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Biomedical subjects

H Kamiya

Publications and source records attributed to H Kamiya.

At least 217 records · Page 12Linked to original sources

Detection of human cytomegalovirus DNA in immunocompromised children by polymerase chain reaction.

BACKGROUND: Cytomegalovirus (CMV) infection is a major complication in immunocompromised patients, such as those with leukemia, acquired immunodeficiency syndrome (AIDS) and allograft recipients. A sensitive and specific diagnostic procedure of CMV infection is required. The polymerase chain reaction (PCR) procedure has been demonstrated to be more sensitive than conventional virus isolation. However, CMV DNA can be detected in patients with latent or asymptomatic infections by PCR because of its extreme sensitivity. OBJECTIVES: In this study, we used PCR and reverse transcription PCR (RT-PCR) to detect CMV DNA and mRNA in immunocompromised patients and evaluated the value of PCR and RT-PCR for diagnosis of active CMV infection. STUDY DESIGN: We examined thirty immunocompromised children for CMV DNA using peripheral blood mononuclear cells (PBMC), polymorphonuclear leukocytes (PMNL). urine and throat swabs from October 1991 to March 1992. Eighteen of the patients had acute lymphoblastic leukemia, four had non-Hodgkin's lymphoma, two had Wilms' tumor and six were recipients of an allogeneic bone marrow transplantation. Four patients developed CMV-related clinical symptoms during the observation period: two patients with pneumonia and two patients with hepatitis. RESULTS: CMV DNA was detected in four patients with clinical symptoms and in five patients without evidence of CMV infection. The incidence of CMV DNA was significantly higher in symptomatic patients than asymptomatic patients. CMV mRNA encoding the late antigen was detected by RT-PCR only in patients with clinical symptoms of CMV infection. CONCLUSIONS: We concluded that detection of CMV DNA by PCR is of little clinical significance for distinguishing latent form active CMV infection. It is suggested that detection of CMV mRNA by RT-PCR is useful for diagnosis of active CMV infection.

Journal Article↗

Detection of varicella-zoster virus (VZV) DNA in throat swabs and peripheral blood mononuclear cells of immunocompromised patients with herpes zoster by polymerase chain reaction.

BACKGROUND: Varicella-zoster virus (VZV) is rarely isolated from throat swabs and peripheral blood leukocytes from patients with herpes zoster by conventional virus isolation methods. The polymerase chain reaction (PCR) is a highly sensitive method to detect VZV genomes. It has been reported that VZV DNA was detected in the cerebrospinal fluid (Puchhammerstockl et al., 1991) and peripheral blood mononuclear cells (PBMC) of patients with VZV-associated neurological symptoms (Gilden et al., 1992) by PCR. OBJECTIVES: We used the nested double PCR to detect VZV DNA in patients with herpes zoster. STUDY DESIGN: Sixteen patients with herpes zoster, ten immunocompromised and six immunocompetent patients, were studied. Throat swabs and PBMC were collected weekly and examined for VZV DNA by the nested double PCR. RESULTS: VZV DNA was detected in 60% (6/10) of throat swabs and in 60% (6/10) of PBMC of immunocompromised patients, and in 16.7% (1/6) of throat swabs and in 33% (2/6) of PBMC of immunocompetent patients within two weeks after the onset of skin rash. VZV DNA was detected in throat swabs or PBMC of two patients 5 and 7 days after cessation of acyclovir. CONCLUSION: VZV DNA was detected in throat swabs and PBMC-associated viremia exist in patients with herpes zoster. It is suggested that VZV spread from sensory ganglia to the skin or pharyngeal area along the nerve fiber or hematogenously and local cutaneous replication of VZV can lead to viremia with subsequent hematogenous dissemination in patients with herpes zoster.

Journal Article↗

Markedly elevated levels of beta2-microglobulin in urine with measles viruria in patients with measles.

