Isolation and characterization of Oct-4+/HLA-G+ mesenchymal stem cells from human umbilical cord matrix: differentiation potential and detection of new markers.

The presence of multipotent cells in several adult and embryo-related tissues opened new paths for their use in regenerative medicine. Extraembryonic tissues such as umbilical cord are considered a promising source of stem cells, potentially useful in therapy. The characterization of cells from the umbilical cord matrix (Wharton's Jelly) and amniotic membrane revealed the presence of a population of mesenchymal-like cells, sharing a set of core-markers expressed by "mesenchymal stem cells". Several reports enlightened the differentiation capabilities of these cells, even if at times the lack of an extensive characterization of surface markers and immune co-stimulators expression revealed hidden pitfalls when in vivo transplantation was performed. The present work describes a novel isolation protocol for obtaining mesenchymal stem cells from the umbilical cord matrix. These cells are clonogenic, retain long telomeres, can undergo several population doublings in vitro, and can be differentiated in mature mesenchymal tissues as bone and adipose. We describe for the first time that these cells, besides expressing all of the core-markers for mesenchymal stem cells, feature also the expression, at both protein and mRNA level, of tolerogenic molecules and markers of all the three main lineages, potentially important for both their differentiative potential as well as immunological features.

Role of environmental and genetic factor interaction in age-related disease development: the gastric cancer paradigm.

The association of Helicobacter pylori (Hp) infection with gastric cancer is well known and might be considered a paradigmatic example of the role that interaction among environmental factors and individual background might play in inducing age-associated disease. To evaluate the role of interaction of Hp infection with genetic background, gastric cancer and chronic gastritis patients as well as random selected controls were typed for five inflammation-related polymorphisms of IL-1 and IL-10 cytokine genes. No association among IL-10 or IL-1 variants with an increased risk of gastric cancer was found, whereas an Hp-independent association of IL-1beta -511T positive genotypes to an increased risk of chronic gastritis was found (Hp-/511T+ OR 1.89, 95% CI: 1.01-3.54; Hp+/-511T+ OR 1.83, 95% CI: 1.05-3.19). Stratification of gastric cancer group according to Hp infection does not allow finding a statistically significant association of Hp+ to the higher histological grading (G3) of gastric cancer (OR 1.54, 95% CI: 0.46-5.11). Our findings seem to confirm that cytokine genetic variants might contribute to determining the background for inflammaging in which H. pylori infection might facilitate cancer development.

[Two cases of actinomycosis]. / Due casi di infezione actinomicotica.

Actinomycosis is a bacterial infection due to Gram-positive bacteria of the Actinomyces genus. The authors describe two cases: one of them occurred in a woman with a clinical presentation of a right submandibular indurative mass. After surgery, histological examination showed a granulomatous tissue with many actinomycetes. The second case was observed in a man admitted to hospital with a diagnosis of pulmonary heteroplastic lesion. Also in this case, histological examination showed many actinomycotic colonies. Actinomycosis is an uncommon disease. Establishment of definite diagnosis requires a high index of suspicion in all cases of relapsing cervical lesions. Pulmonary actinomycosis is a rare diagnosis; respiratory physicians should be aware of this important differential when investigating patients for persistent pulmonary shadowing.

HER2/neu expression in relation to clinicopathologic features of breast cancer patients.

We have evaluated HER2/neu expression in 1,355 breast cancer patients recruited at the Breast Cancer Registry in Palermo between January 1999 and December 2004. In this retrospective study, HER2/neu expression was related to clinicopathologic features of the disease, including tumor size, nodal and menopausal status, estrogen and progesterone receptors. Statistical analysis on all 1,355 patients showed a significant correlation between HER2/neu and nodal status (P < 0.001), and a significant association between HER2/neu overexpression and estrogen and progesterone receptors status (P < 0.001). In 194 patients without metastasis, with an average follow-up > or =5 years, only HER2/neu 3+ and histopathologic grading G3 were significantly associated with overall survival.

