Danuta Skomra

Immunohistochemical demonstration of multiple HPV types in laryngeal squamous cell carcinoma

The aim of this study was to determine the prevalence of human papillomaviruses (HPV) types 6, 11, 16, 18, 31, 33, 42, 51, 52, 56 and 58 in laryngeal squamous cell carcinoma specimens using immunohistochemical reactions and to correlate the presence of HPV with the clinical and pathological characteristics of these patients. Tissue samples were collected from 40 patients with primary laryngeal squamous cell carcinoma (LSCC) and from 33 subjects with non-neoplastic laryngeal lesions or laryngeal nodules, which served as a control group. Human papilloma virus was detected in 6 (15%) of the 40 patients. Five (83.4%) of six patients with HPV positive tumors had G2 (moderately differentiated), one patient (16.6%) had G3 (poorly differentiated), and no patient with HPV positive tumor had a G1 (well-differentiated) tumor. Four (66.6%) of the six HPV positive tumors were in the supraglottic region, one (16.6%) tumor was located in the glottis, and one (16.6%) HPV positive tumor was in the subglotic region. Five (83.4%) of six HPV positive tumors were T3-T4, and one was T2. Three of six HPV positive patients had no clinically evident cervical lymph nodes (N0), and three of the HPV positive patients were N1 or N2. Human papillomavirus was not detected in any of the samples from the control group. The presence of HPV infection in 15% of the cases may suggest a possible role in the etiology of laryngeal squamous cell carcinoma. However, no significant correlation between HPV incidence and histological grading and clinical staging could be demonstrated.

Immunohistochemical Analysis of MIB-1 Proliferative Activity in Human Endometrial Cancer. Correlation with Clinicopathological Parameters, Patient Outcome, Retinoblastoma Immunoreactivity and K-Ras Codon 12 Point Mutations

To test the prognostic utility of MIB-1 in human endometrial neoplasias, the proliferative activities of fifty-two endometrial carcinomas obtained from Polish women were assessed. We also investigated the relationship between the MIB-1 Proliferative Index and the well-known clinicopathological features of cancer (clinical stage, histological type, histological grade, depth of myometrial invasion), patients age, overall survival, retinoblastoma immunostaining and K-ras codon 12 point mutations. The mean MIB-1 Proliferation Index was 43.8%, with a median of 36.0%. Due to the great intratumour heterogeneity of the immunoreaction, the Index ranged from 0% to 98%. A significant relationship was noted between MIB-1 expression and histological grading (p=0.0004) and myometrial invasion of cancer (p=0.01). Multivariate Cox regression demonstrated that the clinical stage was the only independent prognostic factor during follow-up (p=0.025). There was a tendency towards a poorer outcome for women with a Proliferative Index of >31% than for patients whose Index was ≤ 31%; the difference, however, did not reach significance (p=0.25; log-rank test). Interestingly, uterine cancers lacking retinoblastoma protein expression had a mean MIB-1 Proliferation Index that was nearly twice as high as in those neoplasias that stained positively for retinoblastoma (70.33% and 42.14%, respectively; p=0.09; Mann-Whitney-U test). There were no significant differences between K-ras codon 12 point mutation-positive and -negative endometrial carcinomas regarding the proliferative activity of the cancer (mean Indexes 47.6% and 43.8%, respectively; p=0.66, Mann-Whitney-U test). Our data support the view that MIB-1 proliferative activity was significantly increased with a decrease of the histological grading and with the myometrial invasion of human endometrial cancer.

Expression of the cell-cycle regulatory proteins (pRb, cyclin D1, p16INK4A and cdk4) in human endometrial cancer: correlation with clinicopathological features

AbstractIntroduction. Derailments of the control mechanisms in the G1/S phase of the cell cycle play a fundamental role in the initiation and progression of cancer. However, only a few reports have addressed the issue of simultaneously occurring abnormalities of Rb-pathway components in malignant endometrial tumors.Methods. Currently, we assessed the expression of cell-cycle regulatory proteins (pRb, cyclin D1, p16INK4A and cdk4) in 48 sporadic endometrial cancers, and investigated these tumors for a possible relationship between aberrant protein staining and clinicopathological variables of cancer and RB-LOH.Results. There was abnormal pRb, cyclin D1, p16INK4A and cdk4 immunoreactivity in 2%, 50%, 6% and 25% of cases, respectively. Altogether, 33 of 48 (69%) endometrial malignant tumors showed abnormal expression of at least one Rb-pathway protein immunohistochemically. However, there was significant correlation neither between the cell-cycle regulators nor between the frequency of pRb, p16INK4A and cyclin D1 abnormalities and clinicopathological variables of cancer, but a significant correlation did exist between cdk4 staining and the clinical stage of disease (P<0.05, Fishers exact test). Moreover, an inverse relationship was also demonstrated between cdk4 expression and patient age (r=-0.367; P=0.01). However, none of the cell-cycle regulatory proteins, except for pRb, was related to loss of heterozygosity at locus 13q14.Conclusion. As a conclusion, derailments of the Rb-pathway components, cyclin D1 and cdk4 in particular, seems to participate in the endometrial cancer development in humans. Overexpression of cdk4 was related to the progression of neoplastic disease and corresponds with age of onset, suggesting a major role of altered cdk4 immunoreactivity in the progression of endometrial cancer.

