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Biomedical subjects

H Bando

Publications and source records attributed to H Bando.

At least 109 records · Page 6Linked to original sources

Characterizations of the human parainfluenza type 2 virus gene encoding the L protein and the intergenic sequences.

We cloned and determined the nucleotide sequences of cDNAs against genomic RNA encoding the L protein of human parainfluenza type 2 virus (PIV-2). The L gene is 6904 nucleotides long including the intergenic region at the HN-L junction and putative negative strand leader RNA, almost all of which is complementary to the positive strand leader RNA of PIV-2. The deduced L protein contains 2262 amino acids with a calculated molecular weight of 256,366. The L protein of PIV-2 shows 39.9, 28.9, 27.8 and 28.3% homologies with Newcastle disease virus (NDV), Sendai virus (SV), parainfluenza type 3 virus (PIV-3) and measles virus (MV), respectively. Although sequence data on other components of transcriptive complex, NP and P, suggested a closer relationship between PIV-2 and MV, as concerns the L protein, MV is closely related to another group as SV and PIV-3. From analysis of the alignment of the five l proteins, six blocks composed of conserved amino acids were found in the L proteins. The L protein of PIV-2 was detected in purified virions and virus-infected cells using antiserum directed against an oligopeptide corresponding to the amino terminal region. Primer extension analyses showed that the intergenic regions at the NP-P, P-M, M-F, F-HN and HN-L junctions are 4, 45, 28, 8 and 42 nucleotides long, respectively, indicating that the intergenic regions exhibit no conservation of length and sequence. Furthermore, the starting and ending sequences of paramyxoviruses were summarized.

Amino Acid Sequence↗

Hypophysiotrophic TRH-producing neurons identified by combining immunohistochemistry for pro-TRH and retrograde tracing.

To determine hypophysiotrophic thyrotropin-releasing hormone (TRH)-producing neurons in the rat hypothalamus, we employed a combination of the immunohistochemistry for TRH prohormone (pro-TRH) and the retrograde tracing of neurons that project to the median eminence (ME) by injecting biotinylated wheat germ agglutinin (WGA) into the ME. In intact rats, immunoreactive pro-TRH-positive neurons occurred in the parvicellular paraventricular nucleus (parvi-PVN), basal part of the anterior and lateral hypothalamus, perifornical area and dorsomedial nucleus, especially accumulating in the parvi-PVN. Twenty-four hours after injection of the WGA into the middle portion of the ME, we found neurons that incorporated the lectin in the anterior periventricular area, the PVN, and the arcuate nucleus. When we examined serial sections consecutively stained with anti-WGA, anti-pro-TRH, and anti-WGA, most of the pro-TRH-labeled neurons in the medial parvi-PVN and a part of the neurons in the anterior periventricular area and in the anterior, lateral, and dorsal parvi-PVN appeared to incorporate WGA. These neurons may correspond with the hypophysiotrophic TRH-synthesizing neurons in the rat hypothalamus.

Amino Acid Sequence↗

Growth properties and F protein cleavage site sequences of naturally occurring human parainfluenza type 2 viruses.

The growth properties of 24 clinical isolates of PIV-2 obtained from six independent areas in Japan were examined using Vero and primary monkey kidney cells. These viruses could be subdivided into three groups on the basis of the ability of syncytium formation on the two primate cell systems. The distinct correlation between the F protein cleavability and the fusogenic effect was observed in Vero cells, and the importance of consecutive basic residues in the F protein cleavage site for efficient cleavage was suggested by the sequence analyses of their F genes. On the other hand, in PMK cells, their fusogenic activities could not be directly attributed to the F cleavability, fusion peptide sequence, and replication efficiency, indicating that unidentified structural features play an important role in cytopathic activities of naturally occurring PIV-2s.

Amino Acid Sequence↗

Transcripts of simian virus 41 (SV41) matrix gene are exclusively dicistronic with the fusion gene which is also transcribed as a monocistron.

