Biomedical subjects
Publications and source records attributed to H Takagi.
We examined the expression of the I, II and III subtypes of voltage-sensitive Na+ channel mRNA in the rat brain using in situ hybridization histochemistry with oligonucleotide probes. The distribution of cells with strongly positive signals was characteristic for each subtype. Synthesis of each subtype of Na+ channel protein may be regulated by differential mRNA expression.
We studied the distribution of type II Ca2+/calmodulin (CaM)-insensitive adenylyl cyclase mRNA in the adult rat brain by in situ hybridization histochemistry. Although type II adenylyl cyclase mRNA was distributed throughout the rostrocaudal extent of the brain, the labeling intensity varied among regions. Strong expression was seen in fairly restricted regions such as CA1 and dentate-gyrus of the hippocampus, some thalamic nuclei, mammillary nucleus, dorsal raphe nucleus, parabrachial nucleus and locus coeruleus. The distribution of this type of mRNA was significantly different from that of type I Ca2+/CaM-sensitive adenylyl cyclase mRNA. The characteristic localization of type II mRNA suggests that this type of adenylyl cyclase may be involved in some neuronal functions different from the type I enzyme.
The present study attempted to explore the gene expression of the subunits (GluR1-4) of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type receptor, subunit (GluR5) of kainic acid (KA)-type receptor, NR1 [a subunit of N-methyl-D-aspartate (NMDA) receptors] and the possible glutamate-binding subunit of an NMDA receptor complex in the dorsal horn of the rat spinal cords using in situ hybridization histochemistry. These results were compared with those of the spinal motor neurons. Expression of the subunits of the AMPA-type receptor was also examined at the protein level using immunocytochemistry, with reference to the motor neurons. Although all the four subunits of the AMPA-type receptor were expressed throughout the dorsal horn, the pattern of expression was different according to the dorsal horn region and to the subunits. GluR2 showed the strongest expression in the dorsal horn. Huge numbers of strongly labelled cells formed a dense collection in lamina II and superficial parts of lamina III. Many neurons in lamina II and superficial parts of lamina III expressed GluR1 moderately. Scattered neurons moderately expressing GluR3 were also seen in these regions, while the expression of GluR4 was very low. Labelling of the dorsal horn neurons by the GluR5 probe was low, and NR1 probe and a glutamate-binding subunit of an NMDA receptor complex probe labelled them diffusely with low to moderate intensity. These findings show a close relationship between the glutamergic nociceptive primary afferent system and AMPA-type receptors in which GluR2 is especially highly expressed. The present study further showed that the expression pattern of the glutamate receptors in the spinal sensory neurons differs considerably from that of spinal motor neurons. Motor neurons very strongly express GluR3 and 4, while the expression of GluR2 and GluR1 is moderate and low, respectively. Expression of GluR5 is also low in the motor neurons. However, expression of NR1 and the glutamate-binding subunit of an NMDA receptor complex is very strong. These findings indicate that the subunit composition of the AMPA-type receptors regulating motor neurons is different from that of the AMPA-type receptors in the spinal sensory neurons, and that there are at least two kinds of glutamergic systems which regulate motor neurons: via AMPA-type receptors and via NMDA receptors.
We retrospectively examined the effect of HLA-DRB1 matching at the DNA level compared with serological HLA-DR matching on acute rejection and graft survival in patients who underwent primary cadaveric renal transplantation. For patients with serological HLA-DR zero mismatch, the incidence of acute rejection in patients with zero DRB1 mismatch (3/20; 15%) was significantly lower than in those with one or two DRB1 mismatches (10/21; 48%). Five-year graft survival in patients with zero DRB1 mismatch was 100%, whereas that in those with one or two DRB1 mismatches was 76%, although the difference was not statistically significant. The fact that HLA-DRB1 matching at the DNA level influenced incidence of graft rejection after cadaveric renal transplantation is analogous to results in living-related renal transplantation. It is suggested that avoidance of mismatching for DRB1 alleles at the DNA level in recipient selection of cadaveric renal transplantation leads to an improvement of graft outcome.
