Severe endometriosis: low value of AMH did not affect oocyte quality and pregnancy outcome in IVF patients.

OBJECTIVE: The aim of this pilot study was to determine whether the low anti-müllerian hormone (AMH) serum level, due to severe endometriosis, was associated with diminished oocyte yield, poor oocyte/embryo quality and reduced in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) clinical outcomes in young patients (<37 years old). PATIENTS AND METHODS: A total of 50 IVF cycles of patients younger than 37 with severe endometriosis were retrospectively analyzed in a single center between November 2016 and July 2018. The clinical outcome was then compared to a control group of 84 patients with no story of endometriosis and normal AMH value. AMH value was evaluated within three months before the stimulation. In these two groups, number and maturation of retrieved oocytes, embryo quality, and pregnancy outcomes were evaluated and compared using Student's t-test and Fisher's test. RESULTS: The number of oocytes retrieved per cycle and the percentage of mature oocytes (MII) were significantly lower (p < 0.001) in IVF patients with severe endometriosis and AMH value ≤ 1.1 ng/ml (Group A; 3.8±2.6 retrieved oocytes, 70% MII) compared to patients without endometriosis and AMH levels > 1.1 ng/ml (Group B; 6.9±4.6 retrieved oocytes, 83% MII). On the other hand, embryo morphology, implantation rate (31% vs. 33%; p = 0.833) and pregnancy rate (50% vs. 49%; p = 1) were comparable in the two groups. CONCLUSIONS: This study shows that younger patients with an impairment of the ovarian reserve due to severe endometriosis, displayed a diminished oocyte yield but not a reduction in embryo quality and pregnancy outcomes. These results suggest that serum AMH levels should not be adopted as a criterion for discouraging these patients from undergoing IVF/ICSI treatments.

Work related stress and blood glucose levels.

AIM: The aim of the study is to evaluate work-related subjective stress in a group of workers on a major Italian company in the field of healthcare through the administration of a valid "questionnaire-tool indicator" (HSE Indicator Tool), and to analyze any correlation between stress levels taken from questionnaire scores and blood glucose values. MATERIAL AND METHODS: We studied a final sample consisting of 241 subjects with different tasks. The HSE questionnaire - made up of 35 items (divided into 7 organizational dimensions) with 5 possible answers - has been distributed to all the subjects in occasion of the health surveillance examinations provided by law. The questionnaire was then analyzed using its specific software to process the results related to the 7 dimensions. These results were compared using the Pearson correlation and multiple linear regression with the blood glucose values obtained from each subject. RESULTS: From the analysis of the data the following areas resulted critical, in other words linked to an intermediate (yellow area) or high (red area) condition of stress: sustain from managers, sustain from colleagues, quality of relationships and professional changes. A significant positive correlation (p <0.05) between the mean values of all critical areas and the concentrations of glucose values have been highlighted with the correlation index of Pearson. Multiple linear regression confirmed these findings, showing that the critical dimensions resulting from the questionnaire were the significant variables that can increase the levels of blood glucose. CONCLUSION: The preliminary results indicate that perceived work stress can be statistically associated with increased levels of blood glucose.

Effect of transferring a morphologically impaired embryo with a good quality embryo on the pregnancy and implantation rates.

OBJECTIVE: This study aimed to investigate the effect of transferring embryos with different qualities on pregnancy and implantation rates. PATIENTS AND METHODS: In a retrospective multi-center study we analyzed 761 patients aged ≤ 35 years who had an elective transfer of one or two embryos. Embryos were scored morphologically by their developmental stage into good "A" and impaired "B" embryos. Pregnancy and implantation rates were compared between patients who had a transfer of: one grade "A" embryo; two grade "A" embryos, two embryos one grade "A" plus one grade "B" embryos; one grade "B" embryo and two grade "B" embryos. RESULTS: Higher pregnancy and implantation rates were observed in patients who had received one embryo of grade "A" (34.6%) and two grade "A" embryos (45.2%, 25.85% respectively), compared to patients who received two embryos, one of grade "A" plus one of grade "B" (25%, 13.77% respectively). CONCLUSIONS: Transferring a morphologically and developmentally impaired embryo, significantly lower the implantation chance of the good quality embryo.

