Evaluation of the analytical performances of a portable, 18-parameter hemometric system using capillary blood samples for blood donor enrolment.

BACKGROUND AND OBJECTIVES: Blood donor enrolment process is frequently based on the sole capillary haemoglobin (Hb) evaluation while platelet donors by apheresis also requires platelet (Plt) count. The 'sole Hb' approach prevents a complete donor evaluation and does not allow Plt donor enrolment. To extend blood counts before donations, we evaluated the performances of a multiparametric counter using capillary blood. MATERIALS AND METHODS: The ABX Micros 60 (Micros 60) blood analyzer was employed on capillary blood and compared with venous counts by a reference counter (Coulter AcT 5diff) in a first series of 416 donors and in a second series of 136, after a 3-month period of routine use of this study counter. An average of 50 microl of capillary blood was collected whose 10 microl had been aspirated by Micros 60. RESULTS: High correlations were found between capillary counts using Micros 60 and venous counts using the reference counter. Mean Plt counts differed of 37 x 10(9)/l less for capillary approach in the first series of comparisons, but decreased to 10 x 10(9)/l less in the second series due to a greater expertise of operators in capillary sampling. All other parameters were accurate and never reached clinical relevance albeit they showed statistically significant differences. CONCLUSION: Data on Micros 60 demonstrated that capillary predonation counts may represent a feasible and effective approach to realize an accurate enrolment process of blood and Plt donors.

Hepatitis B virus blood screening: impact of nucleic amplification technology testing implementation on identifying hepatitis B surface antigen non-reactive window period and chronic infections.

BACKGROUND: Despite improvements in hepatitis B surface antigen (HBsAg) test sensitivity, post-transfusion hepatitis B virus (HBV) infection still occurs because HBsAg is undetectable during the early window phase (WP) of the infection, in the convalescence core window phase of the infection, or in serologically silent chronic hepatitis or in mutant forms of HBV. HBV-DNA screening using high sensitivity nucleic amplification technology (NAT) assays has recently been introduced to reduce the residual risk of transmission of HBV by transfusion of blood components. MATERIALS: Over 1 year 75 063 donations were individually screened for HBV-DNA by the Ultrio Procleix assay on the Tigris platform. The donations were collected in the Latium region, an area of the central Italy, and they accounted for the 40% of the total blood units collected in this area per year. The initial reactive samples were re-tested and confirmed by the discriminatory HBV assay. Additional HBV serological markers were also performed. Suspected WP infections were followed-up to monitor the development of the immune response. All HBV-DNA-positive donors were called back to check up their infectious status. RESULTS: The results of testing the 75 063 donations are: 33 donations HBsAg positive, 31 out of them HBV-DNA-positive and two HBV-DNA negative; 22 donations HBsAg-negative but HBV-DNA positive with low viral load. Six of the 22 were found to be consistently HBV-DNA reactive whereas the remaining 16 donations showed inconsistent results on multiple NAT retesting. One WP infection was confirmed by the follow-up of the donor for 3 months following the index blood donation. CONCLUSIONS: In the donor population of the Latium region, NAT screening has revealed a higher than expected number of donors who were HBsAg non-reactive but HBV-DNA-positive with three donors showing HBV-DNA as the only marker of infection. The adoption of genome screening has increased the safety of the blood supply and has also contributed to the protection of donor health by identifying either WP or clinically silent infections.

A tetrazolium-based colorimetric assay for quantification of HIV-1-induced cytopathogenicity in monocyte-macrophages exposed to macrophage-colony-stimulating factor.

A sensitive assay was developed for in vitro evaluation of anti-HIV agents in monocyte-macrophage cells (M/M) (a crucial target of HIV in the body). Monocyte-macrophage cells are usually poorly sensitive to the cytopathic effect induced by HIV. However, when fresh adherent monocyte-macrophage cells are cultured at relatively high density in the presence of macrophage-colony stimulating factor (M-CSF), they undergo cytolysis and die in 2-3 weeks. HIV-mediated cell-killing can thus be assessed with a method based on the reduction of the yellow colored 3-(4-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by metabolically active cells to a blue formazan, which can be measured spectrophotometrically. HIV-mediated cytopathic effect of M-CSF-exposed monocyte-macrophage cells was consistently achieved in all experiments performed under the conditions described herein. Anti-HIV activity of zidovudine (AZT) was also comparatively evaluated in M-CSF- and normal monocyte-macrophage cells both using the MTT assay and by measuring HIV-p24 antigen production in supernatants of monocyte-macrophage cells cultures, and similar results obtained with both methods. These results support the use of this colorimetric assay for broad screening of anti-HIV agents in monocyte-macrophage cells.

