Orbital melanocytoma: When a tumor becomes a relieving surprise.

PURPOSE: Melanocytomas are rare pigmented tumors that arise form melanocytes and have been reported in the central nervous system. Orbital melanocytomas "also known as blue nevus" are rarely reported. The occurrence of choroidal melanoma and orbital melanocytomas has never been described. OBSERVATIONS: This is a case of orbital melanocytoma in a 34 year old female who presented with left proptosis and ecchymosis. She has the right eye enucleated to treat a large choroidal melanoma, 6 years earlier. Orbital metastasis was suspected. After orbital imaging and systemic evaluation, incisional biopsy was planned yet the mass could be totally excised and it turned out to be melanocytoma. The condition was not associated with nevus of Ota and the patient is not known to have any predisposing condition for melanocytic lesions. CONCLUSION AND IMPORTANCE: Melanocytoma and malignant melanoma share the same cell of origin. The benign course, the well differentiated cells, absence of anaplasia and the positive reaction to Human Melanoma Black-45 (HMB-45) and S-100 proteins established the diagnosis of the former. Such diagnosis was a relief for this one eyed patient.(HMB-45:human melanoma black-45).

Association of factor V Leiden, Janus kinase 2, prothrombin, and MTHFR mutations with primary Budd-Chiari syndrome in Egyptian patients.

BACKGROUND AND AIM: Budd-Chiari syndrome (BCS) is defined as obstruction of hepatic venous outflow anywhere from the small hepatic veins to the suprahepatic inferior vena cava. The pathogenesis of BCS is still not fully understood. This study aimed to evaluate the association of factor V Leiden (FVL), Janus kinase 2 (JAK2), prothrombin, and methylene tetrahydrofolate reductase (MTHFR) mutations with primary BCS. METHODS: The study was carried out on 35 patients with primary BCS and 15 age and gender matched healthy individuals as a control group. Genotyping of FVL, prothrombin, and MTHFR mutations was determined by GENEQUALITY AB-THROMBO TYPE kit based on the reverse hybridization principle. JAK2 mutation was determined by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: There was a statistically significant difference between patients and controls regarding FVL, MTHFR C677T, and MTHFR A1298C mutations with odds ratio of 1.83, 2.0, and 1.79, respectively. Hetero MTHFR C677T, hetero FVL, and hetero MTHFR A1298C were the most common etiological factors being responsible for 57.1, 42.9, and 42.9% of primary BCS cases, respectively. CONCLUSION: It could be concluded that BCS is a multifactorial disease; in the current study, MTHFR C677T mutation was the most common cause of disease. Identification of one cause of BCS should not eliminate investigations for detection of other etiological factors.

Positive lymph-node breast cancer patients ­ activation of NF-κB in tumor-associated leukocytes stimulates cytokine secretion that promotes metastasis via C-C chemokine receptor CCR7.

Tumor metastasis to lymph nodes is most deadly complication among breast cancer patients. Herein, we investigated the molecular mechanism by which tumor-associated leukocytes (TALs) mediate lymph node metastasis. The density of different leukocyte subtypes infiltrating the tumor microenvironment of negative and positive lymph nodes (nLNs, pLNs) in breast cancer patients was measured using immunohistochemistry. In addition, we isolated TALs from blood drained from the axillary tributaries of nLN and pLN patients during breast surgery. Secretions of TALs were subjected to cytokine profiling using a cytokine antibody array. Our results showed an increase in the number of infiltrated CD45+ cells in the carcinoma tissues of pLN patients with the major proportion being myeloid subsets compared with nLN patients. Furthermore, TALs of pLN patients show a significant fivefold increase in the secretion of interleukin (IL)-1α, interferon-γ, IL-5, IL-3 and tumor necrosis factor-ß, and are characterized by enhanced constitutive NF-κB/p65 signaling compared with TALs isolated from nLN patients. Using an invasion assay, cytokines secreted by TALs of pLN patients were shown to augment the invasive phenotype of breast cancer MCF-7 and SKBR3 cells compared with nLN patients. Using flow cytometry, we found that C-C chemokine receptor 7 (CCR7) is significantly overexpressed in breast carcinoma of pLN patients compared with nLNs patients. Intriguingly, CCR7, a mechanistic clue for metastasis, is upregulated in MCF-7 cells upon stimulation with TAL-conditioned media of pLN patients. Our findings show that the molecular cues secreted by TALs alone or in combination with CCR7 may emerge as future therapeutic targets for lymph node metastasis in breast cancer patients.

Cytokines secreted by macrophages isolated from tumor microenvironment of inflammatory breast cancer patients possess chemotactic properties.

Although there is a growing literature describing the role of macrophages in breast cancer, the role of macrophages in inflammatory breast cancer (IBC) is unclear. The aim of present study was to isolate and characterize tumor associated macrophages of IBC and non-IBC patients and define their role in IBC. Tumor infiltrating monocytes/macrophages (CD14+ and CD68+) were measured by immunohistochemistry using specific monoclonal antibodies. Blood drained from axillary vein tributaries was collected during breast cancer surgery and the percentage of CD14+ in the total isolated leukocytes was assessed by flow cytometric analysis. CD14+ cells were separated from total leukocytes by immuno-magnetic beads technique and were cultured overnight. Media conditioned by CD14+ were collected and subjected to cytokine profiling using cytokine antibody array. Wound healing and invasion assays were used to test whether cytokines highly secreted by tumor drained macrophages induce motility and invasion of breast cancer cells. We found that macrophages highly infiltrate into carcinoma tissues of IBC patients. In addition blood collected from axillary tributaries of IBC patients is highly enriched with CD14+ cells as compared to blood collected from non-IBC patients. Cytokine profiling of CD14+ cells isolated from IBC patients revealed a significant increase in secretion of tumor necrosis factor-α; monocyte chemoattractant protein-1/CC-chemokine ligand 2; interleukin-8 and interleukin-10 as compared to CD14+ cells isolated from non-IBC patients. Tumor necrosis factor-α, interleukin-8 and interleukin-10 significantly increased motility and invasion of IBC cells in vitro. In conclusion, macrophages isolated from the tumor microenvironment of IBC patients secrete chemotactic cytokines that may augment dissemination and metastasis of IBC carcinoma cells.