BACKGROUND: Measles is a highly contagious infectious disease in children. However, the pathogenesis has not yet been fully defined since susceptible cells to measles virus (MV) had not been developed. Recently, B95a cells, which are very susceptible to MV, have been reported. OBJECTIVES: To evaluate measles viruria in patients with measles, isolation of MV from urine was performed using B95a cells. STUDY DESIGN: Isolation of MV from supernates or sediments of urine in patients with measles was performed using B95a cells. The levels of beta2-microglobulin (MG) in urine and serum were also measured. RESULTS: MV was isolated from either supernates or sediments of urine. MV was isolated from 10 of 11 samples (91%) within 2 days of the appearance of a rash. MV was isolated from supernates of urine for up to 4 days after the appearance of a rash, and from sediments for up to 5 days. The levels of urinary beta2-MG were elevated within 2 days of the appearance of a rash. The levels of urinary beta2-MG with measles viruria were significantly higher compared to those without measles viruria. CONCLUSIONS: Measles viruria may occur early in all patients with measles and elevated levels of urinary beta2-MG in patients with measles may be the consequence of tubular injury.

Journal Article↗

Pertussis immunization with acellular vaccines in Ghanaian children.

In the present study, the persistence of antibodies to pertussis antigens was assessed in 51 Ghanaian children immunized with one of two acellular vaccines and one whole cell vaccine in early infancy. The effect of a booster dose 1 year after primary immunization was also examined. Antibody titres to pertussis toxin (PT) and filamentous haemagglutinin (FHA) were measured 1 month and 1 year after primary immunization and 1 month after the booster dose. Although geometric titres (GMTs) to FHA were significantly higher in the two types of acellular vaccinees in the whole cell vaccinees 1 month after primary immunization, GMTs to FHA and PT after 1 year were not significantly different in the three groups. Geometric mean titres to PT and FHA following the booster dose were significantly higher in the acellular vaccinees than in the whole cell vaccinees. Seropositivity rates to PT and FHA in the acellular vaccinees, which were more than 93.3% 1 month after primary immunization, ranged from 50.0 to 77.8% after 1 year. In conclusion, the acellular vaccines did not produce higher antibody levels than the whole cell vaccine 1 year after primary immunization. The booster dose was essential to maintaining sufficient seropositivity to pertussis antigens.

Antibodies, Bacterial↗

8-Hydroxyguanine (7,8-dihydro-8-oxoguanine) in hot spots of the c-Ha-ras gene: effects of sequence contexts on mutation spectra.

We previously reported that 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) at the second position of codon 12 of the c-Ha-ras gene induces many types of mutations in NIH3T3 cells. In this study we incorporated the modified base into the first and second positions of codon 12 in the coding strand and into the first position of codon 61 in the non-coding strand of the gene using a new 8-hydroxyguanine phosphoramidite as a building block during oligonucleotide synthesis. The ras genes with 8-hydroxyguanine were transfected into NIH3T3 cells and the mutations induced were analyzed. 8-Hydroxyguanine residues at the first positions of codons 12 and 61 induced mutations to T at the modified sites almost exclusively. On the other hand, the DNA lesion at the second position of codon 12 induced a G-->A transition in addition to a G-->T transversion, confirming our previous results. Mutations in 5'-flanking sites were observed with 8-hydroxyguanine at the second position of codon 12 or the first position of codon 61. These results indicate that 8-hydroxyguanine in mammalian cells mainly induces a G-->T transversion at the modified site, but that other types of mutations are also elicited.

3T3 Cells↗

Formation of a mutagen, glyoxal, from DNA treated with oxygen free radicals.

Oxygen free radicals cause extensive chemical changes in DNA, including base and sugar modifications and strand breaks. In the present study we found that a mutagen, glyoxal, is produced by exposure of DNA to an oxygen radical forming system (5 mM FeSO4-EDTA, 37 degrees C, 60 min). It was produced with 17 times higher efficiency than 8-hydroxydeoxyguanosine. Thus it is possible that the formation of glyoxal is one of the major types of damage in DNA exposed to oxygen free radicals.

Animals↗

Increased expression of adhesion molecules (CD54, CD29 and CD44) on fibroblasts infected with cytomegalovirus.

