CD5 deletion enhances the antitumor activity of adoptive T cell therapies.

Most patients treated with US Food and Drug Administration (FDA)-approved chimeric antigen receptor (CAR) T cells eventually experience disease progression. Furthermore, CAR T cells have not been curative against solid cancers and several hematological malignancies such as T cell lymphomas, which have very poor prognoses. One of the main barriers to the clinical success of adoptive T cell immunotherapies is CAR T cell dysfunction and lack of expansion and/or persistence after infusion. In this study, we found that CD5 inhibits CAR T cell activation and that knockout (KO) of CD5 using CRISPR-Cas9 enhances the antitumor effect of CAR T cells in multiple hematological and solid cancer models. Mechanistically, CD5 KO drives increased T cell effector function with enhanced cytotoxicity, in vivo expansion, and persistence, without apparent toxicity in preclinical models. These findings indicate that CD5 is a critical inhibitor of T cell function and a potential clinical target for enhancing T cell therapies.

Noninvasive Monitoring of Mantle Cell Lymphoma by Immunoglobulin Gene Next-Generation Sequencing in a Phase 2 Study of Sequential Chemoradioimmunotherapy Followed by Autologous Stem-Cell Rescue.

BACKGROUND: Minimal residual disease (MRD) monitoring has been used to identify early molecular relapse and predict clinical relapse in mantle cell lymphoma (MCL). Few published data exist in MCL on the performance of next-generation sequencing-based assay of immunoglobulin gene rearrangements for MRD assessment. PATIENTS AND METHODS: In a prospective clinical trial (NCT01484093) with intensive induction chemotherapy and autologous stem-cell transplantation, posttreatment peripheral blood samples were collected from 16 MCL patients and analyzed with an earlier version of the Adaptive Biotechnologies MRD assay. RESULTS: Of the 7 patients whose disease remained in remission, the MRD test remained negative in 5 (71%). Of the 9 patients who experienced relapse, the MRD test was positive at least 3 months before relapse in 6 patients (67%) and positive at the time of relapse in 1 patient (11%). All patients with at least 2 positive MRD tests experienced relapse. CONCLUSION: The next-generation sequencing-based MRD assay identified early molecular relapse, and we observed more sensitivity in the cellular (circulating leukocytes) versus acellular (plasma cell-free DNA) compartment. This observation may be due to availability of tumor target or a limitation of the assay.

Produtividade de genótipos de trigo duplo propósito submetidos ao pastejo com vacas em lactação / Productivity of dual-purpose wheat genotypes under grazing with lactating cows

Esta pesquisa foi conduzida com o objetivo de avaliar a produtividade de dois genótipos de trigo de duplo propósito, BRS Tarumã e BRS Umbu, submetidos ao pastejo com vacas em lactação. O delineamento experimental foi o inteiramente ao acaso, com dois tratamentos (genótipos), três repetições (piquetes) e medidas repetidas no tempo (pastejos). Avaliaram-se a precocidade, a composição estrutural dos trigos, as produções de forragem e de biomassa de lâminas foliares, as taxas de acúmulo diário de forragem e de lâminas foliares, a taxa de lotação, as ofertas de forragem e de lâminas foliares, a eficiência de pastejo, o consumo aparente e a produção de grãos. O trigo mais precoce para produção de forragem foi o BRS Umbu. Houve diferença para a produção de forragem (3196 vs. 4143kg MS/ha) e de lâminas foliares (2281 vs. 3205kg MS/ha) para os genótipos BRS Umbu e BRS Tarumã, respectivamente. Valores similares foram encontrados para taxa de lotação (2,26UA/ha); eficiência de pastejo (52,26%), consumo aparente (2,91%) e produção de grãos (1716kg/ha). O genótipo BRS Tarumã é o mais indicado para o manejo de duplo propósito em condições de pastejo com vacas em lactação.(AU)

The objective of this research was to evaluate the productivity of two dual-purpose wheat genotypes BRS Tarumã and BRS Umbu under grazing with lactating cows. The experimental design was completely randomized, with two treatments, three replications (paddocks) and repeated measures (grazing cycles). Studied variables were early growth, the structural composition of wheat, forage production, leaf blade biomass, the stocking rate, the herbage and leaf blade allowance, the grazing efficiency, the herbage intake and grain yield. The earliest genotype for forage production was the BRS Umbu. Differences in herbage yield between BRS Umbu and BRS Tarumã genotypes (3196 vs. 4143kg DM/ha) and leaf blade production (2281 vs. 3205kg DM/ha), respectively, were detected. Similar values between cultivars were found in stocking rate (2,26AU/ha); grazing efficiency (52,26%), herbage intake (2,91 %) and grain yield (1716kg/ha). The BRS Tarumã genotype is the most suitable for dual-purpose under grazing with dairy cows.(AU)

