A novel likely pathogenetic variant p.(Cys235Arg) of the MEN1 gene in multiple endocrine neoplasia type 1 with multifocal glucagonomas.

PURPOSE: Multiple endocrine neoplasia type 1 (MEN1) is a hereditary endocrine syndrome caused by pathogenic variants in MEN1 tumor suppressor gene. Diagnosis is commonly based on clinical criteria and confirmed by genetic testing. The objective of the present study was to report on a MEN1 case characterized by multiple pancreatic glucagonomas, with particular concern on the possible predisposing genetic defects. METHODS: While conducting an extensive review of the most recent scientific evidence on the unusual glucagonoma familial forms, we analyzed the MEN1 gene in a 35-year-old female with MEN1, as well as her son and daughter, using Sanger and next-generation sequencing (NGS) approaches. We additionally explored the functional and structural consequences of the identified variant using in silico analyses. RESULTS: NGS did not show any known pathogenic variant in the tested regions. However, a new non-conservative variant in exon 4 of MEN1 gene was found in heterozygosity in the patient and in her daughter, resulting in an amino acid substitution from hydrophobic cysteine to hydrophilic arginine at c.703T > C, p.(Cys235Arg). This variant is absent from populations databases and was never reported in full papers: its characteristics, together with the high specificity of the patient's clinical phenotype, pointed toward a possible causative role. CONCLUSION: Our findings confirm the need for careful genetic analysis of patients with MEN1 and establish a likely pathogenic role for the new p.(Cys235Arg) variant, at least in the rare subset of MEN1 associated with glucagonomas.

Prevention of the Werther effect: compliance of the Italian newspapers to the 'reporting on suicide' recommendations.

OBJECTIVES: Suicide media coverage could lead to imitation, the so-called 'Werther effect'. The World Health Organization (WHO) published specific recommendations for reporting suicide news. This study aimed to quantify the compliance of Italian newspapers with the WHO suicide reporting guidelines. STUDY DESIGN: This was a quantitative content analysis study. METHODS: All articles published by the three main Italian newspapers from June 2019 to May 2020 describing suicides or attempted suicides were included. The articles were analyzed using a checklist based on the WHO recommendations, which included 18 'negative' items (e.g. 'presence of the word 'suicide' in the title') and nine 'positive' items (e.g. 'the article reports the contacts of a suicide prevention hotline'). Each negative item was scored -1, and each positive item was scored +1. Multivariate linear regressions were performed to identify factors associated with lower adherence to WHO recommendations and higher social media engagement with the articles. RESULTS: A total of 3483 articles were screened, and 110 articles were included in the final analysis. The suicidal was male in 73.6% of cases. The median checklist score was -6 (interquartile range 3). Five percent of the articles had at least one positive item. The word 'suicide' was found in 90% of the titles. Female suicides were associated with a higher checklist score (coefficient 0.81, P = 0.039). No correlation was found between the checklist score and the social media engagement of the articles. CONCLUSIONS: Italian newspapers do not adhere completely to the WHO recommendations on reporting suicide, leading to a potential imitation effect. Public health professionals should make decision-makers and journalists aware of the importance of these recommendations.

Trumpet sounds emitted by male sperm whales in the Mediterranean Sea.

Sperm whale trumpets are sounds only occasionally documented, with a well recognisable and stereotyped acoustic arrangement. This study investigated the acoustic features of the trumpets and the context in which these sounds were recorded, using acoustic data collected over 22 years, in the Pelagos Sanctuary area (North-Western Mediterranean Sea). Analysed trumpets (n = 230), recorded at the beginning of a dive after the whale fluke-up, comprised a series of acoustic units organized in short sequences. Acoustic parameters were derived for the entire trumpet and for each distinguishable unit in a trumpet. Overall, trumpet durations and their initial frequencies were higher in recordings collected when multiple whales were visually or acoustically detected in the observation area. The identity of 68 whales was assessed through photo-identification, with 29 individuals producing trumpets within and between years. The variability of the acoustic parameters appeared to be higher within the same individuals rather than between different individuals, suggesting an individual plasticity in composing and arranging units in a trumpet. Different click patterns were observed before and after the trumpets, with more complex sequences when (1) other whales were visually/acoustically detected, and (2) individuals were in suitable foraging sites (i.e., canyon areas). Trumpets were commonly followed or preceded by click patterns suited for communication, such as codas and/or slow clicks. Significant relations between the trumpet emission and the male-only long-range communication click pattern (i.e. slow clicks) emerged, supporting the hypothesis that a trumpet is a sound emitted by maturing/mature males in feeding grounds. This study provides the first evidence that trumpets were conserved in the sperm whale acoustic repertoire at the decadal timescale, persisting across years and individuals in the same area. This persistence may be functionally specific to foraging activities performed by males in a well-established feeding area.

