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Biomedical subjects

T Kitagawa

Publications and source records attributed to T Kitagawa.

At least 433 records · Page 24Linked to original sources
PubMed1989

Analysis of urinary and circulating FDP subfragments and subunits: application of the western-blot technique.

We applied the Western-blot technique for qualifying fibrin/fibrinogen degradation products (FDP) subfragments and subunits. With this technique we examined urine or serum samples from patients with glomerulonephritis and disseminated intravascular coagulation (DIC), in order to observe how primary or secondary fibrinolysis functioned under these pathological conditions. We also tested the antigenic recognition of several FDP antibodies with this technique. The results obtained were as follows: 1) FDP in serum samples from patients with DIC consisted of both fibrinogenolytic and fibrinolytic products; 2) Fibrinolysis was predominant in the serum from one patient with Henoch-Schönlein purpura nephritis, and its level increased after fibrinolytic therapy with urokinase; 3) Fibrinogen and fibrin polymers were always the major components of urinary FDP, although fibrinolytic products such as subfragment D-D dimer were detected in patients with glomerular disease and increased in acute exacerbation; 4) Anti-FDP D-D dimer monoclonal antibody (DD 3B6), which is usually considered to react specifically with cross-linked fibrin derivatives, also seemed to have cross-reactivity with non-crosslinked fibrin derivatives.

Blotting, Western↗
PubMed1989

Analyses of specific and total antibody responses of rabbits to four kinds of immunogens.

As a basic study to investigate suitable conditions to immunize rabbits with drug-immunogens, two highly sensitive and accurate enzyme immunoassays (EIAs) for specific antibody to viomycin (VM) and blasticidin S (BLS) were developed using the corresponding standard antibody, the solid-phase antigens, and enzyme-labeled goat anti-rabbit immunoglobulin G (IgG) antibody as immunological reagents. The accuracy of the assay results with these newly developed EIAs was demonstrated. The new EIAs as well as two previously developed EIAs, EIA for antibody specific to neocarzinostatin (NCS) and a sandwich EIA for rabbit IgG, were applied for analyses of the changes in contents of total and specific antibodies in rabbit antisera samples collected during immunizations with four antigens. Total IgG levels increased from 7.0-9.9 mg/ml to 30-50 mg/ml in all rabbits immunized under the same immunizing schedule, despite the use of four kinds of antigens. The highest level of specific antibodies, anti-BLS, anti-VM and anti-NCS, was 0.5 mg/ml in each case.

Animals↗
PubMed1989

Comparative study of complete and incomplete Freund's adjuvants for immunization of a drug immunogen using enzyme immunoassay methods.

Highly sensitive and accurate enzyme immunoassays (EIAs), a sandwich EIA for mouse immunoglobulin G (IgG) and an enzyme linked immunosorbent assay for mouse antibody specific to viomycin (VM), were developed. Accuracy and specificity of the assay results were confirmed before their application. The changes of total IgG and antibody specific to VM in mice, immunized with a VM-immunogen with or without two types of Freund's adjuvants under various conditions, were assessed by means of the newly developed EIA methods. Both methods were very useful tools to follow the immunization processes of mice, and complete and incomplete Freund's adjuvant were found to have similar adjuvant activities for production of antibody specific to VM, judging from the amounts of anti-VM antibody formed. It seems to be important that too many booster injections should be avoided in the immunization of mice with a hapten immunogen.

Animals↗
PubMed1989

Studies on the optimal immunization schedule of the mouse as an experimental animal. The effect of antigen dose and adjuvant type.

This study was undertaken to establish the optimal immunogen dose for immunization of mice, using a viomycin-protein conjugate as a hapten immunogen. It was found that specific immunoglobulin G (IgG) formation depends on both the dose of antigen and the type of adjuvant: the optimal antigen dose for an immune response is quite different depending on whether the mice are being treated with Freund's complete adjuvant (FCA) or Freund's incomplete adjuvant (FICA). The total IgG amount depends mainly upon the type of adjuvant used. FCA gave the double the level of IgG compared to that obtained with FICA. The antigen dose was found to have little influence on the total production of IgG. Mice given a primary immunization with 10 micrograms of antigen emulsified in FCA and then given a booster with the same amount of antigen emulsified in FICA produced a strikingly high level of specific anti-viomycin antibody of over 2.5 mg/ml of the antiserum. It was also found that decreases in the weight of the mice were related to the kind of adjuvant used as well as to the level of the specific antibody formed.