Valor nutritivo de pastagens de Coastcross-1 em consórcio com diferentes leguminosas de ciclo hibernal / Nutritive value of coastcross-1 pastures mixed to different cool season legumes

Objetivou-se com esta pesquisa avaliar três sistemas forrageiros constituídos por Coastcross-1 (CC) + 100kg de N/ha/ano + ervilhaca comum; CC + 100kg de N/ha/ano + trevo vesiculoso; e CC + 200kg de N/ha/ano. Durante o período experimental (345 dias), foram realizados treze pastejos. O delineamento experimental utilizado foi o inteiramente ao acaso, com três tratamentos (sistemas forrageiros), três repetições (piquetes) em parcelas subdividas no tempo (valores médios dos pastejos em cada estação do ano). Para avaliação, foram utilizadas vacas em lactação da raça Holandesa. Amostras de simulação de pastejo foram coletadas para análise de proteína bruta (PB), fibra em detergente neutro (FDN) e ácido (FDA), digestibilidade in situ da matéria seca (DISMS) e da matéria orgânica (DISMO) e os nutrientes digestíveis totais (NDT). Os valores médios para PB, FDN, FDA, DISMS, DISMO e NDT foram de 18,1; 16,7 e 17,6%; 57,8; 58,9 e 58,7%; 26,5; 26,5 e 26,7%; 79,6; 78,9 e 80,6%; 79,8; 79,1 e 80,6%; 72,1; 71,4 e 72,7%, respectivamente. Melhores resultados de valor nutritivo foram obtidos no inverno, em especial para o consórcio de Coastcross-1 com ervilhaca.

The aim of this research was to evaluate three grazing systems with Coastcross-1 (CC) + 100kg N/ha/year + common vetch; CC + 100kg N/ha/year + arrowleaf clover; and CC + 200kg N/ha/year. Thirteen grazing cycles were performed during the experimental period (345 days). The experimental design was completely randomized with three treatments (grazing systems), three replicates (paddocks) in completely split-plot time (average values of grazing season). Lactating Holstein cows were used in the evaluation. Forage mass and botanical composition were evaluated. Samples from the hand-plucking method were collected to analyze crude protein (CP), neutral detergent fiber (NDF) and acid (ADF), in situ digestibility of dry matter (ISDMD) and organic matter (ISOMD) and total digestible nutrients (TDN). The averages of CP, NDF, ADF, ISDMD, ISOMD and TDN were 18.1, 16.7 and 17.6 %; 57.8, 58.9 and 58.7 %; 26.5, 26.5 and 26.7 %; 79.6, 78.9 and 80.6 %; 79.8, 79.1 and 80.6 %; 72.1, 71.4 and 72.7 %, respectively. Better results for nutritive value were found during winter, especially on Coastcross-1 mixed with common vetch.

Interleukin-15 administration increases graft-versus-tumor activity in recipients of haploidentical hematopoietic SCT.

Utilizing a clinically relevant haploidentical (HI) murine transplant model, lethally irradiated B6D2F1 (H2K(b/d)) mice were transplanted with T cell-depleted (TCD) BM from B6CBAF1 (H2K(b/k)) mice. We found that administration of IL-15 significantly increases the numbers of CD8+ T and natural killer (NK) cells in spleen and BM after transplantion without GVHD. Graft-versus-tumor (GVT) potency of the graft was evaluated upon tumor challenge using P815 tumor cells (H2(d)). IL-15 administration without T-cell infusion did not result in any survival improvement. However, IL-15 in combination with very low-dose T-cell infusion (1 × 10(4)) significantly increased GVT activity and improved survival in recipients of HI hematopoietic SCT (HSCT). This effect was observed when IL-15 was given at a later time point, rather than immediately following transplantation. IL-15 administration also specifically increased slow-proliferative CD8+ T-cell proliferation and IFN-γ secretion in CD8+ T cells in recipients of CFSE (carboxyfluorescein succinimidyl ester)-labeled HI T-cell infusion, whereas there was no effect on CD4+ T-cell proliferation, suggesting the critical effect of IL-15 on CD8+ T-cell homeostasis in HI host. We conclude that IL-15 can be used for enhancing antileukemia effect of HI-HSCT, which requires presence of donor-derived T cells.

