Andrea Ferrario

Outbreeding and asymbiotic germination in the conservation of the endangered Italian endemic orchid Ophrys benacensis

Abstract The Insubric bee orchid, Ophrys benacensis (Reisigl) O.&E. Danesch & F. Ehrend, occurs in fragmented populations only in northern Italy, and suffers from inbreeding depression. We found that pollen from a depressed population at Monte Barro, Lecco, did not fertilise plants of the same population, nor of a larger population at nearby Valmadrera. Fertilisation was successful at both sites when pollen from Valmadrera was used, although the proportion of seeds containing embryos was almost six times greater at Valmadrera. Embryos produced by both populations were equally likely to develop in vitro. Sowing seed on medium enriched with 50 mL L−1 coconut milk more than doubled the germination rate with respect to a non‐enriched control (i.e. from 14.5% to 39.8%; p = 0.024, Student’s t‐test), whereas other complex organic media inhibited germination. We conclude that both pollen and ovules have inherent developmental problems that can be partially overcome by outbreeding with larger populations. Once seed is produced propagation is relatively easy: sufficient plantlets were produced to enlarge the Monte Barro population to 195 times its current size.

Novel agents in indolent lymphomas

Indolent non-Hodgkin’s lymphomas (iNHLs) include follicular lymphomas (FL), marginal-zone lymphoma, lymphoplasmacytic lymphoma/Waldenström macroglobulinemia and small lymphocytic lymphoma. First-line standard therapy in advanced, symptomatic iNHL consists of rituximab-based immunochemotherapy. The recent rediscovery of the ‘old’ chemotherapeutic agent bendamustine, an alkylating agent with a peculiar mechanism of action, has added a new effective and well-tolerated option to the therapeutic armamentarium in iNHL, increasing response rates and duration. However, patients invariably relapse and subsequent active and well-tolerated agents are needed. In recent years a large number of new targeted agents have been tested in preclinical and clinical experimentation in FL and indolent nonfollicular lymphoma (iNFL), including the new monoclonal antibodies binding CD20 or other surface antigens, immunoconjugates and bispecific antibodies. Moreover novel agents directed against intracellular processes such as proteasome inhibitors, mTOR inhibitors and agents that target the tumour microenvironment, notably the immunomodulatory agent lenalidomide, are under active clinical investigation. The development of these new drugs may change in the near future the approach to iNHL patients, leading to better tolerated and effective therapy regimens.

The clinical and biological features of a series of immunophenotypic variant of B-CLL.

Objectives: To describe the clinical and biological features of a series of immunophenotypic variant of B‐CLL (v‐CLL) characterised by intermediate RMH score, in the absence of t(11;14)(q13;q32) in FISH analysis in comparison with a series of typical CLL. Methods: We studied the clinical and biological features of 63 cases of v‐CLL and 130 cases of CLL. Results: We observed significant differences in terms of age <70u2003yr (Pu2003<u20030.001), lymphocytosis <20u2003×u2003109/L (Pu2003<u20030.001), lymphocyte doubling time ≤12u2003months (Pu2003=u20030.02), high serum β2‐microglobulin levels (Pu2003<u20030.001) and splenomegaly (Pu2003=u20030.002); CD38, CD49d, CD1c were more expressed in v‐CLL, CD43 in CLL (Pu2003<u20030.001). IgVH mutation and trisomy 12 were more frequent in v‐CLL group (Pu2003=u20030.001; Pu2003<u20030.001); del13q14 in CLL (Pu2003=u20030.008). Gene expression profiling of nine v‐CLL and 60 CLL indicated that the atypical group presented a specific molecular pattern. After a median follow‐up of respectively, 55 (4–196) and 60u2003months (6–180), 25/42 patients with v‐CLL (48%) and 55/93 patients with CLL (59%) were treated. Time to treatment was significantly shorter in IgVH‐mutated v‐CLL vs. mutated CLL (Pu2003=u20030.006). The median overall survival was worse in v‐CLL‐mutated cases (Pu2003=u20030.062). Conclusion: v‐CLL should be identified and dealt with separately from classic CLL. In particular, the prognostic markers that are routinely used to characterise classical B‐CLL should not be interpreted as having the same meaning.