The complete nucleotide sequences of the matrix (M) and fusion (F) genes of simian virus 41 (SV41) were determined. Deduced amino acid sequences confirmed the close relationship of SV41 with human parainfluenza type 2 virus (PIV2). Analyses of noncoding regions between the F and the hemagglutinin-neuraminidase (HN) genes suggested the absence of the small hydrophobic gene, which is present between the F and the HN genes of simian virus 5 and mumps virus. It was striking that there was no apparent consensus gene end sequence between the M and the F genes and that the M gene was transcribed exclusively as a dicistron with the F gene. The number of monocistronic transcripts of the F gene was approximately half that of the dicistronic transcripts. However, the F protein of SV41 seemed to be efficiently translated, since viral multiplication and fusion from within were as efficient as in PIV2. These results suggest that the lack of a consensus gene end sequence resulted in the readthrough of viral RNA polymerases between the M and the F genes and that the initiation of F gene transcription could occur by newly entered polymerases independently of the polymerases that started the upstream M gene transcription.

Amino Acid Sequence↗

Studies on the constituents of Aconitum species. IX. The pharmacological properties of pyro-type aconitine alkaloids, components of processed aconite powder 'kako-bushi-matsu': analgesic, antiinflammatory and acute toxic activities.

Eight pyro-type aconitine alkaloids contained in the processed aconite powder 'Kako-bushi-matsu' were studied for their analgesic, antiinflammatory and acute toxic actions. All these compounds showed significant analgesic and antiinflammatory actions. Among the pyro-type alkaloids, 16-epi-pyrojesaconitine and pyrojesaconitine were the most potent analgesics. The analgesic activity of pyro-type aconitine alkaloids was lower than that of each of the parent alkaloids, aconitine, mesaconitine, hypaconitine and jesaconitine. However, pyro-type aconitine alkaloids had very low toxicity, and the decreasing rates of the toxicity in changing from the parent alkaloids to the pyro-type aconitine alkaloids were much larger than those relating to the analgesic activity. Eight pyro-type aconitine alkaloids were found to inhibit the carrageenin-induced hind paw edema at 2 to 6 h after the carrageenin subplantar injection. Consequently, it was demonstrated that the pyro-type aconitine alkaloids produced through the processing of raw aconite roots, 'Bushi', have a role in the medicinal effects of the processed aconite powder 'Kako-bushi-matsu'.

Aconitine↗

Sequence characterization of the matrix protein genes of parainfluenza virus types 4A and 4B.

The complete nucleotide sequences of the matrix protein (M) genes of parainfluenza virus types 4A and 4B (PIV-4A and -4B) were determined from cDNA of the mRNA, and found to be 1548 bases in length, exclusive of poly(A) sequences. The sequences contained a large open reading frame of 1146 nucleotides encoding 362 amino acids. A high degree of identity (96.1%) was observed between the amino acid sequences of PIV-4A and PIV-4B M. These M sequences were compared with those of 10 other paramyxoviruses and a phylogenetic tree was constructed.

Amino Acid Sequence↗

Studies on the constituents of Aconitum species. XII. Syntheses of jesaconitine derivatives and their analgesic and toxic activities.

The role of the substituents at C3 and C8 of jesaconitine (1) on jesaconitine-induced analgesia and toxicity was examined. 3-O-Acetyljesaconitine (2), 3-O-anisoyljesaconitine (3), and 3-deoxyjesaconitine (6) showed dose-dependent analgesic action, and the potency of their compound-induced analgesia and toxicity was lower than those of 1. The most remarkable difference was found in the toxicity. The results indicate that the C3 hydroxy function of 1 participate in the induction of toxicity rather than of analgesia. 8-O-Linoleoyl-14-anisoylaconine (5), 8-O-methyl-14-anisoylaconine (7), 8-O-ethyl-14-anisoylaconine (8), 14-anisoylaconine (4) and 8-deoxy-14-anisoylaconine (9) showed lower activities than jesaconitine-induced analgesia and toxicity. The analgesic activity of 7 was almost the same as that of 8, but the toxicity of 7 was lower than that of 8. The analgesic activity of 9 was lower than that of 4, but the toxicities of both derivatives were not apparent. These facts indicate that the C8 function of 1 is important to the induction of analgesia and toxicity, and also that this function participates differently in the induction of the analgesia and toxicity. Subsequently, it was suggested that substituents at C3 and C8 of 1 played important roles of the induction of the analgesia and toxicity, and that the modes of this participation were not the same in analgesia and toxicity.