Haploidentical hematopoietic SCT increases graft-versus-tumor effect against renal cell carcinoma.

Allogeneic hematopoietic SCT (HSCT) has been shown to be an effective treatment option for advanced renal cell cancer (RCC). However, tumor resistance/relapse remains as the main post transplant issue. Therefore, enhancing graft-versus-tumor (GVT) activity without increasing GVHD is critical for improving the outcome of HSCT. We explored the GVT effect of haploidentical-SCT (haplo-SCT) against RCC in murine models. Lethally irradiated CB6F1 (H2K(b/d)) recipients were transplanted with T-cell-depleted BM cells from B6CBAF1 (H2K(b/k)) mice. Haplo-SCT combined with a low-dose haploidentical (HI) T-cell infusion (1 × 10(5)) successfully provided GVT activity without incurring GVHD. This effect elicited murine RCC growth control and consequently displayed a comparative survival advantage of haplo-SCT recipients when compared with MHC-matched (B6D2F1CB6F1) and parent-F1 (B6CB6F1) transplant recipients. Recipients of haplo-SCT had an increase in donor-derived splenic T-cell numbers, T-cell proliferation and IFN-γ-secreting donor-derived T-cells, a critical aspect for anti-tumor activity. The splenocytes from B6CBAF1 mice had a higher cytotoxicity against RENCA cells than the splenocytes from B6 and B6D2F1 donors after tumor challenge. These findings suggest that haplo-SCT might be an innovative immunotherapeutic platform for solid tumors, particularly for renal cell carcinoma.

A longitudinal study on effects of a six-week course for energy conservation for multiple sclerosis patients.

OBJECTIVE: Fatigue management and energy conservation are effective strategies to minimize fatigue in multiple sclerosis (MS). Sustained results have not yet been reported. METHODS: A fatigue management course was provided for 32 MS patients. They were tested prior to, directly after participation in the course and in a 7-9 month follow-up with the Fatigue Severity Scale, the MS-specific Fatigue Scale, the Modified Fatigue Impact Scale (MFIS), the Pittsburgh Sleep Quality Index and a self-rating scale for depression. The Expanded Disability Status Score (EDSS) and the MS functional composite (MSFC) were evaluated before and after participation in the course. RESULTS: The total score and the Cognitive and Physical subscores of the MFIS showed significant improvements on both points of time. Scores in the Fatigue Severity Scale, MS-specific Fatigue Scale and Psychosocial Fatigue Impact Scale did not improve significantly. MS functional composite and EDSS remained unchanged after six weeks of course participation. Subjective sleep quality improved directly after participation in the course and after 7-9 months. The depression score decreased significantly to a normal level at the end of training and in the 7-9 month follow-up. CONCLUSION: Fatigue management enables MS patients to cope with their fatigue and energy more effectively. Follow-up evaluations showed stable results after 7-9 months.

A meta-analysis for neurobehavioural effects due to electromagnetic field exposure emitted by GSM mobile phones.

BACKGROUND AND OBJECTIVE: Numerous studies have investigated the potential effects of electromagnetic fields (EMFs) emitted by GSM mobile phones ( approximately 900 MHz to approximately 1800 MHz) on cognitive functioning, but results have been equivocal. In order to try and clarify this issue, the current study carried out a meta-analysis on 19 experimental studies. DESIGN: Meta-analysis. METHODS: Nineteen studies were taken into consideration. Ten of them were included in the meta-analysis as they fulfilled several minimum requirements; for example, single-blind or double-blind experimental study design and documentation of means and standard deviation of the dependent variables. The meta-analysis compared exposed with non-exposed subjects assuming that there is a common population effect so that one single effect size could be calculated. When homogeneity for single effect sizes was not given, an own population effect for each study and a distribution of population effects was assumed. RESULTS: Attention measured by the subtraction task seems to be affected in regard to decreased reaction time. Working memory measured by the N-back test seems to be affected too: under condition 0-back target response time is lower under exposure, while under condition 2-back target response time increases. The number of errors under condition 2-back non-targets appears to be higher under exposure. CONCLUSION: Results of the meta-analysis suggest that EMFs may have a small impact on human attention and working memory.

Influence of hydrostatic pressure and sound amplitude on the ultrasound induced dispersion and de-agglomeration of nanoparticles.