The expression of ICAM-1 (CD54), beta 1 integrin (CD29), and CD44 on cytomegalovirus (CMV)-infected human embryonic fibroblasts (HEF) was analyzed by flow cytometry. The expression of these adhesion molecules increased significantly on CMV-infected HEF, on days 2 and 5 after inoculation, compared to uninfected HEF. However, the expression of these adhesion molecules decreased on herpes simplex virus (HSV)-1 and varicella-zoster virus (VZV)-infected HEF. Increased expression was not observed on HEF treated either with inactivated CMV or with supernatant fluid of CMV-infected cells. The addition of anti-cytokine (TNF-alpha, IL-1 beta, or IFN-gamma) antibodies had no effect on the increase of these adhesion molecules. This suggests that the increase in CD54, CD29, and CD44 on CMV-infected cells requires active virus replication and was not mediated by a soluble factor released from CMV-infected cells. Changes in adhesion molecules on CMV-infected fibroblasts may contribute to inflammation induced by CMV infection.

Animals↗

Inhibition of varicella-zoster virus glycoprotein expression by peripheral blood mononuclear cells.

The effect of peripheral blood mononuclear cells (PBMC) on expression of varicella-zoster virus (VZV) glycoproteins (Gps) was analyzed by flow cytometry. PBMC from VZV seropositive and seronegative donors and supernatant of PBMC co-cultured with VZV-infected human embryonic fibroblasts reduced VZV Gp expression. Neutralization of supernatant fluid with mixture of anti-interferons (IFN)-alpha, -beta, -gamma, and tumor necrosis factor (TNF)-alpha partially reduced inhibitory activity of supernatant on VZV Gp expression. Deletion of natural killer (NK) cells and adherent cells from PBMC reduced inhibitory activity of PBMC on VZV Gp expression. These results suggest that IFN-alpha, -beta, -gamma, TNF-alpha and other soluble factors released from NK cells and monocytes by co-cultivation with VZV-infected fibroblasts inhibit VZV Gp expression.

Antibodies, Monoclonal↗

Comparison of incorporation and extension of nucleotides in vitro opposite 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) in hot spots of the c-Ha-ras gene.

DNA templates with 8-hydroxyguanine (7,8-dihydro-8-oxoguanine, oh8Gua) at a site corresponding to the first or second position of codon 12 of the c-Ha-ras gene were prepared, and the nucleotides inserted opposite the modified base were compared. The Klenow fragment (KF) of Escherichia coli DNA polymerase I inserted C opposite oh8Gua at both positions. Taq DNA polymerase incorporated C and A opposite oh8Gua, and the ratio of C to A was higher at the first position than at the second position. DNA polymerase alpha (pol alpha) inserted A and C at the first position, and A at the second position of codon 12, indicating that the ratio of C to A was higher at the first position. Moreover, we studied the extensions of bases paired with oh8Gua by DNA polymerases with or without 3'-5' exonuclease activity. G and T opposite oh8Gua were removed, and subsequently C was inserted by KF. We found that an oh8Gua:A pair was recognized by the exonuclease activity of the enzyme and that A was partially substituted by C. On the other hand, pol alpha extended only C and A opposite oh8Gua. No difference was observed with oh8Gua at the two positions. These results indicate that the ratio of nucleotides incorporated opposite oh8Gua depends on the sequence context, while there is no particular difference in the extension of base pairs involving oh8Gua by DNA polymerases.

Animals↗

Immunogenicity and reactogenicity of the component acellular pertussis vaccine produced by a combination of column purified pertussis toxin and filamentous haemagglutinin.

