The aim of this study was to determine whether the burden of JAK2(V617F) allele correlated with major clinical outcomes in patients with polycythemia vera (PV). To this end, we determined JAK2 mutant ...allele levels in granulocytes of 173 PV patients at diagnosis. The mean (+/-s.d.) mutant allele burden was 52% (+/-29); 32 patients (18%) had greater than 75% mutant allele. The burden of JAK2(V617F) allele correlated with measurements of stimulated erythropoiesis (higher hematocrit, lower mean cell volume, serum ferritin and erythropoietin levels) and myelopoiesis (higher white cell count, neutrophil count and serum lactate dehydrogenase) and with markers of neutrophil activation (elevated leukocyte alkaline phosphatase and PRV-1 expression). As compared to those with less than 25% mutant allele, patients harboring greater than 75% JAK2(V617F) allele were at higher relative risk (RR) of presenting larger spleen (RR 4.7; P<0.001) or suffering from pruritus (RR 3.1; P<0.001). In these patients, the risk of requiring chemotherapy (RR 1.8; P=0.001) or developing major cardiovascular events (RR 7.1; P=0.003) during follow up were significantly increased. We conclude that a burden of JAK2(V617F) allele greater than 75% at diagnosis points to PV patients with high-risk disease.
We investigated the activity of ITF2357, a novel histone deacetylase inhibitor (HDACi) with antitumor activity, on cells carrying the JAK2(V617F) mutation obtained from polycythemia vera (PV) and ...essential thrombocythemia (ET) patients as well as the HEL cell line. The clonogenic activity of JAK2(V617F) mutated cells was inhibited by low concentrations of ITF2357 (IC(50) 0.001-0.01 microM), 100- to 250-fold lower than required to inhibit growth of normal or tumor cells lacking this mutation. Under these conditions, ITF2357 allowed a seven fold increase in the outgrowth of unmutated over mutated colonies. By western blotting we showed that in HEL cells, ITF2357 led to the disappearance of total and phosphorylated JAK2(V617F) as well as pSTAT5 and pSTAT3, but it did not affect the wild-type JAK2 or STAT proteins in the control K562 cell line. By real-time PCR, we showed that, upon exposure to ITF2357, JAK2(V617F) mRNA was not modified in granulocytes from PV patients while the expression of the PRV-1 gene, a known target of JAK2, was rapidly downmodulated. Altogether, the data presented suggest that ITF2357 inhibits proliferation of cells bearing the JAK2(V617F) mutation through a specific downmodulation of the JAK2(V617F) protein and inhibition of its downstream signaling.
In the brain, the spinal cord motor neurones express the highest levels of the androgen receptor (AR). Experimental data have suggested that neurite outgrowth in these neurones may be regulated by ...testosterone or its derivative 5α‐dihydrotestosterone (DHT), formed by the 5α‐reductase type 2 enzyme. In this study we have produced and characterized a model of immortalized motor neuronal cells expressing the mouse AR (mAR) neuroblastoma‐spinal cord (NSC) 34/mAR and analysed the role of androgens in motor neurones. Androgens either activated or repressed several genes; one has been identified as the mouse neuritin, a protein responsible for neurite elongation. Real‐time PCR analysis has shown that the neuritin gene is expressed in the basal condition in immortalized motor neurones and is selectively up‐regulated by androgens in NSC34/mAR cells; the DHT effect is counteracted by the anti‐androgen Casodex. Moreover, DHT induced neurite outgrowth in NSC34/mAR, while testosterone was less effective and its action was counteracted by the 5α‐reductase type 2 enzyme inhibitor finasteride. Finally, the androgenic effect on neurite outgrowth was abolished by silencing neuritin with siRNA. Therefore, the trophic effects of androgens in motor neurones may be explained by the androgenic regulation of neuritin, a protein linked to neurone development, elongation and regeneration.
