Connexin 26 Expression in Mammalian Cardiomyocytes.

Connexins are a family of membrane-spanning proteins named according to their molecular weight. They are known to form membrane channels mediating cell-cell communication, which play an essential role in the propagation of electrical activity in the heart. Cx26 has been described in a number of tissues but not in the heart, and its mutations are frequently associated with deafness and skin diseases. The aim of this study was to assess the possible Cx26 expression in heart tissues of different mammalian species and to demonstrate its localization at level of cardiomyocytes. Samples of pig, human and rat heart and H9c2 cells were used for our research. Immunohistochemical and molecular biology techniques were employed to test the expression of Cx26. Interestingly, this connexin was found in cardiomyocytes, at level of clusters scattered over the cell cytoplasm but not at level of the intercalated discs where the other cardiac connexins are usually located. Furthermore, the expression of Cx26 in H9c2 myoblast cells increased when they were differentiated into cardiac-like phenotype. To our knowledge, the expression of Cx26 in pig, human and rat has been demonstrated for the first time in the present paper.

Morphological features of ovine embryonic lung fibroblasts cultured on different bioactive scaffolds.

Tissue regeneration with autologous cell transplantation is one of the most important goals in clinical research. In this field, the development of bioactive materials that provide microenvironments for cell-matrix interactions mimicking biological conditions is required. In recent years, many synthetic materials have been developed as scaffolds and many procedures for the surface modification of these materials have been applied using biological molecules. In this study, we analyzed the morphology and the molecule production by ovine embryonic lung fibroblasts cultured on three different sponge-like matrices based on poly(L-lactic acid) (PLLA): agarose/PLLA, crosslinked and uncrosslinked gelatin/PLLA. The matrices were produced by using an emulsion freeze-drying method leading to the formation of sponge-like materials with high porosity and with interconnection between the pores. In vitro MTT test demonstrated that transplanted cells were viable and metabolically active. Morphological analysis revealed that fibroblasts adhered to and penetrated the polymeric structures. Moreover, all the different matrices supported fibroblast production of proteoglycans, glycoproteins, and matrix molecules such as elastin, collagen I, and fibronectin. These data suggest that the tested bioactive scaffolds may support the growth and extracellular matrix molecule production of fibroblasts allowing in vitro connective tissue regeneration.

The targets of nephritogenic antibodies in systemic autoimmune disorders.

In situ formation of immune complexes is a well recognized mechanism of renal injury in systemic autoimmune disorders. The identification of intrinsic renal antigens that are targets of nephritogenic antibodies is a field of active investigation. Recently, two proteins expressed in the kidney have been characterized as renal antigens. Alpha-actinin, an actin-binding protein localized in glomerular podocytes, is the major target of nephritogenic anti-DNA antibodies. Alpha-enolase, a glycolytic enzyme, is a target of nephritogenic anti-DNA and non-anti-DNA antibodies.

Naturally-occurring anti-G-CSF antibodies produced by human cord blood B-cell lines infected with Epstein-Barr virus.

INTRODUCTION: Naturally occurring antibodies (auto-Abs) recognizing human granulocyte-colony stimulating factor were detected with high frequency in serum samples obtained from umbilical cord blood of newborns (12 of 65 samples screened) and maternal peripheral blood serum samples from women at the end of gestation (seven of 56 cases tested). The aim of this paper was to demonstrate that auto-Abs anti-G-CSF revealed in the blood of newborns were produced during foetal life. MATERIALS AND METHODS: Mononuclear cells from cord blood samples of different newborns containing high titer anti-G-CSF Abs were infected with Epstein-Barr virus in vitro, and EBV-immortalized B-cell lines were isolated and characterized for specific anti-G-CSF Ab production. RESULTS: Six different, unrelated cell lines of male origin which showed the presence of EBNA-2 antigen in the nucleus, displayed a B-cell phenotype (CD30+, CD5-, CD10-, HLA-DR+, CD19+, CD20+, CD23+, CD38+, CD25+), coexpressed low intensity sIgM and sIgD, and produced only IgM with prevailing lambda clonal restriction and anti-rhG-CSF Ab reactivity. The Ab specificity was proven against either glycosylated or unglycosylated G-CSF by saturable binding in direct enzyme-linked immunosorbent assays, by competition binding and Western immunoblotting assays. CONCLUSION: The secreted Abs did not affect the in vitro generation of granulocyte colonies by human normal adult haemopoietic progenitor cells in soft agar clonogenic assays, suggesting that these Abs were not neutralizing.

Detection and epitope mapping of immunoreactive human endothelin-1 using ELISA and a surface plasmon resonance-based biosensor.