Thirty-five Bacillus brevis strains obtained from culture collections, including protein-producing isolates, were taxonomically studied by using numerical analysis, DNA base composition, and DNA-DNA hybridization. Six DNA relatedness groups were represented, and these groups correlated well with clusters based on the numerical analysis. The B. brevis strains were separated into B. brevis sensu stricto, four new species, and an unidentified species of the genus Bacillus. Bacillus migulanus sp. nov., Bacillus choshinensis sp. nov., Bacillus parabrevis sp. nov., and Bacillus galactophilus sp. nov. are proposed.
1. Intracerebroventricular (i.c.v.) administration of L-arginine (L-Arg), at 10-100 micrograms per mouse, produced antinociception in mice, as assessed by the tail flick test; this antinociception was reversed by pretreatment (s.c.) with naltrindole (NTI), a delta-selective opioid antagonist, and by co-administered L-leucyl-L-arginine (Leu-Arg), a kyotorphin (endogenous Met-enkephalin releaser) receptor antagonist. 2. L-NG-nitroarginine methyl ester (L-NAME), a NO synthase inhibitor, but not D-NG-nitroarginine methyl ester, given i.c.v. at 3-10 micrograms per mouse, exhibited antinociceptive activity that was resistant to naloxone (s.c.), NTI (s.c.) and Leu-Arg (i.c.v.). 3. The L-NAME (i.c.v.)-induced antinociception was not reversed by L-Arg (i.c.v.), which was antinociceptive by itself, but was abolished by combined injection of L-Arg plus Leu-Arg (i.c.v.) or by L-Arg (i.c.v.) after NTI (s.c.). 4. Methylene blue (MB), a soluble guanylate cyclase inhibitor, at 0.1-1 microgram per mouse, produced antinociception by i.c.v. administration. The antinociception induced by MB (i.c.v.) or L-NAME (i.c.v.) was reversed by co-administered dibutyryl cyclic GMP. 5. These findings suggest that L-Arg plays a dual role in nociceptive processing in the brain, being antinociceptive via the kyotorphin-Met-enkephalin pathway and nociceptive via the NO-cyclic GMP pathway.
To assess the effect of Carolina rinse solution on the renal graft function after cold ischemic storage in the University of Wisconsin solution for 48 h, canine renal transplantation was performed. Survival rate was higher in the kidneys rinsed with Carolina rinse solution (4 of 5, 80%) than that of kidneys rinsed with lactate Ringer's solution (0 of 5, 0%). Postoperative serum creatinine level on day 2 was significantly lower in the Carolina group (3.48 +/- 0.47 mg/dl) than that in the Ringer group (5.60 +/- 0.68 mg/dl) (p < 0.01). All of the grafts in the Carolina group functioned immediately. Whereas, in the Ringer group, none of the grafts functioned postoperatively. Histologically, severe renal damage was observed in the kidneys perfused by lactate Ringer's solution. On the other hand, the damage to the grafts was minimal in the Carolina group. With immunohistochemical staining using factor-VIII-related antigen, the glomeruli were more severely damaged in the kidneys perfused by lactate Ringer's solution than by Carolina rinse solution. The present study showed that Carolina rinse solution was effective in the prevention of the kidney graft damage from reperfusion injury.