An immunohistochemical study of cyclin D1 protein expression in laryngeal squamous cell carcinoma.

Conclusion. Contrary to most reports, our study shows that the expression of cyclin D1 is not an independent prognostic factor in patients with laryngeal cancer (LC). No correlation between cyclin D1 expression and survival rates in LC was found in a multivariate analysis. Objectives. The aim of this study was to determine the possible relevance of the expression of cyclin D1 protein in LC as prognostic criteria and to analyse correlation of the expression with clinicopathological features and survival rates. Materials and methods. Immunohistochemistry staining was used to detect the expression of cyclin D1 in 130 samples of laryngeal cancer and in 22 specimens of laryngeal nodules. Results. Cyclin D1 expression was found in 52 (40%) LC samples and in 3 (13.6%) samples of laryngeal nodules. There was no significant correlation between cyclin D1 expression and clinicopathological features of LC. A multivariate analysis of survival confirmed that cyclin D1 expression was not an independent prognostic factor in LC.

Allelic Loss at TP53 Is Not Related to p53 Protein Overexpression in Primary Human Endometrial Carcinomas

We examined loss of heterozygosity (LOH) at the TP53 gene in primary human endometrial carcinomas (EC), and investigated the relationship between allelic loss, p53 protein overexpression, pRb-1 pathway alterations and MIB-1 proliferative activity. Applying the non-isotopic PCR-RFLP/VNTR-silver staining techniques, we investigated TP53 LOH in 46 tumors at four polymorphic loci. Out of 42 informative carcinomas, LOH was found in 19% of the cases studied. In general, there was no significant relationship between LOH and the clinical and pathological variables of cancer, including patient age, clinical stage, histological grade or depth of myometrial invasion. Interestingly, none of 7 tumors associated with hyperplasia revealed allelic imbalance, whereas 8 of 27 (30%) tumors without hyperplasia exhibited LOH (p = 0.312; Fisher’s exact test). Overexpression of nuclear p53 was not correlated with allelic loss at TP53 (p = 0.336, Fisher’s exact test). It is worth pointing out that p53 immunoreactivity was significantly related to proliferative activity of cancer (R = 0.42, p = 0.0037; Spearman’s rank correlation test). A tendency towards a poorer outcome was reported in EC patients displaying TP53 LOH during short-time follow-up (p = 0.093; log-rank test). None of the tumors simultaneously showed LOH at TP53 and RB1 genes(R = –0.211, p = 0.16; Spearman’s rank correlation test). p16INK4A alterations (LOH and gene deletion) occurred concomitantly, with 3 tumors showing the TP53 allelic loss, whereas the cyclin D1/cdk4 complex was overexpressed in a case with TP53 LOH. Altogether, losses at TP53 were not associated with p53 nuclear overexpression, but may affect a subset of EC patients characterized by an unfavorable prognosis at short-time follow-up. Allelic loss at TP53 seems to arise independently of LOH at the RB1 gene in carcinomas of the uterine corpus in humans. Disruptions at p16INK4A and/or cdk4/cyclin D1 concomitantly occurring with TP53 LOH may participate in the development of a subset of endometrioid-type ECs.

Extranasopharyngeal angiofibroma of the infratemporal fossa.

Juvenile angiofibroma (JA) is a benign, vascular neoplasm that accounts for less than 0.5 percent of all head and neck tumors. The characteristic triad of clinical symptoms consists of nasopharyngeal tumor, nasal obstruction, and recurrent epistaxis. JAs of extranasopharyngeal origin have been sporadically reported in the literature. Windfuhr and Remmert in a review of 65 patients with extranasopharyngeal angiofibromas found only one case of tumor located in the infratemporal fossa. We present a rare case of a patient with extensive JA that occupied infratemporal fossa and cheek, without involvement of the nasopharynx. Medical University of Lublin Review Board approved the analysis and description of the patient. A 24-year-old man was referred to our institution in January 2006 with a 9-month history of painless cheek swelling. No signs of infection were present. No lymph nodes were palpated. Biopsy taken previously in another hospital indicated capillary hemangioma. Computed tomography and magnetic resonance revealed an inhomogeneous mass in the left infratemporal fossa that caused displacement of the pterygoid muscles and anterior bowing of the posterior maxillary wall (Fig 1). The tumor spread anteriorly between alveolar process of the maxilla and ramus of the mandible and invaded the cheek. The masseter muscle was displaced posteriorly. The tumor presented intermediate signal intensity on T1and T2-weighted images with focal signal-void areas. After contrast administration, intensive inhomogeneous enhancement of the lesion was visible. There were no signs of bony destruction. A superficial part of the tumor was easily accessible for sonographic evaluation. Color-flow duplex Doppler sonography demonstrated a solid, hypoechoic mass with multiple internal vessels of low-resistance blood flow. Carotid angiography showed abundant vascularity of the tumor with blood supply from the left external carotid artery via internal maxillary artery (Fig 2). Preoperative embolization of the feeding branches with the use of polyvinyl alcohol particles was performed. Final external carotid arteriogram showed occlusion of the feeding vessels. Two days after embolization, the tumor was removed with the use of combined intraoral and infratemporal fossa approach. Apart from cheek hematoma that resolved spontaneously, postoperative course was uneventful. A threeyear follow-up revealed no recurrence and no functional deficits. Histopathologic examination showed findings characteristic for JA: multiple gaping vessels lined by a single layer of endothelial cells embedded in fibrous stroma containing spindle-shaped and stellate cells, collagen fibers, and inflammatory cells (Fig 3, http://www.journal. entnet.org).