Spinal cord motoneurones express high levels of androgen receptor. However, in responsive tissue, the effects of testosterone is often mediated by the more potent androgenic derivative ...5‐alpha‐dihydrotestosterone (DHT). This compound is formed in androgen target cells by the enzyme 5‐alpha‐reductase. Two isoforms of the 5‐alpha‐reductase, with limited degree of homology, have been cloned, type 1 and type 2. The low affinity‐constitutive type 1 isoenzyme is widely distributed in the body; the high affinity‐androgen regulated 5‐alpha‐reductase type 2 is confined to androgen‐dependent structures and shows a peculiar pH optimum at acidic values. We have previously shown that high levels of 5‐alpha‐reductase activity are detectable in rat spinal cord. Here, using reverse transcriptase‐polymerase chain reaction, we show that both isoforms are expressed in the whole spinal cord of the rat. The enzymatic pH optimum measured in immortalized spinal cord motoneurones (NSC34) is typical of the type 2 isoenzyme. Using in situ hybridization technique, we found that 5‐alpha‐reductase type 2 is confined to the motoneuronal cells of the anterior horns of the rat spinal cord, the cells that also are known to express high levels of androgen receptor. Because of the close association of androgen receptor and 5‐alpha alpha‐reductase type 2, motoneuronal cells should be considered as target cells for androgens.
Abstract
Background
Fingolimod is an immunosuppressive drug that prevents the typical autoimmune responses of multiple sclerosis (MS). This drug can cause, even after the first dose, bradycardia and ...atrioventricular conduction block (AVB). According to a decree of the Committee for Medicinal Products for Human Use (CHMP), the first administration of a dose of fingolimod must meet the following conditions: a) 12-lead electrocardiogram (ECG) and blood pressure (BP) evaluations before first dose administration; b) continuous ECG and BP monitoring every hour during the first 6 hours of treatment. The aim of the study was to evaluate the efficacy of our virtual Intensive Care Unit connected to selected Departments of Neurology for cardiology monitoring via web, during the first dose administration of fingolimod in patients with MS.
Methods
Health Telematic Network, in collaboration with our Cardiology Department, has installed in several Neurology Departments in the country intensive care equipment with real time monitoring via the web of ECG, BP and oximetry. The telemonitoring system is composed of an IntelliVue MP2 monitor, for 12-lead ECG recording, and by an IntelliVue Philips M3150 Central, for continuous remote monitoring of ECG and the detection and storage of arrhythmias. The HTN Cardiologist thus can evaluate in real time ECG monitored and each alarm detected.
Results
From Jan 2014 to Dec 2018, 172 sites participated and 6041 patients were enrolled. The telemonitoring system recognized 5116 episodes of bradycardia, 297 first degree AVB, 213 second degree AVB Mobitz I, 48 second degree AVB Mobitz II, 618 pathological lengthening of the QTc interval, single monomorphic ventricular ectopic beats in 266 patients, bigeminy ventricular extrasystoles in 15 patients and polymorphic ventricular extrasystoles in 3 cases. 6 cases of nonsustained ventricular tachycardia occurred and were recognized.
Conclusion
This web-based real time telemonitoring project was an optimal solution for the administration of the first dose of fingolimod, according to current indications of the CHMP. The RITMO project could lead the way for the use of a multidisciplinary approach for the management of new therapies.
U.O. Ematologia, Ospedali Riuniti, Bergamo, Italy
Correspondence: Guido Finazzi, U.O. Ematologia, Ospedali Riuniti, Largo Barozzi 1 24128 Bergamo, Italy. Phone: international +035.269493; Fax: ...international +035.266147. E-mail: gfinazzi{at}ospedaliriuniti.bergamo.it
We compared the laboratory and clinical findings of 179 patients with essential thrombocythemia (ET) and 77 with polycythemia vera (PV) classified according to the presence of the JAK2 V617F mutation. A gradient between patients with JAK2 wild-type ET, JAK2 V617F ET and PV (all carrying the JAK2 mutation) was observed. The rate of thrombotic complications in JAK2-positive ET was significantly higher than in wild-type ET and not statistically different from that of PV patients.
Key words: thrombosis, essential thrombocythemia, polycythemia vera, JAK2 mutation.
The aim of this study was to determine whether the burden of JAK2(V617F) allele correlated with major clinical outcomes in patients with polycythemia vera (PV). To this end, we determined JAK2 mutant ...allele levels in granulocytes of 173 PV patients at diagnosis. The mean (+/-s.d.) mutant allele burden was 52% (+/-29); 32 patients (18%) had greater than 75% mutant allele. The burden of JAK2(V617F) allele correlated with measurements of stimulated erythropoiesis (higher hematocrit, lower mean cell volume, serum ferritin and erythropoietin levels) and myelopoiesis (higher white cell count, neutrophil count and serum lactate dehydrogenase) and with markers of neutrophil activation (elevated leukocyte alkaline phosphatase and PRV-1 expression). As compared to those with less than 25% mutant allele, patients harboring greater than 75% JAK2(V617F) allele were at higher relative risk (RR) of presenting larger spleen (RR 4.7; P<0.001) or suffering from pruritus (RR 3.1; P<0.001). In these patients, the risk of requiring chemotherapy (RR 1.8; P=0.001) or developing major cardiovascular events (RR 7.1; P=0.003) during follow up were significantly increased. We conclude that a burden of JAK2(V617F) allele greater than 75% at diagnosis points to PV patients with high-risk disease.