Adjuvants, Immunologic↗
PubMed1989

Protein binding of quinolonecarboxylic acids. I. Cinoxacin, nalidixic acid and pipemidic acid.

Binding of cinoxain (CINX), nalidixic acid (NA) and pipemidic acid (PPA) to serum proteins was investigated by equilibrium dialysis, ultrafiltration and circular dichroism (CD) spectroscopy. CINX and NA were found to bind mainly to albumin in human serum, the latter interacting with the protein about ten times as strongly as CINX at pH 7.4 and 37 degrees C. PPA showed little or no significant binding to human serum albumin (HSA), alpha 1-acid glycoprotein, and globulins, but showed 20-30% binding to protein in human serum. The CD results were suggestive of some weak interaction of PPA with human apotransferrin. Binding of the three drugs to HSA was found to depend on the lipophilicity of their substituents at the 7-position. The degree of protein binding for human, dog and rat sera at 37 degrees C was in the order of NA (92-97%) greater than CINX (68-90%) greater than PPA (20-30%) at drug concentrations of 10-30 micrograms/ml. CINX showed relatively large species dependence in serum protein binding, which seemed to be due to different affinities of this drug to the respective albumins. CINX was found to bind to rat serum albumin as strongly as NA.

Animals↗
PubMed1989

Protein binding of quinolonecarboxylic acids. II. Spectral changes on the interaction of cinoxacin, nalidixic acid and pipemidic acid with human and rat albumins.

The interaction of cinoxacin (CINX), nalidixic acid (NA), and pipemidic acid (PPA) with human and rat serum albumins (HSA and RSA) was studied by UV difference absorption and circular dichroism (CD) spectroscopy. CINX and NA bound to the albumins and generated difference absorption and induced CD (ICD) spectra. The difference absorption spectral data explained reasonably our previous observations that CINX bound to HSA more weakly than NA, but to RSA as strongly as NA. We used a quantity delta epsilon/epsilon, designated as relative molar difference absorbance, at positions corresponding to the longest wavelength peaks in the difference spectra. The quantity was found to correlate linearly with percent bound to both HSA and RSA, but with different slopes, from which the binding site for CINX and NA in RSA was supposed to provide a much more nonpolar environment than that in HSA. The magnitude of ICD bands observed at 371 nm for CINX and at 342-348 nm for NA corresponded to the binding degrees of these drugs to both albumins. Anisotropy factors for the ICD bands at 350-271 nm for CINX and 320-348 nm for NA were approximately similar between HSA and RSA, suggesting a similar ability to generate the ICD spectra in these wavelength regions upon binding to the albumins. Spectral results for PPA in albumin solutions showed little or no binding of this drug to HSA and RSA. PPA existed as a betaine form in neutral solution and its positively charged group acted as an unfavorable factor for binding to both albumins.

Animals↗
PubMed1989

A supernumerary valvula in the pulmonary semilunar valve.

A supernumerary valvula of the pulmonary semilunar valve was found in a 51-year old, male Japanese. The valve was composed of three equal-sized valvulae and one smaller one with numerous fenestrations present. The supernumerary valvula was located between the anterior and right valvulae. We found this condition in one of 1407 cadavers examined.

Humans↗
PubMed1989

[Liver disorder owing to estrogen therapy in prostatic cancer, examined histopathologically in six autopsy cases].

The parenchymal damage of the liver after estrogen therapy for prostatic cancer, mainly treated with diethylstilbestrol diphosphate (DES-DP), was studied in the six autopsied cases, herein. The parenchymal disorder of the liver was "nonalcoholic steatohepatitis", reported by Ludwig et al., and its degree of disorder was dependent upon the administered dose of estrogen. The acceptable total dose of DES-DP was supposed to be about 150 g at maximum, according to the various degrees of damage examined histopathologically in the six cases who were administered at total doses of DES-DP from 12.6 g to 619 g. Comparison of the histopathologic damage to the liver function tests performed within 10 days before death revealed that only the serum levels of cholinesterase (ChE) were abnormally decreased, suggesting its importance to predict the degree of "nonalcoholic steatohepatitis" by monitoring of ChE.

Aged↗
PubMed1989

Distinct antitumor mechanisms of recombinant interleukin-2 on recombinant interleukin-2-activated killer-sensitive and -resistant murine tumors.

