Biomedical subjects

K Sagisaka

Publications and source records attributed to K Sagisaka.

At least 73 records · Page 4Linked to original sources

PubMed1984

The effect of K+ concentration on the energy metabolism in perfused rat heart.

From experiments at various perfusion pressures in hemoglobin-free perfused rat hearts, oxygen consumption and redox shift of pyridine nucleotide were found to vary linearly with cardiac work. This relation was used for analysis of the energy metabolism associated with ion pumps. Mechanical activities such as left ventricular pressure and heart rate varied with the extracellular K+ concentration. Ion-pump dependent changes in oxygen consumption and redox state of pyridine nucleotide, estimated as the difference of the values at normal (4.7 mM) and various other extracellular K+ concentrations with corrections for the change due to mechanical work, were found to vary linearly with the K+ concentration. The slope for oxygen consumption was about 0.1 mumol/min/g X wet wt per mM K+. Lactate release changed markedly but transiently, about 1 min after changing the extracellular K+ concentration, and its amount varied linearly with the K+ concentration. In the steady state, however, lactate release was almost independent of the extracellular K+ concentration, although oxidized pyridine nucleotide increased with increasing K+ concentration. Coronary flow increased with the extracellular K+ concentration. Heart rate changed little between 1 and 12 mM K+, but decreased sharply above 12 mM K+. At 20 mM K+, heart beat was arrested and approximately 40% of myoglobin was deoxygenated. The intracellular oxygen concentration was estimated to be about 10 microM even during aerobic perfusion. Similarly, Ca2+-free arrested heart was found to be in a hypoxic state. The results showed that oxygen entry into cardiac tissue is facilitated by the cardiac cycle.

Animals↗

PubMed1984

Serological distinction of A antigen between red blood cells and saliva in blood grouping of blood and body fluid stains.

A antigens of red blood cells and body fluids such as saliva, semen and sweat could be serologically distinguished using rabbit or guinea pig immune anti-A. As for antisera specific for red blood cell A, A+ rabbits were intravenously immunized with A group red blood cells. The resulting antisera were absorbed with O and B red cells and with A. Se saliva. The absorbed anti-A reacted with A red cells (titer 1:32) and was not inhibited with A. Se saliva. Guinea pigs were intramuscularly injected with A. Se saliva. Crude antisera contained agglutinins to human red cells which were abolished by absorption with A red cells. After absorption with O. Se saliva, the antisera were proved to have agglutinin activity with A group saliva using latex coated with A. Se saliva. A antigens from blood or body fluid stains could be distinguished by the elution method with these anti-A sera.

ABO Blood-Group System↗

PubMed1984

Immunoelectrophoretic analyses of common antigens between human and monkey sera.

To examine common antigens between human and monkey sera, sera from 19 kinds of monkey were investigated with crossed and rocket immunoelectrophoresis using antisera to human serum proteins. In the crossed immunoelectrophoresis, the number of precipitation lines, 40 in human and 8 in the lower primates, increased according to the order of phylogenetic classification, and the lines due to albumin and gamma-globulin were observed in all the monkey sera examined. Quantitative investigation was performed by rocket immunoelectrophoresis with 8 kinds of mono-specific anti-serum protein. Alpha 2-Macroglobulin, orosomucoid, transferrin, IgG and IgM were not observed in monkeys of Loricidae family. Relatively large amounts of orosomucoid, albumin, IgG and transferrin were detected in monkeys of Cebidae family. It was considered that the distribution of serum proteins possessing antigens common to human is not related with the order of phylogenetic classification.

Animals↗

PubMed1984

On the new secretor status recognized by saliva specific ABH antigens.

The present authors have reported that each of ABH antigens of saliva consisted of saliva specific antigen and common antigen between saliva and red cells. Using saliva specific anti-A and B, 77 samples of A,B and AB groups could be determined as strong secretor (Se) or weak secretor (se). Frequency of Se was 62% for A group saliva, 45% for B, and 76% for A antigen and 75% for B antigen of AB group saliva, respectively. The secretor status of twenty-four O salivas could not be determined by saliva specific anti-H. Compared with the ordinary secretor system, some subjects belonging to Se in the ordinary system were typed as se in the new system, whereas all se saliva (three cases) in the ordinary system were typed as se in the new system.

