Immunologic evaluation of personalized peptide vaccination for patients with advanced malignant glioma.

Purpose: The primary goal of this phase I study was to assess the safety and immunologic responses of personalized peptide vaccination for patients with advanced malignant glioma.

Experimental Design: Twenty-five patients with advanced malignant glioma (8 grade 3 and 17 grade 4 gliomas) were evaluated in a phase I clinical study of a personalized peptide vaccination. For personalized peptide vaccination, prevaccination peripheral blood mononuclear cells and plasma were provided to examine cellular and humoral responses to 25 or 23 peptides in HLA-A24+ or HLA-A2+ patients, respectively; then, only the reactive peptides (maximum of four) were used for in vivo administration.

Expression quantitative trait loci analysis of 13 genes in the rat prostate.

Differential expression of mRNA among animal strains is one of the mechanisms for their diversity. cDNA microarray analysis of the prostates of BUF/Nac (BUF) and ACI/N (ACI) rats, which show different susceptibility to prostate cancers, found 195 differentially expressed genes. To identify loci that control differential expression of 13 genes with diverse expression levels, their expression levels were measured by quantitative RT-PCR in 89 backcross rats, and expression quantitative trait locus (eQTL) analysis was performed.

Whole-genome analyses of loss of heterozygosity and methylation analysis of four tumor-suppressor genes in N-methyl-N'-nitro-N-nitrosoguanidine-induced rat stomach carcinomas.

N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced rat stomach carcinomas are considered to be a good model for differentiated-type human stomach carcinomas. However, as for their molecular basis, only infrequent mutations of Catnb (beta-catenin) and Trp53 (p53) have been observed. Here, we carried out a whole-genome analysis of loss of heterozygosity (LOH) using 21 stomach carcinomas induced by MNNG in F(1) hybrids of ACI and BUF rats, and also analyzed promoter methylation of four tumor-suppressor genes.

Use of lipoplex-induced nuclear factor-kappaB activation to enhance transgene expression by lipoplex in mouse lung.

Background: Although lipofection-induced TNF-alpha can activate nuclear factor kappaB (NF-kappaB), which, in turn, increases the transgene expression from plasmid DNA in which any NF-kappaB responsive element is incorporated, no attempts have been made to use such biological responses as NF-kappaB activation against a vector to enhance vector-mediated gene transfer.

Methods: A lipoplex composed of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium and cholesterol liposome and plasmid DNA encoding firefly luciferase under the control of the cytomegalovirus immediate early promoter (pCMV-Luc) was intravenously injected into mice. Luciferase activity as well as NF-kappaB activation in the lung were evaluated.

Sparse and wavy hair: a new model for hypoplasia of hair follicle and mammary glands on rat chromosome 17.

Mutant animals in the skin and hair have been used to identify important genes in biomedical research. We describe a new mutant rat, sparse and wavy hair (swh), that spontaneously arose in a colony of inbred WTC rats. The mutant phenotype was characterized by sparse and wavy hair, which was most prominent at age 3-4 weeks, and was inherited in an autosomal recessive manner.

Mutation at the Lmx1a locus provokes aberrant brain development in the rat.

A rat short-tail mutation with neurological defects (named queue courte, qc) was discovered. Histopathology in adult qc/qc rats revealed hypoplasia of the cerebellum and hippocampus, maldevelopment of the choroid plexus and corpus callosum. These abnormalities are strongly reminiscent of the phenotypic abnormalities found in the shaker short-tail or dreher (dr) mouse mutation at the LIM homeobox transcription factor 1 alpha locus (Lmx1a).

Cytotoxicity of a GalNAc-specific C-type lectin CEL-I toward various cell lines.

We found that CEL-I was a potent cytotoxic lectin. MDCK, HeLa, and XC cells were highly sensitive to CEL-I cytotoxicity and killed in a dose-dependent manner, whereas CHO, L929, and RAW264.7 cells were relatively resistant to CEL-I, and no significant toxicity was observed up to 10 microg/ml.

Interaction of endogenous compounds in human and rat urine with P-glycoprotein.

The effect of hydrophobic extracts of human and rat urine on in-vitro P-glycoprotein (P-gp) function was examined, in terms of intra-, inter-individual variations, and physiological states. Six urine samples out of 7, obtained from one male subject on different days, suppressed P-gp function with different potencies. Similarly, 11 samples out of 15 different individuals (8 males and 7 females) inhibited P-gp function.

