Parstatin inhibits viability of human retinal pigment epithelium cells: an in vitro cytotoxicity study.

OBJECTIVE: Parstatin, the N-terminal 41-amino-acid peptide cleaved by thrombin from protease-activated-receptor 1, was shown to be highly effective in preventing ocular angiogenesis and as such it has potential therapeutic applications in ocular neovascular diseases. In the frame of a safety program in preclinical development, we investigated whether parstatin exerts any cytotoxic effect in critical ocular cell populations. MATERIALS AND METHODS: Human retinal pigment epithelium cell-19 line (ARPE-19) and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay were used to investigate parstatin's effect in cell cultures. Parstatin 24-41 was used as a control peptide which lacks the hydrophobic domain to demonstrate the specificity and the structure-dependent effect of parstatin. Both peptides were used at concentrations ranging from 0.1-30 µM for 24, 48 and 72 hours. RESULTS: In the presence of parstatin the rate of ARPE-19 cell growth and viability were significantly decreased in a concentration-dependent and time-dependent manner, with an IC50 of approximately 10 µM. When ARPE-19 cells were treated with parstatin 24-41 no inhibitory effect was observed at any concentration or exposure time used. CONCLUSIONS: Parstatin has a clear detrimental effect on the viability of ARPE-19 cells and raises concerns about its use in the eye because of its possible toxic effects.

Expression of estrogen receptors in melanoma and sentinel lymph nodes; a "female" clinical entity or a possible treatment modality?

PURPOSE: The natural history of human malignant melanoma suggests that steroid hormones may affect the biological behavior of this tumor. The purpose of the current study was to investigate the specific immunostaining patterns of estrogen receptors in malignant melanomas and their sentinel lymph nodes (SLNs), as well as to examine any possible association with patients' prognosis and overall survival. MATERIALS AND METHODS: A retrospective analysis of prospectively collected data was conducted during a 12-year period (2001-2012). Sixty patients with mean age of 54.4 ± 14.5 years diagnosed with melanomas of varying depth (Clark) and thickness (Breslow) after excision biopsy of pre-existing melanocytic lesions, were included in the study. All patients underwent wide excision of the primary tumor and SLN identification. Determination of estrogen receptor alpha (ERa) and beta (ERb) status by immunohistochemistry on tumor and nodal paraffin blocks was performed in all feasible cases. RESULTS: ERb but not ERa was the predominant estrogen receptor found in all primary tumors and SLNs examined. The most intense ERb immunostaining was seen in negative SLNs associated with thinner, less invading melanomas. ERb expression in the primary tumor seems to correlate with the cellular microenvironment, possibly altering the process of SLN invasion. CONCLUSIONS: ERb expression is down-regulated in aggressive melanomas with sentinel nodal metastatic disease, suggesting its possible usefulness as a surrogate marker for metastatic potential and prognosis in malignant melanoma.

Good's syndrome and pure white cell aplasia complicated by cryptococcus infection: A case report and review of the literature.

Thymomas can present with a variety of paraneoplastic manifestations, mostly of autoimmune origin, including Good's syndrome when there is associated hypogammaglobulinemia. Although pure red cell aplasia is a recognised complication of thymoma, selective white cell aplasia is very rare, particularly in Good's syndrome. Lethal opportunistic infections are a feature of Good's syndrome, usually occurring in those patients with associated severe T lymphocyte defects. Although the cryptococcus is a recognised fungal pathogen in patients with other causes of CD4+ T cell lymphopenia, surprisingly this complication has not been reported in patients with Good's syndrome. We now describe a 70 year old man with Good's syndrome and pure white cell aplasia who presented with disseminated cryptococcosis, and provide an up-to-date review of the relevant literature. Despite meningeal involvement our patient recovered after combined treatment with intravenous globulin, granulocyte stimulating growth, corticosteroids and antifungal therapy.

Differential expression and cross-talk of peroxisome proliferator-activated receptor γ and retinoid-X receptor α in urothelial carcinomas of the bladder.