From the Hematology and Bone Marrow Transplantation Units of Ospedali Riuniti Bergamo (OS, BP, MT, VG, AS, MCZ, EO, AG, TI, CM, TB, RB, AR); Spedali Civili Brescia (GR); Ospedale S. Maurizio Bolzano ...(PF); Fondazione IRCCS Ospedale Maggiore, Università di Milano (GL-D); Ospedale A. Businco Cagliari (EA) on behalf of Northern Italy Leukemia Group (NILG)
Correspondence: Alessandro Rambaldi, M.D., Divisione di Ematologia, Ospedali Riuniti, Largo Barozzi 1, 24100 Bergamo, Italy. E-mail: arambaldi{at}ospedaliriuniti.bergamo.it
Background and Objectives: The molecular analysis of minimal residual disease (MRD) may provide informaton on the risk of recurrence in patients with acute lymphoblastic leukemia (ALL). The aim of this study was to correlate the kinetics of MRD clearance after allogeneic transplantation with the clinical outcome of adults with ALL.
Design and Methods: MRD was evaluated by real-time quantitative polymerase chain reaction (RQ-PCR) using probes derived from fusion chimeric genes ( BCR/ABL and MLL/AF4 ) (n=22) or rearrangements of the T-cell receptor or immunoglobulin genes (n =21). Forty-three adult patients with ALL were studied to correlate the kinetics of MRD clearance before and after allogeneic hematopoietic stem cell transplantation.
Results: At 36 months, the overall survival of patients who underwent transplantation in hematologic remission (n = 37) was 80% for those who were PCR-negative before transplantation (n = 12) compared to 49% for PCR-positive patients (n = 25)( p =0.17). For the same patients the cumulative incidence of relapse was 0% and 46%, respectively ( p =0.027). Moreover, the relapse rate of patients who were PCR-negative at day +100 after transplantation was remarkably low (7%) compared to that among patients who were PCR-positive (80%, p =0.0006).
Interpretation and Conclusions: The kinetics of MRD clearance may help to identify patients at high risk of leukemia relapse after allogeneic stem cell transplantation. Patients not achieving an early molecular remission after transplantation require prompt and appropriate pre-emptive treatments such as infusions of donor lymphocytes or new experimental drugs.
Key words: MRD, adult ALL, Ph + ALL, allogeneic transplantation.
Introduction
Delirium and sarcopenia are common, although underdiagnosed, geriatric syndromes. Several pathological mechanisms can link delirium and low skeletal muscle mass, but few studies have ...investigated their association. We aimed to investigate (1) the association between delirium and low skeletal muscle mass and (2) the possible role of calf circumference mass in finding cases with delirium.
Methods
The analyses were conducted employing the cross-sectional “Delirium Day” initiative, on patient 65 years and older admitted to acute hospital medical wards, emergency departments, rehabilitation wards, nursing homes and hospices in Italy in 2017. Delirium was diagnosed as a 4 + score at the 4-AT scale. Low skeletal muscle mass was operationally defined as calf circumference ≤ 34 cm in males and ≤ 33 cm in females. Logistic regression models were used to investigate the association between low skeletal muscle mass and delirium. The discriminative ability of calf circumference was evaluated using non-parametric ROC analyses.
Results
A sample of 1675 patients was analyzed. In total, 73.6% of participants had low skeletal muscle mass and 24.1% exhibited delirium. Low skeletal muscle mass and delirium showed an independent association (OR: 1.50; 95% CI 1.09–2.08). In the subsample of patients without a diagnosis of dementia, the inclusion of calf circumference in a model based on age and sex significantly improved its discriminative accuracy area under the curve (AUC) 0.69 vs 0.57,
p
< 0.001.
Discussion and conclusion
Low muscle mass is independently associated with delirium. In patients without a previous diagnosis of dementia, calf circumference may help to better identify those who develop delirium.
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