Potent maturation of monocyte-derived dendritic cells after CD40L lentiviral gene delivery

Dendritic cells (DCs) are being evaluated in immunization protocols to enhance immunity against infectious diseases and cancer. Interaction of T-helper cells expressing CD40 ligand (CD40L) with its cognate CD40 receptor on DCs leads to a mature DC phenotype, characterized by increased capacity of antigen presentation to cytotoxic T cells. The authors examined the ability of third-generation self-inactivating lentiviral vectors expressing CD40L to induce autonomous maturation of ex vivo expanded human monocyte-derived dendritic cells. Transduction with lentiviral vectors achieved a highly efficient gene transfer of CD40L to DCs, which correlated with phenotypic maturation as shown by the expression of immunologic relevant markers (CD83, CD80, MHCI) and secretion of IL-12, whereas DC phenotype was not affected by a control vector expressing only the green fluorescent protein marker. Addition of recombinant IFN-gamma to DCs at the time of CD40L transduction further enhanced IL-12 production, and when co-cultured with allogeneic and autologous CD8+ and CD4+ T cells, a potent activation was observed. Autologous responses against an HLA-A2-restricted influenza peptide (Flu-M1) and a tumor-associated antigenic peptide (gp100 210M) were significantly enhanced when CD40L transduced DCs were used as antigen-presenting cells for in vitro stimulation of CD8+ cytotoxic T lymphocytes. These results demonstrate that endogenous expression of CD40L by lentivirally transduced DCs induced their autonomous maturation to a phenotype comparable to that induced by optimal concentrations of soluble CD40L, providing a novel tool for genetic manipulation of DCs.     

Comparing the rates of urinary tract infections among patients receiving adjuvant pelvic intensity modulated radiation therapy, 3-dimensional conformal radiation therapy,and brachytherapy for newly diagnosed endometrial cancer

PurposeThe purpose of this study was to compare the rates of urinary tract infection (UTI) among patients with endometrial cancer receiving vaginal brachytherapy alone and brachytherapy plus 3-dimensional conformal radiation therapy (3DCRT) or intensity modulated radiation therapy (IMRT).Methods and MaterialsWe retrospectively evaluated the rates of UTI among 581 patients diagnosed with endometrial cancer, treated between 2004 and 2012. A total of 37% (216/581) received brachytherapy alone, 28% (161/581) received brachytherapy plus 3DCRT, and 35% (204/581) received brachytherapy plus IMRT. UTI during the treatment was defined as evidence of pyuria detected by either urine dipstick or urinalysis. All specimens were collected as a clean catch, midstream void to avoid contamination and resultant false positives. The χ² and logistic regression analyses were subsequently employed for statistical analyses.ResultsUTI was diagnosed in 14.6% (85/581) of all patients. Only 2.8% (6/216) of patients receiving brachytherapy were diagnosed with a UTI during treatment, whereas UTI was diagnosed in 37.3% (60/161) of patients receiving brachytherapy plus 3DCRT, and 9.3% (19/204) of patients receiving brachytherapy plus IMRT (P < .0005). Logistic regression analysis found a decreased association between UTI and stage III endometrial cancer (odds ratio [OR], 0.51, 95% confidence interval [CI], 0.26, 0.99; P = .048). When compared with brachytherapy, both types of external beam radiation therapy were associated with an increased risk of UTI, though adjuvant 3DCRT (OR, 47.52, 95% CI, 14.81, 152.47; P < .001) had a more dramatic risk increase than IMRT (7.89, 95% CI, 2.26, 27.62; P = .001).ConclusionsWhen compared with IMRT, 3DCRT is associated with a significantly increased risk of UTI, supporting the use of IMRT as the less toxic external beam radiation therapy for endometrial cancer.     

A hypervariable STR polymorphism in the CFI gene: Mutation rate and no linkage disequilibrium with FGA

A hypervariable short tandem repeat (STR) polymorphism in intron 7 of the human complement factor I gene (CFI) was investigated to estimate the mutation rate in Japanese samples and to test linkage disequilibrium (LD) with an STR in the fibrinogen alpha chain gene (FGA). The expected heterozygosity and the mutation rate of CFI were estimated to be 0.917 and 0.002, respectively. No LD was observed between CFI and FGA. CFI is a useful supplementary marker for forensic science.     