The expression of HSP60 and HSP10 in large bowel carcinomas with lymph node metastase.

BACKGROUND: The involvement of Heat Shock Proteins (HSP) in cancer development and progression is a widely debated topic. The objective of the present study was to evaluate the presence and expression of HSP60 and HSP10 in a series of large bowel carcinomas and locoregional lymph nodes with and without metastases. METHODS: 82 Astler and Coller's stage C2 colorectal cancers, of which 48 well-differentiated and 34 poorly-differentiated, were selected along with 661 lymph nodes, including 372 with metastases and 289 with reactive hyperplasia only, from the same tumours. Primitive tumours and both metastatic and reactive lymph nodes were studied; specifically, three different compartments of the lymph nodes, secondary follicle, paracortex and medullary sinus, were also analysed. An immunohistochemical research for HSP60 and HSP10 was performed and the semiquantitative results were analysed by statistical analysis to determine the correlation between HSPs expression and 1) tumour grading; 2) degree of inflammation; 3) number of lymph nodes involved; 4) lymph node compartment hyperplasia. Moreover, western blotting was performed on a smaller group of samples to confirm the immunohistochemical results. RESULTS: Our data show that the expression of HSP60, in both primary tumour and lymph node metastasis, is correlated with the tumoral grade, while the HSP10 expression is not. Nevertheless, the levels of HSP10 are commonly higher than the levels of HSP60. In addition, statistical analyses do not show any correlation between the degree of inflammation and the immunopositivity for both HSP60 and HSP10. Moreover, we find a significant correlation between the presence of lymph node metastases and the positivity for both HSP60 and HSP10. In particular, metastatic lymph nodes show a higher percentage of cells positive for both HSP60 and HSP10 in the secondary follicles, and for HSP10 in the medullary sinuses, when compared with hyperplastic lymph nodes. CONCLUSION: HSP60 and HSP10 may have diagnostic and prognostic significance in the management of this tumour and their overexpression in tumoral cells may be functionally related to tumoral progression. We hypothesise that their expression in follicular and medullary cells of lymph nodes may be induced by formation of metastases. Further studies based on these observations could lead to a better understanding of the HSPs involvement in colorectal cancer progression, as well as other neoplasms.

Association between single nucleotide polymorphisms in the cyclooxygenase-2, tumor necrosis factor-α, and vascular endothelial growth factor-A genes, and susceptibility to hepatocellular carcinoma.

Cyclooxygenase-2 (COX-2), vascular endothelial growth factor-A (VEGF-A), and tumor necrosis factor-α (TNF-α) are mediators of inflammation and angiogenesis; all of them are produced in liver cirrhosis (LC) and in hepatocellular carcinoma (HCC). It was proposed that there is an association between single nucleotide polymorphisms (SNPs) and HCC. These allelic variants influence the transcriptional activity of these genes, and therefore the proteins levels. The VEGF-A pathway is a potential therapeutic target in HCC, and several antiangiogenic agents have entered clinical trials in HCC. We evaluated the frequency of SNPs of COX-2, TNF-α, and VEGF-A genes in patients with HCC versus LC patients and a control group. The aim of this article was to verify the correlation between the allelic variations and the risk of developing HCC. The study included 96 HCC, 79 LC patients, and 162 healthy subjects. The evaluation of SNPs was performed by the restriction fragment length polymorphism (RFLP-PCR) method. The SNPs analyzed were: -1195 G>A of the COX-2 gene, -308 G>A of the TNF-α gene, and +936 C>T of the VEGF-A gene. Chi-square and Fisher exact tests were used for statistical analysis. Our results confirm that carriers with the C allele in the VEGF-A gene are more frequent in HCC versus LC (p=0.039), suggesting that this SNP may predispose to the development of HCC.

Cystic hydatidosis: a rare case of spine localization.