This is the report on a prospective, single blind, comparative study of a component acellular pertussis vaccine produced by a combination of detoxified, column purified pertussis toxin (PT) and filamentous hemagglutinin (FHA) combined with diphtheria and tetanus toxoids (DTcaP) and the traditional acellular pertussis vaccine produced with essentially the same method as described by Sato with DT (DTaP) of the same manufacturer. A total of 616 infants and children received DTcaP and a total of 289 received DTaP. In all age groups for both vaccines values of serum antibodies to PT and FHA after two doses of the vaccines were comparable to those of convalescent sera. Incidences of systemic and local reactions were, in general, not greatly different between DTcaP and DTaP recipients. In Japan the use of traditional acellular vaccines replaced whole cell vaccines in 1981. Protective antigens of Bordetella pertussis have now been specified and thus component vaccines have become theoretically possible. This is the first component vaccine which has been developed in Japan. Several other component vaccines are now under investigation in the world.

Antibodies, Bacterial↗

Nitric oxide is a mediator of tachykinin NK3 receptor-induced relaxation in rat mesenteric artery.

1. The mechanism of vasodilatation induced by tachykinin peptides was studied in isolated mesenteric arteries of rats. 2. Senktide, a selective NK3 agonist, elicited potent endothelium-dependent relaxation of arteries precontracted with phenylephrine (10(-5) M), but an NK1 agonist did not. 3. A non-peptide NK3 antagonist, SR 142801, inhibited senktide-induced relaxation. However, a non-peptide NK1 antagonist, CP-96,345, and a peptide-based NK2 antagonist, L-659,877, had no effect on senktide-induced relaxation. 4. N omega-nitro-L-arginine (L-NOARG), a nitric oxide synthesis inhibitor, markedly attenuated the relaxant response to senktide. 5. These results suggest that the endothelium of rat mesenteric arteries possesses tachykinin NK3 receptors, and that NK3 agonist-induced vasodilatation is mediated by release of nitric oxide (NO) from the endothelium.

Amino Acid Sequence↗

[Four cases of hemolytic uremic syndrome (HUS) associated with serotype O165 verotoxin producing Escherichia coli (VTEC) identified by LPS-solid phase enzyme-linked immunosorbent assay (ELISA)].

Enzyme-linked immunosorbent assay using LPS derived from newly recognized serotype O165 verotoxin producing Escherichia coli (VTEC) could identify 4 cases of hemolytic uremic syndrome (HUS) associated with O165 VTEC. All 4 cases showed a typical clinical course seen in VTEC-associated HUS. We screened 33 cases of HUS whose pathogen was not identified by culture of serodiagnosis. The O165 serotype was not thought to be important not only as a VTEC but also as an enteropathogenic E. coli. However, the prevalence, 4 cases, was as high as of O111 serotype, which is the second major serotype of VTEC in Japan. We have to be careful for this serotype when we look for the pathogen of the patients with hemorrhagic colitis or with HUS.

Bacterial Toxins↗

Impaired expression of interleukin 2 receptor and CD45RO antigen on lymphocytes from children with acute lymphoblastic leukemia in response to cytomegalovirus and varicella-zoster virus.

The expression of CD25 and CD45RO in lymphocytes from patients with acute lymphoblastic leukemia in response to cytomegalovirus and varicella-zoster virus antigens was analyzed by flow cytometry. It is suggested that the defects in the immune response to cytomegalovirus and varicella-zoster virus in patients with acute lymphoblastic leukemia on chemotherapy are in the expression of CD25 and in the switching of CD45RA cells to CD45RO cells.

CD4 Antigens↗

Inhibitory effect of transforming growth factor beta 1 on cytokine-enhanced eosinophil survival and degranulation.