ABO Blood-Group System↗

PubMed1984

Rh0(D) activity of red blood cells and stroma treated with proteolytic enzymes.

Agglutinability of red blood cells against anti-D increased remarkably when the cells were treated with proteolytic enzymes, such as bromelain, chymotrypsin, ficin, papain, pronase and trypsin. When stroma prepared from normal red blood cells was treated with any of proteolytic enzymes, however, Rh-Hr blood type activities were completely abolished. The similar results were obtained from stroma solubilized with detergents which was treated with enzymes after being prepared. Of all the enzymes, ficin acted more slowly than the others did. Neuraminidase or phospholipase A2 had no effect on Rh-Hr activities at all. SDS-polyacrylamide gel electrophoretic pattern of stroma prepared from bromelain, ficin and pronase-treated red blood cells were quite different from that of normal stroma.

Bromelains↗

PubMed1984

Preparation of anti-Lea and anti-Rh0 (D) sera by immunization with blood group substance trapped in autologous red cell ghost.

Lea substance or immune complex which was prepared with anti-D and D-active Band 3 was trapped in autologous red cell ghost. The trapped immunogens were administered intravenously to rabbits or guinea pigs. Rabbits immunized with Lea substance loading ghost produced antisera of relatively low titers, but they contained specific incomplete anti-Lea antibody (titer 1:128) after absorption procedure, which were higher than that of antisera prepared by the ordinary method. Guinea pig antiserum to the immune complex contained specific incomplete anti-D (titer 1:128). Immunoglobulin class analyses revealed that the anti-Lea consisted mainly of IgM and the anti-D of IgG. It was considered that intravenous injection of the immunogen trapped in ghost is useful for preparing hemagglutinin of sufficient titers.

Animals↗

PubMed1983

Individual difference of seminal plasma detected with crossed immunoelectrophoresis.

Seminal plasma from 120 subjects was examined by crossed immunoelectrophoresis using anti-seminal plasma. The samples were divided into two groups depending on a precipitation line which migrated fast in the first dimension electrophoresis. This precipitation line showed an incidence of 48 percent and was proved to have no acid phosphatase, esterase or lactate dehydrogenase activity and was not stained with Schiff's reagent. Analysis with column chromatography, Sephadex G-200 and Sephadex G-50, revealed that molecular weight of the protein was about 10,000 dolton. The difference was not identified with the other methods; polyacrylamide gel electrophoresis and agarose immunoelectrophoresis.

Blood Proteins↗

PubMed1983

A new method to prepare rabbit immune anti-M and -N sera using blood group substance trapped in autologous red cell ghost as an immunogen.

To prepare anti-M and -N of a sufficient potency, M or N substance trapped in autologous ghost was intravenously administered to rabbits. The potency and specificity of antisera prepared were compared with antisera prepared by other immunization methods. Rabbits immunized intravenously or intramuscularly with M or N substance alone produced no significant antibodies. Immunization with the substance loading ghost produced crude antisera of relatively low titers, but they were proved to contain specific anti-M and -N after absorption procedure which were higher than those prepared by usual intravenous injection of red cells. There was remarkable difference in immunoglobulin class between antisera prepared with the substance loading ghost and that by red cell immunizations; the former consisted of IgM and IgA, and the latter of IgG and IgA. It was considered that intravenous injection of the antigen trapped in ghost is useful for preparing hemagglutinin of sufficient titers.

Animals↗

PubMed1983

Binding sites of seminal acid phosphatase to Canavalia gladiata DC lectin.

Binding sites of seminal acid phosphatase (AcP) to Canavalia gladiata DC lectin (Can G) were examined. Complex of AcP and Can G produced in a test tube was solubilized with alpha-methyl-D-mannoside and was fractionated with Sephadex G-200. The elution pattern of AcP activity of the complex was similar to that of seminal plasma, suggesting that the reaction of AcP and Can G was reversible. Crossed immuno-affinoelectrophoresis with intermediate gel containing free Can G or immobilized one showed that AcP molecule had two or more binding sites per molecule to Can G. AcP activity of frozen sections of the prostate was inhibited by tartaric acid but not by Can G. However, Can G bound to AcP of the sections could be detected with FITC-labeled anti-Can G. These results indicate that the active site of AcP to Can G is not identical with that to the substrate.

