Flow cytometric analysis of phenotypes of canine red blood cells with FITC-labeled Clerodendron trichotomum lectin.

Canine red blood cells were divided into a positive type (C type) and a negative type (c type) by a agglutination test with Clerodendron trichotomum lectin (CTL). Blood cells changed from c to C type after suffering from mammary tumor were named cm. The c type blood cells treated with neuraminidase were named cn.

Criteria for differentiation of normal and abnormal uroflowmetrograms in adult men.

Objective: To study parameters which serve as criteria for differentiation of normal and abnormal uroflowmetrograms.

Subjects And Methods: A total of 105 subjects were investigated, 43 of whom had no symptoms of dysuria, 57 of whom had dysuria demonstrated as an abnormal uroflowmetrogram, subjective symptoms and a small residual urine volume, and five patients who were about to undergo transurethral resection of the prostate. The three parameters used to specify the shapes of the uroflowmetrogram curves were: (i) the ratio of the maximum flow rate (Qmax) and the voiding time (T100), Qmax/T100; (ii) the ratio of the time to peak flow (TQmax) and the voiding time, TQmax/T100; and (iii) the newly introduced voiding efficiency (epsilon), measured by the ratio of the voided volume (VV) and T100 x Qmax, epsilon = VV/(T100 x Qmax), were taken for each patient.

The relationship between self-stimulation and sniffing in rats: does a common brain system mediate these behaviors?

The relationship between brain self-stimulation and brain-stimulation induced sniffing behavior was examined at three brain sites (frontal cortex, hypothalamus and lower brain stem). In the first experiment, sniffing was elicited in the prefrontal cortex and pontine reticular formation (PRF) of anesthetized rats. These sites corresponded to reported self-stimulation sites.

A new genomic polymorphism of Rh-polypeptide genes.

To determine which Rh-polypeptide genes are related to which Rh (rhesus) antigens, genomic DNAs prepared from donors of ten kinds of different Rh phenotypes were analysed by Southern hybridization method using two probes for the 5' moiety and 3' moiety of an Rh-polypeptide cDNA. A polymorphism for the Rh blood-group system was observed with BamHI, HindIII, PstI and EcoRI restriction endonucleases between genomic DNAs from the RhD-positive and RhD-negative individuals as has previously been reported. Further study showed an additional genomic polymorphism.

[A new approach for diagnosis of autoimmune hemolytic anemia].

Sixty four cases of autoimmune hemolytic anemia (AIHA) referred to our laboratory from 1985 to 1993 consisted of 51 warm type AIHA, 9 cold agglutinin disease (CAD), one paroxysmal cold hemoglobinuria, and 3 mixed type AIHA. There were 5 patients who had all clinical features of AIHA except for a positive direct-antiglobulin test (DAT). These patients were diagnosed as DAT-negative AIHA because of the elevation of red blood cell-associated IgG (RBC-IgG).

Molecular analysis of Rh polypeptides in a family with RhD-positive and RhD-negative phenotypes.

To investigate the genetic basis of the Rh polypeptide gene, we attempted the isolation of cDNA clones for Rh polypeptide from a family with the RhD-positive and RhD-negative phenotypes using the reverse transcription (RT)-PCR method for each reticulocyte RNAs followed by subcloning. The isolated cDNAs showed the existence of another Rh-related clone (RhPII-1 cDNA, tentative designation) besides the RhPI and RhPII cDNA clones reported previously by us. The RhPII-1 cDNA had a single nucleotide substitution with one amino acid substitution compared with the RhPII cDNA:substitution C-->T in nucleotide 380, changing codon 127 from GCG to GTG (Ala-->Val).

[A case report of ureteral stump metastasis from renal-cell carcinoma].

This is a case report of the 21st documented case of ureteral metastasis from renal cell carcinoma. A 75-year-old woman was admitted because of asymptomatic gross hematuria. Right radical nephrectomy for renal cell carcinoma had been performed 2 years and 9 months prior to admission.

Molecular analysis of esterase D polymorphism.

