Cyclophosphamide plus vincristine and prednisone in the treatment of severe pemphigus vulgaris refractory to conventional therapy

Five patients with severe pemphigus vulgaris refractory to conventional therapy with azathioprine and corticosteroids were treated with cyclophosphamide, vincristine and prednisone. One patient was not evaluable, while the remaining four patients showed a complete response. Duration of response was in the range of 13-94 months. Toxicity was mainly represented by alopecia, myelosuppression and gastrointestinal side-effects such as nausea/vomiting. Although cyclophosphamide and vincristine may induce severe side-effects, this association may be useful in controlling severe disease resistant to previous conventional therapies.

Quality controls for cell cultures: identification of interspecies cross-contamination by PCR-RFLP analysis of the cytochrome b gene

Cross-contaminations of a cell line with cells of different species represent a potential risk in laboratories handling human and animal cells. Therefore, it is necessary to control such contaminations. Tests based on mitochondrial DNA (mtDNA) are used in forensic analysis, phylogenetic studies and in food authentication. However, the use of mtDNA in quality controls of cell cultures is recent. Mitochondrial sequence differences of closely related animal species are five- to tenfold higher than those of nuclear genes. On the contrary, intraspecies variation in mitochondrial sequences is low in most animal species. Moreover, each cell contains 100–10.000 mitochondrial genomes. The amount of …

Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial-like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD-MSCs) to characterize and differentiate them into endothelial-like cells. AD-MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony-forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM-2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial-like…

Diagnosi di enteriti infettive di origine virale nel cane

Canine Viral Enteritis are widespread in farms and kennels. Responsible viruses are Parvovirus (CPV), Coronavirus (CCoV), Rotavirus (CRV) and Distemper Virus (CDV). Aim of this study was to assess their prevalence in Sicily and to characterize the strains isolated during 2009-2012. For this purpose, samples (stools, rectal swabs, intestine, liver, spleen, heart, lung, brain) collected from dogs were analyzed by PCR, RT-PCR and Real Time RT-PCR. Positive samples were processed for virus isolation on cell lines. Viruses isolated were analyzed by RFLP and sequencing for molecular characterization. Results show an high prevalence of CPV infection in dogs, followed by CCoV, CRV and CDV. CPV prev…

Obtaining Mesenchymal Stem Cells From Adipose Tissue Of Murin Origin: Experimental Study

In vitro production of differentiated Endotelual Cells from Mesenchimal Stem Cells, collected from adult rat Adipose Tissue (AD-MSCs): preliminary results

Mesenchymal Stem Cells (MSCs) are multipotent cells that reside within various tissues, including adipose tissue and bone marrow. Research concerning the potential therapeutic role of MSCs has advanced significantly over the last decade. Many studies have demonstrated that adipose tissue is an excellent source of MSCs (AD-MSCs), easily accessible in experimental animals. AD-MSCs can be expanded in vitro over the short term and they can differentiate toward a variety of cell types of mesodermal lineage. Therefore, they are thought an attractive tool for cell therapy. Selection of appropriate animal models for preclinical evaluations is crucial for optimization and validation of therapeutic p…

Parthenolide generates reactive oxygen species and autophagy in MDA-MB231 cells. A soluble parthenolide analogue inhibits tumour growth and metastasis in a xenograft model of breast cancer.

Triple-negative breast cancers (TNBCs) are clinically aggressive forms associated with a poor prognosis. We evaluated the cytotoxic effect exerted on triple-negative MDA-MB231 breast cancer cells both by parthenolide and its soluble analogue dimethylamino parthenolide (DMAPT) and explored the underlying molecular mechanism. The drugs induced a dose- and time-dependent decrement in cell viability, which was not prevented by the caspase inhibitor z-VAD-fmk. In particular in the first hours of treatment (1–3 h), parthenolide and DMAPT strongly stimulated reactive oxygen species (ROS) generation. The drugs induced production of superoxide anion by activating NADPH oxidase. ROS generation caused…

Analysis of human dysplastic haematopoiesis in long-term bone marrow culture

In this paper we have analysed the behaviour of myelodysplastic marrow in a long-term bone marrow liquid culture system (LTBMC) from eleven patients with myelodysplastic syndromes with regard to cellularity, day-7 and day-14 CFU-GM growth, cluster formation, adherent cells and CFU-F formation. An altered CFU-GM pattern was found in 64% of cases at diagnosis, while normal growth was seen in the remaining cases, all of which were affected by refractory anaemia. The levels of CFU-GM, as well as cellularity, were reduced in myelodysplastic marrows compared to normal controls over the whole duration of LTBMCs. Cases with a normal CFU-GM level at diagnosis also showed pathological behaviour when …

Synthetic scaffolds may be used for the orientation of cardiac stem cells properties, differentiation and extracellular matrix interactions in cardiac tissue engineering: in vitro and in vivo studies.

West Nile Disease (WND) in Sicily

WND is an emerging vector-borne zoonosis caused by an RNA virus included in the Japanese encephalitis group within the Flavivirus genus. In Italy, after the first appearance in Tuscany in 1998, West Nile Virus (WNV) has been continuously circulating since 2008. A National Surveillance Program consisting of monitoring horses, “sentinel poultries”, wild bird mortality and mosquitoes is active in the entire national territory since 2002. Authors describe the outbreaks which occurred in Sicily in 2010 and 2011. Case study: Between September-October 2010, seven horses from the western part of the island, in the Trapani province, showed neurological symptoms. In October 2011, another two horses s…

Canine Mesenchymal Stem Cells from visceral and subcutaneuous adipose tissue for cell-based therapy

This study compared some characteristics of canine Adipose tissue-Derived Mesenchymal Stem Cells (cAD-MSCs) from subcutaneous and visceral fat. These findings were directed to obtain high quantity and quality cAD-MSCs for clinical cell-based therapy.