Correlation between cadmium and blood counts in workers exposed to urban stressor.

The objective of this study was to assess the exposures to cadmium (Cd) in urban workers and the association between Cd exposure and values of blood counts. Urinary Cd, blood Cd, and blood counts were obtained from 355 outdoor workers; a subgroup of 99 subjects were monitored to evaluate personal exposure to airborne Cd. The mean value of personal exposure to Cd was 1.5 ng/m3 for traffic assistants and 1.2 ng/m3 for drivers. Urinary and blood Cd were correlated to the airborne Cd (respectively r=3 and r=4). The multiple linear regression models showed the associations among white blood cell, the percentage of neutrophils (NEU%), the percentage of lymphocytes (LYM%), and the concentrations of blood Cd (respectively R2=27, R2=37, R2=581). The subjects with blood Cd values higher than 1.2 µg/L showed an increase of LYM% mean values and a decrease of NEU% mean values with respect to the group with blood Cd values lower than 1.1 µg/L.

First case of transformation for breast fibroadenoma to high-grade malignant phyllodes tumor in an in vitro fertilization patient: misdiagnosis of recurrence, treatment and review of the literature.

INTRODUCTION: Cystosarcoma phyllodes are very rare tumors and may be difficult to diagnose clinically. BACKGROUND: Fibroadenomas have long been considered benign hyperplastic lesions rather than true neoplastic processes. However, previous clonality studies have shown differing results. AIM: to assess diagnostic and treatment options for phyllodes tumor. MATERIALS AND METHODS: A 41-year-old female patient undergoing assisted fertilization treatment. The patient underwent fine needle aspiration biopsy that confirmed fibroadenoma before the IVF attempt. At 17 weeks of gestation, due to an increase in volume of the fibroadenoma, an excisional biopsy was performer that showed a malignant phyllodes tumor. Then she underwent quadrantectomy and chemiotherapy After 1 year there was a recurrence of phyllodes tumors and she underwent mastectomy and chemotherapy. RESULTS: Fibroadenoma that was transformed into high-grade malignant cystosarcoma after ovarian stimulation, relapsed after one year and it was not immediately diagnosed. The patient underwent mastectomy and chemotherapy. DISCUSSION: it is difficult to diagnose recurrence and to determine tele frequency and the right treatment for such a rare cancer, so it is important to report any case in the literature. CONCLUSIONS: We recommend to remove a fibroadenoma before attempting IVF for the risk of malignant transformation.

Simultaneous administration of human acidic and recombinant less acidic follicle-stimulating hormone for ovarian stimulation improves oocyte and embryo quality, and clinical outcome in patients with repeated IVF failures.

BACKGROUND: Ovarian stimulation is an integral procedure in assisted reproduction treatment. It is achieved by the administration of exogenous gonadotropins to increase follicular recruitment and oocyte yield. Optimization of ovarian stimulation is an essential prerequisite for the success of IVF treatment. AIM: This study aimed to evaluate the effect of a combined stimulation protocol of human FSH and recombinant FSH, simultaneously administered, on oocyte and embryo quality and clinical outcome. PATIENTS AND METHODS: In a prospective randomized study 197 infertile patients with a history of previous IVF failures for at least 3-5 attempts, were enrolled for an in vitro fertilization treatment. All patients had a standard down-regulation with GnRH analog and were then stimulated with FSH. The patients were matched into three groups: group A (no = 66) received human FSH combined with recombinant FSH in equal doses, simultaneously administered; group B (no = 67) received human FSH alone and group C (no = 64) received recombinant FSH alone. RESULTS: There were significantly higher pregnancy (p < 0.04) and implantation rates (p < 0.03) in favor of group A (hFSH/rFSH) compared to groups B (hFSH) and C (rFSH). A significant increase in the proportion of mature metaphase II oocytes (p < 0.002) and grade 1 embryos (p < 0.03) was observed in group A with respect to group B and C. Significantly higher delivery rate (p < 0.01) was achieved in group A compared to groups B and C. No significant differences were observed between groups regarding miscarriage rate and risk of ovarian hyperstimulation syndrome. CONCLUSIONS: The results show that the combination of human and recombinant FSH for ovarian stimulation may produce a positive effect on follicular development as it improve oocyte quality, embryo development, and ultimately clinical outcome.

