A systems genomics approach identifies SIGLEC15 as a susceptibility factor in recurrent vulvovaginal candidiasis.

Candida vaginitis is a frequent clinical diagnosis with up to 8% of women experiencing recurrent vulvovaginal candidiasis (RVVC) globally. RVVC is characterized by at least three episodes per year. Most patients with RVVC lack known risk factors, suggesting a role for genetic risk factors in this condition. Through integration of genomic approaches and immunological studies in two independent cohorts of patients with RVVC and healthy individuals, we identified genes and cellular processes that contribute to the pathogenesis of RVVC, including cellular morphogenesis and metabolism, and cellular adhesion. We further identified SIGLEC15, a lectin expressed by various immune cells that binds sialic acid-containing structures, as a candidate gene involved in RVVC susceptibility. Candida stimulation induced SIGLEC15 expression in human peripheral blood mononuclear cells (PBMCs) and a polymorphism in the SIGLEC15 gene that was associated with RVVC in the patient cohorts led to an altered cytokine profile after PBMC stimulation. The same polymorphism led to an increase in IL1B and NLRP3 expression after Candida stimulation in HeLa cells in vitro. Last, Siglec15 expression was induced by Candida at the vaginal surface of mice, where in vivo silencing of Siglec15 led to an increase in the fungal burden. Siglec15 silencing was additionally accompanied by an increase in polymorphonuclear leukocytes during the course of infection. Identification of these pathways and cellular processes contributes to a better understanding of RVVC and may open new therapeutic avenues.

Defective replication stress response inhibits lymphomagenesis and impairs lymphocyte reconstitution.

DNA replication stress promotes genome instability in cancer. However, the contribution of the replication stress response to the development of malignancies remains unresolved. The DNA replication stress response protein SMARCAL1 stabilizes DNA replication forks and prevents replication fork collapse, a cause of DNA breaks and apoptosis. While the fork regression/remodeling functions of SMARCAL1 have been investigated, its in vivo functions in replication stress and cancer are unclear. Using a gamma radiation (IR)-induced replication stress T-cell lymphoma mouse model, we observed a significant inhibition of lymphomagenesis in mice lacking one or both alleles of Smarcal1. Notably, a quarter of the Smarcal1-deficient mice did not develop tumors. Moreover, hematopoietic stem/progenitor cells (HSPCs) and developing thymocytes in Smarcal1-deficient mice showed increased DNA damage and apoptosis during the proliferation burst following IR and an impaired ability to repopulate the thymus after IR. Additionally, mice lacking Smarcal1 showed significant HSPC defects when challenged to respond to other replication stress stimuli. Thus, our data reveal the critical function of the DNA replication stress response and, specifically, Smarcal1 in hematopoietic cell survival and tumor development. Our results also provide important insight into the immunodeficiency observed in individuals with mutations in SMARCAL1 by suggesting that it is an HSPC defect.

Association of a variable number tandem repeat in the NLRP3 gene in women with susceptibility to RVVC.

Vaginal infections with Candida spp. frequently occur in women of childbearing age. A small proportion of these women experience recurrent vulvovaginal candidosis (RVVC), which is characterized by at least three episodes of infection in one year. In addition to known risk factors such as antibiotics, diabetes, or pregnancy, host genetic variation and inflammatory pathways such as the IL-1/Th17 axis have been reported to play a substantial role in the pathogenesis of RVVC. In this study, we assessed a variable number tandem repeat (VNTR) polymorphism in the NLRP3 gene that encodes a component of the inflammasome, processing the proinflammatory cytokines IL-1ß and IL-18. A total of 270 RVVC patients and 583 healthy controls were analyzed, and increased diseases susceptibility was associated with the presence of the 12/9 genotype. Furthermore, functional studies demonstrate that IL-1ß production at the vaginal surface is higher in RVVC patients bearing the 12/9 genotype compared to controls, whereas IL-1Ra levels were decreased and IL-18 levels remained unchanged. These findings suggest that IL-1ß-mediated hyperinflammation conveyed by the NLRP3 gene plays a causal role in the pathogenesis of RVVC and may identify this pathway as a potential therapeutic target in the disease.

ALK translocation detection in non-small cell lung cancer cytological samples obtained by TBNA or EBUS-TBNA.