Quality and dose optimization in hand computed radiography.

The objective of the present study was to optimize a radiographic technique for hand examinations using a computed radiography (CR) system and demonstrate the potential for dose reductions compared with clinically established technique. An exposure index was generated from the optimized technique to guide operators when imaging hands. Homogeneous and anthropomorphic phantoms that simulated a patient's hand were imaged using a CR system at various tube voltages and current settings (40-55 kVp, 1.25-2.8 mAs), including those used in clinical routines (50 kVp, 2.0 mAs) to obtain an optimized chart. The homogeneous phantom was used to assess objective parameters that are associated with image quality, including the signal difference-to-noise ratio (SdNR), which is used to define a figure of merit (FOM) in the optimization process. The anthropomorphic phantom was used to subjectively evaluate image quality using Visual Grading Analysis (VGA) that was performed by three experienced radiologists. The technique that had the best VGA score and highest FOM was considered the gold standard (GS) in the present study. Image quality, dose and the exposure index that are currently used in the clinical routine for hand examinations in our institution were compared with the GS technique. The effective dose reduction was 67.0%. Good image quality was obtained for both techniques, although the exposure indices were 1.60 and 2.39 for the GS and clinical routine, respectively.

[Repeated bladder metastases from renal cell carcinoma. Report of a case with particular attention to the use of immunomodulators]. / Ripetuta metastatizzazione vescicale da carcinoma a cellule renali (CCR). Descrizione del caso con particolare riguardo all'impiego degli immunomodulatori.

The case of repeated metastases of the bladder from renal cell carcinoma in a 59 year old male patient is presented. Two years after nephrectomy the ureteral stump and the bladder were interested by two different metastases. Afterwards, other nine metastases developed into the bladder during the following 12 months. Biopsy with coagulation were performed in any case. Bladder metastases ranged between 2 and 4 mm. No other metastasis was discovered by MRI and skeletal scintigraphy. Thirteen months after the first bladder metastasis, intracavitary Bacillus Calmette-Guerin was used. No report was found in the literature about BCG in the treatment of superficial bladder metastases from renal cell carcinoma. However 15 months from beginning of BCG treatment no metastasis developed from the bladder that now is disease-free after a follow-up of 19 months.

Measurement of DNA cross-linking in patients on ifosfamide therapy using the single cell gel electrophoresis (comet) assay.

The single cell gel electrophoresis comet assay has become established as a sensitive technique for measuring DNA strand breaks. The technique has been modified to allow the sensitive detection and quantitation of DNA interstrand cross-linking at the single cell level. Cells are irradiated immediately before analysis to deliver a fixed level of random strand breakage. After embedding of cells in agarose and lysis, the presence of cross-links retards the electrophoretic mobility of the alkaline denatured cellular DNA. Cross-links are, therefore, quantitated as the decrease in the comet tail moment compared with irradiated controls. Using this method, a linear response of cross-linking versus dose of chlorambucil over a wide dose range was demonstrated in human lymphocytes after drug treatment ex vivo. The method was also sensitive enough to determine cross-linking in clinical samples after chemotherapy. For example, crosslinking was observed in the lymphocytes of patients receiving ifosfamide (3 g/m2/day) as a continuous infusion for 3-5 days or as a 3-h infusion daily for 3 days. Cross-links were detected in all patients within 3 h, with no evidence of DNA single strand break formation. In patients receiving continuous infusion, a plateau of cross-linking was reached by 24 h. In the patients receiving ifosfamide over 3 h, a clear decrease in the peak level of cross-linking was observed before subsequent infusions.