Cathepsin B: a potential prognostic marker for inflammatory breast cancer.

BACKGROUND: Inflammatory breast cancer (IBC) is the most aggressive form of breast cancer. In non-IBC, the cysteine protease cathepsin B (CTSB) is known to be involved in cancer progression and invasion; however, very little is known about its role in IBC. METHODS: In this study, we enrolled 23 IBC and 27 non-IBC patients. All patient tissues used for analysis were from untreated patients. Using immunohistochemistry and immunoblotting, we assessed the levels of expression of CTSB in IBC versus non-IBC patient tissues. Previously, we found that CTSB is localized to caveolar membrane microdomains in cancer cell lines including IBC, and therefore, we also examined the expression of caveolin-1 (cav-1), a structural protein of caveolae in IBC versus non-IBC tissues. In addition, we tested the correlation between the expression of CTSB and cav-1 and the number of positive metastatic lymph nodes in both patient groups. RESULTS: Our results revealed that CTSB and cav-1 were overexpressed in IBC as compared to non-IBC tissues. Moreover, there was a significant positive correlation between the expression of CTSB and the number of positive metastatic lymph nodes in IBC. CONCLUSIONS: CTSB may initiate proteolytic pathways crucial for IBC invasion. Thus, our data demonstrate that CTSB may be a potential prognostic marker for lymph node metastasis in IBC.

Capturing and characterizing immune cells from breast tumor microenvironment: an innovative surgical approach.

BACKGROUND: In breast cancer patients, venous drainage of the breast may contain cells of immunological importance, tumor cells undergoing dissemination, and other biological factors derived from the tumor microenvironment. Collecting axillary venous blood during modified radical mastectomy and thus before dilution in the circulation may allow us to define biological properties of the tumor microenvironment. Aims were to (1) develop a surgical approach to collect blood from the breast tumor microenvironment through tributaries of the axillary vein and (2) characterize and compare immune cells collected from the axillary vein with those in peripheral blood of breast cancer patients. MATERIALS AND METHODS: We enrolled 17 women aged 30-50 years and diagnosed with breast cancer by mammography, ultrasound, and biopsy (stages II-III). All patients were, preoperatively, treatment-naive. During routine surgical dissection, blood was collected in heparin tubes, 10 mL from tributaries of the axillary vein and 10 mL from peripheral blood. Mononuclear cells were separated, and percentages of different leukocyte populations were determined by flow cytometry. RESULTS: We detected a significant increase in the percentage of total T lymphocytes and T helper cells collected from axillary tributaries, but not in the percentages of cytotoxic T cells, monocytes, natural killer, or B cells compared with peripheral blood. CONCLUSIONS: The present study validated using an intraoperative surgical approach to collect leukocytes drained from the tumor microenvironment through axillary tributaries. Our results showed an increase in the infiltration of total T-lymphocytes and T helper cells in the tumor microenvironment, suggesting that they may contribute to tumor pathogenesis.

Primary yolk sac tumor of the lung.

Yolk-sac tumor mimics the yolk sac of the embryo, and the presence of alpha fetoprotein in the tumor cells is highly characteristic. We present an 18-year-old boy with primary pulmonary yolk-sac tumor diagnosed postoperatively. A computed tomographic scan revealed a huge intrathoracic soft tissue mass 20 x 25 cm occupying most of the left hemithorax. Two trials of computed tomographic-guided needle biopsy were nonconclusive. A left upper lobectomy was performed with a complete tumor resection. Postoperatively, the patient's alpha fetoprotein (AFP) was 10,512 IU/mL with gradual decline under chemotherapy. The patient is alive 10 months after surgery and is disease free.

Pleural mesothelioma: diagnostic problems and evaluation of prognostic factors.

BACKGROUND: Malignant pleural mesothelioma (MPM) in Egypt is mainly attributed to an environmental origin i.e exposure to asbestos, with a high incidence in women and young adults. Immunohistochemistry and ultrastructural features aid in the diagnosis. The p27Kip1 is a kinase inhibitor protein acting as a cell cycle regulator and a putative tumor suppressor gene playing a critical role in the pathogenesis of several human neoplasms. AIM: A clinicopathologic, immunohistochemical and ultrastructural study of mesothelioma in Egyptian patients, with identification of different prognostic factors. MATERIAL AND METHODS: Sixty-one cases of MPM were collected from the department of pathology at the NCI, Cairo. Cases were stained by monoclonal antibodies against CK5/6, calretinin, vimentin, CD15, CEA and p27. RESULTS: More than half (57.4%) of the patients were residents in endemic areas; 50.8% were of epithelioid type. CK5/6 was positive in 45 (73.8%) cases, 39 (63.9%) cases were positive for vimentin, 49 (80.3%) cases were positive for calretinin. One case showed a focal weak positive reaction to CD15. None of the cases stained for CEA. There was a statistically significant relation between p27 expression and the histopathologic type (p=0.02) between overall survival and age (p=0.01), histopathologic type (p=0.02) and stage (p=0.006). CONCLUSION: MPM is an increasing disaster in Egypt which is underestimated and neglected. A panel of immunohistochemical markers should be used for proper evaluation. p27 has proven to be a potential biologic prognostic marker for mesothelioma and more studies as regard its significance are recommended on a larger number.