Acid Phosphatase↗

PubMed1983

Isolation and purification of Rh0(D) antigen of human erythrocyte membrane and its serological property.

To elucidate immunochemical properties of Rh-Hr antigens, influence of various solubilizers on the activity, and isolation of the antigens were investigated. As a very sensitive method to detect Rh-Hr antigens, the agglutination inhibition test was recommended in which diluted antisera (1:400-1:1,600), papainized red cells and microtitrate plates were used. Extraction of D antigen with Brij 35 and Triton X-100 was effective, but SDS was not successful. However, D activity of SDS solubilized stroma was recovered by adding reductants. The activity was slightly demonstrated at the preparation from D negative red cells. The influence of the reductants was discussed. SDS solubilized stroma was fractionated with gel filtrations and activated thiol Sepharose 4B. The final preparation with D activity was proved to correspond with Band 3 which was not stained with PAS reagent. Deoxycholic acid solubilized stroma was fractionated with gel filtrations and affinity chromatography, resulting in the preparation corresponding to Band 3 with potent Rh-Hr blood type activities. The activities were detected without reductants and the preparation from D negative red cells had no D activity. Rh-Hr blood type activities other than D were observed in the same fraction.

Agglutination Tests↗

PubMed1983

Serological distinction of H antigen between saliva and red cell in forensic practice.

Rabbit antisera were prepared by immunization with O red cells or O.Se saliva. Anti-O red cells (anti-Hr) were diluted with O.Se saliva and each of diluents was adopted for elution method on blood or body fluid stains. Anti-Hr gave positive reactions only with red cell stains. On the other hand, anti-O.Se (anti-Hs) absorbed with O red cells kept activity reacting with latex coated with O.Se saliva. Anti-Hs reacted with various body fluid stains but not with red cell stain. It is considered that anti-Hr and -Hs are useful tools for individualization of blood or body fluid stains in forensic practice.

ABO Blood-Group System↗

PubMed1983

Immunochemical investigations of Rh0(D) activity detected in Band 3 of red cell membrane.

Stroma from D positive red cell was solubilized with 1% deoxycholic acid (DOC) and was applied onto Sepharose 4B and 6B, followed by fractionation with activated thiol Sepharose 4B. By this method Band 3 containing no carbohydrate was prepared. The Band 3 was incubated with human anti-D, then immune complex soluble in DOC solution was produced. This complex was not prepared from red cell components other than Band 3, nor from Band 3 from D negative red cell. The complex agglutinated papainized D positive red cells. Molecular weight of the complex was estimated to be 500,000 to 700,000 by the gel filtration method. The molecular weight increased according to the time of incubation. The complex gave a precipitation line in counter immunoelectrophoresis (IEP), a single peak in crossed IEP against anti-Band 3, and a line at gamma-globulin region in IEP against anti-human serum. SDS-polyacrylamide gel disc electrophoresis of the complex showed the pattern consisting of fragments of immunoglobulin G and Band 3 which was not stained with periodic acid Schiff reagent. These support our results that Rh-Hr blood type activities are located in Band 3 almost free of carbohydrate.

Chromatography, Gel↗

PubMed1983

A simple method to detect AB antigen in blood grouping of body fluid stains.

A modified mixed agglutination method is described for detecting AB antigen from body fluid stain. Saline extract of body fluid stain was added to A.Se saliva stain which was beforehand sensitized with anti-A. AB antigen reacting with the stain was detected with the usual mixed agglutination method using anti-B and B red cells.

ABO Blood-Group System↗

PubMed1982

Immunoelectrophoretic analyses of antigenic and human specific proteins of red cell membrane.

