns (5 m) with 0.25 mm inner diameter (I.D.) are applied to the GC analysis of medium complexity samples (up to 30 components) with the aim of shortening analysis time. This approach is complementary to fast GC with narrow-bore columns and is based on compensating the lower efficiency of short columns with conventional I.D.'s (0.25-0.32 mm) by using a stationary phase selectivity suitable to separate the components of the sample under investigation, so that the required resolution power is achieved but, at the same time, the analysis time is shortened. The qualitative and quantitative effectiveness of this approach is demonstrated through the analysis of: essential oils with different compositions (chamomile and rosemary), low-volatility triterpenes in a plant extract (Maytenus aquifolium and M. ilicfolium), thermolabile pyrethrins in a Pyrethrum extract, and a mixture of pesticides applied to protect medicinal plant crops. In all examples, GC analysis was five to ten times faster than with conventional columns.

Characterization of the toxicity of distamycin derivatives on cancer cell lines and rat heart.

The cytotoxicity and cardiotoxicity of benzoyl mustard (FCE 24517) and epoxamido (FCE 24561) synthetic derivatives of distamycin A were reported in the present study. The 50% inhibiting concentration (IC50) of colony formation of FCE 24517 on human SNB-19 glioblastoma, A2780 ovarian cancer and DU 145 prostate cancer was at least three times lower than that of FCE 24561; on the same cell lines the IC50 of DXR was up to 14 and 240 times higher than that of FCE 24561 and FCE 24517, respectively. Isolated rat hearts perfused with concentrations of both derivatives equivalent to their respective IC50 values did not show any significant change in ECG parameters, contractility and coronary flow. Compared to control hearts, FCE 24517 10(-6) M induced a significant increase in PR interval, reduction in + dF/dtmax, heart rate and coronary flow, while FCE 24561 10(-6) M produced a modest but significant increase in S alpha T segment and decrease in + dF/dtmax. Rats treated with FCE 24561 3, 6 or 12 mg/kg, intravenously (i.v.), once weekly for 3 weeks had a modest increase in S alpha T segment and QRS complex duration, while a slight alteration of S alpha T segment and QRS complex duration were observed in rats given FCE 24517 1 or 2 mg/kg i.v. once weekly for 3 weeks. No cardiac histologic alterations were found in hearts from rats receiving FCE 24517 or FCE 24561. For comparison, the cardiotoxicity of doxorubicin (DXR) was evaluated in the same experimental models; perfusion of hearts with DXR 10(-6) M induced severe alterations in all parameters of the isolated hearts; the administration of DXR 3 mg/kg i.v. once a week for 3 weeks was associated with a widening of the S alpha T segment and QRS complex and cardiac histologic picture was markedly altered. In conclusion, distamycin A derivatives display elevated cytotoxicity while no substantial cardiotoxicity was observed.

An uncommon cause of massive intraperitoneal bleeding: leiomyoblastoma of the stomach.

A large exogastric leiomyoblastoma in a 48-year-old male revealed by asymptomatic upper abdominal mass is reported. Abdominal ultrasound, computerized tomography scan and magnetic resonance showed a 20 cm cystic lesion apart from liver and pancreas of undetermined origin. During hospitalization, massive intraperitoneal bleeding due to rupture of the mass was observed. An emergency laparotomy was carried out, and excision of a large, ruptured, cystic mass involving the greater gastric curvature was performed. Microscopy revealed a gastric leiomyoblastoma. Even if infrequent, massive intraperitoneal bleeding is a very serious complication of gastric leiomyoblastoma. Considering the difficulty of an accurate preoperative diagnosis and the risk of intraperitoneal rupture, the authors suggest that similar abdominal masses should be managed by quick diagnostic investigations and early surgical procedures.