Aconitine↗

Comparative study of five preoperative methods for the localization of accessory pathways in the Wolff-Parkinson-White syndrome.

One hundred and thirty-four cases of Wolff-Parkinson-White syndrome were studied to evaluate the relative usefulness of electrocardiography (ECG), electrophysiological studies (EPS), body surface mapping (BSM), gated blood-pool phase analysis (nuclear studies), and vectorcardiography (VCG) in the localization of the accessory pathway (ACP). In comparison with the final localization verified by intraoperative studies, 93.4% in 8-region ACP localization (97.7% in 4-region ACP localization) could be correctly localized by ECG using our criteria, 83.9% (86.8%) by EPS, 82.6% (95.8%) by BSM, 78.8% (87.7%) by nuclear studies, and 67.3% (78.0%) by VCG. It was concluded that: (a) ACP can be localized preoperatively with considerable accuracy by using our simple ECG criteria. (b) The EPS method has some limitation, especially with respect to 8-region ACP localization. (c) Our observation showed no evidence that BSM, VCG, or nuclear studies were superior to ECG in ACP localization. (d) Among the 5 methods studied, ECG and EPS appear to be the appropriate procedures for preoperative ACP identification.

Adolescent↗

Impaired secretion of growth hormone-releasing hormone, growth hormone and IGF-I in elderly men.

The GHRH test and L-dopa test were performed in 12 normal young men (24.1 +/- 1.1 years) and 12 normal elderly men (77.8 +/- 1.4 years) to investigate age-related changes in secretion of GHRH, GH and IGF-I. The basal plasma levels of GHRH and GH were not significantly different in young and elderly men, but the basal plasma level of IGF-I was higher in the young men (159.0 +/- 11.7 vs 86.7 +/- 11.6 micrograms/l). The area under the curve for plasma GH in the GHRH test was less in the elderly group (35.1 +/- 5.9 vs 11.2 +/- 2.1 micrograms.h-1.l-1, p less than 0.001). The AUCs for the plasma GHRH and GH responses in the L-dopa test in young and elderly men were 32.0 +/- 2.7 vs 20.3 +/- 1.8 ng.h-1.l-1 (p less than 0.001), and 21.8 +/- 4.6 vs 5.4 +/- 1.1 micrograms.h-1.l-1 (p less than 0.01), respectively, indicating decreased releases of GHRH and GH in the elderly. Correlations between the AUCs for plasma GHRH and GH responses in L-dopa were found in both groups, but the ratio of the AUCs for GH/GHRH was lower in the elderly group. The elderly group showed a significant correlation between the basal plasma IGF-I level and the AUCs for plasma GH in the GHRH and L-dopa tests. These results suggest that elderly men have a decreased reserve of hypothalamic GHRH, resulting in secondarily impaired GH release, which may lead to a lower level of IGF-I than in young men.

Adult↗

Alternating chemotherapy with cyclophosphamide/adriamycin/vincristine (CAV) and cisplatin/etoposide (PVP) against small cell lung cancer. Eastern Shikoku Lung Cancer Chemotherapy Group.

