Kalliopi Stefanaki

Enhanced mRNA expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in breast carcinomas is correlated with adverse prognosis

Tissue inhibitor of metalloproteinase‐1 (TIMP‐1) has emerged as a multifunctional protein with the contrasting activities of inhibiting tissue‐degrading enzymes and promoting cellular growth. In an attempt to elucidate the clinical significance of TIMP‐1 in breast cancer, the expression of TIMP‐1 mRNA was evaluated in 117 invasive breast carcinomas by mRNA in situ hybridization, in correlation with clinicopathological parameters, immunohistochemical prognostic factors (Ki‐67, c‐erb‐B‐2, bcl‐2) and clinical outcome. TIMP‐1 was detected in stromal cells in areas within the tumours and at the tumour margin. High TIMP‐1 mRNA expression in the marginal portion of the tumours was significantly correlated with lymph node metastasis (p<0.05) and c‐erbB‐2 expression (p<0.05). On the other hand, increased TIMP‐1 mRNA expression within the tumours showed a statistically significant correlation with ER detection (p<0.01). Multivariate analysis revealed worse survival for patients with high TIMP‐1 mRNA expression in the marginal portion of the tumours; the subgroup of these patients co‐expressing high levels of TIMP‐1 mRNA within the tumours as well had even worse survival (p=0.042). In conclusion, our data support the multifunctional role of TIMP‐1, particularly its growth‐promoting activity, on the basis of its significant correlation with lymph node metastasis and adverse prognosis. In addition to the latter property, a probable association of TIMP‐1 with tumour cell differentiation is suggested by its topographical correlation with ER detection. Copyright

Immunohistochemical Expression of Caspase-3 as an Adverse Indicator of the Clinical Outcome in Human Breast Cancer

Objective: Tumor growth is the net result of cell proliferation and cell loss by apoptosis. Caspase-3 (CPP32) has been considered as most directly correlated with apoptosis because of its location in the protease cascade pathway. The aim of this study was to examine the relation of the immunohistochemical expression of caspase-3 in 137 infiltrating breast carcinomas to patients’ clinicopathological parameters and survival. Method: A polyclonal antibody against caspase-3 was applied using a standard immunohistochemical procedure to paraffin sections. Results: By comparison with nonneoplastic breast tissue, caspase-3 appeared to be upregulated in malignant breast tissue, and its overexpression status was detected in 75.2% of the specimens. Significant statistical correlations were observed between overexpression of caspase-3 and low nuclear grade (p = 0.048), lack of p53 protein accumulation (p = 0.039), bcl-2-positive immunostaining (p = 0.027), as well as tissue inhibitor of metalloproteinase-2 immunoreactivity of neoplastic (p = 0.012) and stromal cells (p = 0.0001). Despite the above correlations, multivariate analysis revealed a significant negative influence of caspase-3 expression on patients’ overall survival (p = 0.029). Conclusions: Caspase-3 protein overexpression appears to be involved in the apoptotic pathways influenced by wild-type p53 and bcl-2 proteins. Moreover, the results show that caspase-3 overexpression in breast cancer cells seems to exert an independent adverse effect on patients’ overall survival.

Cutaneous Granulomas with Predominantly CD8+ Lymphocytic Infiltrate in a Child with Severe Combined Immunodeficiency

Background: Combined immunodeficiency disorders comprise a heterogeneous group of diseases characterized by both humoral and cell-mediated immunodeficiency. Cutaneous granulomas manifestations in children with combined immunodeficiency are rare. Objective: We report the case of a 6-year-old boy who presented with disseminated cutaneous granulomas and a history of multiple infections. Methods and Results: Laboratory evaluation revealed severe combined immunodeficiency, and deoxyribonucleic acid (DNA) analysis confirmed mutations on a gene of chromosome 19 that encodes an enzyme called Janus kinase 3 (Jak-3). Immunohistochemistry revealed expression of CD8+ in the perivascular lymphocytic infiltrate Conclusion: Disseminated granulomatous lesions in children with a history of frequent infections should prompt the clinician to initiate detailed immunocompetence evaluation as they might prove to be the first manifestation of immunologic impairment.

In situ evidence of cellular senescence in Thymic Epithelial Cells (TECs) during human thymic involution

Cellular senescence, an age-related process in response to damage and stress, also occurs during normal development and adult life. The thymus is a central lymphoepithelial organ of the immune system that exhibits age-related changes termed thymic involution. Since the mechanisms regulating thymic involution are still not well elucidated, we questioned whether cellular senescence is implicated in this process. We demonstrate, for the first time in situ, that cellular senescence occurs during human thymic involution using SenTraGor™, a novel chemical compound that is applicable in archival tissue material, providing thus further insights in thymus histophysiology.

