A new HLA allele, HLA-B*08:122, described in an unrelated donor of Caucasian origin.

A new human leukocyte antigen-B allele was found in an unrelated Italian donor.

Muscoloskeletal disorders and occupational stress of violinists.

Although musculoskeletal disorders are the most frequent cause of occupational diseases in musicians, very few studies have focused attention on a single category of instruments, in particular on the violin. This involves, in its practice, almost all the areas of the body, besides being in the category of strings which is the most numerous in an orchestra. A specific protocol, investigating postural and clinical profiles of the musculoskeletal apparatus as well as job stress, was utilized in a conservatory on graduates in the tenth year of violin study, who regularly participated in activities of orchestras or string quartets. The investigation revealed target segments of osteoarticular apparatus (jaw, vertebral spine, shoulders, elbows, hands and fingers, lower limbs) electively subjected to overuse, as well as muscle contracture of trapezoids and hyperkeratosis of fingers and clavicle. Although the work environment was comfortable, most violinists claimed to undergo intense rhythms and competitiveness. This study, highlighting subclinical occupational diseases in young musicians (violinists) suggests adequate prevention measures.

The presence of posttransplant HLA-specific IgG antibodies detected by enzyme-linked immunosorbent assay correlates with specific rejection pathologies.

Posttransplant monitoring of anti-HLA antibodies with routine techniques gives unsatisfactory results due to a variety of technical limitations. We investigated how a new alternative technique correlates with posttransplant clinical events. A total of 313 nonselected serum samples from 136 patients were screened by an ELISA utilizing captured soluble HLA class I antigens. We observed the absence of anti-HLA antibody production in acute rejection cases responding to standard antirejection therapy. On the other hand, we showed a clear presence of these antibodies in acute rejection episodes not responding to standard therapy (P<0.0001) and in chronic rejection (P<0.001). We conclude that routine posttransplant monitoring by ELISA offers early risk assessment that is crucial for proper immunosuppression and for antirejection therapy choice.

Biocompatibility of charcoal hemoperfusion. Effects of long-term treatment on lymphocyte characteristics and function.

Biocompatibility of charcoal hemoperfusion was studied in a group of 15 uremic patients, evaluating the effects of long-term treatment on some structural and functional parameters of circulating lymphocytes: in vivo distribution of T-cell subsets; surface T3, T4 and T8 antigen expression, in vivo and in vitro DNA synthesis. A comparative analysis was performed with patients on conventional dialysis using cuprophan membranes.

Fibroblast proliferation over dialysis membrane: an experimental model for "tissue" biocompatibility evaluation.

The present study reports on a biological model based on fibroblast proliferation applied to 3 different types of flat-plate dialysis membrane, in order to ascertain whether the artificial materials currently used in hemodialysis cause in vitro cellular proliferation. The study plan we followed involved plate membrane isolation from non-used dialyzers and used dialyzers, observed through scanning electron microscopy (SEM) both before and after testing with human fibroblasts by means of cell culture. Fibroblast growth was assessed by phase contrast light microscopy examination and cytometric DNA content evaluation. Our investigations proved that the artificial materials we considered interact with fibroblast cultures. Noticeable proliferative response was observed both after contact with unused material and on mediation by the protein layer absorbed on the membrane surface at the end of dialysis sessions. In this last case fibroblast proliferative activity appeared higher than that observed with unused membranes, showing that the soluble molecules entrapped in the protein layer appeared able to exert a biological activity even in vitro tests.

Aorta transplantation in man: clinical and immunological studies.

Aortic transplantation has progressively gained interest over the last few years and it is becoming a first choice indication in the substitution of infected prostheses. The most frequent complication in long-term vascular outcome (wall thickening, aneurysmatic dilation, stenosis), may occur through an immunological mechanism. In this study we investigated nine recipients, aged 48 to 65 years, of aorta segment replacement for anti-HLA antibody production (specificity and Ig class), CD3-CD4-CD8 T cell subpopulation dynamics and aorta wall thickness. Mismatch-specific IgG antibodies to HLA class I and HLA class II antigens were detected 1, 3 and 6 months after transplantation and persisted at a high concentration for at least 1 year. Furthermore, the absolute number of CD3, CD4 and CD8 positive lymphocytes increased progressively after aorta allograft. Tomography scanning showed a progressive thickness of the aorta wall. We can speculate that these anti-HLA antibodies in the recipients have the potential to harm the implant; therefore, aorta allograft should involve the induction of immunological tolerance by appropriate immunosuppressants.

A 3D study of intra and extracellular structures of liver in beta-thalassemia intermedia: an OsO4 maceration analysis.

