Structural rearrangements as a recurrent pathogenic mechanism for SETBP1 haploinsufficiency.

Chromosomal structural rearrangements consist of anomalies in genomic architecture that may or may not be associated with genetic material gain and loss. Evaluating the precise breakpoint is crucial from a diagnostic point of view, highlighting possible gene disruption and addressing to appropriate genotype-phenotype association. Structural rearrangements can either occur randomly within the genome or present with a recurrence, mainly due to peculiar genomic features of the surrounding regions. We report about three non-related individuals, harboring chromosomal structural rearrangements interrupting SETBP1, leading to gene haploinsufficiency. Two out of them resulted negative to Chromosomal Microarray Analysis (CMA), being the rearrangement balanced at a microarray resolution. The third one, presenting with a complex three-chromosome rearrangement, had been previously diagnosed with SETBP1 haploinsufficiency due to a partial gene deletion at one of the chromosomal breakpoints. We thoroughly characterized the rearrangements by means of Optical Genome Mapping (OGM) and Whole Genome Sequencing (WGS), providing details about the involved sequences and the underlying mechanisms. We propose structural variants as a recurrent event in SETBP1 haploinsufficiency, which may be overlooked by laboratory routine genomic analyses (CMA and Whole Exome Sequencing) or only partially determined when associated with genomic losses at breakpoints. We finally introduce a possible role of SETBP1 in a Noonan-like phenotype.

In vitro efficacy of ARQ 092, an allosteric AKT inhibitor, on primary fibroblast cells derived from patients with PIK3CA-related overgrowth spectrum (PROS).

Postzygotic mutations of the PIK3CA [phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha] gene constitutively activate the PI3K/AKT/mTOR pathway in PIK3CA-related overgrowth spectrum (PROS) patients, causing congenital mosaic tissue overgrowth that even multiple surgeries cannot solve. mTOR inhibitors are empirically tested and given for compassionate use in these patients. PROS patients could be ideal candidates for enrolment in trials with PI3K/AKT pathway inhibitors, considering the "clean" cellular setting in which a unique driver, a PIK3CA mutation, is present. We aimed to assess the effects of blocking the upstream pathway of mTOR on PROS patient-derived cells by using ARQ 092, a potent, selective, allosteric, and experimental orally bioavailable and highly selective AKT-inhibitor with activity and long-term tolerability, currently under clinical development for treatment of cancer and Proteus syndrome. Cell samples (i.e., primary fibroblasts) were derived from cultured tissues obtained from six PROS patients [3 boys, 3 girls; aged 2 to 17 years] whose spectrum of PIK3A-related overgrowth included HHML [hemihyperplasia multiple lipomatosis; n = 1], CLOVES [congenital lipomatosis, overgrowth, vascular malformations, epidermal nevi, spinal/skeletal anomalies, scoliosis; n = 1], and MCAP [megalencephaly capillary malformation syndrome; n = 4]. We performed the following: (a) a deep sequencing assay of PI3K/AKT pathway genes in the six PROS patients' derived cells to identify the causative mutations and (b) a pathway analysis to assess the phosphorylation status of AKT [Ser473 and Thr308] and its downstream targets [pAKTS1 (Thr246), pRPS6 (Ser235/236), and pRPS6Kß1 (Ser371)]. The anti-proliferative effect of ARQ 092 was tested and compared to other PI3K/AKT/mTOR inhibitors [i.e., wortmannin, LY249002, and rapamycin] in the six PROS patient-derived cells. Using ARQ 092 to target AKT, a critical node connecting PI3K and mTOR pathways, we observed the following: (1) strong anti-proliferative activity [ARQ 092 at 0.5, 1, and 2.5 µM blunted phosphorylation of AKT and its downstream targets (in the presence or absence of serum) and inhibited proliferation after 72 h; rapamycin at 100 nM did not decrease AKT phosphorylation] and (2) less cytotoxicity as compared to rapamycin and wortmannin. We demonstrated the following: (a) that PROS cells are dependent on AKT; (b) the advantage of inhibiting the pathway immediately downstream of PI3K to circumventing problems depending on multiple classes a PI3K kinases; and (c) that PROS patients benefit from inhibition of AKT rather than mTOR. Clinical development of ARQ 092 in PROS patients is on going in these patients.