PURPOSE: the peroxisome proliferator-activated receptor γ (PPARγ), known to play a key role in homeostatic biological pathways, is also implicated in the process of carcinogenesis. Ligands for PPARγ and its heterodimeric partner, retinoid-X receptor (RXR), have exhibited anticancer effects both in vitro and in vivo. Unexpectedly, some studies suggested that PPARγ ligands may stimulate cancer formation. This study aimed to estimate the signaling of PPARγ-RXRα heterodimer in bladder urothelial carcinomas (BUC). METHODS: we studied PPARγ and RXRα expression in specimens obtained from 97 patients with BUC of various grades and stages using immunohistochemistry. RESULTS: PPARγ expression was significantly downregulated with BUC stage and grade progression, and the dynamics of this phenomenon was significantly influenced by RXRα's level of expression. CONCLUSION: the positive association of PPARγ expression in BUC with more differentiated, non-invasive tumors is strengthened by the presence of RXRα. This knowledge could probably be of use in the development of new chemotherapeutic agents.

Rice-body formation and tenosynovitis of the wrist: a case report.

A 61-year-old woman presented with mild pain and swelling on the volar aspect of the wrist, hand, and little finger. Radiography showed a soft-tissue mass shadow, and magnetic resonance imaging showed acute tenosynovitis of the flexor tendons and an inflammatory mass inside the carpal tunnel. Laboratory test results were normal, except for an elevated erythrocyte sedimentation rate (40 mm/h). The patient had an ambiguous Mantoux test result but no history of mycobacterial exposure. Exhaustive investigations for rheumatic disease were negative. Surgical exploration of the lesion revealed rice bodies in the common flexor tendon synovial sheath, extending distally to the tip of the fifth finger. Removal of the rice bodies and thorough excision of the sheath was performed. The patient regained a full and painless range of motion in about 3 months. One-year follow-up revealed no underlying disorder.

Non-Hodgkin's lymphomas in Greece according to the WHO classification of lymphoid neoplasms. A retrospective analysis of 810 cases.

The purpose of this retrospective study, the largest unselected series in our country, was to illustrate the clinicopathological features of non-Hodgkin's lymphoma (NHL) classified according to the World Health Organization (WHO) classification of lymphoid neoplasms. A retrospective analysis was conducted and clinical features of histological subtypes were established in 810 patients (age > or = 15 years) with NHL who were treated at 8 major centers representative of Greece. There were 435 males and 375 females 95% of them aged >30 years. B symptoms were present in 34% of the patients, while 45.3% had stages I-II and 54.6% had stages III-IV. LDH was increased in 37% of the patients. B cell lymphomas formed 88% of the cases whereas T cell lymphomas formed 12% of the total. Indolent lymphomas accounted for 31.1%, aggressive ones for 66.7% and very aggressive ones for 2.4% of all NHLs. Among indolent lymphomas extranodal ones (MALT B cell lymphoma) were the most common subset while follicular lymphoma grade I and II and small lymphocytic ones presented with equal frequency. Among the aggressive lymphomas diffuse large cell lymphoma (DLCL) was the most common subtype; this entity along with large-cell immunoblastic lymphomas accounted for 45.2% of all B cell lymphomas. Among the T cell lymphomas, peripheral T cell lymphomas and anaplastic large cell lymphomas of the T/null-cell type were the most common subtypes. The most common extranodal presentation was the gastrointestinal tract (GI). Next in frequency were primary extranodal NHL of the head and neck region. MALT B cell lymphomas were found in almost half of the patients with GI tract NHL, whereas in all other extranodal places DLCL was the predominant histological subtype. The median survival for indolent and aggressive NHL was 123.5 and 55.5 months, respectively. This is the first report of a large series of malignant lymphomas in Greece using the WHO classification. It appears that there are no significant differences between NHL in Greece and other large series as far as clinical and extranodal presentation is concerned. The frequency of follicular lymphoma in the current study is comparable to that reported from Asian countries and mainland Europe, but lower than that of US and Northern European series. There were no important differences in the incidence of the remaining histological subtypes between Greece and other European countries.