A scanning electron microscopy study of liver changes has been carried out in three patients affected by beta-thalassemia intermedia (BTI). Applying a new osmium maceration method, recently developed in our laboratory, we had the opportunity to obtain, at SEM, tridimensional images of intra and extracellular structures. Other than the previously reported lesions in BTI, we observed the following pathological findings: disarrangement of the cell structure by a high number of hemosiderin loaded lysosomes; alterations in shape and in diameter of the nuclear pores; presence of apoptotic bodies scattered among the parenchymal cells; deposition of collagen fibers in the space of Disse to form a perihepatocytic dam; enlargement of the sinusoidal endothelial cell fenestrae of the sieve plate. By complete digestion of liver cells, we evidenced a diffuse pericellular fibrosis, made up of interlacing fibrils. Our study evidences some not yet reported morphological lesions in BT. Since patients affected by BTI do not need blood transfusions, these lesions could be considered intrinsic of the disease.

[Collateral effects of 9-alpha-fluor-prednisolone acetate and kanamycin, administered by nasal spray. Clinical experience and experimental data]. / Effetti collaterali da 9-alfa-fluor-prednisolone acetato e kanamicina, somministrati mediante spray nasale. Esperienze cliniche e dati sperimentali.

Iatrogenic pathology due to treatment with steroid drugs used by systemic route is well known. On the contrary iatrogenic pathology due to topical use of these drugs is rarely reported. Two cases of abuse of 9-alpha- fluor-prednisolone and kanamycin administered by endonasal route are reported. The same treatment has been carried out in patients and rabbits. Clinical and bio-humoral data in patients and anatomo-pathological findings in rabbits are reported. The risks, sometimes underestimated, of an overdose of corticosteroid and antibiotic drugs used by endonasal route are pointed out.

Immunologic changes in circulating leukocytes in the presence of atherosclerotic disease.

To assess the numeric and functional changes in circulating white blood cells in the presence of severe atherosclerosis, 25 subjects with marked, angiographically assessed, atherosclerotic lesions and 29 selected controls were studied. Of the differential leukocyte counts, only monocyte count was significantly higher in the atherosclerotic than in the control subjects (449.0 +/- 115.6 (1 S.D.) vs. 344.1 +/- 138.8/mmc; P = 0.0016). By flow-cytometry no significant differences concerning monocyte surface antigens were found, except a feeble decrease in beta 2-microglobulin in the atherosclerotic subjects. As to lymphocytes, an increase in the CD8 population (33.4 +/- 6.8 vs 28.6 +/- 6.5%; P = 0.0144) and decreases in class I HLA antigen (96.6 +/- 7.3 vs 99.4 +/- 0.7%; P = 0.0049), beta 2-microglobulin (97.9 +/- 2.1 vs 99.3 +/- 1.0%; P = 0.0055) and especially in vivo DNA synthesis (3.8 +/- 1.2 vs 5.3 +/- 2.1%; P = 0.0102) percent expressions were found in the atherosclerotic patients with respect to the controls. This study shows that circulating monocytes are increased in atherosclerotic disease, possibly due to their participation in the phagocytosis of lipids in the arterial wall, with no further immunologic involvement. Conversely, the replicative activity of T lymphocytes is decreased, which might be a consequence of or a factor predisposing to atherosclerosis.

Febrile nonhaemolytic transfusion reaction caused by antibodies against human platelet antigen 5a.

Anti-human platelet antigens (HPA) alloantibodies are seldom involved in febrile nonhaemolytic reactions (FNHTRs). We describe a case in which anti-HPA-5a alloantibodies are related to an FNHTR. We studied the specificity of the alloantibodies by flow cytometry, ELISA and MACE. Typing of donors and the patient was performed by sequence-specific polymerase chain reaction. The alloantibodies were found reactive with HPA-5a antigens. The patient was HPA-5b/b, whereas the donor of the platelet apheresis involved in the FNHTR was HPA-5a/a. Despite the low frequency of anti-HPA-5a antibodies, they might be responsible for FNHTR.

Acquired pure megakaryocytic aplasia in course of hepatitis in a patient with immune thrombocytopenic purpura.

An unusual case of selective autoimmune myelopathy, a posthepatitic pure megakaryocytic aplasia, is reported. The disease manifested itself in a patient being treated for thrombocytopenic purpura caused by progressive administration of amitriptyline. The various clinical aspects of this unusual myelo-immunopathy seem to possess the same complexity and polymorphism of immune myelopathies.

Hemoglobin Sabine [beta 91 (F7) Leu-->Pro]: occurrence in a Sardinian individual with hemolytic anemia and inclusion bodies.