The frequency of hepatitis B virus infection in Greek patients with various types of glomerulonephritis

The frequency of hepatitis B surface antigen (HBsAg) was studied in the sera of 622 patients with glomerulonephritis (GN). The prevalence of HBs-antigenemia was 2.8% (18/622; eleven adults and seven children); the difference from 2.6% in the general population of Central and Southern Greece was not statistically significant (χ2=0.01;p>0.50). Two of the 11 HBsAg-seropositive adult patients with GN suffered from IgA nephropathy, one from IgA and membranous glomerulonephritis (MGN), four from diffuse proliferative GN, two from membranous GN and one each from crescentic GN and focal segmental glomerulosclerosis. Five children out of 12 with membranous glomerulonephritis, one out of 24 with IgA nephropathy and one out of 16 with focal segmental glomerulosclerosis had HBs-antigenemia. The frequency of HBs-antigenemia in children with MGN was 41.7%, which is significantly higher than in children with others types of GN (0.9%). All seropositive patients were asymptomatic HBsAg carriers, while one seropositive HBsAg child with MGN suffered from chronic persistent hepatitis. HBsAg was detected by the immunoperoxidase-antiperoxidase (PAP) method in the glomeruli of only 3 children with MGN and HBs antigenemia, while HBcAg was not detected in any case. Our study suggests that in the Greek population there is no increased prevalence of HBs-antigenemia in patients with glomerulonephritis. Moreover, HBsAg was not found to contribute in the pathogenesis of GN in adults but it may be associated with the pathogenesis of membranous GN in children.

Pediatric vasculitis: a single center experience

Existing studies of children with vasculitis are limited. The aim of this study was to assess the epidemiology, clinical manifestations, laboratory findings, course, and outcome of Greek children presenting with vasculitic rash.

The child with vasculitic rash: a 10‐year retrospective study

Editor Vasculitis is defined as the presence of inflammation in the blood vessel walls. The overall incidence of childhood vasculitis is estimated to be 50 cases per 100 000 children per year. Children admitted in our department between 2003 and 2013 with the provisional diagnosis of vasculitis of the skin and who had a follow-up in the ensuing 6 months were included in the study. There were 95 children (58 boys, 37 girls) with a mean age of 5.9 years. In total, 76 (80%) of the children were diagnosed with Henoch–Sch€ onlein Purpura (HSP), 10/95 (10.5%) with Hypersensitivity Vasculitis (HV), 6/95 (6.3%) with Urticarial Vasculitis (UV) and 3/95 (3.1%) with Acute Haemorrhagic Oedema of Infancy (AHOI) (Fig. 1). The mean age of the children was 5.7 years for HSP, 8.5 years for HV, 6.2 years for UV and 0.2 years for (AHOI) respectively. HSP is considered the most common paediatric vasculitis; there is a male predominance worldwide that was also noticed in our study. This finding seems to contradict the sex distribution in other rheumatologic diseases and the reason has not yet been elucidated. Skin rash in HSP seems to be the hallmark of the disease with a nearly 100% involvement, as demonstrated in our study too. Gravity causes immune complexes to accumulate in dependent areas of the body, which explains the predilection of skin lesions over the lower legs and buttocks in ambulatory patients and for the lumbar area, buttocks and ears in infants respectively. Current evidence does not support universal treatment of HSP with corticosteroids, as they do not appear to prevent the onset of renal disease or abdominal complications, nor do they alter recurrence rates. However, corticosteroids do seem to have a role in the symptomatic management of HSP, specifically in treating abdominal pain and arthralgia. In our group, HSP patients with severe abdominal pain who did not respond to analgesics were successfully treated with corticosteroids despite the controversy accompanying the efficacy of their use. Thirty of 76 HSP cases presented with nephritic symptoms and 21 of 30 children were above 5 years of age, a finding consistent with other studies. This might reflect a higher degree of immune stimulation and subsequently a higher severity degree of renal pathology, following the initial trigger factor, in the older age groups though further studies are required to investigate this hypothesis. It is worth noting that 11 of 17 HSP cases who relapsed were above 5 years of age, a finding that parallels the increased incidence of nephritis for the same age group. The HV skin lesions, may be indistinguishable from HSP lesions on clinical grounds but the duration of each lesion and the distribution is quite different. The rash of our HV patients was distributed in a more generalized form compared with the HSP patients and lasted on average 10 days longer than the rash observed in HSP patients. UV patients followed a relapsing course for 3 months on average and manifested in all six children with a characteristic urticarial rash lasting for more than 24 h, similar to other studies. In this group, one patient developed systemic lupus erythematosus and another two patients blood malignancies a few months after the onset of UV. In AHOI cases, the skin lesions were mainly ecchymotic, larger in size compared to HSP and the patients had no other systemic complications, a finding consistent with other studies. In conclusion, the overall prognosis of patients with paediatric skin vasculitides is good, with the exception of those with the UV type. In these patients an underlying disease should be suspected and thoroughly investigated.