Gastric ulcers and risk for cancer. Is follow-up necessary for all gastric ulcers?

The aim of this study was to determine the accuracy of initial endoscopy combined with histology and to define whether there is a point in following-up all gastric ulcers until complete healing. We have studied all patients with gastric ulcers documented at endoscopy during a 6-year period. Ulcers were macroscopically characterised as benign or suspicious for malignancy, and biopsies were taken. A follow-up endoscopy and histology was performed 4-6 weeks and 3 months after an anti-ulcer treatment. Resistant ulcers were treated surgically. All patients were followed-up clinically and endoscopically for a year after complete ulcer healing. 802 patients with gastric ulcers were enrolled. At initial endoscopy, 732 ulcers (91.3%) were macroscopically characterised as benign and 70 ulcers (8.7%) as suspicious for malignancy. In the group of endoscopically benign ulcers, only one (0.1%) had malignancy detected by biopsy in the first examination. None of these ulcers turned out to be malignant on subsequent examinations. From the suspicious for malignancy ulcers, 20 (28.6%) were proven to be malignant. Endoscopy may recognise with great accuracy benign ulcers, but it overestimates the malignant ones. The cost-benefit of serial follow-up endoscopies should be re-evaluated in ulcers that appear benign, and biopsies are negative at the initial examination.

Hepatocellular carcinoma metastasis in the pituitary gland: case report and review of the literature.

Symptomatic pituitary metastases are uncommon and have been reported mainly in autopsy series. Although all types of malignancies can metastasize to the pituitary gland, a review of the literature has indicated that lung and breast carcinomas are the most frequent primary tumors while hepatocellular carcinoma metastasis has only recently been described. A 59-year-old man with abdominal pain and fever was admitted to our hospital. Hepatosplenomegaly was present without signs of ascites. Laboratory tests showed only abnormal hepatic biochemistry while the radiological studies revealed a solid mass occupying the left hepatic lobe. The patient underwent excision of the left hepatic lobe and was closely followed-up. Six months later he readmitted with headache and visual disturbances. MRI revealed a solid mass in the sella region pressing the optic chiasma. Transsphenoidal excision of the pituitary mass was followed and the histological examination of the tumor was compatible with hepatocellular carcinoma. Symptomatic pituitary metastases are uncommon and may be difficult to differentiate from pituitary adenomas. The present case emphasizes on the capricious nature of hepatocellular carcinoma and on the importance of the individualized therapeutic approach.

Corneal complications following abuse of topical anesthetics.

PURPOSE: To describe corneal complications in three patients following abuse of topical anesthetics. MATERIALS AND METHODS: We describe one case with bilateral corneal perforation, another with unilateral corneal perforation and a third case of chronic keratitis following excessive use of topical anesthetics. RESULTS: Two patients continued to instill topical anesthetic drops despite all recommendations to stop. The result was a bilateral corneal perforation in the first case and a large unilateral descemetocele in the second. The third patient who had chronic toxic keratitis discontinued the anesthetic drops and after the appropriate treatment the cornea returned to normal. Corneal grafting and conjunctival flaps were used to seal the corneal perforation but the long-term anatomical and functional results were very poor. CONCLUSIONS: The initial presentation of this rare clinical entity creates difficulties in reaching a correct diagnosis. A presumed acanthamoeba keratitis is the first choice among many similar conditions. Thus abuse of topical ocular anesthetic drops should be included in the differential diagnosis of cases of chronic keratitis as it may masquerade as acanthamoeba keratitis. A current or past history of psychiatric and mental disorders or psychoactive substance abuse is important in the diagnosis. Functional and anatomical results after appropriate treatment are usually poor. Psychiatric counselling is extremely helpful and is in fact mandatory in the management of these patients.

Determination of plasma cell secreting potential as an index of maturity of myelomatous cells and a strong prognostic factor.