Primary cutaneous B-cell lymphoma is associated with somatically hypermutated immunoglobulin variable genes and frequent use of VH1-69 and VH4-59 segments.

BACKGROUND: Accurate assessment of the somatic mutational status of clonal immunoglobulin variable region (IgV) genes is relevant in elucidating tumour cell origin in B-cell lymphoma; virgin B cells bear unmutated IgV genes, while germinal centre and postfollicular B cells carry mutated IgV genes. Furthermore, biases in the IgV repertoire and distribution pattern of somatic mutations indicate a possible antigen role in the pathogenesis of B-cell malignancies. OBJECTIVES: This work investigates the cellular origin and antigenic selection in primary cutaneous B-cell lymphoma (PCBCL). METHODS: We analysed the nucleotide sequence of clonal IgV heavy-chain gene (IgVH) rearrangements in 51 cases of PCBCL (25 follicle centre, 19 marginal zone and seven diffuse large B-cell lymphoma, leg-type) and compared IgVH sequences with their closest germline segment in the GenBank database. Molecular data were then correlated with histopathological features. RESULTS: We showed that all but one of the 51 IgVH sequences analysed exhibited extensive somatic hypermutations. The detected mutation rate ranged from 1.6% to 21%, with a median rate of 9.8% and was independent of PCBCL histotype. Calculation of antigen-selection pressure showed that 39% of the mutated IgVH genes displayed a number of replacement mutations and silent mutations in a pattern consistent with antigenic selection. Furthermore, two segments, VH1-69 (12%) and VH4-59 (14%), were preferentially used in our case series. CONCLUSIONS: Data indicate that neoplastic B cells of PBCBL have experienced germinal centre reaction and also suggest that the involvement of IgVH genes is not entirely random in PCBCL and that common antigen epitopes could be pathologically relevant in cutaneous lymphomagenesis.

Correlation between squamous intraepithelial lesions (SILs) and bacterial vaginosis.

Bacterial vaginosis (BV) is a condition that seldom occurs in prepuberal girls or postmenopausal women, suggesting a hormonal component in its aetiology. The precise mechanisms by which BV arises are not fully understood. One proposed mechanism suggests that carcinogenic nitrosamines act either independently or via human papilloma virus (HPV). Human papillomavirus is known to be associated with the development of squamous intraepithelial lesion (SIL). Still today the relationship between BV and SIL is debated. Many confounding factors regarding the relationship between BV and SIL include the presence of HPV and/or other sexually transmitted diseases. In a case-controlled study the correlation between BV, SIL and the presence of HPV was evaluated. BV was diagnosed according to standard criteria: vaginal pH > 4.5; positive amine test or 'whiff' test; presence of clue cells and abnormal discharge. High risk-HPV testing by PCR was performed. X2 Pearson analysis was applied for statistical evaluation of data. The results of the study have shown that BV is not associated with SIL.

Expression of TCL1 and CD27 in primary cutaneous B-cell lymphomas.

AIMS: To investigate by immunohistochemical analysis the expression of the TCL1 oncogene product and of CD27 in 25 cases of primary cutaneous B-cell lymphomas (PCBCL) classified according to the World Health Organization-European Organization for Research and Treatment of Cancer classification of cutaneous lymphomas. In B-cell ontogenesis TCL1 is mainly expressed by 'naive' B lymphocytes and by a subset of germinal centre B cells, whereas CD27 is expressed by a subset of germinal centre B cells, 'memory' B lymphocytes and plasma cells, suggesting that their expression in physiological conditions is mutually exclusive. METHODS AND RESULTS: Overall, TCL1 was expressed in 5/25 cases (20%) and CD27 in 15/25 cases (60%). Furthermore, 7/25 cases (28%) were TCL1- and CD27- and 2/25 cases (8%) were TCL1+ and CD27+. In particular, primary cutaneous follicle-centre lymphomas (10 cases) showed a variable expression of both TCL1 and CD27, whereas primary cutaneous marginal-zone B-cell lymphomas (eight cases) showed, with the exception of a single case, a definite CD27+/TCL1- profile. CONCLUSIONS: These findings indicate: (i) the TCL1 oncogene product is uncommonly expressed in PCBCL (20% of cases, mainly of the follicle-centre subtype); (ii) in contrast, CD27 is often expressed in PCBCL (60% of cases), mainly of the marginal-zone subtype; (iii) the coexpression of TCL1 and CD27 may be seldom observed in PCBCL (8% of cases); (iv) PCBCL does not seem to show, in terms of either TCL1 or CD27 expression, significant differences compared with its systemic counterparts.