Accuracy of cervical/vaginal cytology in the diagnosis of bacterial vaginosis.

BACKGROUND AND OBJECTIVES: The cytological pattern of bacterial vaginosis (BV) in the Papanicolaou smear has a controversial history (coccoid vs. Gardnerella type) that has not allowed an efficient use of cervical/vaginal cytology in the diagnosis of this condition. Our study is an attempt to clarify this topic. GOALS: To evaluate the accuracy of cervical/vaginal cytology in the diagnosis of BV. STUDY DESIGN: 1,896 separate examinations were performed on 1,471 women attending our sexually transmitted diseases center between 1990 and 1993. Amsel's composite clinical criteria were used as a gold standard. The Pap smear was prepared on two slides with the addition of a vaginal specimen. RESULTS: In the 166 cases of BV, sensitivity is 88.7%, and specificity is 98.8%. Positive predictive value is 89.8, and negative predictive value is 98.7. Kendall's correlation coefficient is 0.88 (p < 0.001). CONCLUSIONS: Our findings support the validity of the vaginal Pap smear in diagnosing BV and suggest the screening of such a disease.

HIV infection increases the risk of squamous intra-epithelial lesions in women with HPV infection: an analysis of HPV genotypes. DIANAIDS Collaborative Study Group.

We assessed the association between different HPV genotypes, HIV infection, and cervical squamous intra-epithelial lesions (SIL) in 236 women with known HIV serostatus enrolled in a longitudinal multicentric study in Italy. Of these women, 135 were HIV-infected, and were not markedly different from HIV-negative women with regard to demographic characteristics, sexual practices, smoking, or intravenous drug use. We obtained 232 cervical smears suitable for cytological examination and HPV-genotype analysis (134 from HIV-positive women and 98 from HIV-negative women). For 86 HIV-positive and 89 HIV-negative women, the smears appeared normal at cytomorphological analysis. Cytological dysplasia of varying degrees was detected in 48 smears from HIV-positive women and in 9 from HIV-negative women. HPV prevalence, assessed using polymerase-chain-reaction analysis, did not significantly differ between HIV-positive and HIV-negative women. The prevalence of HPV-associated SIL was much greater among HIV-infected women. The most frequently detected genotypes in both groups were HPV 16 and HPV 18. The prevalence of HPV 16 among HIV-positive women was similar to that for HIV-negative women; this was also true for HPV 18. However, in the HIV-positive group, most of these genotypes were associated with SIL. HIV-positive women showed a wider spectrum of genotypes, including non-oncogenic and rare types. An association between SIL and HIV infection was confirmed for all HPV genotype classes.

Determinants of squamous intraepithelial lesions (SIL) on Pap smear: the role of HPV infection and of HIV-1-induced immunosuppression. DIANAIDS Collaborative Study Group.

This study aimed to identify risk factors for squamous intraepithelial lesions (SIL) in women with known HIV status and to explore the association between SIL, HPV subtype, and HIV-induced immunosuppression. The study population consisted of women with known HIV serological status who were attending a network of 16 clinical centres in Italy. Detailed behavioural data, clinical and laboratory parameters, and samples for diagnosis of SIL by Papanicolau smear and HPV infection using a polymerase chain reaction (PCR) were obtained from each study participant. The strength of the association between SIL and possible risk factors was assessed calculating crude and adjusted odds ratios derived from univariate analysis and multivariate models. We enrolled 236 women, of whom 135 (57.2%) were HIV-infected. SIL was diagnosed in 57 women (24.1%); of these, 48 (35.6%) were HIV-infected and 9 (8.9%) were HIV-negative. HPV-DNA was detected in 41 (72%) women with SIL and in 45 (25%) women without SIL. HPV-DNA was more often detected among HIV-infected women than among HIV-negative women (40% vs. 32%), but the difference was not statistically significant. Women infected with high-risk types or with low-risk-uncharacterised types of HPV both had a higher risk of SIL compared with HPV-negative women (respectively, AOR: 17.53 and AOR: 2.89). HIV-infected women with severe or moderate immunosuppression were more likely to have SIL than HIV-negative women (respectively, AOR: 7.29 and AOR: 3.09) also independently from HPV infection. Women reporting use of a contraceptive pill had a 2.5 times higher risk of SIL compared with those who never used hormonal contraceptives. The results confirm that high-risk HPV types are strongly associated with SIL, and that HIV infection may strengthen the effect of HPV at cervical level. The use of oral contraceptives may slightly increase the risk of SIL in women with at-risk behaviour for HIV infection.