According to the widely accepted myeloma staging system, the bulk of paraprotein is the main determinant of disease stage. However, myelomatous plasma cells differ considerably in their ability to synthesize and secrete monoclonal paraprotein. We determined plasma cell secreting potential (PCSP) as the amount of M-component, divided by the percentage of marrow plasmacytic infiltration, in 240 patients with myeloma, and correlated our results with chain isotype, plasma cell morphology, severity of bone disease, well-recognized prognostic factors, such as serum LDH, CRP, albumin and beta2-microglobulin, treatment response and overall survival. PCSP was higher in IgG than in other myeloma types, and was an almost constant parameter for each individual patient, in 134/166 cases. A > 10% decrease of PCSP in 26 patients was associated with disease aggressiveness and treatment failure. Patients with MGUS had significantly higher PCSP than those with myeloma of the same chain type. Higher PCSP was associated with stage I, absence of Bence-Jones proteinuria and indolent forms of disease with lower proliferating cell nuclear antigen (PCNA) positivity, serum LDH, alpha2-globulins, CRP and beta2-microglobulin and higher albumin levels. Conversely, patients with immature/plasmablastic morphology and those with severe bone disease had lower PCSP. Good responders to treatment had significantly higher PCSP than moderate and poor responders and PCSP was strongly correlated with overall survival in IgG and IgA myeloma. In conclusion, PCSP reflects the maturation status of myelomatous cells and therefore can be used as a prognostic factor, since patients with high secreting potential represent a lower malignancy group, in comparison to those with a low secreting potential.

Asymptomatic gastrocnemius muscle biopsy: an extremely sensitive and specific test in the pathologic confirmation of sarcoidosis presenting with hilar adenopathy.

OBJECTIVE: To evaluate asymptomatic gastrocnemius muscle biopsy as a tool in the histologic confirmation of the diagnosis of sarcoidosis. METHODS: Twenty-two patients admitted over a 2-year period to our department with bilateral hilar adenopathy and a variety of symptoms compatible with sarcoidosis were studied prospectively. Besides a complete history and physical, serum angiotensin converting enzyme (SACE) determination, pulmonary function, slit lamp eye examination, PPD skin test, gallium 67 scan and gastrocnemius muscle biopsy were performed. RESULTS: The biopsy revealed non-caseating granuloma in all patients, confirming the diagnosis of sarcoidosis. No other patient in our department received this diagnosis over the 2-year period of the study. The procedure was well tolerated by all patients and almost zero morbidity was noted. Erythema nodosum was present in 68.2% of the patients, PPD was negative in all of them, SACE was elevated in 59.1% and pulmonary function was normal in the majority. CONCLUSION: The impressive sensitivity of asymptomatic gastrocnemius muscle biopsy, its safety and ease of performance, along with the extreme rarity of muscle involvement by other granulomatous diseases included in the differential diagnosis, may render it the procedure of choice for the histologic confirmation of sarcoidosis presenting with hilar adenopathy.

Microvessel density, proliferating activity, p53 and bcl-2 expression in in situ ductal carcinoma of the breast.

BACKGROUND: Ductal carcinoma in situ (DCIS) of the breast represents a heterogeneous group of lesions that show important differences in biologic behavior. New vessel formation has been reported as a prognostic indicator in breast carcinoma, but little information is available about its significance in DCIS. This study was planned to examine angiogenesis in DCIS in relation to histologic subtype, proliferation activity, p53 and bcl-2 expression. MATERIALS AND METHODS: Paraffin sections from 24 cases of DCIS (9 comedo and 15 non comedo type) were studied immunohistochemically using polyclonal and monoclonal antibodies to von Willebrand factor, Ki-67, p53 (clone 1801) and bcl-2 proteins. The streptavidine-biotin technique with microwave antigen retrieval was employed. RESULTS: Most cases showed enhanced microvessel formation around ducts with DCIS compared to normal ducts. Comedo carcinomas (CCs) showed enhanced neovascularization compared to non comedo carcinomas (NCCs). Growth fraction determination with Ki-67 antibody showed that 78% of the CCs expressed high proliferating activity compared to 27% of the NCCs. p53 immunoexpression was noted in 78% of the CCs and 20% of the NCCs. Bcl-2 immunoreactivity was observed in 67% of the total cases in 58% of which there was no association with p53 expression. However, an association was found between neovascularization and overexpression of Ki-67 and p53. CONCLUSIONS: This study suggests that neovascularization is an early phenomenon in breast neoplasia and is apparent as early as the in situ stage. CCs express a more aggressive immunophenotype, compared to the other DCIS subtypes, characterized by increased stromal interaction, high proliferating activity, p53 overexpression and a near lack of bcl-2 immunostaining.