[Sentinel node biopsy for malignant melanoma. Technical details and clinical results on 390 patients]. / La biopsia del linfonodo sentinella nel melanoma maligno. Dettagli tecnici e risultati in 390 pazienti.

BACKGROUND: The purpose of this paper is to present personal experience with sentinel node biopsy for the treatment of malignant melanoma. Technical details influencing the efficacy of the procedure are presented and the clinical, therapeutic and prognostic advantages of this technique discussed. METHODS: A total of 390 consecutive patients with primary skin melanoma (T2-3,N0,M0) underwent sentinel node biopsy between March 1996 and May 2001. All patients underwent previous excisional biopsy of the primary lesion and clinical and radiographic examination to exclude lymphatic or systemic macroscopic spreading of the disease. Preoperative lymphoscintigraphy (99mTc nanocoll) was routinely performed in the last 315 patients. Intraoperative detection of the sentinel nodes was performed by perilesional, intradermical, injection of blue dye associated with a g probe (Neoprobe(R) 2000) in the last 315 patients. Sentinel nodes, serially sectioned, were all Haematoxylin-Eosin and immunohistochemically stained. All patients positive for micro-metastasis underwent radical lymphadenectomy. Comparative analysis between the incidence of metastasis in sentinel and non-sentinel nodes, according to the clinical stage of the disease, was done. RESULTS: The overall detection rate of sentinel nodes was 97.4%. Relevant differences were found according to the site of dissection and the use of a g probe. The g-probe makes the procedure more effective, less invasive and less expensive. Timing and accuracy of the preoperative lymphoscintigraphy is a basic step of the procedure. The overall incidence of positive sentinel nodes was 14.7% with differences correlated with thickness of primary lesion (0.75-1.5 mm: 5,8%; 1.5-3 mm:18%; 3-4 mm: 24.6%). Metastasis in other non-sentinel nodes was found only with primary tumour thickness exceeding 2 mm. CONCLUSIONS: Sentinel node biopsy is a procedure requiring a multidisciplinary approach (surgery, nuclear medicine and pathology). A specific learning phase (>30 patients) is recommended to obtain reliable results.

[Immune tolerance in pregnancy. Maternal-fetal interactions]. / La tolleranza immunitaria in gravidanza. Interazioni materno-fetali.

The conceptus may be considered as a sort of semi-allogenic graft for the maternal organism, since it shares a half of genomic complement with the father. Nevertheless, its rejection does not take place physiologically during a pregnancy. The mechanisms resulting in the maternal immune tolerance versus the conceptus are not yet completely clarified. Such mechanisms are probably multiple and interacting with each other. In animal and in vitro studies provide evidence suggesting that the following factors are important in producing the maternal immune tolerance: the anatomical position of the fetus; the absence of expression of the class I and II Major Histocompatibility Complex (MHC) molecules in trophoblast tissues; the activity of blocking antibodies; a modification of the immune response; the fetal-placental production of immunosuppressive hormones and substances. Amongst pregnancy-related changes in the immune response, a reduced Natural Killer (NK) cell activity and an increased synthesis of Th2 cytokines (which inhibit the cell-mediated immunity) with an altered Th1/Th2 balance appear to be remarkably important. With regard to fetal-placental hormones, progesterone seems to exert an important immunosuppressive influence mediated by the protein named "Progesterone Induced Blocking Factor" (PIBF). Nevertheless, the real contribution of each of the above mentioned mechanisms still remains to be elucidated in humans.

Pathogenetic and clinical implications of Bcl-6 and Bcl-2 gene configuration in nodal diffuse large B-cell lymphomas.