Epithelial cells are the major source of biologically active granulocyte macrophage colony-stimulating factor in human endometrium.

Granulocyte macrophage colony-stimulating factor (GM-CSF) has emerged as an important growth factor for trophoblast and other placental cells, leading to improved placental functioning and fetal survival. Recent observations have indicated that GM-CSF is synthesized by epithelial cells in the murine pregnant and non-pregnant uterus. In this study, the production of GM-CSF by cells derived from human endometrium is assessed using a sensitive bioassay and specific neutralization of the cytokine bioactivity with a monoclonal antibody to GM-CSF. Originally, GM-CSF was assayed in the culture supernatants of explant cultures of human endometria. Concentrations of GM-CSF up to 4440 pg/ml were detected. Subsequently, enriched epithelial cell cultures were prepared from glands isolated from human endometrium. The purity of epithelial cultures was demonstrated by the expression of cytokeratin, a weak immunoreactivity for vimentin and a lack of immunoreactivity for leukocyte common antigen, CD68, a macrophage-specific protein and endothelial marker (factor VIII-related antigens). Detected concentrations of GM-CSF were as high as 18,800 pg/ml. Furthermore, pure epithelial cells of a neoplastic endometrial cell line ECC1 secreted GM-CSF, confirming the ability of endometrial epithelial cells to secrete this cytokine. The immunostaining of dated endometria from proliferative and secretory phases showed primarily that epithelial cells, and to a lesser extent stromal cells, exhibited immunoreactivity for GM-CSF. A Western blot analysis, performed to validate the immunohistochemical data, confirmed the presence of an immunoreactive gene product for GM-CSF in human endometrium throughout the menstrual cycle. These findings indicate that human endometrium synthesizes GM-CSF and that epithelial cells are a major contributor to its production.

Ovarian mesothelial and extramesothelial cells in interactive culture.

The ovarian mesothelium (OM) represents the tissue of origin of ovarian epithelial cancer. To gain insight into the regulation of this tissue, OM organoids and submesothelial ovarian stromal cells (SC) were isolated from New Zealand White rabbits by a stepwise tissue dispersal technique, while granulosa cells (GC) were aspirated from mature follicles (14 +/- 4 groups/animal). OM and SC dispersal were sequentially accomplished by: a) 1-h incubation in collagenase type I (300 U/ml), gentle scraping of the ovarian surface, and 1 g sedimentation of OM organoids (equivalent to 0.93 +/- 0.40 x 10(6) cells/animal) on 5% bovine serum albumin (BSA); b) 2-h incubation in pronase-collagenase (0.5%-300 U/ml) under periodical resuspension and gentle scraping of SC (1.40 +/- 0.25 x 10(6)/animal) from OM-denuded ovaries. After a week-long in vitro expansion, OM cells (OMC) were cultured alone and with SC or GC within monocameral vessels or bicameral transfilter vessels in serumless, fibronectinrich (4 micrograms/ml) HL-1 medium. After 7 d of contact cell-cell interaction, cytokeratin-positive OMC became surrounded by fibroblastoid, vimentin-positive SC or by cytokeratin and vimentin-weakly positive GC. Filter-bound OMC humorally interacting with underlying SC or GC displayed a biphasic, epithelioid and spindle, morphology with universal cytokeratin expression. Bromo-2'-deoxyuridine (BrdU) immunoperoxidase revealed mean cell proliferation indices of 14.88% for OMC cultured alone, 11.21% and 19.39% for OMC cultured with GC or SC in monocameral dishes, and 15.25% or 22.47% for OMC cultured in bicameral vessels over GC or SC, respectively. This model provides an experimental tool for investigating the unexplored role of stromal-mesothelial interaction in OM pathobiology.

