An alternative and simple method to consistently prepare viable isolated human islets for clinical transplantation.

We describe a method to consistently prepare human islets for transplantation. By combining a simple collagenase digestion method and a density gradient purification system, we were able to obtain successful isolations (>/=200,000 islet equivalents, >/=50% purity) in 69% of processed glands. No reagent of animal source was used. Isolated islets were morphologically well maintained and functionally competent, with sterility confirmed in 97% of cases. Two patients were transplanted with islets prepared by this method; graft function was demonstrated for a few months. Improved simplicity and consistency, together with adequate quality of the preparations, are the main features of this isolation method.

Lymphocytes bearing the gamma delta T cell receptor in acute Brucella melitensis infection.

A phenotypical analysis carried out by indirect immunofluorescence and two-color cytofluorometry showed that the number of lymphocytes bearing the gamma delta T cell receptor (TcR) heterodimer was dramatically increased in the blood of six children with Brucella melitensis infection. Most in vivo expanded gamma delta T cells reacted with a monoclonal antibody which identifies V delta 2 gene products and a significant proportion expressed CD25 and HLA-DR activation antigens. In addition, whereas only a few gamma delta T lymphocytes were CD8+, nearly all were CD4-. Highly enriched populations of both alpha beta and gamma delta T cells were obtained by negative immunoselection from three subjects with brucellosis sampled during convalescence. Despite the different form of their TcR, the proliferation of these two major T cell subsets in response to a mitogenic anti-CD3 monoclonal reagent (OKT3) was optimal. In contrast, alpha beta, but not gamma delta, T lymphocytes proliferated vigorously in response to the antigenic stimulus elicited by heat-killed Brucella. Further studies are, therefore, needed to determine whether the selective expansion of the gamma delta T cell subpopulation observed during the clinical course of the infection is driven by antigenic determinant(s) borne by the pathogen in vivo or is due to host-derived stimuli, such as autologous heat-shock proteins expressed on the surface of the infected cells.

[Comparison of two methods to determine beta-lactoglobulin antibodies in children with cow's milk protein intolerance]. / Comparazione tra due metodiche di determinazione degli anticorpi anti-beta-lattoglobulina in bambini affetti da intolleranza alle proteine del latte vaccino.

In the last years the evaluation of IgG against beta-lactoglobulin (beta LG-IgG) has been proposed as reliable test to diagnose Cow's Milk Protein Intolerance (CMPI). This test, associated with other diagnostic tools (such as eosinophilia, hemoccult, RAST, challenge, etc.) showed a good sensibility, but less specificity. In fact antibodies against beta-lactoglobulin were found in other gastrointestinal diseases (coeliac disease, inflammatory bowel diseases, cystic fibrosis, etc.). To determine the specificity of the test some Authors attributed great importance to the laboratory method (IFL, ELISA). They retained IFL more specific. In order to verify the behaviour of the two methods and their concordance we carried out a double bind determination of IgG against beta-lactoglobulin (beta LG-IgG) on serum samples from children affected by CMPI. For this study we chose 16 children, aged from 3 months and 6 years, suffering from CMPI of which 11 with gastrointestinal complaints and 5 from extraintestinal ones. All children were diagnosed on the basis of the following criteria: clinical picture, blood eosinophilia, positivity of hemoccult, amelioration after the withdrawal of cow's milk proteins from the diet, positivity of challenge test. All children at the time of the assessment assuming a diet containing cow's milk proteins. beta LG-IgG were measured using simultaneously immunofluorescent and micro-ELISA methods. The obtained data showed that 12 children (75%) had IgG versus beta-lactoglobulin in the serum. This result was obtained both with IFL and ELISA methods, showing a 100% of concordance.(ABSTRACT TRUNCATED AT 250 WORDS)

Vagal afferent innervation of the pylorus and the upper small intestine studied in the rat with the horseradish peroxidase technique.

The neuronal tracer horseradish peroxidase was injected into different segments of the gastrointestinal in the rat, in order to study the vagal afferent innervation. In the nodose ganglia the extent of labeling was greater in the experiments on the gastric antrum and pylorus than in the experiments on the first part of the small intestine. Vagal afferents are scarce in the upper duodenum and originate mainly from the left nodose ganglion.