BACKGROUND: Hemoglobin (Hb) Sabine (beta 91 Leu-->Pro) is an unstable variant detected for the first time in a 16-year-old Scottish-English-German girl affected by moderately severe hemolytic anemia. A second case was described in a patient of Yugoslavian descent. We report another case of this Hb variant arising as a de novo mutation in a Sardinian patient. METHODS: Definition of the mutation was obtained by DNA direct sequencing on amplified beta-globin gene, as well as by structural analysis of the hemoglobin variant. RESULTS AND CONCLUSION: The patient presented a moderately severe hemolytic anemia with red blood cell inclusion bodies. Hemoglobin electrophoresis showed that quantitatively the abnormal fraction represented 9% of the total Hb amount. beta globin gene analysis revealed a single nucleotide substitution, T-->C, at codon 91, which gives rise to a leucine-->proline substitution. Structural analysis of the variant confirmed the amino acid substitution (Leu-->Pro) predicted by DNA sequencing.

Characterization of a new HLA-DRB1*01 allele (HLA-DRB1*010203) in a Caucasian Italian family by using sequence-based typing.

We report the identification of an HLA-DRB1*01 nucleotide sequence variant in three members of a Caucasian Italian family by using sequence-based typing. The nucleotide sequence of exon 2 observed in the new allele is identical to that of HLA-DRB1*010201 except in position 189 (codon 34) where the adenine of the consensus was replaced by a guanine and it was designated officially as HLA-DRB1*010203* by the WHO Nomenclature Committee.

Flow cytometry evaluation of urinary sediment in renal transplantation.

The value of exfoliative urinary cytology for the diagnosis of different pathological conditions in renal transplantation is widely recognized. The method, however, has not yet gained full acceptance, mainly because identification of the different cells is not always possible by means of standard staining techniques. In view of its characteristics, flow cytometry (FC) seems to represent a consistently reliable, rapid and innovative approach for differentialing the various cells present in the urinary sediment and assessing their number. This study gives the examination result of 223 urinary specimens from 127 transplanted patients selected according to pathology. Sediment cells, collected from fresh urine samples, were washed, treated with a lysing solution, resuspended in saline solution and directly analysed in a FACSCAN cytometer. Morphological evaluation showed: a small number of cells in patients with stable renal function; a larger number of cells, with predominance of lymphocytes, during acute rejection episodes; an absolute predominance of neutrophils during bacterial infection; large-sized cellular debris in cases of post-transplant tubular necrosis; and small cell debris in cases of cyclosporine cytotoxicity. Lymphocyte surface-marker evaluation made it possible to differentiate lymphocyte populations observed during acute rejection episodes (cytotoxic T-cell, CD8 and HLA class II and NK cells) from those detected during bacterial infection (T-cell CD4 positive). These results suggest that urinary FC may be a reliable diagnostic tool in clinical renal transplantation.

Value of panel reactive antibodies (PRA) as a guide to the treatment of hyperimmunized patients in renal transplantation.

Patient presensitization represents a considerable problem in candidacy for renal transplantation. While it is well known that hyperimmunized patients--panel reactive antibody (PRA) higher than 60%--create difficulties in donor matching and have a worse outcome than non-hyperimmunized patients, less information is available on patients with an intermediate degree of sensitization (30-60%). In order to evaluate how graft outcome relates to such degrees of sensitization, 241 consecutive transplanted patients were divided into two groups on the basis of their previous year's PRA peak: group A, PRA 0-29%; group B, PRA 30-60%. Group A showed a significantly better survival both in the first year (90% vs 79%, P < 0.05) and in the third year (82% vs 64%, P < 0.01). However, detailed analysis of group B demonstrated that some parameters may significantly influence graft outcome: (1) better compatibility on locus DR; (2) a primary kidney transplant; (3) a dialysis duration of less than 6 months; and (4) the prophylactic use of antilymphocyte globulin (ALG).

Application of flow cytometry in clinical renal transplantation.

Flow cytometry (FC) may be considered as a fundamental technique in studying cell biology and pathology. It combines the quantitative character of biochemical methods with the multiparametric capacities of microscope analysis in a high-precision process for rapid analysis of individual cell characteristics. Three original FC techniques routinely applied in the field of renal transplantation are reported in the present study. They concern the donor-recipient cross-match test, the morphological analysis of urinary sediment and the modulation of the density of various membrane antigens on the lymphocyte surface. A common factor underlies all these methods: they aim to provide the physician with a reliable diagnostic tool in clinical renal transplantation.