Cervical intraepithelial neoplasia and human papillomavirus related lesions of the genital tract in HIV positive and negative women.

Two hundred and twenty one women at high risk for HIV (intravenous drug users and/or those with infected partners) were investigated, through a self-filled questionnaire and gynaecological examination, to define the relationship between genital Human Papilloma Virus (HPV) infections, preneoplastic cervical intraepithelial lesions (CIN) and behavioural risk factors. In the 121 HIV positive women, 58 (47%) had HPV lesions at colposcopic and/or cytologic examination and, out of these 58, 23 (40%) had CIN 1, CIN 2 or CIN 3. Six out of the 16 cases with CIN 1 and CIN 2 (37%) followed-up showed a rapid progression of the lesion to CIN 3; in 3 women the interval was 6 months, in the other 3 about 12 months. Only 5 (7%) of the remaining 66 women without HPV lesions had a CIN lesion, with an obviously significant difference on comparison with HPV positive subjects. Sixty two women out of the 121 (52%) had a previous diagnosis of condylomata. In the 100 HIV negative women, 23 (23%) had HPV lesions and, among these 23, 6 (26%) had CIN 1, CIN 2 or CIN 3; 1 of them had rapid progression from CIN 1 to CIN 3 within a year. Only 5 (3%) without HPV infection showed any kind of CIN. 33 women out of 100 (33%) had a previous clinical history of condylomata. Our findings strongly suggest that HIV infection is associated with HPV lesions and that cervical cytological abnormalities develop in this situation. There is a need for short interval cytological and colposcopic follow-up for women at high risk of HIV infection.

Detection of sexually transmitted diseases by urethral cytology, the ignored male counterpart of cervical cytology.

The detection of sexually transmitted diseases by urethral cytology was investigated in 270 men examined by urethral swabbing smears. Each sample was used to prepare a wet mount smear and smears for staining by the Papanicolaou, Gram and methylene blue techniques. A fifth smear was used for direct staining with fluorescein-conjugated monoclonal antibodies for the detection of Chlamydia trachomatis. The smears were examined for cytoplasmic and nuclear changes as well as for pathogenic organisms and inflammatory changes. Infections with Chlamydia trachomatis, Neisseria gonorrhoeae and human papillomavirus (HPV) produced distinctive cytologic patterns similar to those seen in cervicovaginal smears from women. The patterns in candidiasis, trichomoniasis and herpes simplex virus infection were not as diagnostic. Particularly noteworthy were the nuclear alterations, which appeared to be proplastic in HPV infection but retroplastic in Chlamydia infection. The results of this study indicate that urethral cytology would be an invaluable addition in diagnosing sexually transmitted diseases in men, particularly in the case of Chlamydia and HPV infections. The monomorphic structure of urethral columnar epithelium, as compared to the cervical epithelium, seems to result in a clearer and more constant response to pathogenic infections, as seen in the resulting smears.

Fine needle aspiration cytopathology of phyllodes tumor. Differential diagnosis with fibroadenoma.

The differential diagnosis of benign or borderline phyllodes tumors of the breast in fine needle aspiration biopsy smears was studied. Smears from five histologically proven cases and 20 proven cases of fibroadenoma were evaluated with regard to their cytologic features. The findings indicate that the presence of a high cellularity of stromal fragments (including bipolar naked nuclei), clusters of hyperplastic ductal cells and giant cells and the absence of apocrine metaplasia may suggest a diagnosis of phyllodes tumor, as opposed to fibroadenoma. The borderline phyllodes tumor showed a sarcomatous atypia and/or small intranuclear cytoplasmic invaginations in some cells. The differential diagnosis of this tumor with malignant phyllodes tumor and plasma-cell mastitis is also discussed.