Genetic alteration in gastrointestinal cancer. A molecular and cytogenetic study.

We examined 16 cases of gastrointestinal cancer, of which 11 were from the colon, 1 from the rectum, and 4 of gastric origin, cytogenetically for expression and for loss of heterozygosity (LOH) on chromosome 18 using Deleted Colon Cancer (DCC) gene. LOH on chromosome 18 with DCC probe was detected in 7 out of 11 cases of colon, in 4 out of 4 cases of gastric and in 1 case of rectum cancer. In all gastrointestinal tumors the expression of DCC gene was absent, while it was present in normal tissue. We also found rearrangements of chromosomes 18 (10 cases) and 17 (9 cases), leading respectively to deletions of long and short arms. Other additional abnormalities were observed involving chromosomes 5, 6, 15 and 19. The data recorded in our series differ from other authors' data in three respects: a high incidence of pseudodiploid chromosome number, rearrangements of chromosome 19 and 15, and involvement of DCC gene in the development of gastric cancer, as well as in colorectal cancer as previously reported.

Flow cytometric analysis of DNA content and cell kinetics in colorectal carcinoma.

DNA ploidy and cell proliferation were studied by means of flow cytometry in 98 patients with primary colorectal adenocarcinoma. Multiple samples of tumour burden were pooled and freshly dissociated immediately after surgery for FACS analysis. The relationships between ploidy, proliferative activity, evaluated in terms of S-phase percentages (%S), and some clinico-pathological variables were analyzed. 87 of the 98 tumors yielded evaluable DNA histograms: 32 were diploid (37%) and 55 were aneuploid (63%; median DNA index = 1.6). Multiple aneuploid cell populations were found in 15 tumors (17%). The % S was estimated by means of a mathematical model. Aneuploid tumors showed % S values significantly higher than diploid ones (p < 0.0001). Differences in the distribution of DNA aneuploidy were observed in relation to Dukes' stage and tumor site, left colon, rectum and stage D tumors being more frequently aneuploid. No significant differences in proliferative activity were observed in relation to most of the clinical variables, except for higher % S values observed in tumors of right colon compared to those of left colon and rectum.

Risk management of HBsAg or anti-HCV positive healthcare workers in hospital.

Recommendations are made for controlling the transmission of the hepatitis B and hepatitis C viruses from healthcare workers to patients. These recommendations were based both on the literature and on experts' opinions, obtained during a Consensus Conference. The quality of the published information and of the experts' opinions was classified into 6 levels, based on the source of the information. The recommendations can be summarised as follows: all healthcare workers must undergo hepatitis B virus vaccination and adopt the standard measures for infection control in hospitals; healthcare workers who directly perform invasive procedures must undergo serological testing and the evaluation of markers of viral infection. Those found to be positive for: 1) HBsAg and HBeAg, 2) HBsAg and hepatitis B virus DNA, or 3) anti-hepatitis C virus and hepatitis C virus RNA must abstain from directly performing invasive procedures; no other limitations in their activities are necessary. Infected healthcare workers are urged to inform their patients of their infectious status, although this is left to the discretion of the healthcare worker; whose privacy is guaranteed by law. If exposure to hepatitis B virus occurs, the healthcare worker must undergo prophylaxis with specific immunoglobulins, in addition to vaccination.

Effect of cyclosporin A on B cell maturation and differentiation.

The activation and differentiation of resting B cells into Ig secreting cells are regulated by T cells, macrophages and their secreted factors. The present study evaluated the effect of cyclosporin A (CsA) on this process. Peripheral blood lymphomonocytes (PBMC) drawn from healthy donors were stimulated with protein A (PA) or with lipopolysaccharides plus pokeweed (LPS+PWM) in either the presence or the absence of CsA. Phenotypic B cell changes and immunoglobulin production was then analyzed. The data revealed that CsA decreased the expression of B cell surface receptors of the activation phase, and enhanced the resting phase receptors. Different effects of CsA were found on B cell differentiation, depending on its induction by PA or LPS+PWM. In the first system, CsA decreased the expression of differentiation phase receptors and the secretion of free Ig. In cultures stimulated with LPS+PWM, CsA increased the differentiated phase receptors and Ig secretion. Thus, CsA seemed to act as a blocking agent of the activation phase and as a modulator of the differentiation phase and of IgG secretion, depending upon the antigen used for stimulation.