The authors have reviewed 80 cases of subcortical cerebral hemorrhage, in all of which intraoperative examinations during craniotomy or autopsies were performed. Cases involving trauma and aneurysm were excluded from the study. The diagnosis of subcortical hemorrhage was made by plain computed tomography in all cases. The most common cause of hemorrhage was vascular malformation (68%; 56 cases), in 20 (36%) of which angiographically occult vascular malformations were noted. Hypertension was present in 23 (29%) of 80 cases but was the cause of hemorrhage in only 9 cases (11%). Hypertension was present in 6 (30%) of 20 angiographically occult vascular malformations, all of which were in patients under 65 years. In 12 (75%) of 16 cases of angiographically occult vascular malformations, prolonged high-dose delayed-contrast computed tomography and magnetic resonance imaging were able to provide definite diagnostic information. We recommend that patients with subcortical cerebral hemorrhage, especially those under age 65, with or without hypertension, be carefully examined by prolonged high-dose delayed contrast computed tomography and magnetic resonance imaging for the presence of angiographically occult vascular malformations causing hemorrhage.
Acinetobacter calcoaceticus colonization was observed in the gastrointestinal tracts of C.-B17-scid/scid (SCID) mice, while it was not observed in C.B17-scid/+ and C.B17-+/+ mice with normal immunity housed under the same conditions. A. calcoaceticus and other viable enteric bacteria were not isolated from any organs other than gastrointestinal tract in SCID mice. The mice colonized with this organism were apparently healthy and no significant visceral lesion was observed.
Of 170 cows with ovarian cysts which were detected by palpation of the ovaries per rectum, 88 cows (51.8%) were diagnosed as having follicular cysts on the basis of milk progesterone concentrations. Eighty-three of the 88 cows were used for treatment trials. A group of 56 cows injected subcutaneously with fenprostalene, prostaglandin F2 alpha analog, 14 days after treatment with fertirelin, an analog of gonadotropin-releasing hormone, compared with another 27 cows treated only with fertirelin (controls), showed a higher pregnancy rate within 100 days after the initial treatment (66.1 vs 48.1%) and shorter intervals from the treatment to conception (30 +/- 21 vs 43 +/- 27 days). It was concluded that the administration of fenprostalene following fertirelin treatment is effective in shortening the interval from treatment to conception in cows with follicular cysts.
The cattle with adequate alpha-tocopherol (Vit E) and marginally deficient selenium (Se) status manifested significantly lower anti-Haemophilus somnus antibody titer than the cattle supplemented with Se in the later stage of an 8-week trial. However, in the early stage no difference was observed in magnitude of anti-H. somnus antibody development between them. These results suggested that Se may contribute to anti-H. somnus antibody production, and that Vit E can make up for Se deficiency to a certain degree.
We studied 2 types of a cementless total hip prosthesis in dogs. Both were coated with titanium plasma-spray. In both components, the pores in the deep layer of 1 group were further coated with apatite and wollastonite containing glass-ceramic (AW glass-ceramic). 50 dogs underwent unilateral total hip replacements, and were killed at 1, 3, or 6 months postoperatively. We evaluated the femoral and the acetabular components mechanically and histologically. At 1 month, the detaching load and bone ingrowth of the AW glass-ceramic-coated femoral and acetabular components were higher than those of the control implants. At 3 and 6 months there were no differences between the 2 types of components. Thus, AW glass-ceramic enhanced the early phase of cementless implant fixation.
One-hundred-and-twenty patients underwent first cadaveric kidney transplantation from the non-heart beating donors. All of the organs were procured with the use of double balloon catheter for in situ cooling. The mean warm ischemic time and cold ischemic time were 10.7 +/- 17.0 minutes and 18.9 +/- 11.4 hours, respectively. One- and 5-year graft survival rates were 85.0 and 72.7%, respectively. Among 120 recipients, 30 (25%) grafts functioned immediately (immediate function), 82 (68.3%) grafts functioned after varying length of oliguric periods (delayed function) and 8 (6.7%) grafts never functioned (non-function). The mean age of the donors in the group of immediate function (31.5 +/- 16.1 yr) was significantly lower than those of other two groups. The mean warm ischemic time in the group of immediate function (6.0 +/- 11.2 min) was significantly shorter than that of delayed function. However, there was no significant difference in donor hypotensive episode, types of preservation fluid and cold ischemic time between the groups. The conclusion is that the ultimate result of cadaveric kidney transplant from the non-heart beating donors with the use of double lumen catheter is acceptable despite a relatively high incidence of delayed graft function.
