Increasing evidence suggests that impaired cytomegalovirus (CMV)-specific cell-mediated immunity (CMV-CMI) is a major cause of uncontrolled CMV reactivations and associated complications in ...hematopoietic stem cell transplantation (HSCT). No reliable test exists to predict patients at risk of primary and/or recurrent CMV reactivations following HSCT. Accurately assessing CMV-CMI might therefore improve the risk stratification of patients and allow optimizing and individualizing patient care.
This study aimed to evaluate the suitability of a novel IFN-γ ELISpot assay (T-Track® CMV), based on the stimulation of peripheral blood mononuclear cells with T-activated® pp65 and IE-1 CMV proteins, to predict protection from recurrent CMV reactivation following the resolution of a treatment-requiring primary CMV reactivation.
A prospective, longitudinal, observational, multicenter study was conducted in 175 intermediate- and high-risk (Donor (D)+/Recipient (R)+, D+/R-, D-/R+) HSCT recipients (ClinicalTrials.gov ID: NCT02156479). Patients underwent preemptive antiviral therapy per institutional guidelines. CMV DNAemia was analyzed by quantitative PCR. CMV-CMI was measured at day 45, 60, 80, 100 and 120 post-transplantation, as well as at onset and following the end of preemptive treatment. Occurrence of recurrent CMV reactivation was monitored up to 7.5 months post-transplantation. 154/175 patients fulfilling the inclusion/exclusion criteria and having at least one valid T-Track® CMV test result were included in the final analysis.
101/154 (66%) patients experienced at least one (treatment-requiring) CMV reactivation during the observational period. Out of 74 patients (24 D+/R+, 3 D+/R-, 47 D-/R+) who experienced a first CMV reactivation and had a valid ELISpot test result at the end of this primary reactivation, 30 (41%) faced a recurrent CMV reactivation during the observational period. Interestingly, 41/44 patients free of recurrent reactivation had a positive test result (i.e. positive for at least one of pp65- and/or IE-1-specific result) after resolution of the primary CMV reactivation, resulting in a 93% specificity in diagnostic accuracy. Accordingly, a time-to-event analysis indicated a significantly lower incidence of recurrent CMV reactivation in patients with a positive test result (Figure 1; Hazard ratio=5.68; Log-Rank Test, p<0.001). A ROC analysis also demonstrated that pp65-specific response measured following a primary CMV reactivation is a good negative predictor for future CMV reactivation (AUC=0.840 95% CI 0.741-0.939; p<0.001).
Altogether, this novel standardized IFN-γ ELISpot assay allows an improved risk stratification of CMV-related clinical complications, and can support clinicians in the identification and management of patients with increased risk of recurrent CMV reactivation following HSCT.
In the recent years T-ALL/LBL has turned to a favourable subtype of adult ALL due to intensive chemotherapy and/or SCT in first CR. At relapse, however, the disease is highly refractory and rapidly ...progressive. In the GMALL study 05/93 in ‘early' relapse during therapy the CR rate with HD regimens was 29% and the survival 8%. The major problem was achievement of CR in order to proceed to SCT. Therefore the T-cell specific purine analogue compound GW506U78 (NSC 686673, Nelarabine) was evaluated. The compound was provided by the National Cancer Institute and administered as single drug (1.5 g/m2 day 1,3,5) in 53 adult pts. The median age was 31 (19–81) yrs. 47 (89%) had T-ALL and 6 T-LBL. 44 presented BM and 9 only extramedullary involvement. All pts had heavily pretreated, refractory disease. 36 (68%) were included in 1st ‘early', 7 (13%) in 2nd relapse, 7 (13%) in relapse after SCT. 3 (6%) had never obtained CR. 32/36 pts in 1st relapse were refractory to >= one HD salvage therapy (FLAG-IDA 5, Cladribine/VP16/HDAC 18, other HDAC/HDMTX based 9).
25/53 evaluable pts (47%) achieved CR, 7 PR (13%) and 21 (40%) were refractory. The highest CR rate was achieved in pts with thymic T-ALL (76%). 19/25 CR pts (76%) were rapidly transferred to SCT (4 sibling, 14 MUD, 1 auto). Median time to SCT was 41 (20–83) d. 7/25 CR pts are in continuous CR at median 13 (1–36) mo. 4 pts died in CR (1 sepsis/liver failure, 3 transplant related). 14 pts relapsed, 10 after SCT. Time to relapse was median 5 (1–8) mo. 2 pts developed AML in relapse after SCT. 10 pts were included in the programme a second time, 8 in relapse after SCT. 4 CRs, 1 subjective improvement and 5 failures were observed. The probability of survival in the whole cohort is 16%, but significantly higher in pts who achieved CR (27%). Moderate bone marrow suppression and elevated liver enzymes were the most frequent toxicities. Neurotoxicity was encountered in only two pts (reversible psychosyndrome with agitation and somnolence).
We conclude that the compound has a impressive single drug activity in highly resistant relapsed T-ALL and is well tolerated even in heavily pretreated pts. Exploration in earlier stages e.g. molecular relapse, in front-line therapy and in combination is warranted. Since a durable remission cannot be expected with chemotherapy a high proportion of CR pts was transferred to SCT. Importantly no unexpected mortality or morbidity was observed after SCT. Long-term relapse free survival was achieved in some pts. The major problem were relapses. Therefore a better remission quality and lower tumor load before SCT should be obtained e.g. by consolidation cycles with the compound. After SCT close monitoring should aim for early detection of beginning relapse by MRD analysis in order to decide on interventions e.g. reduction of GvHD prophylaxis, donor lymphocyte infusions or additional cycles with the compound.Partly supported by Deutsche Krebshilfe (M84/92Ho1), NCI/CTEP and GlaxoSmithKline
Allogeneic stem cell transplantation is the most effective therapy for myeloid leukemias. Still, especially patients with refractory disease, unfavourable cytogenetics or blast transformation of a ...myeloproliferative disorder (mpd) have a poor prognosis as a result of a high rate of treatment related mortality and relapse with conventional conditioning regimens. Recently, Kolb et al. have introduced a sequential protocol utilizing early TBI-based non-myeloablative transplantation following conventional induction therapy in high-risk patients.
