Transfer of minimally manipulated CMV-specific T cells from stem cell or third-party donors to treat CMV infection after allo-HSCT.

Cytomegalovirus (CMV) infection is a common, potentially life-threatening complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT). We assessed prospectively the safety and efficacy of stem cell-donor- or third-party-donor-derived CMV-specific T cells for the treatment of persistent CMV infections after allo-HSCT in a phase I/IIa trial. Allo-HSCT patients with drug-refractory CMV infection and lacking virus-specific T cells were treated with a single dose of ex vivo major histocompatibility complex-Streptamer-isolated CMV epitope-specific donor T cells. Forty-four allo-HSCT patients receiving a T-cell-replete (D+ repl; n=28) or T-cell-depleted (D+ depl; n=16) graft from a CMV-seropositive donor were screened for CMV-specific T-cell immunity. Eight D+ depl recipients received adoptive T-cell therapy from their stem cell donor. CMV epitope-specific T cells were well supported and became detectable in all treated patients. Complete and partial virological response rates were 62.5% and 25%, respectively. Owing to longsome third-party donor (TPD) identification, only 8 of the 57 CMV patients transplanted from CMV-seronegative donors (D-) received antigen-specific T cells from partially human leukocyte antigen (HLA)-matched TPDs. In all but one, TPD-derived CMV-specific T cells remained undetectable. In summary, adoptive transfer correlated with functional virus-specific T-cell reconstitution in D+ depl patients. Suboptimal HLA match may counteract expansion of TPD-derived virus-specific T cells in D- patients.

The proteome pattern cGvHD_MS14 allows early and accurate prediction of chronic GvHD after allogeneic stem cell transplantation.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) may be curative, but is associated with significant morbidity and mortality. Chronic graft-versus-host disease (cGvHD), characterized by inflammation and fibrosis of multiple target organs, considerably contributes to the morbidity and mortality even years after allo-HSCT. Diagnosis of cGvHD is based on clinical features and histology of biopsies. Here, we report the generation of a urinary cGvHD-specific proteome-pattern (cGvHD_MS14) established by capillary electrophoresis-mass spectrometry to predict onset and severity of cGvHD as an unbiased laboratory test. cGvHD_MS14 was evaluated on samples from 412 patients collected prospectively in four transplant centers. Sensitivity and specificity was 84 and 76% by cGvHD_MS14 classification. Sensitivity further increased to 93% by combination of cGvHD_MS14 with relevant clinical variables to a logistic regression model. cGvHD was predicted up to 55 days prior to clinical diagnosis. Acute GvHD is not recognized by cGvHD_MS14. cGvHD_MS14 consists of 14 differentially excreted peptides, six of those have been sequenced to date and are fragments from thymosin ß-4, eukaryotic translation initiation factor 4γ2, fibrinogen ß-chain or collagens. In conclusion, the cGvHD_MS14-pattern allows early, highly sensitive and specific prediction of cGvHD as an independent diagnostic criterion of clinical diagnosis potentially allowing early therapeutic intervention.

Stereoscopic pyrometer for char combustion characterization.

For many pulverized fuels, especially coal and biomass, char combustion is the time determining step. Based on intensified ICCD cameras, a novel setup has been developed to study pulverized fuel combustion, mainly in a laminar flow reactor. For char burning characterization, the typical measurement parameters are particle temperature, size, and velocity. The working principle of the camera setup is introduced and its capabilities are discussed by examination of coal particle combustion under CO(2)-enriched, so-called oxy-fuel atmospheres with varying O(2) content.

MiRNome and transcriptome aided pathway analysis in human regulatory T cells.

