Association between endometrial senescent cells and immune cells in women with repeated implantation failure.

PURPOSE: The aim of this study was to compare women with recurrent implantation failure (RIF) and control group in terms of the associations between p16-positive senescent cells and certain types of immune cells in human endometrium during the mid-luteal phase METHODS: Immunohistochemical staining was performed in 116 endometrial biopsies taken from 57 women presenting RIF, and control group of 59 women who became pregnant after the first intracytoplasmic sperm injection. Endometrial tissue sections were stained immunohistochemically for p16 (Senescent cells), CD4 (T-helpers), CD8 (T-killers), CD14 (Monocytes), CD68 (Macrophages), CD56 (Natural killers), and CD79α (B-cells). The percentage of positively stained cells for each marker was calculated by HALO image analysis software. The quantity and the relationship between senescent cells and immune cells were assessed and compared between the two groups. RESULTS: The correlation coefficient was highest between senescent cells and CD4+ cells and was lowest between senescent cells and CD14+ cells in RIF women, similarly to the control group. However, most of the observed correlations among senescent and immune cells weaken notably or disappear in the RIF group. When comparing senescent cell-to-immune cell quantitative ratios, only p16+/CD4+ cell ratio was significantly higher in RIF women as compared with patients from the control group. CONCLUSION: Our study indicates that the quantity of senescent cells in human endometrium during the mid-luteal phase has the strongest association with the amount of T helpers. Moreover, the specificity of this association might have an important impact on the occurrence of RIF.

Interviewing children: the impact of the COVID-19 quarantine on children's perceived psychological distress and changes in routine.

BACKGROUND: The COVID-19 outbreak has resulted in governments implementing disease containment measures such as school closures, social distancing, and home quarantine. To date, only a few studies have drawn attention to the psychological impact of lockdown on Italian children's mental health. The present study aimed to investigate the psychological distress (anxiety and mood symptoms) and perceived changes in routine among Italian primary and middle school students during the COVID-19 quarantine. METHODS: This interview study was performed between the 18th of May and 7th of June 2020: it involved a sample of 82 children and adolescents living in Milan (Italy), attending primary and middle school (aged 6 to 14 years), and their parents. RESULTS: Almost 30 % of the subjects reported having struggled to adjust to home learning. 36 responders completely changed their dietary habits during the lockdown: they were not eating the same amount of food and were consuming more junk food. Sleep habits were also affected by the lockdown measures: 28 % of the sample had difficulties sleeping and wished to sleep in their parents' bed. Concerning psychological distress, 64 (78 %) children and adolescents had anxiety symptoms; 43.9 % of the students reported significant mood symptoms. CONCLUSIONS: Children are not indifferent to the dramatic impact of the COVID-19 epidemic: our data confirm their difficulties in adapting to the quarantine measures. The effects of stress exposure may not manifest later on during the children's development, and, for this reason, it would be interesting to follow up on these participants to improve our understanding of how long these outcomes may last.

Use of granulocyte colony-stimulating factor for the treatment of unexplained recurrent miscarriage: a randomised controlled trial.

BACKGROUND: Recurrent miscarriage (RM) is defined as the occurrence of three or more clinically detectable pregnancy losses in the first trimester. In most cases of RM, its aetiology remains unexplained. Granulocyte colony-stimulating factor (G-CSF), a cytokine, and its receptor are expressed in placental tissue. To investigate the effectiveness of G-CSF in preventing embryo demise, we administered G-CSF to women with RM. METHODS: A randomised controlled trial in women with RM treated with G-CSF or placebo was conducted in one private reproductive medicine clinic. Sixty-eight women with unexplained primary RM, all with at least four consecutive miscarriages and negative for all clinical investigations, were selected. Patients were randomized for s.c. treatment with G-CSF (n = 35) (1 microg/kg/day) starting on the sixth day after ovulation, or with placebo (n = 33). Patients were randomized using a computer-generated randomization number sequence. Pregnancy outcome (delivery of a healthy baby without major or minor malformations) was the primary outcome measure. RESULTS In the group treated with G-CSF, 29 out of 35 (82.8%) women delivered a healthy baby, whereas in the placebo group, this figure was only 16 out of 33 (48.5%) (P = 0.0061, odds ratio = 5.1; 95% confidence interval 1.5-18.4). Significantly higher beta-hCG levels were found in gestation weeks 5-9 in women treated with G-CSF versus placebo (P < 0.001). CONCLUSIONS: Our data show that G-CSF may be effective in the treatment of unexplained RM. However, further studies are needed to confirm the effectiveness of this treatment in women with unexplained RM, refractory to conventional treatment. The study was registered with a ICMJE recognized registry, the Clinical Trial.gov Protocol Registry System, with the number NCT00772122.

