Leonardo Resta

Tissue remodelling in breast cancer: Human mast cell tryptase as an initiator of myofibroblast differentiation

Mangia A, Malfettone A, Rossi R, Paradiso A, Ranieri G, Simone G & Resta L
(2011) Histopathology58, 1096–1106
Tissue remodelling in breast cancer: human mast cell tryptase as an initiator of myofibroblast differentiation

Chronic Endometritis Due to Common Bacteria Is Prevalent in Women With Recurrent Miscarriage as Confirmed by Improved Pregnancy Outcome After Antibiotic Treatment

Recurrent miscarriage (RM) is defined as 3 or more miscarriages before 20 weeks’ pregnancy. In recent years, interest has been focused on chronic endometritis (CE), a subtle inflammation thought to be associated with RM. We aimed to evaluate the relationships between CE and RM. The records of 360 women with unexplained RM were retrospectively analyzed. Data from hysteroscopy, endometrial histology, endometrial culture, and polymerase chain reaction for chlamydia, performed before and after antibiotic treatment for CE, were analyzed. The occurrence of successful pregnancies within 1 year after treatment was also evaluated. Results showed that 208 (57.8%) women with RM showed CE at hysteroscopy; 190 (91.3%), positive at hysteroscopy, were also positive at histology, and 142 (68.3%) had positive cultures. Common bacteria were found in 110 (77.5%) patients. Mycoplasma and Ureaplasma were found in 36 (25.3%) patients and Chlamydia in 18 patients (12.7%). In 102 (71%) women, antibiogram-based antibiotic treatment normalized hysteroscopy, histology, and cultures (group 1); while in 40 (28.2%) patients, CE was still present at hysteroscopy (group 2). In 16 of the 66 patients positive at hysteroscopy, but not at cultures, the hysteroscopy becomes normal (group 3) after a Centers for Disease Control and Prevention-based therapy; while in 50 women, CE was still present (group 4). One year after treatment, group 1 showed a significantly higher number of pregnancies (78.4%) compared to group 2 (17.5%; P < .001) and group 4 (15.3%; P = .005). The CE is frequent in women with RM. Antibiotic treatment seems to be associated with an improved reproductive outcome.

AT1 angiotensin II receptor subtype in the human larynx and squamous laryngeal carcinoma

The presence of the angiotensin type 1 receptor (AT1 Ang II-R) was investigated in normal and diseased human larynx using a specific monoclonal antibody (6313/G2). When tissue AT1 content was studied by SDS electrophoresis with immunoblotting, the receptor was detected in 10/10 laryngeal tumours, and in 7/10 samples of normal tissue from the same patients. Two immunostaining bands, approximately 75 kDa, were present in all cases. Immunocytochemistry performed on sections of 45 formalin-fixed, paraffin-embedded laryngeal tissue samples showed that the receptor was expressed in normal respiratory epithelium only in a perinuclear pattern, above the nucleus toward the cell apex. In addition, the antigen was invariably present in skeletal muscle cells and in the columnar duct epithelium of minor salivary glands. The secretory cells were negative, but the antibody stained the adjacent myoepithelial cell layer. As expected, smooth muscle cells of the vessel walls also expressed Ang II-R. In metaplastic epithelium deriving from respiratory epithelium, the receptors were distributed diffusely throughout the cytoplasm of basal and parabasal cells. In dysplastic epithelium, cells of all layers were strongly positive. Finally, squamous cell tumours showed varying numbers of immunoreactive cells, which stained in a diffuse cytoplasmic and membranous pattern. Computer-assisted image analysis of the stained sections showed that the positivity for Ang II-R dramatically increased in dysplastic and well-differentiated cancer cells (3- and 5.5-fold higher than in normal epithelium, respectively), but there was less in poorly and very poorly differentiated cancer. Receptor abundance was not correlated with tumour size nor lymph node involvement. These results suggest a possible role of Ang II in the growth or function of normal and neoplastic larynx tissue, which is especially significant in early neoplastic change.

Placing mitochondrial DNA mutations within the progression model of type I endometrial carcinoma.