We have analyzed the esterase D (EsD) polymorphism at the nucleic acid level. Two common alleles, EsD1 and EsD2, are characterized by the substitution of one amino acid (Gly-to-Glu), which is caused by the point mutation of one nucleotide (G-to-A). Individuals exhibiting the EsD1 and EsD 2 phenotypes are homozygotes for EsD 1 and EsD 2 cDNAs, respectively.

Identification of two Rh mRNA isoforms expressed in immature erythroblasts.

Two isoforms of RhPI cDNA encoding an Rh polypeptide, RhPI-alpha and RhPI-beta, were isolated from mRNAs of immature erythroblasts cultured in a selective two-phase liquid culture system for human erythroid progenitors in peripheral blood. The RhPI-alpha and RhPI-beta cDNA clones encoded the hydrophobic membrane polypeptides of 242 and 244 amino acids which were missing in amino acid residues 227 to 383 and 212 to 384 of RhPI polypeptide, respectively. These peptide deletions produce unexpected structural properties resulting in the generation of a new open reading frame with a new stop codon caused by frameshift translations in RhPI-alpha cDNA and in the reverse orientation of topology in RhPI-beta cDNA.

An exon 28 mutation resulting in alternative splicing of the glycoprotein IIb transcript and Glanzmann's thrombasthenia.

The alternatively spliced from mRNA of platelet glycoprotein IIb (GPIIb) with a deletion of exon 28 (GPIIb-28) has been isolated from the HEL cell cDNA library. The defective expression on the surface of DNA cotransfected COS-1 cells with GPIIb-28 and GPIIIa cDNAs was described in an earlier report. We studied siblings with Glanzmann's thrombasthenia who expressed only the GPIIb-28 mRNA in their platelets.

A patient with Tn syndrome associated with myelodysplastic syndrome showed abnormal glycophorin B.

A Japanese male patient with myelodysplastic syndrome (MDS) was shown to have associated Tn syndrome; the first report of Tn syndrome with MDS. The Tn expression was demonstrated on erythrocytes, granulocytes, monocytes, platelets, and lymphocytes by flow cytometric analysis using a lectin and an antibody. Electrophoresis of erythrocyte membrane proteins revealed slower mobility of glycophorin B from the patient than that from normal individuals, suggesting a glycophorin B molecular abnormality.

Rapid detection of mitochondrial DNA polymorphism by photoactivatable biotin.

A new method for the detection of mitochondrial (mt)DNA polymorphism by agarose gel electrophoresis is presented. In this method, after mtDNA digestions with endonucleases, the mtDNA fragments are labeled directly with photoactivatable biotin in the digestion buffer, separated by agarose gel electrophoresis, transferred to a nylon membrane and detected enzymatically using streptavidin-alkaline phosphatase, Nitroblue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolylphosphate (BCIP). This method is useful for the detection of mtDNA digestion patterns by agarose gel electrophoresis, especially when enough purified mtDNA cannot be obtained and the radioisotope cannot be handled because of several restrictions.

Genetic Polymorphisms of Human Parotid Saliva and Their Application to Forensic Science.

Human saliva is one of the valuable resources for the identification of genetic markers. A number of polymorphisms are found in salivary proteins and enzymes. They could provide the important information for genetic studies and forensic science.

Point mutation in the band 4.2 gene associated with autosomal recessively inherited erythrocyte band 4.2 deficiency.

A patient who represented acute hemolytic crisis was studied. Analysis of the erythrocyte membrane proteins by SDS-PAGE revealed a deficiency of band 4.2.

Mitochondrial DNA polymorphism in Jindo dogs.

Mitochondrial DNA (mtDNA) polymorphism was studied in 21 Jindo dogs inhabiting Jin Island off the Korean peninsula. The polymorphism was analyzed with 10 restriction endonucleases that recognize six base pairs. The sizes of the mtDNA fragments produced by digestion using each endonucleases were separated by agarose gel electrophoresis, and the polymorphisms were detected with Japanese mongrel dog mtDNA as a probe.

Isolation of a new cDNA clone encoding an Rh polypeptide associated with the Rh blood group system.