Owing to their manifold immune regulatory functions, regulatory T cells (Treg) have received tremendous interest as targets for therapeutic intervention of diverse immunological pathologies or cancer. Directed manipulation of Treg will only be achievable with extensive knowledge about the intrinsic programs that define their regulatory function. We simultaneously analyzed miR and mRNA transcript levels in resting and activated human Treg cells in comparison with non-regulatory conventional T cells (Tcon). Based on experimentally validated miR-target information, both transcript levels were integrated into a comprehensive pathway analysis. This strategy revealed characteristic signal transduction pathways involved in Treg biology such as T-cell receptor-, Toll-like receptor-, transforming growth factor-ß-, JAK/STAT (Janus kinase/signal transducers and activators of transcription)- and mammalian target of rapamycin signaling, and allowed for the prediction of specific pathway activities on the basis of miR and mRNA transcript levels in a probabilistic manner. These data encourage new concepts for targeted control of Treg cell effector functions.

MR-based coronary artery blood velocity measurements in patients without coronary artery disease.

To evaluate the feasibility of MR-based coronary blood velocity measurements (MRvenc) in patients without coronary artery disease (CAD). Eighty-three patients with angiographically excluded CAD received MRvenc of the proximal segments of both coronary arteries (CAs). Using a retrospectively ECG-gated breath-hold phase-contrast FLASH sequence with high temporal resolution, flow data were technically acquirable in 137/166 (83%) CAs. Quantification and analysis of blood velocities in systole and diastole of both CAs were performed. Biphasic velocity profiles were found in 83/100 CAs. Median systolic and diastolic velocities differed significantly in LCA (19 cm/s, 24 cm/s; P<0.0001) and RCAs (14 cm/s, 16 cm/s; P<0.01). The diastolic/systolic velocity ratio was calculated in LCAs and RCAs with a median of 1.3 and 1.1, respectively. The velocity profiles of the remaining CAs were monophasic (17 CAs) or revealed severe alterations of the physiologic velocity profile with reduced flow undulations and steady velocities (37 CAs). Optimized clinical MRvenc is feasible to quantify blood velocities in the CAs. Potential indications are (1) non-invasive monitoring of patients after aortic valve reconstruction as well as (2) detection of asymptomatic CAD patients.

MR imaging characteristics of osteoradionecrosis of the pelvis after radiation therapy on gynecological tumors.

PURPOSE: To describe MR imaging characteristics of osteoradionecrosis (ORN) of the pelvis as a result of radiation therapy (RT) on gynecological tumors. MATERIAL AND METHODS: Radiography, computed tomography (CT) and magnetic resonance imaging (MRI) were performed on 9 women (mean age 67.5 years) with gynecological tumors to identify ORN. T1- and T2-weighted sequences and contrast-enhanced t1-weighted sequences with and without fat saturation were used. The patients began developing pain after the completion of RT indicating a possible ORN a which time MRI was performed. MR images were correlated with the results of clinical examinations. RESULTS: Depending on the time elapsed after RT, ORN presented with different signal intensities. The acquired images suggested that signal changes in T2-weighted images as well as the different enhancement behaviour of ORN could be dependent on the time elapsed after RT. Visualisation of the affected regions was best achieved with fat-saturated T1-weighted sequences. CT showed increased density in the affected regions corresponding to osteosclerosis. In all cases the sacroiliac joint was affected, some times bilaterally. CONCLUSION: MRI is helpful in detecting and characterizing ORN. Changes in signal intensity, based on histopathological tissue changes could make a chronological classification possible.

[Role of radiotherapy in the treatment of multiple myeloma]. / Wertigkeit der Radiotherapie in der Behandlung des multiplen Myeloms.