OBJECTIVE: As the diagnosis of non-small cell lung cancer (NSCLC) is based on cytology in around 70% of cases, it is important to use the same material for molecular analyses. Fluorescence in situ hybridization (FISH) is the only approved test for the detection of the translocation and inversion of anaplastic lymphoma kinase (ALK), but the optimal procedures for the fixation or staining of the sample before FISH evaluation have not been established. We investigated whether ALK gene status determined by FISH in a prospectively enrolled case series of patients was affected by fixation and staining. METHODS: One hundred and fifteen cytological samples were obtained by transbronchial needle aspiration (TBNA) or endobronchial ultrasound (EBUS)-TBNA from 109 patients with NSCLC. All samples were evaluated for epidermal growth factor receptor (EGFR) mutation by pyrosequencing and for ALK rearrangement by FISH. Specimens for ALK determination had been fixed with Cytofix(®) and/or Carnoy's solution or 10% formalin (cell blocks) and variously stained. RESULTS: Sixteen (14%) of the 115 samples were mutated for EGFR and 99 (86%) showed wild-type EGFR status. Of these 115 samples, 79 (69%) were negative for echinoderm microtubule-associated protein like 4 (EML4)-ALK translocation, nine (8%) were positive and 27 (23%) were unevaluable. In particular, 19 (26%) of the 72 Papanicolaou-stained smears fixed with Cytofix were unevaluable because of inadequate samples or cell overlapping; neither of the two May-Grünwald-Giemsa-stained samples were evaluable. Ten of 17 smears used for rapid on-site evaluation (ROSE) and immediately post-fixed in Carnoy's solution or 80% alcohol were evaluable. CONCLUSIONS: In this series, smears were unevaluable as a result of inadequate samples, cell overlapping or lack of fixation performed immediately after FNA.

Fine-tuning of Th17 Cytokines in Periodontal Disease by IL-10.

Periodontitis (PD) is a chronic disease caused by the host inflammatory response to bacteria colonizing the oral cavity. In addition to tolerance to oral microbiome, a fine-tuned balance of IL-10 levels is critical to efficiently mount antimicrobial resistance without causing immunopathology. Clinical and animal studies support that adaptive T-helper (Th) cytokines are involved in the pathogenesis of alveolar bone destruction in PD. However, it remains unclear what type of Th response is related to human PD progression and what role IL-10 has on this process. We addressed the contribution of IL-10 in limiting Th1 and Th17 inflammatory response in murine and human PD. Through a combination of basic and translational approaches involving selected cytokine-deficient mice as well as human genetic epidemiology, our results demonstrate the requirement for IL-10 in fine-tuning the levels of Th17 (IL-17A and IL-17F) cytokines in experimental and human PD. Of novelty, we found that IL-17F correlated with protection in murine and human PD and was positively regulated by IL-10. To our knowledge, this is the first demonstration of the protective role for IL-17F in PD, its positive regulation by IL-10, and the potential differential role for IL-17A and IL-17F in periodontal disease.

Role of BRAF molecular analysis in the management of papillary thyroid carcinoma: analysis of cytological and histological samples.

BACKGROUND: Although fine needle aspiration (FNA) is the standard diagnostic test for the characterization of a suspicious thyroid nodule, in some cases cytological evaluation is inconclusive. The aim of this study was to determine the role of BRAF mutation in aiding diagnosis and to verify whether archival cytological samples could be suitable for molecular analysis. METHODS: Eighty-five patients with suspicious (Thy4) or follicular (Thy3) lesions on cytology were resubmitted to a second FNA for BRAF mutation analysis. Of these, 56 subsequently underwent surgery. The usefulness of archival samples for molecular analysis was also studied in a second cohort of 42 patients with a confirmed diagnosis of papillary thyroid carcinoma for whom both archived paraffin-embedded histological samples and cytological smears were available. A further 15 patients with paired fresh FNA and archived cytological and histological samples were recruited. RESULTS: BRAF mutation was found in the fresh FNA samples from 10 of 56 patients who had surgery with previous inconclusive cytology (4/45, 9%, Thy3 and 6/11, 55%, Thy4). The BRAF test showed a specificity and positive predictive value of 100% (26/26 and 10/10, respectively), sensitivity of 33% (10/30) and negative predictive value of 57% (26/46). There was absolute concordance between the BRAF results obtained with 42 histological and cytological archived samples. BRAF analysis on 15 archived cytological samples showed absolute concordance with histology, whereas there was one false negative on the matched fresh FNA. CONCLUSION: BRAF analysis is a highly specific test that can facilitate cytological diagnosis in some cases and can also be performed on archived cytological samples.

New biomarkers to predict the evolution of in situ breast cancers.

