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BACKGROUND: Normal B lymphoid maturation occurs in bone marrow (BM) throughout life, but immature B-cell progenitors (BCPs) are more numerous in children than in adults. To assess the normal values according to age became important as BCPs are decreased in myelodysplastic syndromes and have been considered an important diagnostic and prognostic feature in these clonal disorders. METHODS: in a multicenter retrospective study from the Brazilian Group of Flow Cytometry we analyzed the variation of BCPs in normal BM according to age and technical peculiarities of each laboratory. We analysed of 45 BM donors and 89 cases examined for elucidation of transitory reactive cytopenias presenting a normal BM immunophenotyping. BCPs were enumerated as CD19+ /CD34+ /CD45dim /CD10+ cells (panel 1) or CD19+ /CD34+ /CD45dim cells (panel 2) among the total nucleated non-erythroid cells and as percentage of CD34+ cells. RESULTS: we included 134 cases. Panel 1 was applied in 88 cases and panel 2 was used in 46. Age range: 10 months to 89 years. In a multiple regression, % BCPs/total nucleated cells was an exponential function of age. Age explained alone 49.4% of the variance, while 'panel used' explained 1.8% and 'laboratory' explained 0.7%. Age explained only 24.9% of the variance of BCPs/CD34+ cells. CONCLUSIONS: in normal individuals, BM B-cell precursors varied mainly according to age, but were also dependent on technical peculiarities of operators and equipments. Analysis by phenotype and as percentage of total nucleated cells was more accurate and less susceptible to variation than evaluating % BCPs/total CD34+ cells. © 2017 International Clinical Cytometry Society.

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Propionibacterium acnes has been described as a potent adjuvant to immune responses in vitro and in vivo. Presently, we analysed the modulation of peritoneal exudate cells (PEC) by heat-killed P. acnes or its purified soluble polysaccharide (PS), both injected intraperitoneally in C57Bl/6 mice, aiming at their recruitment and cytotoxicity. Both treatments induced an increase in macrophages, immature dendritic cells, B1a lymphocytes and NK1.1(+) CD3(+) cells. The bacterium caused a remarkable increase in a NK1.1(+) CD3(+) CD4(-) CD8(-) cells subpopulation, whereas the PS component seemed responsible for the recruitment of mainly macrophage cells. To assess P. acnes and PS adjuvant effect on PEC cytotoxicity we evaluated their in vitro effect on murine B16F10 melanoma cells. The effector cells from the heat-killed bacteria and PS-treated groups lysed melanoma cells in co-cultures with PEC. Mice genetically deficient in IFN-gamma, when stimulated with P. acnes or PS, had reduced PEC cytotoxicity, and the cytotoxic effect was completely abrogated in PEC from iNOS(-/-) mice. The tumoricidal activity of PEC from P. acnes-treated mice was mediated by macrophages and NKT cells stimulated with IL-12. In PS-treated mice the cytotoxicity was mediated mainly by macrophages. Moreover, both treatments increased IL-4 and IFN-gamma production by NKT cells. In conclusion, we show that P. acnes act mainly by recruiting and activating NKT double-negative cells in PEC, which were shown to be tumoricidal in vitro when induced by IL-12. Macrophages induced by both P. acnes and PS have their antitumour effect dependent on NO production.

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AIM: The purpose of this study is to verify the role of gastroesophageal reflux as responsible for otolaryngological symptoms; it is often difficult to identify, both because of its subtle and aspecific clinical-endoscopic picture and because the traditional distal esophageal pHmetry may show readings which are still within the norm at this level. The esophageal pHmetry carried out at proximal level gives greater diagnostic accuracy. METHODS: We use a catheter with 3 antimony electrodes, 2 of them placed at the esophageal level, respectively at 5 and 20 cm above the inferior esophageal sphincter, and 1 in the stomach. We studied 3 groups of patients: 41 asymptomatic no-refluxer patients (group I, control), 59 refluxer patients with only typical esophageal symptoms (group II) and 68 patients with laryngo-pharyngeal symptoms suffering from acid reflux (group III). RESULTS: At both the proximal and distal esophageal levels, the reflux, with reference to control, turned out to be significantly higher in groups II and III, both as regards the number of episodes (p<0.01), the time of acid exposition (p<0.01), the length of the longer reflux (p<0.01), whereas no differences were found between groups II and III. The proximal esophageal acid refluxes in both groups II and III occurred mainly in the upright position: alone (57.2% and 62.5% respectively) or combined with supine position (23.8% and 34.0% respectively). CONCLUSION: Our experience suggests that the esophago-gastric pHmetry with 3 electrodes represents an effective procedure for the correlation between otolaryngological symptoms and gastro-esophageal reflux.

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The best treatment of bile duct lithiasis in the era of the laparoscopic cholecystectomy has not yet been defined, as we can choose between different kinds of therapies which range from the traditional laparotomic treatment to the more modern endoscopic and laparoscopic techniques. Although in some centres a wholly laparoscopic access to the bile duct is used, we believe that the fastest, safest and most reliable method is still the sequential treatment, endoscopic and laparoscopic. In our experience the endoscopic cleaning of the choledochus was performed in 87.1% of the cases, with no mortality and 4.2% morbidity.

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We evaluable the efficacy and reliability of 24-hour pH monitoring studing the gastro-esophageal reflux. 119 patients with gastro-esophageal reflux disease (MRGE) and 60 normal subjects were tested with this method ambulatorially. The study time started at January 1991 and finished at May 1994. Symptoms of MRGE showed high specificity but a low diagnostic sensitivity; moreover they are not able to foresee oesophageal damage. On the contrary the pH monitoring showed a 95% sensitivity. DeMeester's parameters identify refluxer subjects effectively, linking it to oesophageal damage severity. Symptoms and mucoesophageal damages are distributed equally on every kind of reflux patients including upright refluxers.