Sjögren's syndrome in patients with newly diagnosed untreated non-Hodgkin's lymphoma.

The purpose of the study was to detect cases of Sjögren's syndrome among newly diagnosed untreated patients with non-Hodgkin's lymphoma and, furthermore, to identify in such cases clinical and serologic features known to occur more frequently in Sjögren's syndrome patients who evolve into lymphoma. Accordingly, thirty-three cases of newly diagnosed non-Hodgkin's lymphoma, prior to any treatment administration, were thoroughly studied for evidence of Sjögren's syndrome. Immunophenotyping for T and B cells and kappa and lambda light chains was concomitantly performed on both lymphomatous tissues and minor salivary glands. There were 5 patients with T cell and 28 with B cell lymphoma of various histologic subtypes and grades. Two of the latter (7.1%) had a positive for Sjögren's syndrome minor labial salivary gland biopsy, positive responses to the specific questionnaires for both eye and mouth dryness and abnormal Schirmer's and rose Bengal eye tests, substantiating the diagnosis of Sjögren's syndrome. Both were male with lung and stomach non-Hodgkin's lymphoma respectively, enlargement of the lacrimal glands, monoclonal gammapathy of the IgM kappa type and, one of them had high titer and of fine speckled pattern positive antinuclear antibodies and anti-Ro(SSA) and anti-La(SSB) antibodies in his serum. A monotypic infiltrate with kappa light chain restriction, identical to that in the lymphomatous tissue of these two patients, was present in their minor salivary gland biopsy as well. Such a finding was not encountered in any of the remaining patients. Although our sample is relatively small, our results confirm the relationship between Sjögren's syndrome and non-Hodgkin's lymphoma, looked at from the opposite direction. Obviously, studies involving larger populations would be more definitive, regarding the issue of what percentage of this lymphoma patients originates from Sjögren's syndrome.

Oral ornithine a-ketoglutarate accelerates healing of the small intestine and reduces bacterial translocation after abdominal radiation.

The effect of dietary ornithine a-ketoglutarate (OKG) on intestinal mucosal integrity and bacterial translocation was studied in rats following administration of a single dose of abdominal radiation (1100 cGy). Following the radiation injury the rats were randomized to receive a nutritionally incomplete diet which contained only water and OKG or a control diet with water and the non-essential amino-acid glycine. Four days after radiation, rats were anaesthetized and a laparotomy was performed. Cultures from mesenteric lymph nodes were taken and two tissue samples from the terminal ileum were also taken for light microscopy, protein and DNA determination. We examined the following parameters: number of villi per cm (V/cm), villus height (Vh), number of mitoses per crypt (M/c) and we measured the mucosal protein and DNA content. Nine of 16 rats who received the OKG-free diet had positive cultures but only 3 of 18 rats who received the OKG-enriched diet (P= 0.002). The group on the OKG-enriched diet had a better intestinal mucosal architecture than the group on the OKG-free diet and the studied parameters of the gut mucosa were significantly better: (V/cm: 130 +/- 8.1 vs 99 +/- 7.9, P = 0.001. Vh(mm): 0.40 +/- 0.03 vs 0.24 +/- 0.05, P= 0.002. M/c: 1.71 +/- 0.03 vs 0.34 +/- 0.2, P= 0.001, Protein (mg/cm): 2.300 +/- 0.033 vs 1.207 +/- 0.014, P = 0.002. DNA (microg/cm): 203 +/- 6.41 vs 130 +/- 4.94, P = 0.001. We conclude that OKG-enriched diet prevents the deleterious effects of radiation on intestinal mucosal morphology and integrity, abolishing thus, the increased bacterial translocation observed after abdominal radiation.