A surface plasmon resonance-based biosensor (BIA technology) and enzyme-linked immunosorbent assays (ELISA) have been used for detecting and characterizing human endothelin (ET), a potent vasoactive 21 amino acid polypeptide. Antibodies produced against the isoform ET-1 and its C-terminal eptapeptide ET-1(15-21) have been characterized with respect to their binding capacity to the two isoforms ET-1 and ET-3, the non-secreted portion of the precursor molecule Big.ET-1(22-38), the C-terminal of ET-1, six analogues of ET-1(16-21) each containing a substitution with Ala of a single amino acid in positions 16-21, respectively, and three synthetic cyclic peptides mimicking the N-terminal portion of ET-1. Antibodies reacting with ET-1 also bound to ET-1(16-21) and, with less affinity, to ET-3 but did not cross-react with Big.ET-1(22-38). Ala substitution in positions 16, 17 and 19 of ET-1(16-21) hardly affected the antibody binding capacity of ET-1(16-21), whereas Ala substitution of Asp18, Ile20 and, in particular, Trp21, inhibited its immunoreactivity. The C-terminus thus represents an immunodominant epitope in ET-1 and is important for antibody binding. Epitope mapping using as antibody pairs polyclonal anti-ET-1 and monoclonal anti-ET-1(15-21) antibodies indicated the presence of another immunogenic domain in the N-terminal portion of the molecule. There was excellent agreement between the epitopes determined using ELISA and BIA analyses.

Natural and therapy-induced anti-GM-CSF and anti-G-CSF antibodies in human serum.

Serum samples were obtained from patients with lymphoid and plasma cell malignancies who received after chemotherapy human recombinant GM-CSF or G-CSF. Sera from some patients revealed the presence of anti-cytokine antibodies, particularly after repetitive cytokine injections. Antibody Fab binding in a saturable manner by ELISA and Western immuno-blotting confirmed antibody specificity. Anti-cytokine antibodies were detected before the exogenous cytokine injections in some patients, but increasing antibody levels were found after one or subsequent treatments. Low levels of anti-GM-CSF and anti-G-CSF antibodies were also detected in a relatively large proportion (about 10-30%) of normal sera from different adult healthy volunteers who had never been treated before with exologous cytokines as well as from cord blood. EBV-immortalized cord blood derived B-cell cultures were also found to produce anti GM-CSF and/or anti-G-CSF antibodies with high frequency.

Alpha-enolase is a renal-specific antigen associated with kidney involvement in mixed cryoglobulinemia.

OBJECTIVE: To study the specificity of antibodies reactive with renal antigens in mixed cryoglobulinemia. METHODS: Sera from mixed cryoglobulinemia (MC) patients were tested on human kidney extracts by immunoblot. A partially purified renal antigen was subjected to N-terminal sequencing. RESULTS: Antibodies reactive with a renal antigen of 48 kD were detected in 7 out of 11 patients with MC and renal involvement. N-terminal sequencing of this antigen showed that it was identical with alpha-enolase. This result was confirmed by the reactivity of the renal antigen with a rabbit anti-serum specific for alpha-enolase. CONCLUSIONS: The results indicate that antibodies specific for alpha-enolase are frequently produced by mixed cryoglobulinemia patients with renal involvement.

Immunohistochemical demonstration of the small GTPase RhoA on epoxy-resin embedded sections.

The purpose of the present study was to establish a method for light microscopical immunohistochemical localization of the small G protein RhoA on specimens treated and embedded for routine transmission electron microscopy. There are advantages in antigen immunolocalization on resin semi-thin sections compared to cryostat or paraffin sections: the preservation of morphological details, the well-defined immunoprecipitate localization and the possibility to correlate the immunohistochemical results with those obtained by electron microscope on neighbouring sections. These advantages are particularly useful for the subcellular localization of low molecular weight proteins such as RhoA, a small G protein able to cycle from the inactive cytoplasmic form to the plasma membrane-bound active form.

Evaluating a fear appeal message to reduce alcohol use among "Greeks".

OBJECTIVE: To evaluate the impact of a fear appeal message on college students' drinking behavior using the extended parallel process model. METHOD: A survey was administered to a random sample of undergraduates (n=224) in 38 national fraternal organizations. RESULTS: Both perceived efficacy and perceived threat were significantly correlated with drinking behavior. There was a significant difference both in drinking behavior and attendance at alcohol-free events between those who heard and those who did not hear the message. CONCLUSIONS: Theoretically based fear appeal messages may be a useful way to promote responsible drinking among college students.

[Anesthesiologic problems in abdominal mini-invasive surgery]. / Problemi anestesiologici in chirurgia mininvasiva addominale.