To clarify the clinico-pathological characteristic of poorly differentiated adenocarcinoma of gastric cancer with medullary growth pattern, the immunohistochemical study was performed using antibodies against blood group related antigens (Lewis(a), Lewis(b), Lewis(x), Lewis(y)) and gastrointestinal tissue related antigens (CEA, AFP and NSE). The following results were obtained. 1. Foveolar epithelium of normal gastric mucosa has type 1 blood group associated antigen (Lewis(a), Lewis(b)) and deep gland has type 2 antigen (Lewis(x), Lewis(y)) respectively, as the differentiation antigens. 2. Expression of Lewis(a) and Lewis(b) are more often observed in poorly differentiated adenocarcinoma with medullary and scirrhous growth pattern than in well and moderately differentiated adenocarcinoma. Poorly differentiated adenocarcinomas postulated to have closer relation with normal gastric deep gland in terms of antigen expression. 3. Poorly differentiated adenocarcinoma with medullary growth pattern is characterized as more differentiated type cancer than scirrhous type with respect to expression of blood group related antigen. 4. Poorly differentiated adenocarcinoma with medullary growth pattern may be divided into 3 subgroups by each antigen expression of CEA, NSE, and AFP.
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To assess exocrine pancreatic function in patients before and after pancreatoduodenectomy (PD), we used the breath test, with nonradioactive 13C-labeled trioctanoin, in 14 patients before pancreatic resection because of localized pancreatic mass (preop-PD group), and in 13 patients who had undergone pancreatoduodenectomy more than 5 yr before (post-PD group). The results were compared with those of the secretin test, N-benzolyl-L-tyrosyl-p-amino benzoic acid (BT-PABA) test, and fecal chymotrypsin. Means +/- SD and frequencies of low values of the recovery of the breath test were 42.0 +/- 3.4%, 0/5 in the control; 24.2 +/- 10.5%, 14/14 in the preop-PD group; and 18.6 +/- 8.0, 13/13 in the post-PD group. The overall sensitivities in the preop- and post-PD groups were 100% for the recovery and 93% for the maximal mass ratio of the breath test, 89% for the secretin test, 67% for the BT-PABA test, and 64% for fecal chymotrypsin. The recovery of the breath test correlated significantly with the duodenal outputs of lipase, amylase, and chymotrypsin, and was not affected in patients with obstructive jaundice or with low D-xylose absorption. The breath test is as sensitive as the secretin test, more reliable than the conventional tubeless tests, and is available to follow up the exocrine pancreatic function before and after pancreatoduodenectomy.
Proliferating cell nuclear antigen (PCNA) appears in the cell nuclei during the late G1 to S phases of the cell cycle and is thought to be closely related to cellular proliferation. The authors have conducted an immunohistochemical study in order to investigate the tissue expression of PCNA and its clinical significance in breast cancer. Excluding cases with absolutely no positive cells on the section specimen, the mean value (%) for the PCNA labeling index (LI) was 30.4 in 187 cases of invasive ductal carcinoma. No correlations between PCNA LI and any clinicopathological factors such as tumor diameter and tumor stage were observed. Also, no significant correlation was observed with Ki-67 LI. A positive correlation was, however, observed with the tissue expression of c-erbB-2 protein. We divided 82 patients with stage II invasive ductal carcinoma into PCNA LI of < 10, PCNA LI of 10-50 and PCNA of > 50, and analyzed the specimens for any correlation with prognosis. The group with PCNA LI of > 50 had significantly poorer prognoses than the other groups. From the above, we concluded immunostaining for PCNA to be useful as a prognostic factor and as an indicator of the degree of malignancy in breast cancer.
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