Seventy-four confirmed small cell lung cancer (SCLC) patients received alternating combination chemotherapy with CAV and PVP. The CAV comprised of cyclophosphamide 800 mg/m2 on day 1, adriamycin 50 mg/m2 on day 1 and vincristine 1.4 mg/m2 on day 1, administered every 3-4 weeks. The PVP comprised cisplatin 80 mg/m2 on day 1 and etoposide 75 mg/m2 on day 1-5 administered every 3-4 weeks. Of these 74 patients, 63 (85.1%) achieved complete or partial responses with 16 (21.6%) obtaining a complete response. The median survival time was 13.2 months: 10.4 months in patients with extensive disease (ED), 16.3 months in those with limited disease (LD). A three-year disease-free period was achieved in eight patients (11.2%: 4.8% with ED, 16.8% with LD). The median duration of response was 28.3 weeks: 20.1 weeks with ED and 44.0 weeks with LD. The most commonly encountered side effects were nausea, vomiting, alopecia and myelosuppression but all were tolerable. We consider CAV-PVP to be an effective combination regimen for treating SCLC.

Adult↗

Correlation between plasma levels of ACTH and cortisol in basal states and during the CRH test in normal subjects and patients with hypothalamo-pituitary disorders.

Using a new ACTH-immunoradiometric assay (IRMA), we measured plasma ACTH levels in the basal states and during CRH test in normal subjects and the patients with hypothalamo-pituitary disorders. The basal levels of plasma ACTH in 76 normal young (25-45 yr) and 140 elderly (60-85 yr) subjects were 23.1 +/- 13.6, and 17.5 +/- 11.2 pg/ml, respectively. The plasma ACTH levels were less than detection limit (5 pg/ml) in 3 patients with isolated ACTH deficiency, and less than 10 pg/ml in 6 of 7 patients with hypopituitarism. A significant correlation was observed between the basal levels of plasma ACTH and of cortisol in two age groups, with almost the same regression line, showing no age-related decline in the plasma levels of ACTH and cortisol. In 2 normal subjects and 2 patients with Cushing's disease, synchronized secretions of ACTH and cortisol were observed between 0800h and 1800h. In normal subjects and the patients with pituitary disorders, a significant correlation was observed between the Area Under the Curve's for plasma ACTH and cortisol during the CRH test. The correlation constant was higher in normal subjects, but lower in the patients with acromegaly, non-functioning pituitary tumor, and Cushing's disease in this order, suggesting low sensitivity of the pituitary-adrenal axis in these patients. These results suggest that the ACTH-IRMA kit provide reliable data for clinical investigation, and that the secretions of ACTH and cortisol correlate each other in basal states and during the CRH test in the patients with pituitary disorders as well as in normal subjects.

Adolescent↗

Sequencing analyses and comparison of parainfluenza virus type 4A and 4B NP protein genes.

The nucleotide sequences of the cDNA copies of the mRNA coding for the nucleocapsid proteins (NPs) of human parainfluenza viruses type 4A (PIV-4A) and type 4B (PIV-4B) were determined. The copy of PIV-4A NP mRNA contained 1885 nucleotides encoding a protein with a calculated molecular weight of 62,561. The same number of amino acids with a similar molecular weight (62,425) were predicted for the PIV-4B NP protein. Comparisons on the nucleotide sequence and the amino acid sequence of NP protein between these two subtypes revealed extensive homologies in the nucleotide sequence (87%) and in the amino acid sequence (93%). Furthermore, a conserved region with about 100 amino acids was observed between PIV-4s and other paramyxoviruses, Newcastle disease virus (NDV), Sendai virus, mumps virus (MuV), PIV-3, BPIV-3, measles virus (MV), and canine distemper virus (CDV), indicating a common ancestor for these nine viruses. Our data also indicated that the PIV-4 NP proteins were more closely related to MuV and NDV than to other parainfluenza viruses, PIV-3, BPIV-3, and Sendai virus. Interestingly, the NP protein homology between PIV-4s and the morbillivirus group, MV and CDV, was slightly higher than that between PIV-4s and the parainfluenza viruses, PIV-3, BPIV-3, and Sendai virus.