In most applications, nanoparticles are required to be in a well-dispersed state prior to commercialisation. Conventional technology for dispersing particles into liquids, however, usually is not sufficient, since the nanoparticles tend to form very strong agglomerates requiring extremely high specific energy inputs in order to overcome the adhesive forces. Besides conventional systems as stirred media mills, ultrasound is one means to de-agglomerate nanoparticles in aqueous dispersions. In spite of several publications on ultrasound emulsification there is insufficient knowledge on the de-agglomeration of nanoparticulate systems in dispersions and their main parameters of influence. Aqueous suspensions of SiO2-particles were stressed up to specific energies EV of 10(4) kJ/m3 using ultrasound. Ultrasonic de-agglomeration of nanoparticles in aqueous solution is considered to be mainly a result of cavitation. Both hydrostatic pressure of the medium and the acoustic amplitude of the sound wave affect the intensity of cavitation. Furthermore, the presence of gas in the dispersion medium influences cavitation intensity and thus the effectiveness of the de-agglomeration process. In this contribution both, the influence of these parameters on the result of dispersion and the relation to the specific energy input are taken into account. For this, ultrasound experiments were carried out at different hydrostatic pressure levels (up to 10 bars) and amplitude values (64-123 microm). Depending on the optimisation target (time, energy input,...) different parameters limit the dispersion efficiency and result. All experimental results can be explained with the specific energy input that is a function of the primary input parameters of the process.

[Unilateral acral necrosis as a minor form of hand-foot syndrome. Patient with unilateral acral necrosis secondary to capecitabine therapy for metastatic breast cancer]. / Unilaterale akrale Nekrosen als Abortivform einer palmoplantaren Erythrodysästhesie. Patientin mit unilateralen akralen Nekrosen unter Capecitabine-Therapie bei metastasiertem Mammakarzinom.

Hand-foot syndrome is a common toxic side-effect, particularly during chemotherapy, with an incidence of 6-64% overall and 10-24% for capecitabine (Xeloda) monotherapy. Consequently, differential diagnostic considerations should include hand-foot syndrome when acral skin lesions appear, particularly during capecitabine monotherapy. The dosage should be reduced or therapeutic regimen changed. We present a unique patient who while on capecitabine developed unilateral acral necrosis which should be classified as a minor form of hand-foot syndrome.

Sleep spindle-related activity in the human EEG and its relation to general cognitive and learning abilities.

Stage 2 sleep spindles have been previously viewed as useful markers for the development and integrity of the CNS and were more currently linked to 'offline re-processing' of implicit as well as explicit memory traces. Additionally, it had been discussed if spindles might be related to a more general learning or cognitive ability. In the present multicentre study we examined the relationship of automatically detected slow (< 13 Hz) and fast (> 13 Hz) stage 2 sleep spindles with: (i) the Raven's Advanced Progressive Matrices (testing 'general cognitive ability'); as well as (ii) the Wechsler Memory scale-revised (evaluating memory in various subdomains). Forty-eight healthy subjects slept three times (separated by 1 week) for a whole night in a sleep laboratory with complete polysomnographic montage. Whereas the first night only served adaptation and screening purposes, the two remaining nights were preceded either by an implicit mirror-tracing or an explicit word-pair association learning or (corresponding) control task. Robust relationships of slow and fast sleep spindles with both cognitive as well as memory abilities were found irrespectively of whether learning occurred before sleep. Based on the present findings we suggest that besides being involved in shaping neuronal networks after learning, sleep spindles do reflect important aspects of efficient cortical-subcortical connectivity, and are thereby linked to cognitive- and memory-related abilities alike.

Effects of macromolecular impurities and of crystallization method on the quality of eubacterial aspartyl-tRNA synthetase crystals.

Although macromolecular purity is thought to be essential for the growth of flawless protein crystals, only a few studies have investigated how contaminants alter the crystallization process and crystal quality. Likewise, the outcome of a crystallization process may vary with the crystallization method. Here, it is reported how these two variables affect the crystallogenesis of aspartyl-tRNA synthetase from the eubacterium Thermus thermophilus. This homodimeric enzyme (Mr=130,000) possesses a multi-domain architecture and crystallizes either in a monoclinic or an orthorhombic habit. Minute amounts of protein impurities alter to a different extent the growth of each crystal form. The best synthetase crystals are only obtained when the crystallizing solution is either enclosed in capillaries or immobilized in agarose gel. In these two environments convection is reduced with regard to that existing in an unconstrained solution.