Physiological and morphological identification of photosensitive neurons in the opisthosomal ganglia of Limulus polyphemus.

The motor outputs of the isolated opisthosomal ventral nerve cord in Limulus polyphemus are modulated by light. We have identified the photosensitive neurons and examined their physiological and morphological properties using intracellular recording and staining techniques. We found that photosensitive neurons are present in each ganglion of the opisthosomal ventral nerve cord.

Rat Phenome Project: the untapped potential of existing rat strains.

The National Bio Resource Project for the Rat in Japan collects, preserves, and distributes rat strains. More than 250 inbred strains have been deposited thus far into the National Bio Resource Project for the Rat and are maintained as specific pathogen-free rats or cryopreserved embryos. We are now comprehensively characterizing deposited strains as part of the Rat Phenome Project to reevaluate their value as models of human diseases.

Immunohistochemical and morphometrical studies on myelin breakdown in the demyelination (dmy) mutant rat.

The demyelination (dmy) rat is a unique mutant exhibiting severe myelin breakdown in the central nervous system (CNS). In this study, we conducted immunohistochemical and morphometrical investigations in the dmy rat. From around 6 weeks of age, the affected rats developed ataxia especially in the hindlimbs.

Photosensitivity of the central nervous system of Limulus polyphemus.

The photosensitivity of the central nervous system (CNS) of the horseshoe crab, Limulus polyphemus, was investigated by analyzing changes in motor nerve activity in the segmental nerves of prosomal and opisthosomal ganglia. Spontaneous efferent impulses were recorded in the dark from all the investigated segmental nerves. Impulse trains from the 7th dorsal nerve in the prosomal CNS were inhibited in response to illumination of the whole CNS.

The rat Ruby ( R) locus is Rab38: identical mutations in Fawn-hooded and Tester-Moriyama rats derived from an ancestral Long Evans rat sub-strain.

Hermansky-Pudlak syndrome (HPS) is a group of rare, recessive disorders in which oculocutaneous albinism, progressive pulmonary fibrosis, bleeding diathesis, and other abnormalities result from defective biogenesis of multiple cytoplasmic organelles. Seven different HPS genes are known in humans; in mouse, at least 16 loci are associated with HPS-like mutant phenotypes. In the rat, only two HPS models are known, Fawn-hooded (FH) and Tester Moriyama (TM), non-complementing strains in which HPS-like hypopigmentation and platelet storage pool deficiency result from a mutation of the Ruby (red eyed dilution; R) locus on Chromosome (Chr) 1.

Selected single blastocyst transfers maintained pregnancy outcome and eliminated multiple pregnancies.

Transfer of more than one embryo following fertilization/intracytoplasmic sperm injection cycles have increased pregnancy rate at the cost of increasing the incidence of triplets and twins. It has been proposed that prolonged culture to the blastocyst stage would automatically result in the selection of good quality embryos for transfer and minimize the incidence of triplets and twins. The objectives of the present retrospective analysis were to examine the pregnancy outcome, multiple pregnancy and related data following: (i) single blastocyst transfer (BT) and double BT; (ii) single BT in patients belonging to different age groups; and (iii) good, fair or poor quality of BT.

Rat neurological mutations cerebellar vermis defect and hobble are caused by mutations in the netrin-1 receptor gene Unc5h3.

Rats homozygous for the spontaneous cerebellar vermis defect mutation (cvd) or hobble mutation (hob) exhibit cerebellar and midbrain defects, possibly as a result of abnormal neuronal migration. Both mutant rats demonstrate laminar structure abnormalities in the fused cerebellar hemispheres and ectopic cerebellar tissues in the cerebello-pontine junction. Previous genetic studies showed that cvd and hob were allelic and suggested that Unc5h3, the causative gene of the mouse rostral cerebellar malformation (rcm) mutation, was a strong candidate for both cvd and hob.

PCR-based genotyping of the rat Atrn(mv) mutation.

Rat myelin vacuolation mutation at the Attractin locus (Atrn(mv)) is a genomic deletion including the whole exon 1 of the Atrn gene. The precise size and location of the deleted region has not yet been identified because of poor information on genomic organization of the rat Atrn gene. Here, we identified the breakpoints of the Atrn(mv) mutation, using a draft sequence of the rat genome.

Global expression analysis of N-methyl-N'-nitro-N-nitrosoguanidine-induced rat stomach carcinomas using oligonucleotide microarrays.