Amino Acid Sequence↗

Sequence determination of the hemagglutinin-neuraminidase (HN) gene of human parainfluenza type 2 virus and the construction of a phylogenetic tree for HN proteins of all the paramyxoviruses that are infectious to humans.

The nucleotide sequence of the hemagglutinin-neuraminidase (HN) gene of human parainfluenza type 2 virus (PIV-2) was determined. The PIV-2 HN gene was 2112 nucleotides excluding poly(A) tail. There was a single large open reading frame in the mRNA which encoded a protein of 571 amino acids with a calculated molecular weight of 63,262. Analysis of the deduced amino acid sequence revealed that there were fourteen potential glycosylation sites and a major hydrophobic region near the N-terminus, which would anchor the protein in the viral membrane. Comparisons of the HN protein sequences of PIV-2 with those of Simian virus 5 (SV5), Sendai virus (SV, parainfluenza virus type 1), human parainfluenza virus type 3 (PIV-3), type 4 (PIV-4), bovine parainfluenza virus type 3 (BPIV-3), mumps virus (MuV), and Newcastle disease virus (NDV) showed definite amino acid sequence relatedness, indicating a common ancestor for these viruses. Furthermore, statistical analysis of the protein sequences suggested a possible evolutionary relatedness among the paramyxoviruses. This is the first time that a phylogenetic tree has been constructed for all the parainfluenza viruses and mumps virus which are infectious to humans. In addition, amino acid sequences involved in hemagglutinating and neuraminidase activities of paramyxovirus were discussed.

Amino Acid Sequence↗

Antigenic and structural properties of a paramyxovirus simian virus 41 (SV41) reveal a close relationship with human parainfluenza type 2 virus.

Seven structural component proteins of a paramyxovirus simian virus 41 (SV41) were identified with the aid of monoclonal antibodies prepared against SV41 and human parainfluenza type 2 virus (PIV2). The nucleoprotein is antigenically very close to that of PIV2, while it is comparatively far from that of simian virus 5 (SV5). The hemagglutinin-neuraminidase (HN) protein showed no immunological relationship to either of the HN proteins of PIV2 or SV5. The amino acid sequence of the SV41 HN protein was deduced from the nucleotide sequence of its HN gene and revealed that the SV41 HN is unexpectedly close to the PIV2 HN (61.2% identity in amino acid sequence), while the SV5 HN showed only 48.3% identity with the PIV2 HN. The SV41 HN is also related to the SV5 HN (51.3% identity); thus, the SV41 HN is phylogenetically situated between the PIV2 and SV5 HNs. These results indicate that SV41 is the virus closest to PIV2 at present.

Amino Acid Sequence↗

Sequence analyses of the 3' genome end and NP gene of human parainfluenza type 2 virus: sequence variation of the gene-starting signal and the conserved 3' end.

We cloned and determined the nucleotide sequences of cDNAs against nucleocapsid protein (NP) mRNA and the genomic RNA of human parainfluenza type 2 virus (PIV-2). The 3' terminal region of genomic RNA was compared among PIV-2, mumps virus (MuV), Newcastle disease virus (NDV), measles virus (MV), PIV-3, bovine parainfluenza type 3 virus (BPIV-3), Sendai virus (SV), and vesicular stomatitis virus (VSV), and an extensive sequence homology was observed between PIV-2 and MuV. Although no significant sequence relatedness was observed between PIV-2 and other viruses, the terminal four nucleotides were identical in the viruses compared, implying a specific role of these nucleotides on the replication of paramyxoviruses. A primer extension analysis elucidated the major NP mRNA initiation site with the sequence UCUAAGCC, which showed a moderate homology with the gene-starting consensus sequences of other paramyxoviruses. On the other hand, the NP mRNA was terminated at the nucleotide stretch AAAUUCUUUUU, and this sequence was conserved in all the PIV-2 genes, indicating that the oligonucleotides will form a part of the gene attenuation signal of PIV-2. Comparisons of NP protein sequence indicated a possible subgrouping of the paramyxoviruses into two groups, one of which is a group including PIV-2, PIV-4, MuV, and NDV, and another is a group including PIV-3, BPIV-3, and SV. This result supports an idea from our previous studies using polyclonal and monoclonal antibodies. Furthermore, our data indicated that the PIV-2 NP protein sequence was more closely related to MV and CDV than to other parainfluenza viruses, PIV-3 and SV.