[Myoepithelial differentiation markers in salivary gland neoplasia]. / Marcatori di differenziazione mioepiteliale nelle neoplasie delle ghiandole salivari.

Salivary gland tumors frequently present myoepithelial cell differentiation that is not always easily identified on routinely stained sections. Recently novel markers of myoepithelium have been studied, such as calponin (CALP), caldesmon (CALD), and smooth muscle myosin heavy chain. These markers, together with smooth muscle actin may be useful tools for identifying myoepithelial cells. We immunohistochemically studied a series of 23 benign and malignant salivary gland tumors using antibodies to these four markers. The tumors were classified as follows: pleomorphic adenoma (n = 8), basal cell adenoma (n = 3), myoepithelioma with plasmacytoid cells (n = 2), epithelial-myoepithelial cell carcinoma (n = 6) and adenoid cystic carcinoma (n = 4). All tumors were positive for at least one of the four markers. CALP and smooth muscle actin were the markers more frequently expressed. Positivity was mostly located in the myoepithelial cells that constitute the external layer of the glandular or tubular neoplastic structures. In poorly differentiated epithelial myoepithelial carcinomas, composed of solid sheets of neoplastic cells and sometimes of clear cells, immunohistochemical staining for myoepithelial markers evidenced rudimentary glandular structures. CALP and smooth muscle actin were positive in the two cases of myoepithelioma with plasmacytoid cells. In conclusion, the combined staining with four markers helps to disclose myoepithelial cell differentiation and can be a useful tool for the correct histopathological diagnosis of salivary gland tumors. Among the four markers studied, CALP and smooth muscle actin were the most useful to identify myoepithelial cell differentiation.

Progesterone receptor expression in orbital cavernous hemangiomas.

Orbital cavernous hemangiomas (OCH) have thick and highly cellular vascular walls. Ultrastructural studies have demonstrated the smooth muscle nature of these cells. Vascular neoplasms can modify their morphological and clinical features under hormonal stimulation. The purpose of the present study was to investigate the presence of smooth muscle markers and sex steroid receptors in 12 cases of OCH. Orbital cases were compared with cutaneous hemangiomas and subcutaneous angioleiomyomas. Smooth muscle actin (SMA) and desmin were localized in spindle cells of the vascular walls of all 12 cases studied. OCH showed immunohistochemical positivity with progesterone receptor (PR) antibody both in smooth muscular and in endothelial cells. For comparison, sex steroid receptors were studied in 10 cases of cutaneous cavernous hemangioma and in 10 cases of subcutaneous angioleiomyoma. PR was found in smooth muscle and endothelial cells of 6 out of 10 cases of subcutaneous angioleiomyoma and in none of the cases of cutaneous cavernous hemangioma. No positivity was obtained with estrogen receptor (ER) antibody in any of the cases tested. The present data suggest that OCH share morphological and immunohistochemical features with subcutaneous angioleiomyomas. Furthermore, immunohistochemical positivity with PR antibody indicates that OCH have to be added to the list of mesenchymal lesions that express sex steroid receptors.

[Malignant myoepithelioma associated with in situ and invasive ductal carcinoma. Description of a case and review of the literature]. / Mioepitelioma maligno associato a carcinoma duttale in situ ed invasivo NAS. Descrizione di un caso e revisione della letteratura.

A case of malignant myoepithelioma of the breast, associated with in situ and invasive carcinoma NOS is described. Myoepithelial differentiation was demonstrated with immunohistochemistry and electron microscopy. The tumour affected the left breast of a 72 year old lady. The patient had been treated with quadrantectomy with axillary dissection, followed by radiotherapy. At the time of diagnosis no local or distant metastases were found. Bone, pulmonary and cerebral metastases appeared 28 months after treatment. Malignant myoepitheliomas share histological and immunohistochemical features with monophasic sarcomatoid carcinomas. Comparison and and possible relationship with monophasic sarcomatoid carcinomas is discussed.

[Subcutaneous myolipoma with bizarre cells: morphological, immunohistochemical and ultrastructural study of a case and review of the literature]. / Miolipoma sottocutaneo con cellule bizzarre: studio morfologico, immunoistochimico e ultrastrutturale di un caso e revisione della letteratura.