The effects of transforming growth factor (TGF) beta 1 on cytokine-enhanced eosinophil survival and degranulation were investigated in vitro to determine whether it is an inhibitory regulator of allergic inflammation. Peripheral blood eosinophils purified by Percoll density gradient centrifugation and the CD16 negative selection technique were incubated in the presence of eosinophil-activating cytokines (interleukin-5 (IL-5), IL-3, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon (IFN)-gamma) with and without TFG-beta 1 for 1-3 days. On day 1, eosinophil protein X release was measured by radioimmunoassay. Eosinophil viability on day 3 was determined by staining the cells with fluorescein diacetate and propidium iodide, and on the same day DNA was extracted and subjected to gel electrophoresis to test for fragmentation. TGF-beta 1 significantly inhibited eosinophil survival enhanced by IL-5, IL-3, GM-CSF and IFN-gamma in a dose-dependent manner. The inhibitory effects of TGF-beta 1 on IL-5-enhanced survival was partially reversed by high concentrations of IL-5 and was completely neutralized with anti-TGF-beta antibody. IL-5 inhibited DNA fragmentation of eosinophils in vitro. TGF-beta reversed the effect of IL-5, indicating that TGF-beta 1 activates the pathway of apoptosis. TGF-beta 1 significantly suppressed eosinophil protein X release induced by IL-5. These results suggest that TGF-beta 1 may play a role in the modulation of allergic inflammation.

Apoptosis↗

Changes in 3H-quinuclidinyl benzilate binding and protein synthesis in the striatum following chronic administrations of muscarinic agonist.

Injection of the muscarinic agonist oxotremorine into rat striatum for seven consecutive days resulted in a 44% reduction in 3H-quinuclidinyl benzilate (3H-QNB) binding, and it caused decreases in binding affinity to agonists. Furthermore, it eliminated the shift from the high affinity site to the low affinity site that occurs in the presence of 5'-guanylyl imidodiphosphate (Gpp (NH)p). In addition, oxotremorine caused a 1.5-fold increase in the incorporation of 3H-leucine into the striatum, suggesting that it increased the syntheses of proteins other than the muscarinic receptor protein. The present results show that chronic treatment of the striatum with oxotremorine causes alterations in not only the quantity, but also the sensitivity of muscarinic receptors to guanine nucleotide.

Animals↗

[Studies on SY5555 in the field of pediatrics].

SY5555, a new oral penem, in the form of dry syrup (powder which is dissolved before use) was evaluated for its pharmacokinetics and clinical efficacy in pediatric patients. Oral administration of 5 mg/kg and 10 mg/kg of SY5555 in dry syrup resulted in respective maximum plasma concentrations of 1.08 +/- 0.38 micrograms/ml (n = 4) and 2.50 +/- 1.81 micrograms/ml (n = 4), half-lives (T 1/2) of 2.72 +/- 1.86 hours (n = 3) and 1.14 +/- 0.88 hours (n = 4), and urinary excretion until 6 hours of 4.7% (n = 1) and 3.86 +/- 2.01% (n = 4). Clinical efficacy was evaluable in 22 patients, and the overall efficacy rate was 100%. As for bacteriological efficacy, all 5 strains of pathogenic organisms identified were eradicated (eradication rate, 100%). No remarkable adverse reactions or abnormal laboratory values were observed.

Administration, Oral↗

Role of epidermal Langerhans' cells in the induction of protective immunity to Schistosoma mansoni in guinea-pigs.

Percutaneous exposure of guinea-pigs to attenuated or normal larvae of Schistosoma mansoni induced proliferative T-cell responses in the skin-draining lymph nodes (SLN). The responses elicited by attenuated larvae were stronger and more prolonged [2-12 days post-infection (p.i.)] than those by normal larvae (3-8 days p.i.). The former were coincident with greater and more sustained increases in numbers of SLN dendritic cells. During this event, epidermal Langerhans' cells (LC) showed marked changes in their distribution and morphology. Resident LC were similarly exhausted by either attenuated or normal larvae between 12 hr and 1 day p.i., but thereafter more blood-borne LC were recruited around the former, since reaggregation of LC around these persisted larvae was more frequent and intensive, and enhanced replenishment of epidermal LC was achieved by 8 days p.i. When the skin depleted of epidermal LC by short-wavelength ultraviolet (UVC) irradiation was exposed to attenuated larvae, consequent T-cell responses were delayed. Excision of the whole exposed skin on day 4 p.i. also reduced T-cell responses to marginal levels. These results indicate that during the afferent phase of immunity to S. mansoni, efficient T-cell responses in the SLN need an active involvement of not only resident LC but also blood-borne LC as immunostimulatory cells.

Adenosine Triphosphatases↗