Marija Petković

Nuclear EGFR in ductal invasive breast cancer: correlation with cyclin-D1 and prognosis

The epidermal growth factor receptor (EGFR)-family and cyclin-D1 have been extensively studied in breast cancer; however systematic studies that examine protein expression and gene status in the same cohort of patients are lacking. Also emerging evidences suggest existence of a direct EGFR-signaling pathway, which involves cellular transport of EGFR from cell membrane to the nucleus, and transcriptional regulation of the target genes. Thus, we examined the protein expression of membrane EGFR, nuclear EGFR, cyclin-D1 and the corresponding gene status in 113 breast carcinomas by immunohistochemistry and fluorescence in situ hybridization using tissue microarrays. Membrane EGFR overexpression and EGFR gene amplification were detected in 2% cases, while nuclear EGFR was detected in 40% of cases, with 12% having high nuclear EGFR staining. Nuclear EGFR correlated with tumor size (P=0.0005), lymph node metastasis (P=0.0288), Nottingham prognostic index (P=0.0011) and estrogen receptor (ER) expression (P=0.0258) but the letter correlation was observed only in premenopausal group of patients. Strong cyclin-D1 expression and cyclin-D1 gene (CCND1) amplification were found in 64 and 13% of the cases, respectively. Cyclin-D1 expression showed positive correlation with ER (P=0.0113) and inverse correlation with Nottingham prognostic index (P=0.0309) and membrane EGFR (P=0.0201). CCND1 amplification also showed inverse correlation with membrane EGFR (P=0.0420). A strong correlation between membrane EGFR expression and gene amplification (P=0.0035), as well as cyclin-D1 overexpression and gene amplification (P=0.0362), was demonstrated. On univariate analysis cyclin-D1 expression showed a correlation with longer overall survival in the premenopausal group and nuclear EGFR correlated with shorter overall survival in whole cohort as well in the premenopausal group of patients. Multivariate analysis revealed nuclear EGFR to be an independent prognostic factor and showed 3.4 times greater mortality risk for nuclear EGFR+++ patients as compared with nuclear EGFR negative patients (hazard ratio =3.402; P=0.0026).

The first report of extraosseous Ewing's sarcoma in the rectovaginal septum.

Aims and Background To report an extremely rare case of Ewings sarcoma located in the rectovaginal septum. Ewings sarcoma is a highly malignant neoplasm of bone, which usually occurs during childhood. Common extraosseous localizations of Ewings sarcoma include the trunk, extremities, uterus, cervix and vagina. Methods A 45-year-old woman presented to us with a six-month history of pain in the lower abdomen during intercourse. Pelvic examination was performed and a palpable mass was found. The mass had a size of 9 × 6 cm, a soft tissue consistency, was partially movable and the patient felt the pain during palpation. Examination of the inguinal lymph nodes revealed no signs of inguinal adenopathy. The results of laboratory tests, rectoscopy, chest X-rays, barium enema and bone scan were normal. Computed tomography (CT) showed an inhomogeneous expansive mass in the rectovaginal septum measuring 8.7 × 6.1 cm, without any signs of rectum or bladder invasion. The vascular structures of the pelvis were normal. At laparotomy the process was judged inoperable and only biopsy of the tumor mass was carried out. Histology showed a neoplasm with small, round to oval cells with scarce cytoplasm. Immunohistology with the monoclonal antibody CD99 (MIC-2 gene product, Ewings sarcoma marker, clone 12E7, DAKO A/S, Glostrup, Denmark) revealed an extraosseous Ewings sarcoma. The patient was treated with chemotherapy followed by whole-pelvis external beam radiation and intracavitary brachytherapy. Results A residual mass measuring 3.5 × 2.5 cm was visible on a control CT scan 18 months after treatment; however, the patient was feeling well and refused surgery to remove the residual mass. Conclusions To our knowledge this is the first reported case of extraosseous Ewings sarcoma in the rectovaginal septum.

Tributaries of the Human and Canine Coronary Sinus

The coronary sinus is an anatomic landmark and conduit for many diagnostic and therapeutic procedures. We studied 40 human and 40 canine coronary sinuses of the heart in order to ascertain various functional anatomic features of the coronary sinus and its tributaries. We performed a comparative analysis of human and canine cardiac veins. We also studied the formation, tributaries, length, diameter and the ostium of the coronary sinus. The tributaries of the coronary sinus were highly variable in human and dog. The main and constantly present tributaries of both the human and canine coronary sinus were the great and the middle cardiac veins. We also found a rare case of a small cardiac vein connecting two cardiac venous systems--the coronary sinus and its tributaries and the anterior cardiac veins. In two cases we found a common trunk of the small cardiac vein and middle cardiac vein which opened into the coronary sinus. An intramyocardial course of the anterior interventricular vein was found in 2 cases and in 1 case the vein curved twice around the anterior interventricular branch of the left coronary artery. The Thebesian and Vieussens valves and ostial valves of other cardiac veins were also found.