RB protein expression in human endometrial carcinomas--an immunohistochemical study.

The aim of the current study was to investigate the immunohistochemical expression of the retinoblastoma protein (pRB) in formalin-fixed, paraffin-embedded specimens obtained from 62 patients suffering from endometrial cancer. The avidin-biotin-peroxidase detection system with microwave pretreatment and the mouse anti-human NCL-RB1 monoclonal antibody were used. Heterogeneous nuclear immunostaining for the pRB was generally observed in the glandular cells in 59 out of 62 (95%) endometrial carcinomas, while stromal components were unreactive. In one case of stage Ic endometrioid adenocarcinoma, a small percentage of glandular cells (5%) stained positively with the anti-RB antibody, while two other tumors (stage IIa adenosquamous carcinoma and stage IIIa endometrioid adenocarcinoma) were pRB negative. In the cases with concomitant hyperplastic and neoplastic endometrial lesions, pRB immunoreaction was heterogeneous in the hyperplastic endometrial cells and in the adjacent neoplastic endometrium. Moreover, eight cases of endometrial carcinoma harboring K-ras codon 12 gene point mutation overexpressed pRB (more than 80% of glandular endometrial cells were positive) immunohistochemically, while none of three pRB negative slides had a K-ras gene alteration. Our data support the view that the pRB is expressed in most of the human endometrial neoplasms, but the lack of pRB immunoreactivity may correspond with the retinoblastoma gene rearrangements in a subset of advanced endometrial carcinomas.

Immunohistochemical analysis of carcinomatous and sarcomatous components in the uterine carcinosarcoma: a case report.

Uterine carcinosarcoma (malignant mixed Mullerian tumor) is an uncommon female genital tract neoplasm characterized by an admixture of epithelial and stromal malignant cells. We report a case of 50-year-old peri-menopausal woman diagnosed to have early-stage (IB due to FIGO) uterine carcinosarcoma of the homologous type with superficial (3mm) myo-invasion. The patient showed no clinical symptoms of the disease and had no family history of female genital tract malignancies. Positive immunostaining for steroid receptors (estrogen-alpha and progesterone receptors), cytokeratin, and EGFR was detected only in the carcinomatous area, whereas beta-catenin, BCL-2, COX-2, p16(INK4a), PTEN, RB-1, and vimentin were immunoreactive in both components. Androgen receptor, CD10, desmin, HER-2/neu, and P53 were found to be negative either in the carcinomatous or in the sarcomatous area. Tumor proliferative activity was higher in the carcinomatous (25%) than in the sarcomatous (2%) component. Based on these findings, immunohistochemical evaluation of multiple receptor status in the carcinomatous and sarcomatous areas of carcinosarcoma may provide a clue to the pathogenesis and hormonal receptor status of this uncommon uterine malignancy.

Gaucher disease diagnosed after bone marrow trephine biopsy — a report of two cases

The hematologist is at the forefront of specialists to whom patients with Gaucher disease present because of cytopenia and hepatosplenomegaly. Usually, patients with such symptoms have undergone trephine biopsy. We present the cases of two patients in whom Gaucher disease was suspected because of the discovery of Gaucher cells in trephine biopsy, and subsequently confirmed via enzymatic and molecular investigations.

Endometrial carcinoma with pleural metastasis: A case report.

BACKGROUND There have been a limited number of studies giving the incidence of pleural metastasis from female genital tract tumors. CASE An unusual case occurred of recurrent pleural malignant effusion associated with disseminated serous papillary endometrial adenocarcinoma (EC). A total abdominal hysterectomy with bilateral salpingo-oophorectomy, pelvic lymphadenectomy, appendectomy and omentectomy was performed. Treatment of the pleural malignant effusion consisted of thoracotomy with partial decortication, systemic chemotherapy and radiotherapy. The patient died of circulatory failure 8 months after the primary diagnosis. CONCLUSION Although the pleura is a rare site of widespread EC, one should recognize the possibility of pleural spread from female genital tract neoplasms presenting with associated symptoms. Cytopathologic examination of the pleural effusion and the finding ofcarcinoma cells mandate an investigation for the primary site of the neoplasm by a multidisciplinary group.