The large diffusion of minimally invasive surgery offers to surgeons and anaesthesiologists the opportunity of controlling new technologies and different problems. The goal of this paper is assess the physiological modifications during the operation with minimally invasive laparoscopic technique in references to risks and complications.

[Anatomo-pathologic study of parenchymal segments with bullous pneumopathy. Mini-invasive surgical technic]. / Studio anatomo-patologico di segmenti di parenchima affetti da pneumopatia bollosa. Tecnica chirurgica mini-invasiva.

The treatment of bullous pneumopathy using minimally invasive surgery gets available "new" specimens for the pathologist. The change of the specimen's features is related to the peculiarity of the analyzed elements and the systematicity of the treatment. Observing 41 cases, we have noticed modifications in 26 cases of the vascular wall 25 cases of the elastic and 23 of the muscular tissues and 21 cases inflammatory features.

[Minimally invasive surgery in abdominal emergencies]. / La chirurgia mininvasiva nelle urgenze addominali.

The authors report 34 cases of abdominal emergencies treated by means of Minimally Invasive Surgery. The value of treatment by this method is discussed on the ban of diagnostic evaluation of the severity of the lesion.

[The minimally invasive treatment of bullous pneumopathy. The anatomicopathological and pneumological findings in 18 months of experience]. / Trattamento mininvasivo della pneumopatia bollosa. Considerazioni anatomopatologische e pneumologische in 18 mesi di esperienza.

The authors present 18-month experience in the minimally invasive treatment of bullous pneumopathy and review of the relative literature. Results of pneumologic and pathologic studies are correlated with the advantages of the technique. Tobacco smoking, positive family history, associated disease and long standing treatments have been considered. Special attention is drawn to macro and microscopic findings. The authors concluded that: (a) There is a positive correlation between surgical technique of pleural cavity exploration and recidival pneumopathy. (b) There is no correlation between family history and pneumological and pathological findings. (c) A wider experience on bullous pneumopathy may offer new interesting pneumological knowledge.

[Middle lobe syndrome]. / Sindrome del lobo medio.

The authors, based on their actual experience with four cases and using previously published articles, state the pathological and clinical features of the middle lobe syndrome. Considering the physio-pathological processes, the authors determine the therapeutic program: the first approach would be pharmacological, second, if necessary, surgical-seeking to be as conservative as possible.

[Role and limitations++ of the urease test on gastric biopsy for Helicobacter pylori infection. Study of 50 cases]. / Ruolo e limiti del test all'ureasi su biopsie gastriche per l'infezione da Helicobacter pylori. Studio su 50 casi.

The authors after a review of the international literature, analyzed 50 cases of gastroduodenal pathology submitted to endoscopy and biopsy. The biopsy was analyzed with the CP test. The analysis of results of CP test and local endoscopic status offers useful indications for therapy. The authors consider and assess the data of this epidemiological study.

[Drainage of the pleural cavity]. / Il drenaggio della cavità pleurica.

The authors, from their personal experience, demonstrate the goals, the indications, the techniques and complications of performing a drain of the pleural cavity (DCP). DCP has an important role as a single operation or at the end of every thoracic operation. For this reason it is mandatory to respect the principles and the concepts reported by the authors.

[Laser therapy of esophageal tumors. Technical notes and personal experience]. / Il trattamento dei tumori esofagei con laser. Note di tecnica ed esperienza personale.

In Italy the incidence of oesophageal cancer is very low: 4 cases every 100,000. More than 60% of patients are observed with dysphagia and oesophageal stenosis. For these patients their is no surgical indication and prognosis is less than 1 year. We have applied the recanalization of oesophagus with laser in 137 patients (range 47-88 years). One year survival was 53.1%, three years 5.8%. The aim of this paper is to describe our techniques and analyze 5 years of activity.

Preparation of stable dispersion of barium titanate nanoparticles: Potential applications in biomedicine.

Nanoscale structures and materials have been explored in many biological applications because of their extraordinary novel properties. Here we propose a study of cellular interactions with barium titanate nanoparticles, an interesting ceramic material that has received a lot of interest in the nanotechnology research, but without any attention about its biological potential. We introduced for the first time an efficient method for the preparation of stable aqueous dispersions of barium titanate nanoparticles, characterized with FIB, TEM and AFM imaging, light scattering, Z-potential and UV/vis analysis. Finally, we presented a systematic study of short-term cytotoxicity of the prepared dispersion based both on quantitative (metabolism, proliferation) and qualitative (apoptosis, viability, differentiation) assays.

Investigation of interactions between poly-L-lysine-coated boron nitride nanotubes and C2C12 cells: up-take, cytocompatibility, and differentiation.

Boron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, myotube formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription - polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.