Flow cytometric minimal residual disease assessment in multiple myeloma

W świetle wysokiej skuteczności nowych metod leczenia szpiczaka plazmocytowego oraz zwiększającego się odsetka osiąganych całkowitych remisji (CR) ocena minimalnej choroby resztkowej (MRD) jest obecnie istotną metodą określającą głębokość odpowiedzi. Wieloparametryczna cytometria przepływowa (MFC) jest obecnie najczęściej wykorzystywaną metodą monitorowania MRD w szpiku kostnym pacjentów ze szpiczakiem plazmocytowym, jednak w tym celu można również stosować metody molekularne. Rodzaj protokołu stosowanego przy badaniu MRD metodą MFC może istotnie wpływać na uzyskiwane wyniki, jednak obecnie jest już dostępny wystandaryzowany i wysoce czuły protokół cytometrii następnej generacji (NGF). Wykazano, że głębokość odpowiedzi oceniona na podstawie pomiaru MRD koreluje z przeżyciem wolnym od progresji (PFS) oraz przeżyciem całko¬witym (OS) chorych na szpiczaka plazmocytowego. Ponadto ujemny wynik badania w kierunku MRD jest lepszym czynnikiem prognostycznym w odniesieniu do PFS oraz OS niż osiągnięcie CR. Z tych względów wynik oceny MRD, uzyskany wysoce czułą oraz powtarzalną metodą MFC, jest potencjalnie użytecznym klinicznie biomarkerem do oceny skuteczności różnych strategii leczniczych i może być podstawą do podejmowania decyzji terapeutycznej oraz użytecznym czynnikiem prognostycznym w szpiczaku plazmocytowym.Minimal residual disease (MRD) assessment in light of the effectiveness of new multiple myeloma (MM) treatment modalities and related to it increasing ratios of achieved complete remissions (CR), becomes an important tool in recognition of the depth of the response. Multiparametric flow cytometry (MFC) is currently the most popular method for monitoring of MRD presence in bone marrow of MM patients; however, molecular techniques may also be used for MRD assessment. The choice of protocol utilized for MFC-MRD measurement can significantly affect the obtained results, nevertheless standardized and highly sensitive approach of next generation flow (NGF) is already available. The depth of response based on MRD assessment was shown to be an inde-pendent predictor of progression-free survival (PFS) and overall survival (OS). Furthermore, the MRD-negative status surpasses the prognostic value of CR achievement for PFS and OS. Thus, MRD status detected by highly sensitive and reproducible MFC is potentially a clinically applicable biomarker for evaluation of different treatment strategies efficacy potentially influencing treatment decisions, and acting as prognostic factor in MM patients

Citizen journalism is as old as journalism itself: An interview with Stuart Allan

Professor Stuart Allan from Cardiff University in the UK is one of the leading scholars in contemporary journalism studies. He has made a significant contribution to the development of this research field, having authored or edited seventeen books to date (many of which have been translated into multiple languages), as well as a wide range of journal articles and book chapters. He is a co-founder of the peer-reviewed journal Journalism Education, and serves on the editorial board of ten journals, including Digital Journalism, Journalism: Theory, Practice and Criticism, and New Media & Society. Although he is a man of many interests, Allan’s personal scholarship in journalism studies revolves around four themes: 1) journalism and democracy, where his attention has focused on the evolving role of the journalist in public life (Allan, 2010, 2012; Carter, Branston, and Allan, 1998; Fowler-Watt and Allan, 2013); 2) online news, with a particular interest in citizen journalism and what he terms citizen witnessing (Allan, 2006, 2013; Thorsen and Allan, 2014); 3) the changing nature of war, conflict and crisis reporting (Allan and Zelizer, 2004; Matheson and Allan, 2009; Zelizer and Allan, 2013); and 4) science journalism, with a special interest in how it is evolving in digital contexts (Allan, Adam and Carter, 2000; Allan, 2002; Anderson, Petersen, Wilkinson and Allan, 2009). Further research interests include journalism and human rights, media history, photojournalism, and young people’s civic engagement with digital media

Citizen journalism is as old as journalism itself: An interview with Stuart Allan

