In this study, methylation-specifi c polymerase chain reaction was used to investigate the potential prognostic signifi cance of the methylation status of p15 , p16, MGMT , and DAPK genes in 51 specimens of diffuse large B-cell lymphoma (DLBCL). Hypermethylation of p15 gene was signifi cantly more prevalent in patients without relapse ( p = 0.001) and there was a trend toward more frequent presence of p15 methylation in patients without death outcome within 5-year follow-up period ( p = 0.086) Also, there was a trend toward accumulation of p15 methylation with favorable clinicopathological parameters including: age ≤ 60 years ( p = 0.091), normal levels of lactate dehydrogenase ( p = 0.090), Eastern Cooperative Oncology Group performance status < 2 ( p = 0.095), and low/intermediate low International Prognostic Index ( p = 0.076). In the female group and group of the patients without bulky tumor mass, treated with chemotherapeutic regimens including rituximab, methylation of p15 was signifi cantly related to longer overall survival ( p = 0.036 and 0.027, respectively). Our results suggest that promoter methylation of p15 gene could have prognostic value in DLBCL patients treated with rituximab when used in combination with gender and tumor size. Clin Trans Sci 2014; Volume 7: 384-390Keywords: diffuse large B-cell lymphoma , methylation, p15 , prognosis , rituximab 385 VOLUME 7 • ISSUE 5 WWW.CTSJOURNAL.COM Krajnović et al. ■ Hypermethylation of p15 Gene in DLBCL considered positive for Bcl-2, Bcl-6, or CD10 when at least 50%, 10%, or 20% of tumor cells expressed Bcl2, Bcl6, or CD10 protein, respectively. Expression of Ki67 protein in 0-30% of tumor cells was considered as weak, in 30-60% of tumor cells as moderate, and in more than 60% of the cells as strong. All procedures were carried out with the prior informed consent of the patients and with the approval of the local Ethic Committee. Treatment consisted of CHOP/R-CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone with or without rituximab) regimen. Some patients had adjuvant radiotherapy and/or surgery, and some of them underwent autologous stem-cell transplantation. Response criteria and survival outcomes were defi ned according to the recommendation of Cheson et al. 17Analysis of methylation status of the p15 , p16 , MGMT , and DAPK genes DNA methylation patterns in the promoter CpG islands of the p15 , p16 , MGMT , and DAPK genes were determined in all of 51 samples by methylation-specifi c polymerase chain reaction (MSP) following the bisulfi te modifi cation of isolated genomic DNA, as described earlier.18 Briefl y, DNA was isolated from deparaffi ned tumor specimens using standard proteinase K, phenol/chloroform/isoamyl alcohol extraction, and ethanol precipitation.19 Two micrograms of isolated DNA were denatured by NaOH (fi nal 0.3 mol/L) at 42°C for 30 minutes and modifi ed by sodium bisulfi te (5.20-5.69 mol/L, pH 5.0, Sigma, St Louis, MO, USA) for 18 hours at 50°C . Aft er incubation, DNA was purifi ed using the DNA extraction...

In accordance with our findings and recommendations provided by other authors, cladribine can be considered an effective alternative treatment for ECD, especially with CNS involvement and BRAF V600E-mutation-negative status, when interferon-alpha as the first-line therapy fails.

We analyzed 158 patients, women/men ratio 64/94 median age 29 (range 9-57), who underwent allogeneic SC transplantation between 1989 and 2009. All included patients had diseases as follows: acute myeloid leukemia (AML)--39, acute lymphoblastic leukemia (ALL) 47, chronic myeloid leukemia (CML)--32, myelodysplastic syndrome (MDS)--10, Hodgkin's lymphoma (HL)- 2, multiple myeloma (MM) 3, granulocytic sarcoma (GrSa) 3, severe aplastic anemia (sAA)--22. The patients underwent transplantations were divided into two groups: BMT group (74 patients) and PBSCT group (84 patients). Each recipient had HLA identical sibling donor. SCs from bone marrow were collected by multiple aspirations of iliac bone and from PB by one "Large Volume Leukapheresis" (after recombinant human granulocyte colony stimulating factor, rHuG-CSF) application (5-12 microg/kgbm, 5 days). Conditioning regimens were applied according to primary disease, GvHD prophylaxis consisted of combination of a cyclosporine A and methotrexate. Results. Engraftment, according to the count of polymorphonuclear and platelets, were significantly (p < 0.001) faster in the PBSCT vs BMT group. The needs for transfusion support were significantly (P < 0.01) higher in the BMT group. Those patients had more frequently oropharingeal mucositis grade 3/4 (33.3% vs 10.0%, p < 0.05). There were no significant differences in the incidence of aGvHD and cGvHD between the two groups. The patients who underwent PBSCT had more frequently extensive cGvHD in comparison with the BMT group (29.1% vs 11.29%, p < 0.05). SC source (SCS) had no significant influence on the TRM (21.62% vs 23.8%, p = 0.64) and the incidence of relapses (21.6% vs 29.7%, p = 0.32). Finally, the patients treated by BMT hd a significantly better OS (logrank 2.33, p < 0.05). Conclusion. SCs harvesting from PB resulted in improved cell yield, faster engraftment, as well as in a decrease of immediate transplantation related complications with a reduced treatment cost. Allogeneic PBSCT were associated with more frequent extensive cGvHD, while the influence of SCS in TRM and relapses was not observed. Finally, the long-term OS was better in the patients treated by BMT. To verify impact of SC source on transplantation (PBSCT vs BMT) overall efficacy, more larger randomized clinical studies are needed.

Background Clostridioides difficile infection (CDI) is one of the most common healthcare-associated (HA) infections. Cancer patients, particularly haemato-oncological patients, have an increased risk for CDI due to more risk factors compared with non-cancer patients. The aim of this study was to investigate differences in outcomes associated with HA CDI in patients with solid and haematological malignancies compared with patients with no underlying malignant disease in a tertiary healthcare centre in Serbia.MethodsA prospective cohort study was conducted including adult patients diagnosed with an initial episode of HA CDI. Their demographic and clinical characteristics associated with risk factors for CDI were documented. Outcomes such as all-cause 30-day mortality, cure of infection, diarrhoea relaps and recurrence of disease were followed. Patients were assigned to cancer and non-cancer groups. Within the cancer group, patients were divided into the solid tumour subgroup and haematological malignancy subgroup.ResultsDuring a 7-year period, HA CDI was observed in 28 (5.1%) patients with haematological malignancy, 101 (18.3%) patients with solid tumours and 424 (76.7%) non-cancer patients. Older age (OR 1.04, 95% CI 1.02 to 1.07, p<0.001), admission to the intensive care unit (ICU) (OR 2.61, 95% CI 1.37 to 4.95, p=0.003), mechanical ventilation (OR 5.19, 95% CI 2.78 to 9.71, p<0.001) and use of antibiotics prior to CDI (OR 1.04, 95% CI 1.02 to 1.06, p=0.02) were associated with increased mortality. Compared with patients with solid tumours, patients with haematological malignancy were younger (65 vs 57 years, p=0.015), did not require ICU admission (25.0% vs 0%) or mechanical ventilation (8.9% vs 0%) and were treated longer with antibiotics prior to CDI (14 vs 24 days, p=0.002).ConclusionsPatients with haematological malignancy were exposed to different risk factors for CDI associated with mortality compared with patients with solid tumours and non-cancer patients. Older age, ICU stay and mechanical ventilation, but not presence or type of cancer, predicted the all-cause 30-day mortality.