Everolimus in diffuse large B-cell lymphomas

Satisfactory treatment of relapsed/refractory diffuse large B-cell lymphoma (DLBCL) is not currently available and novel therapies are needed. mTOR is an intracellular kinase that is part of an aberrantly activated pathway in DLBCL. Preclinical studies in DLBCL cell lines demonstrated that everolimus, an oral selective mTOR inhibitor, induces cell cycle arrest and is synergistic with rituximab. Phase I studies indicated 10 mg daily to be the best dosing of everolimus in DLBCL. A large Phase II study in relapsed/refractory DLBCL confirmed the substantial activity (overall response rate: 30%) and good tolerability of everolimus in DLBCL, with thrombocytopenia being the main toxicity. The combination of everolimus and rituximab showed encouraging results (objective response rate: 38%; complete response: 13%), without increasing toxicity. Combination studies of everolimus with novel agents or with immunochemotherapy are underway.

Investigational therapies targeting lymphocyte antigens for the treatment of non-Hodgkin's lymphoma.

Introduction: The advent of the anti-CD20 mAb rituximab has opened a new era in the treatment of non-Hodgkin’s lymphomas (NHL), markedly altering standard treatment strategies. Moreover, the proof-of-concept that targeting a specific lymphocyte surface antigen may induce a highly effective and safe targeted killing of malignant cells has opened the door to the development of a plethora of novel mAbs directed towards different B- and T-cell-specific antigens. Areas covered: This review discusses the recent available clinical data about new-generation anti-CD20 mAbs characterized by increased antibody- (obinutuzumab) or complement-dependent cyotoxicity (ofatumumab) as well as novel investigational agents targeting other lymphocyte antigens (e.g., CD19, CD22, CD30, CD40, CD52, CCR4), which are currently under investigation for B- and T-cell NHL treatment. In addition, antibody–drug conjugates (inotuzumab ozogamicin, brentuximab vedotin, polatuzumab vedotin), bispecific T-cell engagers (blinatumomab) and a new class of antibodies targeting cytotoxic T-lymphocyte-associated antigen 4, programmed death 1 or programmed death ligand 1 (immune checkpoint inhibitors) are specifically considered. Expert opinion: Among the novel mAbs challenging rituximab, obinutuzumab seems to be in the most advanced phase, with the results of randomized trials awaited shortly. Brentuximab vedotin is increasing its role in T-cell NHL. Furthermore, immune checkpoint inhibitors have the potential to acquire a great relevance in NHL therapy.

Emerging resistant bacteria strains in bloodstream infections of acute leukaemia patients: results of a prospective study by the Rete Ematologica Lombarda (Rel).

Multiresistant bacterial infections are a potentially life-threatening condition in acute leukaemia (AL) patients. We aimed to better define the very recent epidemiology and outcome of bloodstream infections (BSIs) in a real-life setting. We prospectively collected all consecutive febrile/infectious episodes occurring in AL patients admitted to 9 haematology units. In 293 AL patients, 433 BSIs were diagnosed. Gram-positive (GP) bacteria were isolated in 44.8xa0% BSI and Gram-negative (GN) in 38.3xa0%, while polymicrobial aetiology- or fungi-related events were identified in 15.7 and 1.1xa0% of the cases, respectively. GP was observed more frequently in patients not in complete remission (pu2009=u20090.04), while GN during consolidation cycles (pu2009=u20090.003). Extended spectrum β-lactamase-producing strains accounted for 23.2xa0% of enterobacteria. They were associated with previous antibiotic exposure, including fluoroquinolones prophylaxis (pu2009=u20090.01). Carbapenem-resistant (CR) strains occurred in 9xa0% of enterobacteria. Among Pseudomonas aeruginosa strains, 21.6xa0% were multiresistant. Overall 30-day mortality was 8.5xa0%. CR GN and multiresistant P. aeruginosa BSIs were independent predictors of death (pu2009=u20090.002), as well as relapsed/resistant AL (18.3xa0%; pu2009=u20090.0002) and the presence of pulmonary infiltrates (26.6xa0%; pu2009<u20090.001). Although GP still predominate over GN BSI, the percentage of antibiotic resistant GN strains is considerable in AL patients and it is associated with poor prognosis.