An innovative method for the detection of contaminant viral genome in cell cultures

The use of cell cultures involves different fields of biology, from diagnosis to research. Moreover, technologies based on animal cells represent a useful tool to the development of biological products for the prophylaxis and therapy in humans and animals. Therefore, it is necessary to perform quality controls, including virological tests. Several tests performed in research laboratories are able to discriminate one or more viral species, but it is not possible to demonstrate the presence of contaminant viral genome with one non-specific method. The aim of this work consisted on the realization of a biomolecular method able to detect and to identify by sequencing extraneous viral genome in …

Detection of bovine Papillomavirus and efficacy of treatment using saponinDetection of bovine Papillomavirus and efficacy of treatment using saponin

Papillomas of teats and udders in cows may cause skin lesions, thelitis, mastitis and poor milking resulting in economic losses. We describe an outbreak of bovine papillomavirus (BPV) and successful treatment with a purified extract of Quillaja saponaria bark (Quil-A).

Ovine Catarrhal fever (bluetongue): Analysis of Culicoides species in seropositive farms

Bluetongue (BT) is an orbiviral disease of wild and domestic ruminants, mainly sheep. In Sicily, the first Bluetongue outbreak occurred in October 2000; there have been 76 recorded outbreaks so far. The National Surveillance Plan, based on European Union Commission Decision 138/2001/CE, establishes serological and entomological surveys. This plan consists of controls of seronegative cattle, called 'sentry' as indicators for the presence and circulation of virus in defined areas. To check the seroconversions, the regional territory has been subdivided in 400 km2 areas including 58 seronegative cattle, periodically checked by serological tests. All positive sera have been tested to detect the…

Biologic relevance of elevated red cell adenosine deaminase activity in myelodysplastic syndromes and paroxysmal nocturnal hemoglobinuria.

Red cell adenosine deaminase (ADA-RBC) activity in patients with myelodysplastic syndromes and paroxysmal nocturnal hemoglobinuria is significantly increased compared to that observed in normal controls. ADA-RBC activity is not related to fetal hemoglobin concentration, but it is significantly correlated with hemoglobin concentration at diagnosis and with the degree of morphologic dysplasia in the erythroid lineage. The results of our study suggest that the observed enzymatic abnormality may constitute a non-specific manifestation of the stem cell alteration that determines these disorders.

Haemorragic syndrome in cattle by subgenotype BVDV-1h

Bovine viral diarrhoeaemucosal disease (BVD-MD) is caused by a Pestivirus. Two genotypes are recognized: BVDV-1 and BVDV-2. BVDV-1 causes diarrhoea, respiratory disorders and reproductive disorders. BVDV-2 causes a haemorrhagic syndrome. We describe several hypervirulent outbreaks characterized by severe haemorrhagic signs caused by BVDV-1 in Sicily.

Messa a punto di un protocollo sperimentale per la ricerca di Enterovirus in sedimenti marini

In riferimento al Decreto Ministeriale del 24/01/1996, il controllo dei fondali marini utilizzati per il ripascimento dei litorali, costituisce un passo importante per la valutazione della salubrità dell’ambiente marino e dei prodotti ittici da allevamento, con particolare riferimento alla coltivazione di molluschi eduli lamellibranchi. Diversi studi, infatti, hanno dimostrato come i sedimenti, per le loro caratteristiche chimico-fisiche, rappresentano dei “serbatoi” di agenti microbici di varia natura che, da qui possono essere risospesi nelle acque, compromettendo la salubrità dell’ambiente. Poiché i molluschi eduli lamellibranchi sono organismi filtratori e trattengono agenti patogeni ca…

CUTANEOUS NEOFORMATIONS ASSOCIATED WITH COINFECTION BY ORF VIRUS AND ORTHOPOXVIRUS IN GOATS IN SICILY

Parapoxvirus - ORF virus (ORFV) is the etiological agent of contagious ecthyma, a severe zoonotic exanthematic disease that affects small ruminants. It is usually characterized by highly infectious pustules and papillomatous lesions on the skin of the lips, tongue, and around the mouth. In more severe cases, the skin of the eyes, feet, vulva or udder may also be affected. Orthopoxvirus has been sometimes associated with naturally spreading disease in domestic and wild animals. Parapoxvirus infection can be complicate by opportunistic agents, mainly Fusiformis necrophorus, Dermatophilus congolensis or be invaded by the larvae of flies. Little is reported on co-infection with other viruses. T…

Foreign body response to subcutaneously implanted scaffolds for cardiac tissue engineering.

Genotyping of bovine diarrhea virus (BVDV) isolated from cattle in Sicily

Bovine Viral Diarrhoea - Mucosal Disease (BVD-MD) is a widely spread infectious disease that cause economically important losses in farms. Several epidemiological studies indicate a high genetic heterogeneity among Bovine Viral Diarrhoea Virus (BVDV) strains circulating in Italy. The aim of this study was to investigate the genotypes of BVDV in Sicily, a region in the South of Italy. For this purpose, 17 BVDV strains collected from cattle breed in Sicily between 2005 and 2008 were genetically typed by sequencing of 5'-untraslated region (5’-UTR) of the viral genome. In this study, phylogenetic analysis showed that all 17 examined strains were clustered within the BVDV genotype 1. Particular…

Fibroin and poly-lactic acid scaffolds may be used in cardiac tissue engineering to drive the differentiation of cardiac progenitor cells: in vitro and in vivo studies.