Molecular analysis of HTLV-I and HTLV-II isolates from Italian blood donors, intravenous drug users and prisoners.

Using molecular methods three or five major variants of HTLV-I have been identified; moreover two subtypes of HTLV-II defined as HTLV-IIa and HTLV-IIb with six variants within each of these groups have been described. In the present study we analysed proviral DNA obtained from the peripheral blood mononuclear cells (PBMCs) of a significant group of Italian intravenous drug users (IVDUs), prison inmates and blood donors (BDs) who were HTLV antibody positive. Restriction fragment length polymorphism (RFLP) of amplified LTR region with ApaI, NdeI, DraI, SacI and MaeIII endonucleases was used to define the HTLV-I subtypes, while the different variants of HTLV-IIa and -IIb were defined by RFLP of the LTR region with the AvaII, BglI, SauI, XhoI and BanII endonucleases. The four HTLV-I isolated from BDs were characterized as C type. All the 11 HTLV-II detected in the IVDUs were HTLV-IIb4, while among the prisoners one HTLV-IIb5 and five HTLV-IIb4 were found. Interestingly, in the BDs group two HTLV-IIa0 and one HTLV-IIb4 were detected. It should also be noted that 82% of the IVDUs and 50% of the prisoners were coinfected with HIV, while all the BDs were HIV negative. These data indicate that HTLV-IIb4 is the predominant genotype in Italian IVDUs and prisoners, while the significant variability observed in the BD HTLV-II isolates could be due to the different source of infection among this group.

Elevated alanine aminotransferase in blood donors: role of different factors and multiple viral infections.

Many different aetiological agents stimulate alanine aminotransferase (ALT) production. Viral markers and other aetiologies were investigated in 2166 individuals, randomly selected from 10,000 consecutive blood donors. Elevation of ALT was found in 10.8% of subjects. Grouping donors according to ALT level and correlating with, respectively, hepatitis B core antibody (HBcAb), cytomegalovirus antibody alone, or associated with HBcAb, showed similar findings (high ALT 11.1%, normal 11.6%; high 85.4%, normal 81.4%; high 10.2%, normal 11.0%, respectively). Hepatitis C virus (HCV) antibody was found to be significantly associated with elevated ALT levels (high 1.7%, normal 0.26%). Other causes of ALT elevation were alcohol abuse (17%), obesity (25%) and dyslipidaemia (38%), but in 11% there was no obvious aetiology. Although HCV is a rare cause of elevated ALT in blood donors, it seems to be the only virus, among those tested, to account for liver damage. This may be due to the non-protective role of HCV antibody, the low specificity of ALT, or the pathogenic role of uninvestigated viruses.

[Vesical hernias. Considerations apropos of a case]. / Ernie vescicali. Considerazioni a proposito di un caso.

A case of inguinoscrotal hernia involving the bladder is presented. This pathology is so frequent in males over 50 that preoperative diagnostic investigations should include retrograde cystography and if necessary urography.

[The use of an ultrasonic lithotripter during video laparoscopic cholecystectomy]. / Uso del litotritore ad ultrasuoni in corso di colecistectomia videolaparoscopica.

The video laparoscopic cholecystectomy, a new technique recently introduced in surgical surgery, includes, among other complications, also the dimension of stones. Our intention in the present work is to remove this limitation using an ultrasound lithotripter to reduce the dimension of lithiasic formations, avoiding, after all, to resort to minilaparotomy or to the use of dilators which are in contrast at least with two of the principles of methodology, the aesthetic and functional one for the patient.

The role of the administration time of prophylactic antibiotic therapy in colorectal cancer surgery: a review of 6,069 patients from 36 randomized clinical trials.