TISSUE ZINC DYNAMICS DURING THE IMMUNE REACTION IN MICE

Owing to the importance of zinc for the functioning of the immune system, the role of endogenous Zn, located both in lymphoid and nonlymphoid organs, was investigated during the standard humoral and cellular types of immune response. For this purpose, the dynamics of hepatic, thymic, splenic, and renal Zn content was determined in mice sensitized with (a) sheep red blood cells and (b) semiallogeneic lymphocytes during the local host vs graft reaction (HVGR). The data obtained by ion-coupled plasma spectrometry revealed that the humoral type of immunity is characterized by a significant increase of Zn concentration in the liver and in the thymus. Simultaneously, linear regression analysis showed that the generation of plaque-forming cells in the individual mouse was highly positively correlated with Zn concentration in the liver (r = 0.897), and spleen (r = 0.833), and negatively with Zn concentration in the thymus (r = -0.624). Similar relationships between the intensity of local immune reaction and tissue Zn levels were found in local HVGR at the fifth day in the liver and spleen (r = 0.861 andr = 0.695, respectively), at the seventh day in the thymus (r = -0.797), and at the tenth day in the liver (r = -0.859). The data emphasize the necessity of Zn for the development of normal immune response and point to the existence of a Zndependent hepato-thymic axis during the humoral and cellular types of immune reactivity.

EGFR and cyclin D1 in nodular melanoma: correlation with pathohistological parameters and overall survival.

Considering that nodular melanoma (NM) has the potential to show an early distant metastasis, there is an urgent need for the discovery and evaluation of new diagnostic and prognostic biomarkers. We aimed to investigate the protein expression of membrane and nuclear epidermal growth factor receptor (EGFR), cyclin D1, and the corresponding gene status in NM samples and correlate the results obtained with clinicopathological parameters and overall survival of patients. Immunohistochemical and fluorescence in-situ hybridization analyses were carried out on tissue microarrays constructed from 110 NM samples, 30 compound nevi, and 38 dysplastic nevi. NM samples showed 24% strong cyclin D1 and 37% strong Ki67 protein expression compared with 3 and 0% strong cyclin D1 and Ki67 expression in the control group. Membrane EGFR expression was detected in 50% of NM cases, whereas EGFR gene amplification was detected in only 4% of NM cases. Multiple NM samples presented simultaneous membrane and nuclear EGFR expression. We found a negative correlation between tumor thickness and membrane EGFR expression. It was also observed that membrane EGFR 3+ NM samples presented ulceration significantly more often than membrane EGFR-negative (0) NM samples. In univariate analysis, carried out on 44 patients with follow-up data, both nuclear and membrane EGFR overexpression showed a correlation with a shorter overall survival. Nuclear EGFR (++, +++) showed 3.06 and membrane EGFR (2+, 3+) showed 2.76 higher risk of mortality compared with patients with low and negative nuclear and membrane EGFR expression (P<0.05).

CD30-positive cutaneous anaplastic large cell lymphoma with ichthyosis acquisita.

Anaplastic large cell lymphoma (ALCL) is a high-grade non-Hodgkins lymphoma recognized by the expression of the CD30 marker and by its morphology. We report an unusual case of ALCL in a 42-year-old woman. For ten years the patient only had pruritic erythematous skin lesions, then a plaque with nodules and ulcers on the right thigh and leg developed, followed by ichthyosis acquisita. The development of ALCL in women and the association of ichthyosis acquisita with this type of lymphoma are uncommon.

Sternal metastasis as a first manifestation of adenocarcinoma of the pancreas

A 63-year-old woman noticed a bump growing on her anterior chest wall (Fig. 1), but she did not present to the hospital until jaundice and abdominal pain occurred. A thorough examination disclosed that the patient was suffering from metastatic tumor of the pancreatic head. She had suffered from an insulin dependent diabetes mellitus for years. Laboratory tests showed mild alterations in glucose, alkaline phosphatase, GGT, AST and ALT levels. Ultrasound of the abdomen revealed the hypoechogene lesion in the position of uncinate processus with an intensive pneumobilia. A radioisotope bone scan with Technetium- 99m methylene diphosponate revealed the only pathological accumulation of activity in the upper part of the sternum. An X-ray of the sternum demonstrated an expansive osteolitic lesion of the manubrial region as a result of the destruction of the original cortex in the anterior part. This was confirmed by a C large part, the bone was expanded as a result of the anterior cortical destruction (Fig. 2). Histopathological examination scan, the lesion was necrotic in a of a biopsy revealed small bone fragments and the connective tissue infiltrated with unspecific gland formations and typical signed ring cells of an adenocarcinoma were seen. The patient underwent choledochoduodenostomy to relieve her of jaundice and pain. The biopsy of the pancreas confirmed the diagnosis of the pancreatic adenocarcinoma. Although the patient was treated with a chemotherapy regimen with gemcitabine and irradiated with megavoltage photons of 60Co afterwards, the sternal metastasis continued to grow. There are only few reports of bone metatases originating from carcinoma of the pancreas, all of them affecting temporal bones and presenting with hearing loss [1]. We are not aware of a previous report of an “ osteophobic” tumor, such as pancreatic adenocarcinoma, presenting with a sternal metastasis as a first manifestation.