Mitochondrial DNA (mtDNA) mutations have been described in almost all types of cancer. However, their exact role and timing of occurrence during tumor development and progression are still a matter of debate. A Vogelstein-like model of progression is well established for endometrial carcinoma (EC), however, mtDNA has been scarcely investigated in these tumors despite the fact that mitochondrial biogenesis increase has been shown to be a hallmark of type I EC. Here, we screened a panel of 23 type I EC tissues and matched typical hyperplasia for mutations in mtDNA and in four oncosupressors/oncogenes, namely PTEN, KRAS, CTNNB1 and TP53. Overall, mtDNA mutations were identified in 69% of cases, while mutational events in nuclear genes occurred in 56% of the cases, indicating that mtDNA mutations may precede the genetic instability of these genes canonically involved in progression from hyperplasia to tumor. Protein expression analysis revealed an increase in mitochondrial biogenesis and activation of oxidative stress response mechanisms in tumor tissues, but not in hyperplasia, in correlation with the occurrence of pathogenic mtDNA mutations. Our results point out an involvement of mtDNA mutations in EC progression and explain the increase in mitochondrial biogenesis of type I EC. Last, since mtDNA mutations occur after hyperplasia, their potential role in contributing to genetic instability may be envisioned.

Poor reliability of vaginal and endocervical cultures for evaluating microbiology of endometrial cavity in women with chronic endometritis.

Background: Chronic endometritis (CE) is a subtle pathology causing infertility and abnormal uterine bleeding. We evaluated the reliability of vaginal and cervical cultures for detecting infectious agents at the endometrial level. Methods: In a prospective diagnostic study, 181 women diagnosed with CE and 100 controls underwent vaginal, endocervical and endometrial sampling. Cultures for common bacteria, Neisseria gonorrhoeae, yeast and Ureaplasma urealyticum and PCR for Chlamydia trachomatis were performed. Results: The prevalent infectious agents at the endometrial level were common bacteria(59.7% of cases); U. urealyticum was detected in 11.0% and C. trachomatis in only 2.8%. The concordance rate between endocervical and endometrial specimens for common bacteria was 48.3%; 100% for C. trachomatis and 58.3% for U. urealyticum. The concordance rate between vaginal and endometrial cultures for common bacteria was 50.2%, only 16.7% for C. trachomatis and 48.8% for U. urealyticum. For common bacteria both vaginal and cervical cultures showed low sensitivities of 0.30 and 0.19, respectively. Conclusion: Common bacteria and U. urealyticum were the prevalent infectious agents in the uterine cavity of women diagnosed with CE. Both vaginal and endocervical cultures had low concordance with endometrial cultures. Only C. trachomatis test at cervical level had high concordance with endometrial findings.

Human larynx expresses isoforms of the oestrogen receptor

Commercially available enzyme immunoassays (EIAs) were used for oestrogen (ER) and progesterone (PR) receptor determination in the cytosol fraction of 118 human larynx cancer specimens and in the corresponding histologically proven non-malignant tissues. Fifty-one ER positive cancerous samples had corresponding non-cancerous tissues also expressing the receptor. A high resolution isoelectric focusing (IEF) technique followed by immunoblotting with the H222 anti-ER monoclonal antibody was used to evaluate the presence of ER isoforms in the 51 ER positive human larynx cancer specimens and in their corresponding non-malignant tissues. In both tissues, four ER isoforms were detected, with isoelectric points (pI) similar to those obtained in breast and endometrium carcinomas (6.1, 6.3, 6.6 and 6.8). A significant difference in the expression of ER isoforms between cancerous and non-cancerous tissue was found; precisely, the 94.1% of the ER positive non-malignant specimens co-expressed the four isoforms while they were detected in only the 35.5% of the malignant specimens (P < 0.0001 by Fishers exact test). In larynx cancer, the concentration values of ER and PR did not correlate, nevertheless tumours co-expressing the four ER isoforms had PR levels significantly higher than those which did not (P = 0.02 by Mann-Whitney Wilcoxon sum rank test). To investigate the possibility that the isoforms of the monomeric 4S form of the ER (those with pI 6.3, 6.6, and 6.8) could dimerise, a cold agarose gel electrophoresis technique was used on IEF-separated ER isoforms. In summary, the evidence shows that all the isoforms are able to form homodimers and that the isoforms at pI 6.3 and 6.8 are able to dimerise with that at pI 6.6 but, under the same experimental conditions, they do not form the 6.3/6.8 heterodimer. It was concluded that: (1) the four isoforms of the ER are co-expressed by the non-malignant human larynx and the cancer loses the capacity to express some of them; (2) the complete complement of ER isoforms (all four) is needed for PR expression; (3) the monomeric 4S isoform with pI 6.6 has the capacity to form homo- and heterodimers, while the remaining two are only able to homodimerise.

A rare case of choroidal metastasis presented after conservative management of endometrial cancer

Hematogenous dissemination from endometrial cancer is quite rare. We report a 31-year-old woman who developed choroidal metastasis following conservative management of early-stage endometrial carcinoma. She had received kidney transplantation and was taking steroids and cyclosporine. Three years after hysterectomy for persistent endometrial carcinoma, she developed multiple metastatic disease (to both lungs and right pelvis), and while on treatment with paclitaxel and carboplatin, she complained of a rapid visual deterioration. Ophthalmologic evaluation revealed a metastatic choroidal tumor associated with multiple central nervous system metastases. The patient refused further treatment and died 1 month after diagnosis of choroidal involvement. In conclusion, this is the first reported case of choroidal metastasis from endometrial cancer and highlights the need to consider immunosuppressive treatment as an absolute contraindication to conservative fertility-sparing treatment in gynecological malignancies.