Factors influencing quality of life in multiple sclerosis patients: disability, depressive mood, fatigue and sleep quality.

OBJECTIVES: In a series of 504 patients with multiple sclerosis (MS), quality of life (QOL) and its main clinical and demographic determinants were assessed in comparison with healthy individuals. MATERIALS AND METHODS: A postal questionnaire with self-completed measures of disability (Expanded Disability Status Scale, EDSS), QOL (Quality of Life Index, QLI), depressive mood (Self-rating Depression Scale, SDS), fatigue severity (Fatigue Severity Scale, FSS) and sleep quality (Pittsburgh Sleep Quality Index, PSQI) was sent to this sample of MS patients. RESULTS: Most patients were severely disabled; almost half were mildly to severely depressed, suffering from reduced sleep quality and/or fatigue. The multiple sclerosis patients had significantly lower QLI scores than healthy controls. EDSS and SDS scores were found to be predictors of global QLI score. Regarding the different QLI domains, mean SDS scores remained predictive for all QLI items, while mean EDSS, PSQI and FSS scores were only predictive for physical domains. CONCLUSION: Our study clearly demonstrates that depressive mood is the main factor influencing QOL. The disability status, fatigue and reduced sleep quality have an impact mainly on physical domains of life quality.

From conventional crystallization to better crystals from space: a review on pilot crystallogenesis studies with aspartyl-tRNA synthetases.

Aspartyl-tRNA synthetases were the model proteins in pilot crystallogenesis experiments. They are homodimeric enzymes of Mr approximately 125 kDa that possess as substrates a transfer RNA, ATP and aspartate. They have been isolated from different sources and were crystallized either as free proteins or in association with their ligands. This review discusses their crystallisability with emphasis to crystal quality and structure determination. Crystallization in low diffusivity gelled media or in microgravity environments is highlighted. It has contributed to prepare high-resolution diffracting crystals with better internal order as reflected by their mosaicity. With AspRS from Thermus thermophilus, the better crystalline quality of the space-grown crystals within APCF is correlated with higher quality of the derived electron density maps. Usefulness for structural biology of targeted methods aimed to improve the intrinsic physical quality of protein crystals is highlighted.

Structure of tetragonal hen egg-white lysozyme at 0.94 A from crystals grown by the counter-diffusion method.

Very high quality crystals of tetragonal hen egg-white lysozyme were grown in the Advanced Protein Crystallization Facility (APCF) on board the Space Shuttle using a modified free-interface diffusion (FID) reactor designed ad hoc to have a longer diffusion path. This design allows the performance of true counter-diffusion experiments. Crystals were obtained under the classical chemical conditions defined 50 y ago with NaCl as a crystallizing agent and acetate pH 4.5 as a buffer. Counter-diffusion crystallization allows a "physical" instead of chemical optimization of growth conditions: indeed, this method screens for the best supersaturation conditions in a single trial and yields crystals of very high quality. A complete diffraction data set was collected at atomic resolution from one of these crystals using synchrotron radiation at the DESY-EMBL beamlines. The overall R(merge) on intensities in the resolution range 31-0.94 A was 5.2% and the data were 98.9% complete. Refinement was carried out with the programs CNS and SHELX97 to a final crystallographic R factor of 12.26% for 72 390 reflections. A mean standard uncertainty in the atomic positions of 0.024 A was estimated from inversion of blocked least-squares matrices. 22 side chains show alternate conformations and the loop 59-75 adopts in the same crystal packing two conformations that were observed for either triclinic or tetragonal lysozyme in previous high-resolution studies. In addition to 255 water molecules, the crystallizing agent (one hexacoordinated sodium ion and five chloride anions) participates in the ordered lysozyme hydration shell.

Growth kinetics, diffraction properties and effect of agarose on the stability of a novel crystal form of Thermus thermophilus aspartyl-tRNA synthetase-1.

Growth kinetics and diffraction properties of monoclinic crystals of eubacterial Thermus thermophilus aspartyl-tRNA synthetase-1 (AspRS-1) prepared in the presence of polyethylene glycol and agarose are studied. Their solubility and two-dimensional phase diagram are compared with those of orthorhombic crystals which grow in the presence of sodium formate or ammonium sulfate. The growth mechanism of the novel crystals was monitored by atomic force microscopy. The gel stabilizes the crystal lattice under the cryogenic conditions used for structure determination at high resolution.