On the Role of T Lymphocytes in Stimulation of Humoral Immunity Induced by Peptidoglycan–Monomer Linked with Zinc

Effects of peptidoglycan linked with zinc (PGM–Zn) were investigated on plaque–forming cell (PFC) generation to sheep red blood cell (SRBC) and SRBC–unrelated antibody production in primary and secondary immune response in mice depleted in vivo of CD4+ and/or CD8+ T lymphocytes. PGM–Zn in nondepleted mice stimulated the PFC generation and IgM or IgG and IgG1 production in primary and secondary reaction. Single depletion of CD4 or CD8+ T cells did not change this ability. The effects of PGM–Zn after CD8+ depletion were even greater than those in nondepleted mice. Depletion of both T cell subsets, however, completely abrogated immunostimulatory effects of PGM on PFC generation (primary and secondary response), as well as on primary SRBC–unrelated antibody production, leaving only the increase of IgG in secondary response unchanged. Immunostimulatory effects and isotype switching to IgG1 and IgG2a correlated with the changes in splenic CD4+, CD8+, CD5+ cells, pointing to the regulatory role of these cells and/or their cytokines in PGM–Zn–induced immunostimulation. Altogether the data suggest that PGM–Zn may potentiate the costimulatory signals coming from activated T cells and act on B cells without the T cell help.

Immunoregulating effects of peptidoglycan monomer linked with zinc in adult mice.

Since it is well known that both zinc ions and bacterial immunostimulants influence the function of the immune system, in the present study we investigated the immunomodulating activity of a new analog of peptidoglycan monomer (PGM), in which the basic molecule was linked to zinc (PGM-Zn). Its effects in BALB/c mice, aged 10-12 months, were compared with the effects of equimolar doses of PGM and ZnCl2. The treatment lasted 26 days (one i.p. injection every fifth day). The results showed that PGM-Zn may markedly enhance antibody production to sheep red blood cells, as well as spontaneous and concanavalin A (ConA)-induced blastogenesis. The generation of plaque-forming cells in individual mouse was positively correlated with the expression of class II antigens in the liver and negatively correlated with the total quantity of hepatic proteins. PGM-Zn also induced the appearance of peritoneal macrophages, which in cocultures with syngeneic splenocytes were less able to enhance the spontaneous, and particularly the ConA-induced blastic transformation. The enhancing activities of PGM-Zn were in some respects more closely correlated with the action of PGM, whereas the induction of suppressive macrophages resembled more the activity of ZnCl2. The data emphasize that PGM-Zn may both stimulate and inhibit immunoregulative pathways by mechanisms which are not identical to those of PGM or ZnCl2.

Primary large-cell neuroendocrine carcinoma of the scrotum

ZusammenfassungDie vorliegende Arbeit berichtet über einen 78-jährigen Mann mit einem schnell wachsenden Knötchen am Skrotum und einer Schwellung in der Inguinalregion. Es bestanden sonst keine subjektiven Symptome. Die Familieanamnese für erbliche Tumoren war negativ. Sexuell übertragbare Krankheiten, Traumata, Pilzinfektionen oder chronische Reizung der Region wurden negiert. Ebenso hatte keine Radiotherapie, chemische oder Arsen-Exposition statt gefunden. Beide Hautveränderungen wurden 1,2 cm im Gesunden exzidiert. Die histologische Untersuchung zeigte ein umschriebenes, undifferenziertes großzelliges neuroendokrines Karzinom mit Metastasen in den inguinalen Lymphknoten. Drei Monate nach der Exzision fanden wir ein lokales Rezidiv. Die Histopathologie zeigte, dass das Rezidiv mit dem primären Tumor identisch war. Bisher ist unserem Wissen nach in der medizinischen Literatur noch nicht über einen neuroendokrinen Tumor des Skrotums berichtet worden.SummaryNeuroendocrine tumors (NETs) mostly develop from the neural crest cells but a few arise from neuroectoderm. They are common in the lungs and gastrointestinal tract but rare in the genitourinary tract. A 78-year-old man with no family history of malignant or hereditary diseases presented with a 3-month history of a rapidly growing asymptomatic scrotal nodule and swelling in the groin. He had a negative history of sexually transmitted disease and of trauma, fungal infection or chronic irritation in the scrotal area; there was no history of radiotherapy or exposure to chemicals or arsenic. Both the scrotal and groin lesions were excised with a minimum of 1.2 cm of normal skin. Examination of the specimen revealed a confined poorly differentiated large-cell neuroendocrine carcinoma with a metastasis to the inguinal lymph nodes. Three months after the excision we found a local recurrence. The recurrent tumor revealed tumor tissue concurrent with the primary lesion. To the best of our knowledge, there have been no previously published case reports on neuroendocrine tumor of the scrotum.