In this study, in dependence to Kolb, we have treated 21 patients (median age 51 years, range 17–63, 4 female, 17 male) with advanced leukemias according to an intensified sequential high-dose protocol. Diagnoses were AML (primary refractory 2, relapsed 5), MDS (refractory 7, untreated 1) and blast transformation of a MPD (refractory 6, untreated 0). The median blast count at induction was 22% (range 1–90) and 9 patients (43 %) had unfavourable cytogenetics. Induction therapy consisted of fludarabine, high-dose cytarabine with or without idarubicine, amsacrine or topotecan. ATG, high-dose melphalan with or without high-dose thiotepa was used for conditioning while patients were still cytopenic from the preceeding induction therapy. Following transplantation of a median of 6.1x10E6 CD34+ cells/kg (range 0.85–12.0) from related (6) or unrelated (15) donors, the median time to a wbc>1000/μl and plt>20000/μl was 15 days (range 10–24) and 17 days (range 8–30), respectively. No toxic death was observed during the first 30 days and 95% of patients achieved CR by day 28, 100% by day 50. Twelve patients (57%) developed aGvHD (12I-II°, 0III-IV°) and 7 patients (33%) at risk have developed chronic GvHD (5 limited, 2 extensive) so far. After a median time of 181 days (range 3 – 596 days) fifteen patients (72%) are alive and all are in complete remission. Six patients died, one of infection (5%), two of a combination of a chronic GvHD and infections (9%), two of disease progression (9%) and one patient from a second malignancy (SCLC).
We conclude that early sequential transplantation during induction therapy induced cytopenia using high-dose melphalan and thiotepa based conditioning is feasible and highly effective in patients with advanced myeloid leukemias and poor prognosis. Although these early results are promising, a longer follow up and comparison with standard regimens are required.
The blood-brain barrier (BBB) prevents the passage of plasma-protein bound chemotherapeutic agents into the CNS, presenting a major obstacle for effective chemotherapy of malignant gliomas. It has ...previously been shown that Nd:YAG Laser-Induced Thermo Therapy (LITT) is associated with BBB permeability changes. In this paper we briefly review the anatomy of the BBB and pharmacological consequences for the delivery of chemotherapeutic agents to the brain and discuss approaches for opening the BBB for therapeutic purposes. We then analyze the basic mechanisms of LITT-induced BBB changes and address specifically the question of whether LITT-induced disruption of the BBB facilitates a locoregional passage of chemotherapeutic agents into the brain tissue. Finally we review the feasibility of this method as an adjuvant therapeutic modality of adjuvant glioma therapy.
The purpose of this study was to investigate the development of pre-service chemistry teachers' (PCTs') argumentation skills in a laboratory course throughout a semester. A case study design was ...utilised. A total of six PCTs participated in the study. To investigate the development of PCTs' argumentation skills, the laboratory reports that were in the form of science writing heuristic and pre and post semi-structured interviews were employed. Results showed that PCTs' argumentation skills developed over time as they started to generate more powerful arguments which also included deeper conceptual knowledge during the argumentation-based laboratory. The other result of the study revealed that PCTs become successful in explaining chemistry concepts at the sub-microscopic level throughout the course.
In this study, a direct assay, a modified CUPRAC (Cupric Ion Reducing Antioxidant Capacity) method, is developed to determine transition metal ion (Cu(II))-catalyzed pro-oxidant activity of ...polyphenolic compounds, vitamins C and E, and herbal samples in the presence of proteins containing thiol groups. Since transition metal ion-catalyzed pro-oxidant activity of phenolics is usually initiated with the reduction of the metal to lower oxidation states (as a prerequisite of Fenton-type reactions), this method involves the reduction of copper(II) ions to copper(I) by polyphenolic compounds (simultaneously giving rise to reactive species), binding of the formed Cu(I) to egg white protein –SH groups, and liberation of copper(I)-neocuproine (Cu(I)-Nc) chelate (showing maximum absorbance at 450nm) by treating the incubation product with a neocuproine–ammonium acetate mixture. The proposed method is validated against atomic absorption spectrometric (AAS) determination of protein-bound copper and protein carbonyl assay of oxidative stress. The proposed assay is faster and more specific than the carbonyl assay, and uses low-cost reagents and equipment. Pro-oxidant activity (i.e. proportional to absorbance) varies linearly over a relatively wide range with concentration, as opposed to the reciprocal correlations (i.e. linear regression of 1/(pro-oxidant activity) versus 1/concentration) of other similar assays. The pro-oxidant activity order of the tested antioxidant compounds in terms of ‘Quercetin Equivalent Pro-oxidant Activity' (QREPA) coefficients is: gallic acid>epicatechin>quercetin≈catechin>α-tocopherol>rosmarinic acid>trolox>caffeic acid>ascorbic acid.
•A direct colorimetric pro-oxidant activity assay was developed for polyphenols.•Cu(II)-induced pro-oxidant action of polyphenols produces reactive species and Cu(I).•Cu(I) binds to egg white protein thiols, and is liberated by buffered neocuproine.•Pro-oxidant activity of phenolics is proportional to copper(I)-neocuproine absorbance.•Pro-oxidant activity varies linearly with antioxidant concentration, and is additive.
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