The antitumor mechanism of recombinant human interleukin-2 (rIL-2) was studied using two murine tumor systems. Meth 8 tumor cells were easily lysed in vitro by rIL-2-activated killer (AK) cells, which mainly consisted of Thy1.2+, Lyt2.2+, L3T4- T cells, and asialo GM1+ natural killer (NK) cells; on the other hand, X5563 tumor cells were only slightly lysed in vitro by AK cells under the same conditions. One of these two tumors was inoculated i.d. into C3H/HeN mice and then rIL-2 (5 X 10(4) J.U./mouse/day) was repeatedly injected s.c. For AK-sensitive Meth 8-bearing mice, rIL-2 therapy starting 1 day after tumor inoculation was more effective for the growth than the therapy starting 7 days later and the therapeutic effect was abrogated by in vivo treatment with anti-asialo-GM1 serum. In contrast, for mice bearing AK-resistant X5563 tumor cells, delayed administration starting on day 7 or later was more beneficial than earlier administration on day 1 or 4. This treatment schedule resulted in complete tumor regression in a dose-dependent manner including significant inhibition of metastases in the spleen and/or lymph nodes. These therapeutic effects of rIL-2 on X5563 were not seen in T-depleted mice with anti-mouse thymocyte serum but were found in NK-depleted mice upon treatment with anti-asialo-GM1 serum. The results of these studies showed that the growth of AK-sensitive Meth 8 tumor was inhibited by AK cells, while the growth and metastases of AK-resistant X5563 tumor was inhibited by tumor-specific T cells, which were generated after tumor development and activated by rIL-2 therapy, rather than AK cells.

Animals↗
PubMed1989

[Determination of the blood pressure level in mild hypertension. Significance of 20 minute resting blood pressure measurement comparing to 24 hour ambulatory blood pressure monitoring].

Determination of the blood pressure (BP) level in patients with mild hypertension (MHT) is quite difficult, since ulcerations of BP are tremendously exaggerated in the doctor's office. It has been well known that casual BP is less reliable to estimate LVH than BPs obtained at home or work-site. Although 24 hour ambulatory BP monitoring (ABPM) has been widely accepted to overcome this problems, it is still controversial whether this method is applicable to all hypertensive subjects with special regards to its cost and effect. Therefore, our study has dealt mainly with the development of more convenient and less expensive method to get reasonable BPs. Twenty two nonmedicated patients with MHT were selected for the study. After taking casual BP in the office, the resting 20 minute BP measurements at every 2 minute interval were performed with Dynamap 950. Ten BP values thus obtained were divided into two categorical phases; early and late. The mean systolic and diastolic pressures (Ps & Pd) in the early phase were significantly higher than those in the late phase. Beside mean Ps and Pd obtained from 24 hour ABPM, 4 categorical phases based on the time of a day were defined; morning (from awaking to noon), afternoon (from noon to 6 pm), evening (from 6 pm to bed time) and night (during sleeping). Mutual correlation coefficients of these categorical BPs were calculated and compared to identify reasonably high significant correlations. The results revealed the highest BP at the office and the lowest one during sleeping. The office BPs closely resembled to the ones during afternoon period.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

[Clinicopathological correlation of young onset chronic glomerulonephritis].

In a retrospective analytical study involving 98 children with primary glomerulonephritis who were seen by us at our hospital during a 2-year period from 1984 through 1985 and who had renal biopsy performed previously, attempts were made to correlate pathological findings with both clinical findings and prognosis. The results are summarized as follows: 1) Of 87 patients with asymptomatic chronic glomerulonephritis, glomerular findings were those of minimal change lesion, mesangial proliferative nephritis, MPGN, membranous nephropathy and FGS or sclerosing nephritis in 29.9%, 51.7%, 13.8%, 1.1% and 3.5%, respectively. Among the other 11 patients in whom the diagnosis was made after manifesting the nephritic symptoms, minimal change was noted less frequently and MPGN was detected more frequently than in the aforementioned asymptomatic group. IgA nephropathy was estimated to account for 44.2% of cases of asymptomatic chronic nephritis. 2) Mild mesangial proliferation was observed relatively frequently and severe mesangial proliferation or MPGN rather infrequently in hematuria cases without proteinuria while in those with severe proteinuria minimal change lesion was uncommon and severe mesangial proliferative changes, MPGN or FGS were relatively frequent. 3) In 22 patients with IgA nephropathy and 11 with non-IgA nephritis the severity of glomerular changes was related to the intensity of proteinuria at the time of renal biopsy. 4) A 3 to 5 years' follow-up study of patients with mesangial proliferative nephritis inclusive of IgA nephropathy disclosed that 26-28% of patients became free from urinary abnormalities, 27-37% had persistent hematuria without proteinuria and 24-32% still had proteinuria of 2 plus or above. Patients with milder glomerular changes had a definitely better prognosis than those with severe glomerular lesions.