BACKGROUND: Genomic studies have shown that gene expression profiles are similar in in situ (CIS) and invasive breast cancers, suggesting that several biofunctional modifications of the transformation process occur before or during the development of CIS lesion. METHODS: We investigated 3 biomarkers in 44 patients with CIS: TG2 (transglutaminase 2), HJURP (Holliday junction recognition protein), and HIF-1α (hypoxia inducible factor-1 alpha). RESULTS: TG2 was more highly expressed than the other two markers and significantly more so in stromal than in tumor cells. HIF-1α evaluation showed a higher expression in both tumor and stromal cells in patients with relapsed G3 tumors, indicating a potential role of this marker in CIS evolution. A greater than sevenfold higher risk of relapse (P = 0.050) was observed in patients highly expressing HJURP in stroma and a tenfold higher recurrence risk (P = 0.026) was seen in those with a higher stromal HIF-1α expression. An important increase in risk accuracy (AUC 0.80) was obtained when HIF-1α and HJURP were evaluated together. CONCLUSIONS: Despite the limited number of relapsed patients, we formulated some hypotheses on the factors responsible for malignant evolution and recurrence which are now being tested in a large case series with a longer follow-up.

Differences in biological features of breast cancer between Caucasian (Italian) and African (Tanzanian) populations.

Information on hormone receptor and human epidermal growth factor receptor-2 (HER2) expression in breast cancer is acknowledged as mandatory for prognostic stratification and treatment planning. Data on the biological features of African breast cancers are poor. We decided to compare histopathological and biomolecular characteristics (estrogen and progesterone receptor­ER, PgR, and HER2) of Tanzanian and Italian breast cancers. Differences in proliferating index and androgen receptor (AR) expression in triple-negative patients from the two case series were also assessed. Of the 103 consecutive patients seen at the Bugando Medical Center (Mwanza, Tanzania) from 2003 to 2010, who underwent biopsy or surgical resection of primary breast cancer, 69 patients had tissue samples that were evaluable for estrogen receptor (ER), progesterone receptor (PgR), and HER2. Histopathological assessment and biomolecular determinations were performed at the Cancer Institute of Romagna (IRST IRCCS, Meldola, Italy). Caucasian breast cancers were randomly extracted from an electronic database and matched (1:2 ratio) for year of diagnosis and age at diagnosis. Median age of both populations was 51 years (range 27­84). With respect to Caucasian tumors, Tanzanian breast cancers at diagnosis more frequently showed high histological grade (mainly grade 3) (P = 0.03), advanced clinical stage (III or IV) (P\0.001), ER negativity (52.2 %, P\0.001) and high proliferation (P = 0.0002). Triple-negative tumors were over-represented in Tanzanian women. AR was positive in 38.5 and 38 %of triple-negative Tanzanian and Italian breast cancers, respectively. Our results show that histopathological and biomolecular characteristics in Tanzanian and Italian breast cancers differ substantially. The high frequency of poorly differentiated, ER-negative, highly proliferating tumors, together with advanced stage at presentation, could be considered as the main prognostic factors linked to the high mortality rates for breast cancer in the African population.

Charcoal tattoo localization for differentiated thyroid cancer recurrence in the central compartment of the neck.

Recurrence of differentiated thyroid cancer can often require further surgical options. Reoperations may carry significant risk of surgical complications; additionally, as the anatomy is subverted, there is the possibility of leaving residual neoplasm. In order to avoid such problems during reoperation for differentiated thyroid cancer recurrence, we have introduced the technique of preoperative ultrasound-guided tattooing localization of the lymphatic structure to be removed with a 4% solution of active charcoal. Using ultrasound guidance, the lesion is identified and 0.5-2 ml of colloidal charcoal is injected near the lesion. The extraction of the needle is accompanied by injection at constant pressure of other charcoal as to leave a trace of colouring along the path of the needle up to the skin. The preoperative injection was well tolerated in all cases. In the last 5 years, we have used this technique in 13 patients with suspected recurrence in the central compartment (all from papillary carcinomas). Postoperative ultrasound and histological examination confirmed the removal of the lesion in all patients; in one case, the lesion was a parathyroid cyst. Complications were observed in two of 13 (15.4%) cases (one transitory hypoparathyroidism, and one transitory vocal cord paresis). Considering our experience, charcoal tattoo localization can be considered a safe, low-cost technique that is extremely useful for facilitating surgical procedures, and reduces the risk of iatrogenic damage.

Human anatomy in wax during Florentine enlightenment.