Rat stomach carcinomas induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are widely used as a model for differentiated-type human stomach carcinomas. Here, we analyzed expression profiles in five MNNG-induced rat stomach carcinomas by the high-density oligonucleotide microarray containing approximately 8000 probe sets. 244 and 208 genes were up- and down-regulated, respectively, by 3-fold and over in four or five carcinomas.

Therapeutic effect of intravenous interferon gene delivery with naked plasmid DNA in murine metastasis models.

Interferons (IFNs) exhibit pleiotropic biological activities that are considered to play important roles in tumor suppression and rejection. Therefore, IFN genes are promising for in vivo cytokine gene therapy targeted against tumors. In the present study, we evaluated the efficacy of hydrodynamics-based IFN gene transfer for tumor treatment, in which the naked pDNA encoding IFN-beta or IFN-gamma was administered into the tail vein of mice following portal vein (liver metastasis), tail vein (lung metastasis), or subcutaneous injection (subcutaneous tumor) of mouse colon carcinoma CT-26 cells.

Cloning of the 5' upstream region of the rat p16 gene and its role in silencing.

Hypermethylation of the 5' upstream region (5' region) of the human p16(CDKN2A) (p16) gene is known to cause silencing, which is involved in a wide range of human cancers. For the rat p16 gene, its 5' region has not been cloned, and it is uncertain whether surrogate use of exon 1 alpha is adequate for analysis of p16 silencing. In this study, we observed that methylation analysis of exon 1 alpha gave false positive results in three samples of normal rat mammary epithelia and in two of six primary mammary carcinomas.

The myelin vacuolation (mv) rat with a null mutation in the attractin gene.

We recently found a spontaneous tremor mutant in an outbred colony of Sprague-Dawley rats. The tremor behavior was exhibited from around 3 weeks of age and inherited as an autosomal recessive trait. The mutant rats had variously sized vacuoles in the neuropil and white matter throughout the central nervous system, especially in the brain stem, cerebellum, and spinal cord.

Etiology-specific gene expression profiles in rat mammary carcinomas.

Identification of etiology of human cancers is important for effective cancer prevention, and attempts to estimate the roles of a variety of environmental carcinogens in human cancers are being made. Here, we applied cDNA microarray technology to estimate whether gene expression profiles of cancers would reflect their etiology. Using rat mammary carcinoma models, expression profiles were analyzed in two groups of carcinomas induced by distinct carcinogens but with the same histological classification.

Insertional mutation of the Attractin gene in the black tremor hamster.

The hamster black tremor (bt) mutation induces a black coat color and a defective myelination in the central nervous system (CNS) that manifests as a tremor. On the other hand, loss-of-function mutations of the Attractin (Atrn) gene, such as Atrnmg, Atrnmg-L, and Atrnmg-3J in mice, and Atrnzi in rats, induce both darkening of coat color and hypomyelination and vacuolation in the CNS. The close resemblance of the mutant phenotypes led us to postulate that the bt/bt hamster also might harbor a mutation in Atrn.

Urinary bile alcohol profiles in healthy and cholestatic children.

Background: Bile alcohols are normal constituents of urine.

Methods: To better understand bile alcohol profile in childhood, urinary specimens from 41 healthy children and 10 children with cholestasis, and 3 healthy adults, were analyzed by GLC and GC-MS.

Results: Five bile alcohols, 27-nor-5beta-cholestane-3alpha,7alpha,12alpha,24S,25R-pentol, 5beta-cholestane-3alpha,7alpha,12alpha,24S, 25-pentol, 5beta-cholestane-3alpha,7alpha,12alpha,24S,26-pentol, 5beta-cholestane-3alpha,7alpha, 12alpha,25,26-pentol, and 5beta-cholestane-3alpha,7alpha,12alpha,26,27-pentol were identified in all specimens.

Hepatic uptake and gene expression mechanisms following intravenous administration of plasmid DNA by conventional and hydrodynamics-based procedures.

Hepatic uptake and gene expression mechanisms following intravenous administration of naked plasmid DNA (pDNA) by conventional and hydrodynamics-based procedures were studied in mice. After conventional (normal) intravenous injection, (32)P-labeled pDNA was rapidly eliminated from the circulation and predominantly taken up by the liver nonparenchymal cells while no significant gene expression was observed in this organ. The hepatic uptake process was saturable.