Incidental metastases of well-differentiated thyroid carcinoma in lymph nodes of patients with squamous cell head and neck cancer: eight cases with a review of the literature

The examination of a large series of cervical lymph nodes in patients with head and neck cancer revealed the presence of incidental metastases of occult thyroid carcinoma in eight patients, of which six cases were squamous cell carcinoma of glottic and supraglottic sites of the larynx and two cases were pyriform sinus and tongue carcinomas. Three patients had two lymph nodes and the remaining patients had one lymph node each involved. The nodal chains affected were the jugular (n=5; level IV), Kuttner (level II), supraomohyoid (level III) and supraclavicular (level VI). In four cases, a subtotal thyroidectomy or unilateral lobectomy was performed during laryngectomy (for surgical reasons) or after histologic nodal examination; a minimal focus of thyroid papillary carcinoma was detected in one patient. Three of eight patients died from recurrence of the squamous cell carcinoma; no case presented clinical evidence of thyroid malignancy. The differential diagnosis from benign thyroid heterotopia was based on the presence of minimal nuclear atypia. The choice of treatment of patients with a coexisting neoplasm characterized by poor prognosis is difficult, and contrasting opinions exist regarding the use of radical thyroidectomy and the subsequent management. As reported in the literature (66 cases), the more aggressive squamous cell carcinoma will determine the prognosis of these patients; in fact, only one of the referred cases died of cerebellar metastases of the thyroid cancer. Our results emphasize the importance of an accurate re-evaluation and follow-up of patients with incidental occult metastases for detection of a primary thyroid tumor. In the general population, this incidental nodal involvement may be related to a minimal occult thyroid carcinoma.

Prognostic value of the prelaryngeal node in laryngeal and hypopharyngeal carcinoma.

The role in tumor spread and the prognostic value of metastasis in a prelaryngeal node were investigated in 124 cases of laryngeal and hypopharyngeal carcinoma selected from about 900 total laryngectomies and pharyngolaryngectomies. Metastasis in a prelaryngeal node was noted in 26 cases: 22 without capsular rupture (N+) and 4 with capsular rupture (N+R+). Vascular embolism (E+) was never noted. Patients with a metastatic prelaryngeal node had a tumor of the piriform sinus in 13 cases (50 %), a transglottic tumor in 6, glotto-subglottic in 3, glottic in 2 and supraglottic in 2. They also more frequently had involvement of the cervical nodes (81 %) and of the thyroid gland. The 5-year survival in these patients is very poor: only 3 patients (11.5 %) were alive and free of disease. These data demonstrate the role of prelaryngeal nodes in tumors that affect the hypopharynx and the antero-inferior portion of the larynx and that directly reach this node through the anterior lymphatic peduncle.

Prognostic implications of NUMB immunoreactivity in salivary gland carcinomas.

The gene numb encodes for a protein (Numb) involved in cell fate decisions in Drosophila, with proposed endocytic and developmental functions in mammalians. The distribution pattern of Numb in human tissues however, has not been fully characterized. We set out to explore the immunohistochemical expression of Numb in normal and neoplastic (28 adenoid cystic and 34 mucoepidermoid carcinomas) salivary glands, and correlated the results with the clinico-pathologic features of the neoplasms. Intense Numb immunoreactivity was detected in normal ductal cells and in a subset of acinar cells. In salivary carcinomas, we detected diffuse and intense Numb immunostaining in 5 adenoid cystic and 8 mucoepidermoid carcinomas. By contrast, the majority of adenoid cystic and mucoepidermoid cancers showed only moderate (14 and 5 cases) or focal staining (9 and 21 cases), respectively. The corresponding expression of Numb mRNA was documented in normal parotid gland and adenoid cystic carcinoma. Numb immunoreactivity was inversely correlated with the histological grade and Ki-67 immunoreactivity of both adenoid cystic and mucoepidermoid carcinomas. In addition, while tumor grade, stage, Ki-67 and Numb immunoreactivity were associated with disease-free survival in univariate analysis, only Numb and Ki-67 immunoreactivities retained independent prognostic significance in multivariate analysis. These data suggest that loss of Numb is implicated in aberrant differentiation programs of salivary gland carcinomas and may serve as a prognostic indicator in patients treated for these neoplasms.