Adolescent↗
PubMed1988

Generalized lymph node metastasis of early uterine cancer in an HTLV-I carrier.

Generalized lymphadenopathy due to metastases of keratinizing squamous cell carcinoma developed in a 68-year-old woman who was a carrier of human T-cell leukemia Type I (HTLV-I). On her 74th hospital day, she died of massive metastases of the superficial and deep-seated lymph nodes, thyroid, lungs, pleura, liver, spleen, pancreas, kidneys, and retroperitoneum. At autopsy, the primary tumor was found in the uterine cervix. The depth of stromal invasion was approximately 4.0 mm. Such an extensive dissemination usually does not occur in cervical cancer with this type of early stromal invasion. It is conceivable that the chronic HTLV-I infection compromised the immunosurveillance against cancer and accelerated progression of the disease in this patient.

Carrier State↗
PubMed1988

Enzyme-linked immunosorbent assay for the quantification of actinomycin D using beta-D-galactosidase as a label.

An enzyme-linked immunosorbent assay (ELISA) for actinomycin D (AMD) has been developed, which allowed us to measure accurately as little as 50 pg of AMD per assay well. Anti-AMD sera was obtained by immunizing rabbits with an AMD derivative, 7-aminoactinomycin D (7AMD), conjugated with mercaptosuccinyl bovine serum albumin via N-maleoylaminobutyric acid chloride as a coupling agent. An enzyme marker was similarly prepared by coupling 7AMD with beta-D-galactosidase (EC 3.2.1.23) via N-maleoylaminobutyric acid. The ELISA with anti-AMD immunoglobulin G fraction as a solid phase and 7-aminoactino-mycin beta-D-galactosidase conjugate was specific to AMD as well as 7 AMD and showed 30% cross-reaction with actinomycin V, while no cross-reactivity was seen with drugs commonly used with AMD in combination chemotherapy for cancer treatment. The sensitivity of the ELISA was about 1000 times higher than high performance liquid chromatography in detecting AMD in lower concentrations. Using this assay, drug levels were easily measured in the blood and urine of rats following administration of AMD in a single dose of 0.25 mg/kg i.v. These results indicate that the ELISA provides a nonradioactive, inexpensive, sensitive, and rapid method applicable for pharmacological analyses of the drug.

Animals↗
PubMed1988

Sandwich enzyme immunoassay of tumor-associated antigen sialosylated Lewisx using beta-D-galactosidase coupled to a monoclonal antibody of IgM isotype.

Enzyme conjugates with antibody of IgG type have been used extensively in immunohistochemistry, but conjugates with antibody of IgM type have not been reported. This paper describes the beta-D-galactosidase (Gal) labeling of a monoclonal IgM antibody designated CSLEX1 (for cytotoxic sialosylated Lewisx), which is directed against a tumor-associated antigen sialosylated Lewisx (S-Lex). The antibody was first acylated with a heterobifunctional agent N-(gamma-maleimidobutyryloxy)succinimide (GMBS) to introduce the maleimide groups into the molecule; excess reagent was removed by gel filtration and then the activated antibodies were crosslinked to the thiol groups of Gal. The conjugates were partially purified of free Gal by DEAE-Toyopearl column chromatography with an increasing linear concentration of NaCl. The conjugates thus prepared retained almost full enzyme activity and were demonstrated to be free of CSLEX1 by affinity chromatography using anti-galactosidase antibody bound to Sepharose 4B. The conjugates were used as a label in a sandwich enzyme immunoassay (SEIA) to detect the antigen at concentrations as low as 0.2 U/well. The SEIA was used to measure serum S-Lex levels in both healthy subjects and lung cancer patients and mean concentrations of 70 U/ml and 198.6 U/ml were detected respectively.

Animals↗