Bcl-6 (LAZ-3) and Bcl-2 gene rearrangements have been respectively reported in 20-35 per cent and 10-25 per cent of diffuse large B-cell lymphomas (DLBCLs). Although these genetic lesions have been associated with different clinical outcomes (i.e., more favourable in Bcl-6 rearranged cases and poorer in Bcl-2 rearranged cases), their prognostic significance is still controversial. In the present study, we have investigated by Southern blot analysis the Bcl-6 and Bcl-2 gene configuration in a series of 80 lymph nodes involved by well-characterized DLBCLs, histologically defined according to the REAL and the updated Kiel classifications. The molecular findings have been correlated with the clinical features at presentation and with response to therapy. The majority of cases (57/80 = 71.2 per cent) had a centroblastic morphology. Bcl-6 rearrangements were detected in 23/80 cases (28.8 per cent), and were similarly associated with centroblastic (18/57 = 31.6 per cent) or immunoblastic (3/11 = 27.3 per cent) histotypes. In contrast, Bcl-2 was found to be rearranged in only three cases of centroblastic lymphoma (3.8 per cent). No significant differences were found between Bcl-6 rearranged and germline cases, as far as the clinical features at presentation are concerned. Forty-one patients, in whom the lymph node biopsy was performed at diagnosis, could be evaluated for response to treatment and clinical outcome. Most of these cases (30/41 = 73.2 per cent) were nodal DLBCL, without extranodal site involvement. Analysis of the clinical outcome showed no statistically significant differences between Bcl-6 rearranged and Bcl-6 germline cases (actuarial overall survival 50 per cent vs. 48 per cent, event-free survival 45 per cent vs. 46 per cent, at 4 years). These findings confirm that Bcl-6 rearrangements are the most frequent genetic lesion in DLBCL. The incidence of Bcl-2 involvement in our series is significantly lower than the figures reported in other studies, mainly from North American countries, probably reflecting heterogeneous patient selection and/or epidemiological variability. Finally, our results suggest that no relevant clinical differences are observed between Bcl-6 rearranged and Bcl-6 germline cases, when nodal DLBCLs are considered.

Simultaneous occurrence of large B-cell non-Hodgkin lymphoma and myelodysplastic syndrome rapidly evolving into acute myeloblastic leukemia.

We describe a case of simultaneous occurrence of large B-cell non-Hodgkin lymphoma and myelodysplastic syndrome in the absence of previous chemotherapy or radiotherapy. After initiation of steroid treatment, the myeloid clone showed a rapid increase in both the bone marrow and peripheral blood with transformation into acute myeloid leukemia. The diagnosis were confirmed by immunophenotypic studies performed in the histologic sections of the lymph node, as well as in bone marrow and peripheral blast cells. This case may be indicative of potential down-regulation of a malignant myeloid clone induced by the malignant lymphoid clone.

Analysis of the BCL-6 gene configuration in diffuse B-cell non-Hodgkin's lymphomas and Hodgkin's disease.

BCL-6 is a novel proto-oncogene that codes for a zinc-finger protein sharing homologies with many transcription factors. It has recently been shown that BCL-6 is involved in chromosome band 3q27 aberrations in non-Hodgkin's lymphomas (NHLs) and BCL-6 rearrangements have been detected in 34-45 per cent of diffuse large cell lymphomas with B immunophenotype. We have studied the BCL-6 gene configuration by Southern blot analysis in 60 cases of B-cell NHL and in 17 cases of Hodgkin's disease (HD). BCL-6 was rearranged in 15/46 (32.6 per cent) diffuse B-large cell lymphomas, mainly with centroblastic morphology, and in 2/11 (18.2 per cent) follicular (centroblastic-centrocytic) lymphomas. Conversely, all cases of HD, including four cases of lymphocyte predominant, nodular type (nodular paragranuloma), had a germline configuration. These findings confirm that BCL-6 is rearranged in a significant percentage of diffuse B-large cell lymphomas, suggesting that this proto-oncogene might have a pathogenetic role in this subset of NHLs, but our preliminary analysis suggests that BCL-6 lesions are not involved in the pathogenesis of HD. However, further investigations using more sensitive techniques are required to confirm these findings.