New uses for brentuximab vedotin and novel antibody drug conjugates in lymphoma

ABSTRACT Introduction: Brentuximab vedotin (BV) is a potent anti-CD30 antibody drug conjugate (ADC) that has been approved in relapsed or refractory Hodgkin lymphoma (HL) after autologous stem cell transplantation (ASCT) and anaplastic large-cell lymphoma (ALCL). Beyond these consolidated indications, BV has been tested in a number of different settings with promising results, leading for example to the recent approval as a consolidation after ASCT in high-risk HL patients. Areas covered: Main emerging areas of clinical investigation of BV include the use as a single-agent or in combination with bendamustine in first-salvage therapy of HL (bridge to ASCT), in the frontline setting in combination with AVD chemotherapy in HL and with CHP in ALCL, in relapsed or refractory cutaneous T-cell lymphomas and finally in diffuse large B-cell lymphomas (DLBCL) expressing CD30. Moreover, many new ADCs are currently under clinical evaluation, as for example the anti-CD79A polatuzumab vedotin in DLBCL. Expert commentary: In few years BV changed the therapeutic scenario of relapsed or refractory HL and ALCL and is rapidly moving toward first-line approval in combination with standard chemotherapy if ongoing randomized trials will demonstrate improved results. Combination strategies with bendamustine in first-salvage HL and with R-CHP in first-line DLBCL appear very promising.

Pea seed extracts stimulate germination of the terrestrial orchid Ophrys apifera Huds. during a habitat restoration project

Novel methods are required to break the seed dormancy of temperate-zone orchids and aid the conservation of rare species. Zeatin is produced in increasing concentrations during the development of pea (Pisum sativum) seeds. We hypothesised that hot water extracts of pea seeds stimulate germination of the orchid Ophrys apifera in vitro, particularly for extractions made during later pea development. Pea seeds, exposed to 0, 3, 6, 9, 12 or 15 days of periodic wetting, were extracted in hot water and the extracts were added to Malmgrens growth medium. Germination of O. apifera on this medium was quantified after 7 months, stimulated by a range of pure hormones. Pea seed extract collected later in pea development (at 6–15 days) inhibited germination of O. apifera. However, extracts taken at 0 and 3 days significantly increased germination from 3.8 ± 0.32% in the control to 9.1 ± 1.84% and 7.6 ± 0.79%, respectively: increases comparable to the most effective of the pure hormones. Dried peas therefore provide an economical alternative source of germination stimulants for orchids. We briefly report how sufficient mature plants of O. apifera were produced to allow a population to be multiplied to 15 times its original size during a habitat restoration project.

Clinical Predictors of Outcome in MPN

Myeloproliferative neoplasms include 3 diseases: polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). PV and ET are dominated by a high risk of thrombosis and a late risk of clonal evolution into secondary MF and acute myeloid leukemia. Patients with PMF may encounter many complications associated with disease progression or with PMF evolution. This article defines factors that determine prognosis in these 3 diseases.

Rituximab plus Bendamustine as front line treatment in frail elderly (>70 years) patients with diffuse large B-cell non-Hodgkin's lymphoma: a phase II multicenter study of the Fondazione Italiana Linfomi

We conducted a phase II study to assess activity and safety profile of bendamustine and rituximab in elderly patients with untreated diffuse large B-cell lymphoma (DLBCL) who were prospectively defined as frail using a simplified version of the Comprehensive Geriatric Assessment (CGA). Patients had to be over 70 years of age, with histologically confirmed DLBCL. Frail patients were those younger than 80 years with a frail profile at CGA or older than 80 years with an unfit profile. Treatment consisted of 4-6 courses of bendamustine [90 mg/m2 days (d)1-2] and rituximab (375 mg/m2 d1) administered every 28 days. Other main study end points were complete remission rate and the rate of extra-hematologic adverse events. Forty-nine patients were enrolled of whom 45 were confirmed eligible. Overall, 24 patients achieved a complete remission (53%; 95%CI: 38-68%) and the overall response rate was 62% (95%CI: 47-76%). The most frequent grade 3-4 adverse event was neutropenia (37.8%). Grade 3-4 extra-hematologic adverse events were observed in 7 patients (15.6%; 95%CI: 6.5-29.5%); the most frequent was grade 3 infection in 2 patients. With a median follow up of 33 months (range 1-52), the median progression-free survival was ten months (95%CI: 7-25). The study shows promising activity and manageable toxicity profile of BR combination as first-line therapy for patients with DLBCL who are prospectively defined as frail according to a simplified CGA, as adopted in this trial (clinicaltrials.gov identifier: 01990144).