Attachment styles and sexual dysfunctions: a case-control study of female and male sexuality.

The aim of this study was to investigate attachment styles in a group of women and men with sexual dysfunction. We recruited 44 subjects (21 women and 23 men) with sexual dysfunction and 41 subjects (21 women and 20 men) with healthy sexual function as the control group. Validated instruments for the evaluation of male and female sexual dysfunctions (M/F SD) and a psychometric tool specifically designed to investigate attachment style were administered. In women, significant differences were found between subjects with sexual dysfunction and healthy controls. The scales indicating an insecure attachment showed: discomfort with closeness (FSD = 42.85 ± 11.55 vs CTRL = 37.38 ± 8.54; P < 0.01), relationship as secondary (FSD = 26.76 ± 2.60 vs CTRL = 18.42 ± 7.99; P < 0.01), and need for approval (FSD=26.38 ± 3.61 vs CTRL = 20.76 ± 7.36; P < 0.01). Healthy women also had significantly higher scores in secure attachment (confidence: FSD = 24.57 ± 3.89 vs CTRL = 33.42 ± 5.74; P < 0.01). Men with sexual dysfunctions differed from healthy men in confidence (MSD = 30 ± 6.33 vs CTRL = 36.05 ± 5.26; P < 0.01) and in discomfort with closeness (MSD = 39.08 ± 8 vs CTRL = 34.25 ± 7.54; P < 0.05). These results suggest that particular aspects related to insecure attachment have a determinant role in people with sexual dysfunctions. It is therefore fundamental to identify the attachment styles and relational patterns in patients receiving counselling and psychological treatments focussed on sexual problems.

Missense mutations in exon 2 of the MED12 gene are involved in IGF-2 overexpression in uterine leiomyoma.

Uterine leiomyoma (UL), the most common benign tumour found in females, is associated with many recurrent genetic aberrations, such as translocations, interstitial deletions and specific germline mutations. Among these, mutations affecting exon 2 of the mediator complex subunit 12 (MED12) gene are commonly detected in the majority of ULs. Mutational analysis of the MED12 gene, performed on 36 UL samples, revealed that 12 leiomyomas (33.4%) exhibited heterozygous missense mutations in codon 44 of exon 2 of the MED12 gene, four leiomyomas (11.1%) showed internal in-frame deletions, and two leiomyomas (5.5%) exhibited deletions involving intron 1-exon 2 junction, which caused a predicted loss of the splice acceptor. No mutations were detected in uterine myometrium (UM) and pseudocapsule (PC) samples, including those from women with a MED12 mutation in UL. These data showed that the PC is a healthy tissue that surrounds the UL to maintain UM integrity. Analysis of insulin-like growth factor 2 (IGF-2) and collagen type IV alpha 2 (COL4A2) mRNA expression levels in the same set of ULs revealed that only those with MED12 missense mutations expressed significantly higher levels of IGF-2 mRNA. In contrast, MED12 gene status does not appear to affect mRNA expression levels of the COL4A2 gene. On the basis of this finding, we suggest that the MED12 status stratifies the ULs into two mutually exclusive pathways of leiomyoma genesis, one with IGF-2 overexpression and the other with no IGF-2 activation. The occurrence of IGF-2 overexpression could be therapeutically targeted for the non-surgical treatment of leiomyomas.

Alexithymia and vaginismus: a preliminary correlation perspective.