The high mortality rate of mothers and children during delivery was not taken into consideration by science before the end of the XVII Century; gynaecology became an official discipline in the Schools of Medicine at the beginning of the next century. The knowledge of anatomy was fundamental for doctors but human cadavers were still difficult to obtain; thus the need of models for the teaching. Anatomical models were obtained by injection of salts and alcohol on bodies or part of bodies: the procedure was difficult and the preparation did not last for a long time. Models in wax proved to be scientifically reliable, and everlasting. Schools of anatomical wax modelling were established in Italy: first in Bologna and later in Florence at the Museum "La Specola", a laboratory which soon became famous all through Europe. Description follows of the technique and the material used for the construction of the models; the scientists and the workers in the laboratory. Short notes on Felice Fontana, Paolo Mascagni, Giuseppe Galletti, Clemente Susini, Luigi Calamai, Egisto Tortori. A last chapter is devoted to the XVII Century Sicilian wax modeller Gaetano Giulio Zumbo, mainly renouned for the "Plague waxes" and some anatomical heads: most of his works are exhibited in the Museum La Specola in Florence.

[Dirofilaria (Nochtiella) repens in the spermatic cord: a new human case in Italy]. / Dirofilaria (Nochtiella) repens dans le cordon spermatique: un nouveau cas humain en Italie.

A case of human dirofilariasis associated with Dirofilaria (Nochtiella) repens, localised in the spermatic duct, is reported in a 52-year-old man from Lugo di Romagna (Ravenna province, Northern Italy). It represents the 22nd case of human dirofilariasis occurring until now in the same region. According to the relevant world literature this appears to be the 2nd case with this rare localisation the first one occurring in Italy as well.

[The role of measures of personal hygiene in the prevention of infections in autologous bone marrow transplantation. Results of a randomized prospective study]. / Ruolo delle pratiche igieniche personali nella prevenzione degli episodi infettivi nel trapianto di midollo osseo autologo. Bisultati di una sperimentazione prospettica randomizzata.

The paper reports on the results of a prospective, randomized trial in which two different regimens of personal hygiene procedures have been compared in 30 patients undergoing autologous bone marrow transplantation. The main aim of the study was to evaluate the incidence of infectious episodes in patients enrolled in the trial arm with the simplified hygiene protocol as compared to the standard one, used in our Unit for patients undergoing allogenic transplant. No significant differences between the two study groups in the incidence of infectious episodes or of positive skin cultures were found. Therefore, we suggest that a simplified hygiene protocol is as effective as the standard one in preventing infectious complications in patients undergoing autologous bone marrow transplantation.

Effectiveness of hardiness, exercise and social support as resources against illness.

The effects of the resistance resources of personality hardiness, exercise, and social support, taken singly and in combination, on concurrent and prospective levels, and probability of illness were studied. In 1980, 85 male business executives identified as high in stressful events were tested for the three resistance resources. Predicting their illness scores in 1980 formed the concurrent aspect of the study. For the prospective aspect, illness scores in 1981 were available on 70 of the subjects. With regard to resistance resources, when there are none, one, two or three, the level and probability of both concurrent and prospective illness drop in a regular and marked fashion. These results highlight the importance of multiple resistance resources. Estimates of relative effectiveness indicate that hardiness is the most important of the resistance resources studied.

Personality and social resources in stress resistance.

This study examined personality, social assets, and perceived social support as moderators of the effects of stressful life events on illness onset. In a group of 170 middle and upper level executives, personality hardiness and stressful life events consistently influenced illness scores, the former serving to lower symptomatology, the latter to increase it. Perceived boss support had its predicted positive effect. Executives under high stress who perceived support from their supervisors had lower illness scores than those without support. Perceived family support, on the other hand, showed a negative effect on health when reported by those low in hardiness. Finally, social assets made no significant impact on health status. These results underscore the value of differentiating between kinds of social resources, and of monitoring the effects of two or more stress-resistance resources in a single study.

Personality and exercise as buffers in the stress-illness relationship.

This study examined exercise and personality-based hardiness as independent buffers of the stressful event-illness relationship. Self-report measures of exercise, hardiness, stressful events and illness were obtained from 137 male business executives. Hardiness and exercise each interact with stressful events in decreasing illness. Further, subjects high in both hardiness and exercise remain more healthy than those high in one or the other only. These additive effects are consistent with the view that hardiness buffers by transforming the events themselves so as to decrease their stressfulness, whereas exercise buffers by decreasing the organismic strain resulting from experiencing stressful events.