Rabbit antisera were raised for red cell stroma extracted with each of the following solutions; 5 mM phosphate buffer pH 8.0 (A antigen). 5 mM EDTA containing 5 mM 2-mercaptoethanol (B), 1/15 M phosphate buffered saline (C), 0.05% SDS (D), 50 mM Tris-HCl buffer pH 8.0 containing 1% SDS, 1 mM EDTA and 1% 2-mercaptoethanol (E) and 1% Triton X-100 (F). Crossed immunoelectrophoresis showed that E antigen produced relatively numerous precipitation lines, 7-9, against each of the antisera, suggesting that E solution was superior to the others for membrane solubilization. However, the Ouchterlony test demonstrated potent precipitin activity in anti-C, and the antiserum absorbed with monkey stroma which was extracted with F solution was proved to have strict human specificity. By adopting the antiserum to affinity chromatography, human specific antigens were isolated. The antigens gave four bands moving faster than Band 5 at SDS-polyacrylamide gel electrophoresis none of which was stained by Schiff's reagent. Using absorbed anti-C, human specificity of blood stains kept us to for two years could be identified.

Animals↗

PubMed1982

A family study of an extremely rare Rh-Hr variant (-D-/-D-) in two generations.

A rare variant (-D-/-D-) in two generations due to consanguineous marriage was reported. The propositus (T.M., 27 years of age) delivered her second baby who suffered from severe jaundice. She had agglutinin of titer 1:64 in saline and 1:1,024 in Coomb's test against red cells of her husband (CDe/cDE). Family study revealed that T.M.'s father and his two siblings were grouped as -D-/-D-. Each of the siblings (both females, 53 and 46 years of age) had three children who were delivered normally without hemolytic disease. No agglutinin concerning the Rh-Hr system was detected in the sera of the siblings.

Adult↗

PubMed1982

An esterase isozyme in non-hemoglobin proteins of cell lysate reacting with mushroom lectin.

Of 20 kinds of plant lectins examined, agglutinin activity of only mushroom lectin was inhibited with non-hemoglobin proteins of the red cell lysate. Chromatography of non-hemoglobin proteins on DEAE-cellulose and on hydroxylapatite resulted in isolating a protein which gave a single precipitation line with esterase activity in immunoelectrophoresis. SDS-polyacrylamide gel disc electrophoresis demonstrated that the protein consisted of two subunits molecular weight of which was approximately 100,000 each.

Agglutination↗

PubMed1982

An interaction between human seminal acid phosphatase and Canavalia gladiata DC lectin.

Interactions between human seminal acid phosphatase (AcP) and five kinds of lectins were studied. Seminal plasma was mixed with the lectins at various ratios. The mixtures were centrifuged and the supernatants were assayed for AcP activity. The activity was effectively reduced only with Canavalia gladiata DC (CG) lectin. CG lectin purified by Sephadex G-200 gel filtration was added to two fractions of AcP separated by Sephadex G-200 gel filtration. There was no difference in the rate of reduction of the two fractions. To isolate seminal AcP by affinity chromatography, CG lectin was coupled to agarose gel. AcP bound with the agarose gel could be eluted with alpha-methyl-D-mannoside. Specific activity of the purified AcP was elevated to as much as 54-fold.

Acid Phosphatase↗

PubMed1981

The specificity of anti-A agglutinins from immunized rabbits and plant lectins.

A+ and A- rabbits were immunized with human A red cells, saliva of A+ or A- rabbits or human A.Se. A- rabbits responded well against A.Se saliva and A cells and slightly against A+ saliva, whereas A+ rabbits produced high-titered antibody against A cells which was abolished by absorption with O cells. Agglutination inhibition tests showed that anti-A from A- rabbits immunized with A.Se or A cells was not inhibited with A+ saliva, and anti-A from rabbits immunized with A cells was not absorbed with A.Se saliva. Hydroxylapatite chromatography separated each of A.Se and A+ saliva into three fractions of which the 1st and 3rd fractions of A.Se and only the 3rd of A+ had A activity. The 3rd fraction of A.Se and A+ did not inhibit anti-A from any rabbits immunized but the 1st of A.Se inhibited anti-A from all rabbits except for A+ immunized with A cells. The 1st and 2nd fractions reacted slightly with anti-A from A- immunized with A+ and from A+ immunized with A cells. D. biflorus anti-A distinguished human A from rabbit one but Mushroom anti-A could not. These results suggested qualitative differences between human and rabbit saliva A antigens.

ABO Blood-Group System↗