The aim of this study was to measure the prevalence of alexithymia and emotional dysregulation in women with vaginismus not associated with other organic or psychopathological disorders. The study involved the psychometric assessment of 41 patients with vaginismus and 100 healthy women, all of childbearing age. Alexithymia was evaluated by TAS-20 (Toronto Alexithymia Scale). Sexual function was assessed by FSFI (Female Sexual Function Index). In patients with vaginismus, the primary diagnosis of dyspareunia was excluded and an expert psychologist evaluated patients and controls according to DSM IV (Diagnostic and Statistical Manual of Mental Disorders: 4th edition) criteria to exclude mental disorders. Over half (51.1%) of the patients with vaginismus were classified as alexithymic or borderline (alexithymic trend), compared with just 18% of the control group. In addition, there was a significant difference in the TAS-20 total scores between the two groups (P<0.0001). In terms of relative risk, women suffering from vaginismus thus have a 3.8 times higher probability of showing alexithymia than do healthy women. Vaginismus is a complex syndrome and alexithymia is far from being its only characteristic. However, we found a significant correlation between vaginismus and alexithymia. In theory, alexithymia could thus be a risk factor for vaginismus, although future studies are required to demonstrate any chain of causation between these two conditions.

Gene expression analysis reveals an angiogenic profile in uterine leiomyoma pseudocapsule.

The pseudocapsule (PC) of the uterine leiomyoma (UL) is an anatomic entity that surrounds the myoma separating it from the myometrium (UM). Although a number of microarray experiments have identified differences in gene expression profile in the UL when compared with the UM, there is a lack of systematic studies on the PC. In this study, quantitative RT-PCR analysis was performed on 18 matched PC, UL and UM specimens and results showed that the PC displays a specific gene expression profile. The low expression level of insulin-like growth factor (IGF-2), a fibroid specific marker, that we found in the PC and the UM when compared with the UL, clearly indicates that the PC is in structural continuity with the UM. However, the significant increase in endoglin expression level in PC with respect to the UL and UM indicates that an active neoangiogenesis is present in PC. Conversely, other angiogenic factors such as von Willebrand factor (vWF) and vascular endothelial growth factor A (VEGF-A) seem to have little influence on the PC angiogenesis. Because the endoglin is preferentially expressed in proliferating endothelial cells, whereas the vWF and VEGF-A are preferentially expressed in preexisting endothelial cells, our idea is that the angiogenic activity in the PC is linked to wound healing. The angiogenic activity is also sustained by intermediate expression level of cystein-rich angiogenesis inducer 61, connective tissue growth factor and collagen 4α2 genes all involved in the neoangiogenesis, that we detected in the PC. Taken together our data demonstrate that the specific expression pattern observed in the PC could be the response of the uterine wall's smooth cells to the tension imposed by the tumor. As a consequence, a neovascular structure is generated involving regenerative processes. For these reasons, we suggest that the laparoscopic intracapsular myomectomy (LIM), a new surgical technique that preserves the PC during the UL removal, should always be preferred, to favor a faster and proper uterine healing.

Full and partial PTSD among young adult survivors 10 months after the L'Aquila 2009 earthquake: gender differences.

BACKGROUND: Earthquakes are one of the most frequently occurring natural disasters and extensive research has been conducted on mental disorders on exposed populations, particularly on post-traumatic stress disorder (PTSD). On April 6th 2009, the town of L'Aquila (Abruzzo), in central Italy, was struck by an earthquake with a strength of 5.9 on the Richter scale. In the town of L'Aquila many buildings collapsed and large parts of the town were destroyed. Overall, 309 people were killed, 1600 injured among which 200 severely injured and hospitalized, more than 65,000 people were displaced. METHODS: The aim of the present study was to investigate prevalence rates of PTSD, either full-blown or partial PTSD, among 512 students attending the last year of high school in L'Aquila about 10 months after the earthquake. According to the literature, partial PTSD was defined as the presence of symptoms in the DSM-IV Criterion B and C or D for PTSD diagnosis. Gender differences in the symptoms reported were investigated. Assessments included the Trauma and Loss Spectrum-Self Report (TALS-SR) and the Impact of Event Scale (IES). RESULTS: The results of the present study showed the presence of a diagnosis of PTSD in 192 (37.5%) of the students examined, with significantly (p=.000) higher rates in women than men (N=120, 51.7% and N=72, 25.7%, respectively). Moreover, 153 (29.9%) students reported partial PTSD (75, 32.3% women and 78, 27.9% men respectively). Significantly higher PTSD symptoms were reported by women with respect to men. LIMITATIONS: The lack of information on the impact of the earthquake on subjects and on the presence of Axis I psychiatric comorbidities are two major limitations besides the use of self-report instruments. CONCLUSIONS: Our results show high rates of full or partial PTSD in adolescents who survived the April 2009 L'Aquila earthquake, with women being the most affected. Thus, these results highlight the need to carefully explore these conditions.