The aim of the present report was to establish the effectiveness of different prophylactic antibiotic regimens and administration times in colorectal cancer surgery. Six thousand and sixty nine patients from 36 selected randomized clinical trials, published between 1980 and 1989, were reviewed. The occurrence of septic events, isolated bacterial strains, fever and postoperative hospitalization times were also analyzed. The therapeutic schedules that included the perioperative administration of antibiotics provided better results that those that did not (p. less than .0001 for infections both specifically related and unrelated to colorectal surgery). The number of postoperative administrations did not affect the clinical results, even if the predominant choice was to give more than one administration of antibiotics. A factorial design demonstrated that prolonging the perioperative administrations up to the postoperative period provided statistically significant benefits (p less than .0001) only with regard to the risk of infections that were not specifically related to colorectal surgery.

Inverse relationship between spontaneous interleukin-1 production and mitogen-driven proliferation in patients with pulmonary tuberculosis.

The blastogenic response to mitogens of peripheral blood mononuclear cells (PBMC) obtained from healthy volunteers and patients with chronic or acute tuberculosis (TB) was evaluated. Cells derived from TB patients showed a reduced proliferative capacity compared to that of healthy individuals. Three possible causes of such an impairment were investigated, namely: 1) a change in the proportion of lymphocyte subpopulations; 2) an altered ratio between monocytes and lymphocytes and 3) a reduction in the state of monocyte-macrophage activation, with an impaired production of interleukin-1 (IL-1). We observed no significant modification of lymphocyte subsets from TB patients and normal individuals. However, the relative number of monocytes in the patients was always higher than the controls. Furthermore, circulating monocytes from the patients with TB exhibited a decreased phagocytosis of latex beads, a normal expression of DR antigens, and an increased spontaneous production of IL-1. The possibility that the hyperactivation of macrophages may be responsible for the observed low blastogenic response is discussed.

[Bifid ureter with a blind branch. Apropos of a case]. / Uretère bifide à branche borgne. Au sujet d'un cas.

The authors write about a case of blind-ending, bifid and symptomatic ureter diagnosed by urography and excised surgically.

Complement-dependence of platelet serotonin release test in polytransfused patients.

The research of platelet isoantibodies in patients with Cooley's anaemia was performed by simultaneous determination of the platelet-complement fixation test, platelet factor 3 availability assay and 14C-serotonin release test. In 93% of the examined patients we obtained positive results with the 5HT-release test, which appeared to be a complement-dependent reaction in most of the sera-containing isoantibodies, different from sera of patients affected by autoimmune thrombocytopenia.

The lymphocyte stimulation test: a sensitive technique for the detection of circulating platelet antibodies.

Platelet autoantibodies are unable to react in standard immunological reactions and this makes it difficult to detect their presence in the serum. The present work shows that peripheral blood lymphocytes from normal donors are stimulated in vitro by autologous platelets sensitized with sera from patients with different types of chronic thrombocytopenia. Comparison of the lymphocyte stimulation test with the platelet factor 3 (PF3) availability assay and the serotonin (5HT) release test demonstrated that the former method was the more sensitive one. Some sera from patients with SLE and thrombocytopenia also induced both a slight lymphocyte stimulation in the absence of platelets and a complement-dependent release of serotonin, probably due to the presence of immune complexes.

Platelet antibodies in different forms of chronic thrombocytopenia.

Three techniques have been employed for the in vitro detection of circulating platelet antibodies in thrombocytopenic patients affected by 'idiopathic' form or by lupus erythematosus (SLE), the complement fixation test, the platelet factor 3 availability test and the serotonin release test. 29 of the 35 sera tested (82.8%) gave positive results for antiplatelet activity. In particular the serotonin release test allows to distinguish 4 groups of patients: a first group affected by idiopathic form; two groups with autoimmune thrombocytopenia and various degrees of serotonin release, and finally a fourth group which comprises subjects affected by SLE, with circulating immunocomplexes.

Persistence of HIV-1 silent infection in seronegative subjects at high risk.

Twenty regular sexual partners of HIV-1 infected subjects, without detectable human immunodeficiency virus (HIV-1) antibody and positive for HIV-1 genome by in situ hybridization (ISH), were selected and studied longitudinally for 6-36 months to estimate the duration of silent infection. During the follow-up period, 10 showed atypical Western Blot (WB) patterns. Two seronegative partners seroconverted. Rapid progress to AIDS was observed in 7 seropositive subjects.