BACKGROUND: Chemotherapy is the treatment of choice in multiple myeloma; but there are no curative options. Therefore, the treatment rationale is characterized by reduction of symptoms and inhibition of complications. Regarding reduction of pain, treatment of (impending) fractures, and spinal cord compression radiation is an important part of palliative treatment. In our retrospective study we report the effect of radiotherapy on reduction of pain, recalcification and the reduction of neurological symptoms and evaluate factors which have an impact on therapeutic outcome. PATIENTS AND METHODS: From 1, Jan 1988 to 31, Dec 1998, 42 patients (19 women, 23 men; range of ages 46 to 85 years, median age 64.9 years) with 71 target volumes were irradiated (median dose 36 Gy, 2 to 3 Gy 5 times/week) because of symptomatic disease (67/71: osseous pain, 45/71: fractures/impending fractures, 13/71: spinal cord compression) (Tables 1 and 2). The median time from diagnosis to the first course of radiotherapy was 11.9 months (0.3 to 90 months). At the time of first irradiation, 5 and 37 patients were in tumor Stage II and III (Salmon/Durie), respectively. The median value of the Karnofsky performance was 70% (40 to 90%). RESULTS: During follow-up (at least 6 months) in 85% of target volumes complete and partial pain relief (measured by patients' perception and the use of analgetic medication) was achieved; recurrences were seen in 8.8%. In 26/56 (46.4%) lesions evaluable a recalcification was seen whereas 17.9% showed progressive disease (comparison of radiographs before and after radiation). In 22.3% of all lesions initially with impending fracture (4/18) radiotherapy failed because of fracture after treatment (Tables 3 and 4). Simultaneous chemotherapy and a Karnofsky performance > or = 70 had a significant impact on a positive response to treatment, respectively. Spinal cord compression symptoms were reduced in 7/13 (53.8%) of patients (scaled due to the classification by Findlay 1987). The median survival from diagnosis for the entire group was 34.9 months (7.5 to 119.3 months), after irradiation 13.1 months (0.2 to 105.3 months) (Figure 1). CONCLUSION: When adequately indicated radiotherapy has shown to be an effective palliative treatment. Taking under consideration that the results are retrospective we suppose that in multiple myeloma the local response to radiation is supported by a favorable performance status and simultaneous chemotherapy. Irradiation treatment does not change prognosis regarding overall survival.

[Prognostic factors and the effect of radiotherapy in the treatment of differentiated thyroid carcinomas]. / Prognostische Faktoren und Einfluss der Strahlentherapie bei Behandlung differenzierter Schilddrüsenkarzinome.

PURPOSE: In the treatment of differentiated thyroid carcinomas the indication for adjuvant radiotherapy is discussed contradictory. The following study analyzes the long-term survival rates of patients with follicular and papillar thyroid carcinomas after percutaneous radiotherapy. PATIENTS AND METHOD: Records of 178 patients with differentiated thyroid carcinomas (132 female, median age 46 years; 46 male, median age 47 years) were evaluated. Following thyroid-resection and radioiodine therapy external beam irradiation was performed with a telecobalt device and high energy electrons (mean reference dose 54.7 Gy). Hundred and twenty patients (67.4%) had a histologically confirmed papillary carcinoma, 58 (32.6%) patients had follicular carcinoma. In the group with papillary carcinoma 57 patients (47.5%) were classified as stage I, 11 patients (9.2%) as stage II, 48 patients (40.0%) as stage III, 4 patients (3.3%) as stage IV, respectively, in the group with follicular carcinoma 21 patients (36.2%) were classified as stage I, 4 patients (6.9%) as stage II, 28 patients (48.3%) as stage III and 5 patients (8.6%) as stage IV. Survival, recurrence rate and prognostic factors were analyzed.

Deoxycholate and cholate modulate the source of cholesterol substrate for bile acid synthesis in the rat.