18-fluorine fluorodeoxyglucose positron emission tomography with computerized tomography versus computerized tomography alone for the management of solitary lung nodules with diameters inferior to 1.5 cm.

OBJECTIVE: Aim of the study was to establish a noninvasive method for the preoperative characterisation of a pulmonary nodule when biopsy of the small mass is impossible. METHODS: From 1 January 2006 to 31 December 2008, we observed 124 asymptomatic patients with a noncalcified single lung nodule highlighted by computerised tomography (CT) of the thorax. Patients were divided into 2 groups: Group A consisted of 57 patients with lesion diameters between 0.5 cm and 0.99 cm; Group B consisted of 67 patients with lesion diameters between 1.0 cm and 1.5 cm. Fibreoptic bronchoscopy was negative for endobronchial neoformation in all patients. The topographic distribution of the lesions advised against CT-guided transthoracic needle biopsy or video-assisted thoracoscopy. All patients had preoperative 18-fluorine fluorodeoxyglucose (18F-FDG) positron emission tomography (PET) associated with CT of the thorax, which was compared with CT for evaluation of the mass. RESULTS: Postoperative histological diagnosis revealed 54 primary lung cancers, 47 lung metastases and 23 benign lesions. In Group A the sensitivity of 18F-FDG PET/CT and CT was 95 % and 73 % and the specificity was 72 % and 64 %, respectively ( P = 0.000001 for 18F-FDG PET/CT; P = 0.000177 for CT). In Group B the sensitivity of 18F-FDG PET/CT and CT was 95 % and 97 %, and the specificity was 80 % and 87 %, respectively ( P = 0.000001). CONCLUSIONS: Our study shows that 18F-FDG PET/CT improves the identification and characterisation of potentially malignant pulmonary nodules with a diameter < 1 cm. This technique could be a valid alternative to a surgical approach, currently the main method to investigate indeterminate lung nodules.

Ethmoidal adenocarcinoma with lung metastases: diagnosis and multimodal treatment.

We report a case of 68-year-old patient underwent a magnetic resonance imaging (MRI) of the skull and a computed tomography (CT) of the thorax for rhinorrhea and dyspnea. The MRI showed an irregular ethmoidal lesion and the CT of the thorax underlined a solid nodular neoformation in the upper right pulmonary lobe. The patient underwent rhinoscopy with biopsies that showed an ethmoidal adenocarcinoma; excision of the tumour was carried out via trans-sphenoid. After one month the patient underwent wedge-resections in video-thoracoscopy (VATS). Perioperative histologic examination revealed a lung metastases due to an adenocarcinoma of the ethmoid. The patient was treated with chemotherapy and did not show relapses after 12 months from VATS.

Dimensional, non-taxonic latent structure of psychotic symptoms in a student sample.

INTRODUCTION: We examined the construct of psychosis using the self-report instrument CAPE (Community Assessment of Psychic Experiences) in a sample of 1,323 students. MATERIALS AND METHODS: Taxometric analysis was carried out using Taxometric Programs for the R Computing Environment. The MAXCOV CCFI was 0.34, indicating a dimensional latent structure. All other taxometric analysis yielded very similar results indicative of dimensional structure. CONCLUSION: In this study, using powerful analytic techniques designed expressly for the purpose, i.e. taxometric analysis, the latent construct of psychosis in a sample of young students appeared to be consistent with a dimensional, non-taxonic latent structure.