The iCub humanoid robot: An open-systems platform for research in cognitive development

We describe a humanoid robot platform — the iCub — which was designed to support collaborative research in cognitive development through autonomous exploration and social interaction. The motivation for this effort is the conviction that significantly greater impact can be leveraged by adopting an open systems policy for software and hardware development. This creates the need for a robust humanoid robot that offers rich perceptuo-motor capabilities with many degrees of freedom, a cognitive capacity for learning and development, a software architecture that encourages reuse & easy integration, and a support infrastructure that fosters collaboration and sharing of resources. The iCub satisfies all of these needs in the guise of an open-system platform which is freely available and which has attracted a growing community of users and developers. To date, twenty iCubs each comprising approximately 5000 mechanical and electrical parts have been delivered to several research labs in Europe and to one in the USA.     

Decreased expression of reelin receptor VLDLR in peripheral lymphocytes of drug-naive schizophrenic patients

Reelin, a secretory protease that plays major roles in neurodevelopment and synaptic plasticity, may also play a role in the pathogenesis of schizophrenia. The present study was undertaken to examine whether the expression of two receptors for reelin, very low-density lipoprotein receptor (VLDLR) and apolipoprotein E receptor type 2 (ApoER2), were abnormal in peripheral blood lymphocytes of schizophrenic patients. In this study, we measured the mRNA levels of VLDLR and ApoER2 in blood lymphocytes from patients with schizophrenia (drug-naive patients (n=20) and medicated patients (n=20)) and age-and gender-matched healthy controls (n=40) using quantitative real-time RT-PCR. Furthermore, we examined the correlation between mRNA levels and clinical variables in patients. Levels of VLDLR mRNA in drug-naive, unmedicated patients with schizophrenia were significantly lower than those of controls. In contrast, levels of ApoER2 mRNA in drug-naive patients did not differ from those of controls, although the levels of ApoER2 mRNA in medicated patients were significantly lower than those of controls. Interestingly, levels of VLDLR mRNA in drug-naive patients showed significant increases with respect to baseline after six months of antipsychotic treatment, whereas levels of ApoER2 mRNA were significantly lower than baseline after six months of treatment. In all patients, there was a negative correlation between VLDLR mRNA levels and the severity of clinical symptoms. Our findings suggest that peripheral VLDLR mRNA levels may serve as a reliable peripheral biological marker of schizophrenia, and that the reelin-VLDLR/ApoER2 signaling pathway plays a role in the pathophysiology of schizophrenia.     

Neuroprotective Effect of Endogenous Pituitary Adenylate Cyclase-Activating Polypeptide on Spinal Cord Injury

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuroprotective peptide expressed in the central nervous system. To date, changes in the expression and effect of endogenous PACAP have not been clarified with respect to spinal cord injury (SCI). The aim of this study was to elucidate the expression pattern and function of endogenous PACAP on the contusion model of SCI using heterozygous PACAP knockout (PACAP(+/-)) and wild-type mice. Real-time polymerase chain reaction methods revealed that the level of PACAP mRNA increased gradually for 14?days after SCI and that PAC1R mRNA levels also increased for 7?days compared with intact control mice. PACAP and PAC1R immunoreactivities colabeled with a neuronal marker in the intact spinal cord. Seven days after SCI, PAC1R immunoreactivity was additionally co-expressed with an astrocyte marker. Wild-type mice gradually recovered motor function after 14?days, but PACAP(+/-) mice showed significantly impaired recovery from 3?days compared with wild-type mice. The injury volume at day?7 in PACAP(+/-) mice, and the number of single-stranded DNA-immunopositive cells as a marker of neuronal cell death at day?3 were significantly higher than values measured in wild-type mice. These data suggest that endogenous PACAP is upregulated by SCI and has a neuroprotective effect on the damaged spinal cord.