Cystic hydatidosis is a zoonosis endemic both to Sicily and other Mediterranean areas. Generally, Echinococcus granulosus tapeworms develop in the liver, lung and less frequently in the peritoneum, spleen or kidney. We present a rare case of spinal hydatid disease. The patient was a 38-year-old housewife with a vertebral echinococcosis revealed by acute paraplegia of the legs. Medical treatment with albendazole and surgical intervention improved the clinical symptoms. This case is emblematic both for the unusual localization and for the need of a multidisciplinary approach for diagnosing and monitoring suspected hydatid lesions. Patients with suspected abdominal or lung echinococcosis should also be investigated for other localizations such as the brain, spine and heart. Furthermore, in endemic areas hydatidosis must be suspected in the presence of lesions occupying space in these districts.

Genetic determined downregulation of both type 1 and type 2 cytokine pathways might be protective against pancreatic cancer.

Many cytokine polymorphisms have been studied for associations with susceptibility to breast, gastric, liver, lung, prostate, and ovarian cancer without conclusive results. The cytokine network, indeed, is characterized by complex interactions, and the final biological effect of a single genetic variation depends on the balance among different molecular signals. As is well known, Th1/Th2 cytokine unbalanced production might predispose to different pathologies, cancer included. In general, a prolonged type 1 inflammatory response might allow that cells accumulating enough "genetic hits" are promoted to neoplastic transformation. On the other hand, IL-13-producing cells through the IL-13/IL-4 receptor-alpha (R-alpha) pathway might facilitate escape from tumor immunosurveillance. Here are reported data on the evaluation of the influence of some type 2 and type 1 cytokine genetic polymorphisms as risk factors for pancreatic cancer. There was no overall association between pancreatic cancer risk and single cytokine SNPs. On the other hand, in evaluating the influence of combined cytokine genotypes we found that the combined IL-10-1082GA heterozygous and IL-4 Ralpha-1902AA homozygous genotype is underrepresented in the pancreatic cancer subject group. As is well known, the IL-10-1082GA genotype is associated with an intermediate production of this regulatory cytokine, whereas the IL-10-1902AA genotype of the IL-4Ralpha gene is associated with a reduced efficiency in signal transduction when the receptor is engaged by IL-13 or IL-4. These results strongly suggest that a genetic background associated to a mild downregulation of type 1 and type 2 inflammatory signals might be protective against pancreatic cancer.

Hsp60 expression, new locations, functions and perspectives for cancer diagnosis and therapy.

Hsp60 in eukaryotes is considered typically a mitochondrial chaperone (also called Cpn60) but in the last few years it has become clear that it also occurs in the cytosol, the cell surface, the extracellular space, and in the peripheral blood. Studies with prokaryotic models have shown that Hsp60 plays a role in assisting nascent polypeptides to reach a native conformation, and that it interacts with Hsp10 (which also resides in the mitochondria and is also named Cpn10). In addition to its role in polypeptide folding in association with Hsp10, other functions and interacting molecules have been identified for Hsp60 in the last several years. Some of these newly identified functions are associated with carcinogenesis, specifically with tumor cell survival and proliferation. Thus, assessing the levels of Hsp60 in tumor cells and in sera of cancer patients is becoming an attractive area of investigation aiming at the development of means for practical applications in clinical oncology. Since Hsp60 participates in extracellular molecular interactions and cell signalling and also in key intracellular pathways of some types of tumor cells, the idea of using Hsp60 in anti-cancer therapy (chaperonotherapy) is being investigated. The Hsp could be used either as an anticancer agent alone or in combination with tumor antigens, or as target for anti-chaperone compounds. In this article, a brief review is presented of representative research efforts aimed at assessing Hsp60 in a variety of tumors with the purpose of illustrating possible implications and applications for making early and differential diagnoses, assessing prognosis, monitoring response to treatment, and for developing novel anti-cancer strategies.