The polymerase chain reaction (PCR) was used to amplify Rh-related cDNAs from erythroid cells cultured by the selective two-phase liquid culture system for human erythroid progenitors in peripheral blood. Direct sequencing based on PCR presents heterozygous bands. Two Rh polypeptide cDNAs have been isolated from the PCR products and tentatively designated RhPI cDNA and RhPII cDNA.

Two cases of urothelial tumor with high serum level of carcinoembryonic antigen and TA-4.

Two rare cases of urothelial tumor with high serum carcinoembryonic antigen (CEA) and TA-4 levels are presented. One is a case of lymph node metastasis due to bladder tumor: pathological diagnosis revealed mucoepidermoid carcinoma. The other is a case of left renal pelvic tumor: pathological diagnosis revealed transitional cell carcinoma with glandular and squamous differentiation.

Combined effect of interleukin 2 and cyclophosphamide in therapy of mice with transitional cell carcinoma.

We investigated the effect of combination therapy with interleukin 2 (IL-2) and cyclophosphamide (CPM) in C3H/HeN mice implanted with mouse bladder tumor cells (MBT2). MBT2-bearing mice were treated with a single intraperitoneal injection of 120 mg/kg CPM on day 10 and/or subcutaneous administration of 5 x 10(4) units IL-2/day from day 11 to day 20. As a result, the growth of tumor in mice treated with IL-2 alone was slightly suppressed.

Expression of the Tn antigen on erythroid cells from a patient with Tn syndrome.

In order to examine expression of the Tn antigen on erythroid cells from a patient with Tn syndrome, we applied a selective two phase liquid culture system for human erythroid progenitors in peripheral blood. The cells were analyzed with flow cytometry employing an anti-Tn antibody and a lectin of Vicia villosa which recognizes only the Tn determinant. In the second phase, the Tn antigen was expressed on the cultured cells from the patient on day 3 and Tn-positive cells reached 62.

Mutagenic activation of aflatoxin B1 by pulmonary, renal, and hepatic cytochrome P450s from rats.

The genotoxic and mutagenic activation of aflatoxin B1 (AFB1) by hepatic, renal, and pulmonary microsomes and purified cytochrome P450s was investigated in Salmonella typhimurium TA1535/pSK1002 cells in which an umu response shows DNA damage. The activity of the hepatic microsomes was greatest. Pulmonary microsomes had moderate activity and renal microsomes had low activity.

Expression of hepatic microsomal cytochrome P450s as altered by uremia.

The proportions of different hepatic microsomal cytochrome P450s expressed in uremic rats were studied with specific antibodies and with a steroid hydroxylase assay. In male uremic rats, the hepatic levels of P450 2C11, a male-specific form, and 3A2, a male-dominant form, were decreased to about 30% at 5 weeks after the induction of uremia. These changes were paralleled by decreases in the activities of testosterone 2 alpha-, 16 alpha-, and 6 beta-hydroxylation.

Further study of human salivary alpha-amylase polymorphism.

Genetic variants of salivary alpha-amylase were studied using isoelectric focusing in a pH gradient of 6-8 and silver staining methods. Five phenotypes which were tentatively named Amy1 N, Amy1 SN, Amy1 V1N, Amy1 V2SN and Amy1 V3N were detected. The phenotype frequencies in 371 unrelated Japanese were: Amy1 N = 94.

Mitochondrial DNA polymorphism in dogs.

Mitochondrial DNA (mtDNA) polymorphism was studied in 20 mongrel dogs using 14 restriction enzymes. The polymorphism was observed in the cleavage patterns of Apa I, EcoR I, EcoR V, Hinc II and Sty I. Three morphs using EcoR I and Apa I and two morphs using EcoR V, Hinc II and Sty I were found.

The effects of early social isolation on the motivation for social play in juvenile rats.

Fifteen-day-old rats were subjected to one of three housing conditions: mother-and-peer (family), peer, and isolation conditions. At 24 days of age, all subjects were rehoused individually. In Experiment 1, play behaviors were monitored in like-raised pairs.