Professor Stuart Allan from Cardiff University in the UK is one of the leading scholars in contemporary journalism studies. He has made a significant contribution to the development of this research field, having authored or edited seventeen books to date (many of which have been translated into multiple languages), as well as a wide range of journal articles and book chapters. He is a co-founder of the peer-reviewed journal Journalism Education, and serves on the editorial board of ten journals, including Digital Journalism, Journalism: Theory, Practice and Criticism, and New Media & Society. Although he is a man of many interests, Allan’s personal scholarship in journalism studies revolves around four themes: 1) journalism and democracy, where his attention has focused on the evolving role of the journalist in public life (Allan, 2010, 2012; Carter, Branston, and Allan, 1998; Fowler-Watt and Allan, 2013); 2) online news, with a particular interest in citizen journalism and what he terms citizen witnessing (Allan, 2006, 2013; Thorsen and Allan, 2014); 3) the changing nature of war, conflict and crisis reporting (Allan and Zelizer, 2004; Matheson and Allan, 2009; Zelizer and Allan, 2013); and 4) science journalism, with a special interest in how it is evolving in digital contexts (Allan, Adam and Carter, 2000; Allan, 2002; Anderson, Petersen, Wilkinson and Allan, 2009). Further research interests include journalism and human rights, media history, photojournalism, and young people’s civic engagement with digital media

Increased T regulatory cells are associated with adverse clinical features and predict progression in multiple myeloma.

Background: Regulatory T (Treg) cells play an important role in the maintenance of immune system homeostasis. Multiple myeloma (MM) is a plasma cell disorder frequently associated with impaired immune cell numbers and functions. Methods: We analyzed Treg cells in peripheral blood (n = 207) and bone marrow (n = 202) of pre-malignant and malignant MM patients using flow cytometry. Treg cells and their subsets from MM patients and healthy volunteers were functionally evaluated for their suppressive property. A cohort of 25 patients was analyzed for lymphocytes, CD4 T cells and Treg cells before and after treatment with cyclophosphamide, thalidomide plus dexamethasone (CTD). Results: We found elevated frequencies of Treg cells in newly diagnosed (P<0.01) and relapsed MM patients (P<0.0001) compared to healthy volunteers. Also, Treg subsets including naive (P = 0.015) and activated (P = 0.036) Treg cells were significantly increased in MM patients compared to healthy volunteers. Functional studies showed that Treg cells and their subsets from both MM and healthy volunteers were similar in their inhibitory function. Significantly increased frequencies of Treg cells were found in MM patients with adverse clinical features such as hypercalcemia (.10 mg/dL), decreased normal plasma cell (<5%) count and IgA myeloma subtype. We also showed that MM patients with >5% of Treg cells had inferior time to progression (TTP) (13 months vs. median not reached; P = 0.013). Furthermore, we demonstrated the prognostic value of Treg cells in prediction of TTP by Cox regression analysis (P = 0.045). CTD treatment significantly reduced frequencies of CD4 T cells (P = 0.001) and Treg cells (P = 0.018) but not Treg cells/CD4 T cells ratio compared to pretreatment. Conclusions: Our study showed immune deregulation in MM patients which is evidenced by elevated level of functionally active Treg cells and patients with increased Treg cells have higher risk of progression

The cyanobacterial protoporphyrinogen oxidase HemJ is a new b-type heme protein functionally coupled with coproporphyrinogen III oxidase

Protoporphyrinogen IX oxidase (PPO), the last enzyme that is common to both chlorophyll and heme biosynthesis pathways, catalyzes the oxidation of protoporphyrinogen IX to protoporphyrin IX. PPO has several isoforms, including the oxygen-dependent HemY and an oxygen-independent enzyme, HemG. However, most cyanobacteria encode HemJ, the least characterized PPO form. We have characterized HemJ from the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis 6803) as a bona fide PPO; HemJ down-regulation resulted in accumulation of tetrapyrrole precursors and in the depletion of chlorophyll precursors. The expression of FLAG-tagged Synechocystis 6803 HemJ protein (HemJ.f) and affinity isolation of HemJ.f under native conditions revealed that it binds heme b. The most stable HemJ.f form was a dimer, and higher oligomeric forms were also observed. Using both oxygen and artificial electron acceptors, we detected no enzymatic activity with the purified HemJ.f, consistent with the hypothesis that the enzymatic mechanism for HemJ is distinct from those of other PPO isoforms. The heme absorption spectra and distant HemJ homology to several membrane oxidases indicated that the heme in HemJ is redox-active and involved in electron transfer. HemJ was conditionally complemented by another PPO, HemG from Escherichia coli. If grown photoautotrophically, the complemented strain accumulated tripropionic tetrapyrrole harderoporphyrin, suggesting a defect in enzymatic conversion of coproporphyrinogen III to protoporphyrinogen IX, catalyzed by coproporphyrinogen III oxidase (CPO). This observation supports the hypothesis that HemJ is functionally coupled with CPO and that this coupling is disrupted after replacement of HemJ by HemG