Relapsing bilateral uveitis and papilledema in sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease).

Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) is a non-hereditary histiocytic proliferative disorder affecting young people, with extranodal manifestations in 28-43% of cases. Eye involvement is infrequent. Lymphoproliferation in the soft tissues of the orbit and in the lids has been reported in 12% of cases but intraocular involvement is rare. We describe the case of a 12-year-old boy affected by Rosai-Dorfman disease with bilateral relapsing uveitis and papilledema that appeared four years before the onset of lymphadenopathy.

Multinucleated giant cells generation induced by interferon-gamma. Changes in the expression and distribution of the intercellular adhesion molecule-1 during macrophages fusion and multinucleated giant cell formation.

BACKGROUND: Multinucleated giant cells (MGC), interferon-gamma (IFN-gamma) production, and increased expression of adhesion molecules are features of granulomatous reactions. IFN-gamma induces the fusion of macrophages and the subsequent MGC generation in vitro. Moreover, IFN-gamma increases ICAM-1 expression on lymphoid cells and an important role for adhesion molecules in MGC generation has been proposed. EXPERIMENTAL DESIGN: The time course of the IFN-gamma-driven MGC generation was investigated in slide-chamber cultures of adherence-purified human monocytes. The fusion index, the monocytes clustering the total number of MGC were determined. The expression of intercellular cell adhesion molecule-1 (ICAM-1), LFA-1 and HLA-DR was investigated by immunohistochemistry. The effect of anti-ICAM-1, anti-LFA-1 and anti-HLA-DR monoclonal antibodies on IFN-gamma-induced MGC generation was also examined. RESULTS: IFN-gamma enhanced the generation of MGC in a dose- and time-dependent fashion. In all experiments, MGC formation was preceded by a sequence of changes in the morphology of cultured monocytes. Cell clustering occurred as early as 3 days after IFN-gamma stimulation and was followed by the adhesion of cells that eventually fused. Immunohistochemistry showed that ICAM-1 was increased by IFN-gamma and constantly polarized on a cell uropode. When monocytes clustered, ICAM-1 was localized on the membrane where the cell-to-cell contact occurred. In newly formed MGC, ICAM-1 stained in the center of the giant cell. The cellular distribution of LFA-1 on cultured monocytes was not modified by IFN-gamma. HLA-DR expectedly enhanced by IFN-gamma was mostly cytoplasmic and tended to disappear when MGC formed. Finally, anti-LFA-1 and anti-ICAM-1 monoclonal antibodies variably inhibited IFN-gamma-induced MGC generation. CONCLUSIONS: Taken together, these data add support to the concept that IFN-gamma is essential for MGC generation by promoting cell clustering and cell-to-cell adhesion. The present data also indicate that among the mechanisms by which IFN-gamma exert such a promoting effect, changes in the ICAM-1 expression and cellular distribution are included. The observation that ICAM-1 appears to be trapped in the cytoplasm of IFN-gamma-driven MGC and that HLA-DR tend to disappear from macrophages undergoing MGC formation, also suggest changes in the functional properties of these cells.

Peripheral neuroectodermal tumor of the chest (Askin tumor) as secondary neoplasm after Hodgkin's disease: a case report.

The authors present the case of a 20-year-old woman who developed a peripheral neuroectodermal neoplasm of the thoracopulmonary region (Askin tumor) 7 years after achieving complete remission of stage-IV Hodgkin's disease. The previous treatment had consisted of eight courses of alternating MOPP/ABVD combined with involved-field 20-Gy radiotherapy. The second neoplasm appeared in a nonirradiated area of the chest wall, with erosion of the ribs as shown by sonography and computed tomography. The histological pattern was in accordance with a generic diagnosis of a malignant small cell tumor; the immunostaining positivity of the neoplastic cells for the neuron-specific enolase allowed us to make the diagnosis of a tumor with a neuroectodermal origin. Partial resection of the neoplasm and four courses of chemotherapy including adriamycin, cisplatin, and ifosfamide induced a complete remission, confirmed by surgical restaging. She is alive and well 10 months after the completion of therapy. The clinical, radiological, and microscopic features of this tumor occurring as a secondary neoplasm after Hodgkin's disease are described.