Necrotizing mediastinitis linked to Boerhaave's syndrome: a surgical approach.

We report 5 cases of Boerhaave's syndrome with necrotizing mediastinitis and pleural empyema. We chose a surgical treatment characterized by manual suture and heterologous tissue to repair esophagus rupture. This option allows a better result in terms of morbidity and mortality.

Organization, structure and evolution of 41kb of genomic DNA spanning the D-J-C region of the sheep TRB locus.

A genomic region of 41,045 bp encompassing the 3'-end of the sheep T cell receptor beta chain was sequenced. Extensive molecular analysis has revealed that this region retains a unique structural feature for the presence of a third D-J-C cluster, never detected in any other mammalian species examined so far. A total of 3 TRBD, 18 TRBJ and 3 substantially identical TRBC genes were identified in about 28kb. At 13kb, downstream from the last TRBC gene, in an inverted transcriptional orientation, lies a TRBV gene. Sequence comparison and phylogenetic analyses have demonstrated that the extra D-J-C cluster originated from an unequal crossing over between the two ancestral TRBC genes. Interspersed repeats spanning 22.2% of the sequence, contribute to the wider size of the sheep TRB locus with respect to the other mammalian counterparts, without modifying the general genomic architecture. The nucleotide and predicted amino acid sequences from peripheral T cells cDNA clones indicated that the genes from cluster 3 are fully implicated in the beta chain recombination machinery. Closer inspections of the transcripts have also shown that inter-cluster rearrangements and splice variants, involving the additional cluster, increase the functional diversity of the sheep beta chain repertoire.

Foreign bodies aspirated in children: role of bronchoscopy.

OBJECTIVE: To examine the diagnostic and therapeutic role of fiber-optic and rigid bronchoscopy in pediatric patients with foreign body inhalations. METHODS: From January 1986 to December 2004, we observed 128 young patients with suspicion of foreign body aspiration. Patients were divided into 3 groups: group I, patients with negative chest X-ray; group II, patients with radiological direct signs; group III, patients with radiological indirect signs. RESULTS: Removal of the foreign body was effected in 105 patients by rigid bronchoscopy and in 13 patients by fiber-optic bronchoscopy. In 3 group II patients a thoracotomy with a bronchotomy was necessary. CONCLUSIONS: Fiber-optic bronchoscopy showed a diagnostic accuracy rate of 100 % but played a poor therapeutic role with a case resolution of 10.7 %. Rigid bronchoscopy was the main technique, permitting the removal of the tracheobronchial foreign body in 97.2 % of patients.

CO2 Activation by Nb(+) and NbO(+) in the Gas Phase. A Case of Two-State Reactivity Process.

Gas-phase carbon dioxide activation by Nb(+) and NbO(+) was studied at the density functional level of theory using the hybrid exchange correlation functional B3LYP. Three reaction profiles corresponding to the quintet, triplet, and singlet multiplicities were investigated in order to ascertain the presence of some spin inversion during the CO2 reduction. Carbon dioxide activation mediated by metal cations was found to be an exothermic spin-forbidden process resulting from a crossing between quintet and triplet energetic profiles. The endothermic reaction of NbO(+) with carbon dioxide was a barrierless process involving spin inversion. Geometries of minima along potential energy surfaces and reaction heats were in agreement with those from experimental studies carried out by using a guided ion beam tandem mass spectrometer.

Pulmonary and parotid hydatid cyst: an unusual double localization.

We described a rare case of pulmonary and parotid cyst for echinococcus granulosus in an eighty-one-year-old patient. Lesion was discovered in the lung owing to a clinical presentation. A swelling was the only sign in the parotid gland. Time between lung and parotid excision was 14 days. Time of hospitalizations was 4+/-1 day.