A new case of subcutaneous myolipoma is reported. Grossly it appeared as a yellow-whitish nodule of 5.5 cm, in its major axis and showed well defined borders. Histologically the tumour was constituted by two components: areas of mature fat tissue were interminged with more cellular areas composed of bundles of spindle shaped eosinophilic cells, reminiscent of smooth muscle cells. In these latter component, cells showing multilobated, bizarre nuclei were also focally evident. No areas of necrosis, nor mitosis were found in both the components of the lesion. The immunohistochemical and ultrastructural findings revealed the myofibroblastic nature of the cellular myoid component of the lesion. The presence of myoid cells with bizarre, multilobated nuclei was considered expression of a regressive phenomenon, as observed in the so called "bizarre" leiomyoma of the uterus. Two years after the initial diagnosis the patient is well and free of disease. Soft tissue myolipoma is a benign lesion which has to be distinguished from lesions with malignant or uncertain biologic behaviour. The clinicopathologic features of the present case are discussed and compared to those of the previously reported cases of soft tissue myolipoma.

Differential expression of IGF-I and IGF-II in eutopic and ectopic endometria of women with endometriosis and in women without endometriosis.

PROBLEM: Factors regulating the development, growth, and differentiation of endometrial cells of endometriotic lesions are poorly understood. To investigate the paracrine-autocrine regulation of ectopic endometrial cell growth, the expression of IGF-I and IGF-II were studied. METHOD: Tissue specimens of eutopic and ectopic endometria were obtained from eight patients with endometriosis at laparoscopy and from the endometria of 14 women without endometriosis as controls. They were tested for the expression of IGF-I and IGF-II by immunohistochemical analysis. RESULTS: Immunohistochemical study for IGF-I in controls showed a more intense staining during the proliferative phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis a reduction in the staining was observed, whereas in epithelial cells of fibrotic peritoneal adhesions an intense immunostaining for IGF-I was observed. Immunohistochemical study of IGF-II in controls showed a more intense staining during secretory phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis, a reduction in the staining was observed, whereas in epithelial cells of fibrotic peritoneal adhesions an intense immunostaining for IGF-I was observed. Immunohistochemical study of IGF-II in controls showed a more intense staining during secretory phase both in stromal and epithelial cells. In eutopic endometria of women with endometriosis, a reduction in the staining was observed, whereas in epithelial cells of ovarian lesions and fibrotic peritoneal adhesions, no immunostaining for IGF-II was observed. CONCLUSIONS: In endometriosis there is an alteration of mechanisms regulating cell proliferation and differentiation.

Systemic methotrexate treatment in early unruptured ectopic pregnancy.

Ectopic pregnancy is one of the most common and dangerous complications of the early pregnancy period. Until now diagnosis has been late because major symptoms occur after tubal rupture and so only demolitive surgery has been possible. At present, with the appearance of ultrasound in obstetrics an earlier diagnosis of this pathology can be made before tubal rupture so medical treatment has become possible. We treated a series of twelve patients with early ectopic pregnancy (9 tubal and 3 with no localized site of implantation) with intramuscular 0.5 mg/kg methotrexate and oral 0.1 mg/kg of folic acid (Citrovorum Factor) on alternate days, in the attempt to reduce hospitalization and obtain more effective and safer medical management. We observed a fall in serum beta-HCG levels after one cycle of treatment in 11 out of 12 patients and after two cycles of therapy in the remaining case. Minimal side-effects were observed in four cases. Three pregnancies occurred after treatment before the advised interval time and ended in blighted ovum. Methotrexate systemic therapy can be considered an elective treatment and a sufficiently safe management in early unruptured ectopic pregnancy when a good clinical selection of patients is performed.

[Sarcoid-like hepatic granulomas, associated with gastric neoplasia. Description of a case]. / Granulomi epatici sarcoido-simili, associati a neoplasia gastrica. Descrizione di un caso.

Giant cell granulomas in liver biopsies is a relative common finding. Among the many causes of granulomatous lesions of the liver primary biliary cirrhosis and sarcoidosis are the most frequently diagnosed. On the other hand sarcoid-like granulomatous reaction can be encountered associated to malignant tumours. Purpose of the present paper is to describe a case of a sarcoid-like reaction of the liver associated to gastric adenocarcinoma. The patient was a 66 yr old man who underwent gastrectomy for a signet-ring cell adenocarcinoma. Pathological anamnesis was unremarkable. Liver function tests were within normal limits. Chest x ray was normal. A liver biopsy was performed during surgery as the liver presented an irregular surface. On histology giant cell granulomas with sarcoid-like features were seen in the hepatic parenchyma. Same reaction was present in the perigastric lymph nodes. The patient died immediately after surgery due to massive pulmonary embolism. No autopsy was performed. Among the possible diagnoses primary biliary cirrhosis, sarcoidosis and paraneoplastic sarcoid-like granulomatous reaction were considered. Primary biliary cirrhosis and sarcoidosis were excluded on the basis of the past clinical history of the patient, that was unremarkable; furthermore liver function tests performed preoperatively were within normal ranges. Thus paraneoplastic sarcoid-like reaction involving the liver was regarded as the most likely diagnosis.