In the current study, the role of the supply of preformed and newly synthesized cholesterol for the feedback control of the synthesis of different bile acids and the secretion of biliary cholesterol was investigated. To define these cholesterol fluxes and the possibility of a different modulation by bile acids with different suppressive capacities, a continuous labeling with tritiated water was used in rats with an extracorporeal bile duct receiving intraduodenal infusions of taurocholate or taurocholate plus deoxycholate. After bile acid pool depletion (6 to 9 hours) total muricholate, cholate, and chenodeoxycholate synthesis was variably increased (24% to 93%) during an infusion of 304 mumol taurocholate/kg per hour. The increase in bile acid synthesis and biliary cholesterol output was predominantly due to the utilization of preformed (unlabeled) cholesterol. The addition of 52 mumol/kg per hour of deoxycholate to 258 mumol/kg per hour of taurocholate had a comparable effect. In the late period (30 to 54 hours), the taurocholate infusion had little impact on total muricholate and chenodeoxycholate synthesis but caused by a significant increase of the proportion from performed cholesterol. Both total cholate production and its synthesis from de novo (labeled) cholesterol was inhibited by 30% (P < .05) and 64% (P < .01), respectively. The secretion rate of total and de novo biliary cholesterol was higher (65% and 72%; P < .01) compared with controls. In comparison, the combined bile acid infusion led to a further increase of total muricholate synthesis (P < .05), which was again due to an enhanced synthesis from performed cholesterol (P < .001). Similar changes were observed in chenodeoxycholate. The more pronounced suppression of total cholate synthesis by 81% (P < .05) was due to a diminished cholate synthesis from both de novo cholesterol by 72% (P < .001) and preformed cholesterol by 91% (P > .05). We conclude that the modulation of the synthesis of the various primary bile acids in the rat differs and feedback regulation of cholate synthesis by taurocholate and deoxycholate is mediated by different mechanisms of control, including inhibition of cholesterol 7 alpha-hydroxylase, HMG-CoA reductase, and uptake of lipoprotein cholesterol.

Bile acid synthesis from newly synthesized vs. preformed cholesterol precursor pools in the rat.

The present study defines the origin of cholesterol subserving bile acid synthesis in male rats with an extracorporal bile duct by labeling newly formed cholesterol with tritiated water. Within 6 hr after interruption of the enterohepatic circulation, the bile acid pool was depleted. At this early time point the proportion from de novo cholesterol was 8% and 12% for biliary cholesterol and cholate, but 18% and 19% for muricholate and chenodeoxycholate, respectively. This proportion gradually rose to 40%, 34%, 51% and 51%, respectively, at 15 to 30 hr. At 78 hr after bile diversion, 64% of cholate was labeled, compared with 84% to 88% of the other biliary lipids and 71% of plasma cholesterol. Total and labeled bile acid secretion exhibited the same diurnal rhythm. To allow differentiation between direct hepatocytic de novo synthesis of bile acids from acetate and recycling of labeled plasma cholesterol, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase (pravastatin) was infused from 54 to 78 hr. It suppressed total synthesis of primary bile acids by 60% to 80% but decreased the tritium label of bile acids only from a range of 74% to 92% (54 hr) to a range of 54% to 63% (78 hr), which was in the range of plasma cholesterol (58%). We conclude that bile acids and biliary cholesterol are synthesized mostly from preformed (i.e., plasma) cholesterol, both immediately after depletion of the pool in enterohepatic circulation and after derepression. Moreover, the hepatic cholesterol pools subserving the synthesis of different bile acids and biliary cholesterol secretion are not identical.

The influence of disease activity on the number of blood cells of multiple sclerosis patients.

The influence of disease activity on blood cell counts was studied in over 100 multiple sclerosis patients examined at regular intervals of 4-8 weeks over an 18-month period. T-lymphocytes were assessed by the cytochemical alpha-naphthyl-acetate-esterase (ANAE) stain in addition to conventional differentials. In all patients as a group as well as in individual patients studied sequentially, an exacerbation was marked by a striking reduction in both the relative and absolute numbers of ANAE-positive cells. Simultaneously, a strong increase in granulocytes and a slight augmentation of ANAE-negative cells occurred. Similar changes related to disease activity in leucocytes and lymphocytes were also detected in conventional blood smears. Fluctuations in the number of blood cells with disease activity persisted under therapy with corticosteroids and to a lesser degree also with azathioprine.