Characterization of Bordetella pertussis strains of recent isolation.

During the clinical trial conducted in Italy to evaluate the efficacy of new acellular pertussis vaccines, the most favorable conditions for the recovery and characterization of Bordetella pertussis strains, isolated from children with cough, were adopted. The nasopharyngeal aspirates were collected and sent to the laboratory in refrigerated conditions within 24 hours. Charcoal agar selective and non selective plates were used, and most of the isolates were recovered after 3-4 days of incubation. Confirmation of all suspected colonies included the use of biochemical tests and specific agglutination reaction with B. pertussis antiserum. Serotyping of fimbriae, susceptibility to erythromycin and DNA fingerprinting by Pulsed Field Gel Electrophoresis (PFGE), were performed to characterize B. pertussis isolates and to determine relatedness among different strains. Serotype 1,3 was the most represented in the bacterial population examined. A predominant pulsetype (PTA) characterized most of the isolates accounting for 71.4% of the strains examined. Eight subclones (23.5%) and three unrelated pulsetypes were also found. No resistant strains to erythromycin were detected.

Antibody kinetics and long-term sero-prevalence in the Italian clinical trial of acellular pertussis vaccines.

In the Italian Pertussis Vaccine Trial, data were collected to evaluate the persistence of anti-pertussis antibodies. A sub-cohort of 1275 children was followed for this purpose until a mean age of 21 months. An additional evaluation included pooled cross-sectional analysis of serum specimens collected for analysis of cough illnesses. Antibodies to PT, FHA and PRN were measured by ELISA using a standardized technique. With both acellular vaccines in the study (the Chiron Biocine three-component and SmithKline Beecham three-component vaccines) there was a fast and steep decrease of mean geometric titres for PT, FHA and PRN in the months immediately following vaccination. Titres were close to the detection limit 15 months after the end of primary immunization. The immunogenicity of the whole-cell study vaccine (produced by Connaught Laboratories, Inc. Swiftwater, USA) was poor as determined one month after the third dose and no antibody was detected in nearly all children 15 months after the end of vaccination.

Iron deficiency anemia in 1-year-old children of disadvantaged families in Montreal.

OBJECTIVES: To determine the prevalence of iron deficiency anemia among 1-year-old infants of disadvantaged families in Montreal as well as certain predictors of this condition. DESIGN: Cohort study. SETTING: Five poorest health districts in Montreal. PARTICIPANTS: Infants 10 to 14 months of age were identified from registration lists of births from May 1988 to August 1989. Those whose mother had less than 11 years of schooling and a family income below the government-defined low-income cutoff point were eligible. INTERVENTION: During a home visit capillary blood samples were obtained from the child, and the mother answered a questionnaire about infant-feeding practices. Infants with a serum ferritin level of 10 micrograms/L or less and either a hemoglobin level of 115 g/L or less or a mean corpuscular volume of 72 fL or less were considered as having iron deficiency anemia. RESULTS: Of the 299 mothers who were eligible and could be located 220 (74%) agreed to participate; 218 blood samples were available. Iron deficiency anemia was found in 25% of the infants (95% confidence interval [CI] 19% to 31%). The mean hemoglobin level was 115 (standard deviation 11) g/L. The serum ferritin level, assessed routinely in the last 62 infants, was 10 micrograms/L or less in 37% of the infants. The factors that were found to be predictors of iron deficiency anemia included the use of whole cow's milk before 6 months of age (odds ratio [OR] 3.56 [95% CI 1.07 to 11.26]) and the use of iron-fortified infant cereal for less than 6 months (OR 3.15 [95% CI 1.25 to 7.96]). A low birth weight and the use of iron-fortified formula for less than 6 months were associated with iron deficiency anemia. CONCLUSIONS: Despite a decrease in the prevalence of iron deficiency anemia among children of disadvantaged families in the United States socioeconomically disadvantaged infants in Montreal are at risk. Preventive measures must be taken to ensure adequate iron status in the first year of life.