Amino Acid Sequence↗

Complete nucleotide sequence of the matrix gene of human parainfluenza type 2 virus and expression of the M protein in bacteria.

The sequence of the M gene of human parainfluenza virus type 2 (PIV-2) has been determined. The sequence contained a large open reading frame with 1131 nucleotides encoding a protein with a calculated molecular weight of 42,312. Comparison of M protein sequence indicated that PIV-2 was more closely related to mumps virus and Newcastle disease virus than to other parainfluenza viruses, Sendai virus (SV), and parainfluenza virus type 3 (PIV-3), indicating a possible subdividing of the Paramyxovirus into two groups. This grouping is consistent with that obtained from analysis of the HN gene. Measles virus and canine distemper virus definitely belong to the subgroup composed of SV and PIV-3. No homology region was found in all the paramyxoviruses compared. However, a tertiary structure may be conserved in each subgroup of paramyxovirus. The M protein of PIV-2 was expressed in bacteria, and the product was recognized by a monoclonal antibody specific for the PIV-2 M protein. The bacterial-expressed protein, however, was heterogeneous and smaller in size.

Amino Acid Sequence↗

Molecular cloning and sequence analysis of human parainfluenza type 4A virus HN gene: its irregularities on structure and activities.

We cloned the cDNA of human parainfluenza type 4A virus (PIV-4A) HN gene by reverse-transcription of virus-specific mRNAs and genomic RNA, and determined the complete nucleotide sequence of the HN gene. The predicted HN protein sequence of PIV-4A showed significant relatedness with those of other paramyxoviruses, SV5, NDV, MuV, PIV-3, BPIV-3, indicating a common ancestor. The homologies between the viruses suggested that PIV-4A is more closely related to NDV, SV5, and MuV than to the parainfluenza viruses, PIV-3, bovine parainfluenza type 3 virus (BPIV-3), and Sendai virus (SV). Sixty amino acids were commonly conserved among the viruses, other than PIV-4A. Two of these amino acids were substituted in PIV-4A HN and are predicted to be located near the active site of the neuraminidase. The analysis of neuraminidase of PIV-4 revealed that the activity is hardly detectable, suggesting the significant effect of the substituted amino acid sites on neuraminidase activity.

Amino Acid Sequence↗

Terminal structure of a Densovirus implies a hairpin transfer replication which is similar to the model for AAV.

We cloned the complete sequence of Bombyx DNV (Ina isolate; Bm DNV-1) genome in a bacterial plasmid pUC 119 and determined the nucleotide sequences of both termini, resulting in elucidation of the nucleotide sequence of the complete genomic DNA of DNV. The complete sequence of the DNV DNA (5048 nucleotides) has inverted repeats of 225 nucleotides and the terminal 153 nucleotides are palindromic. The palindromes can fold back on themselves to form a hairpin structure but, unlike AAV, the small internal palindrome which forms a T-shaped conformation was not observed. End-label analysis demonstrated that the palindromic sequences at both termini can exist in either of two orientations (flip or flop) in virion DNA with different frequencies. These data suggest that the hairpin transfer model for AAV replication must be modified to explain the DNV replication. Additionally, a comparison study on the terminal structures of insect, human, and rodent parvoviruses allowed a prediction on the ancestral terminal structure of parvovirus genome.

Amino Acids↗