Decrease in glutamate in the jugular vein after head injury.
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Craniocerebral Trauma↗
Biomedical subjects
A Kudo
Publications and source records attributed to A Kudo.
In situ detection of gonadotropin-releasing hormone (GnRH) receptor mRNA expression in the rat ovarian follicles.
To clarify the target cells of GnRH in the ovary, in vivo expression of ovarian GnRH receptor mRNA was examined histologically by in situ hybridization in immature rats treated with PMSG only or in combination with hCG. Strong hybridization signals were observed in the granulosa cells of atretic follicles. However, no significant signals were found in the granulosa cells of healthy small, preantral, or early antral follicles. Healthy Graafian and preovulatory follicles also showed intense signals in their mural granulosa cells, but no signals were detected in the cumulus oophorus cells. Corpora lutea showed only weak signals, but luteinizing follicles probably after atresia exhibited signals of moderate intensity in their luteinized and remaining granulosa cells. No signals were detected in the theca cells and oocytes in all the follicles. Interstitial cells sometimes exhibited hybridization signals of moderate intensity, when the cells were eosinophilic. Pretreatment with different combinations of gonadotropins yielded different ovarian histology, but this had no influence on the localization of hybridization signals. These results, showing that the authentic GnRH receptor mRNA was expressed in a certain cell population in the rat ovary, suggest that the receptor is involved in the control of various ovarian functions including follicular development, atresia, ovulation, and luteinization after ovulation and follicular atresia.
Animals↗
Expression and characterization of murine osteoblast-specific factor 2 (OSF-2) in a baculovirus expression system.
Osteoblast-specific factor 2 (OSF-2) is a approximately 90-kDa protein selectively expressed in bone. OSF-2 cDNA was recently isolated from mouse and human cDNA libraries and shows limited sequence homology with fasciclin I, a cell adhesion protein expressed in insect nerve cells. Here we describe the expression of recombinant murine OSF-2 (rmOSF-2) in a baculovirus/insect cell system. Western blotting analysis employing polyclonal antiserum raised against a C-terminal synthetic OSF-2 peptide detected a protein of approximately 90-kDa as early as 2 days after infection of Sf9 cells with the recombinant virus. Tunicamycin treatment of infected cells resulted in a mobility shift of OSF-2 (approximately 90-kDa band) on Western blots. N-Glycanase digestion resulted in the same mobility shift of OSF-2, indicating that rmOSF-2 expressed in insect cells is N-glycosylated. However, OSF-2 was insensitive to endoglycosidase H digestion while a major fraction of this protein had affinity for concanavalin A. Finally, it was demonstrated that rmOSF-2 was able to bind to heparin. This finding suggests that OSF-2 might be associated with the bone extracellular matrix after secretion by osteoblasts and participate in cell adhesion and/or cell communication. The establishment of the baculovirus expression system with a high productivity of recombinant OSF-2 (around 40 micrograms/ml at maximum) and its heparin binding properties should allow us to obtain large amounts of rmOSF-2.
Amidohydrolases↗
Immunocytochemical localization of estrogen and progesterone receptors (ER & PgR) in breast cancer.
The localization of estrogen receptors (ER) and progesterone receptors (PgR) in human breast cancer was examined light and electron microscopically by using specific monoclonal antibodies. Among the breast cancer patients, ER-positive (ER[+]) cases were shown by enzyme immunoassay (EIA) to account for 58.9% (63/107), and 63.5% of these cases were women in the postmenopausal state. The PgR-positive (PgR[+]) rate was 50.5% (54/107), with 44.4% of these positive cases being postmenopausal women. The rates of ER(+) and PgR(+), ER negative (ER[-]) and PgR negative (PgR[-]), and ER(+) and PgR(-) cases were 43.9% (47/107), 34.6% (37/107) and 15.0% (16/107), respectively. Although ER(-) and PgR(+) cases were in few number, they were found (6.5%, 7/107). These results correlated well with those obtained by the immunocytochemical method. In either case, i.e anti-ER or anti-PgR reaction, positive nuclei and negative nuclei were found intermingled with each other in a given visual field. In electron microscopy, both anti-ER and anti-PgR antibodies bound to sites in the euchromatin area of the nucleus.
Antibodies, Monoclonal↗
B cell development in mice with a defective lambda 5 gene.
The surrogate light chain encoded by the two pre-B cell-specific genes VpreB and lambda 5 plays a critical role in B cell development of the mouse. It has been shown that targeted disruption of the lambda 5 gene results in a depletion of B220+ CD43- IgM-pre-B cells in bone marrow, and in a delayed appearance both of CD5+ as well as CD5- surface immunoglobulin (sIg)+ B cells in the periphery. In this report we show that DHJH-rearranged B220- and B220+, CD43+, c-kit+, sIgM- pro- and pre-B-I cells with long-term capacity to proliferate in vitro on stromal cells in the presence of interleukin-7 are present in normal numbers in the bone marrow of lambda 5 T/lambda 5 T mice at various ages. They express normal levels of VpreB mRNA but, in contrast to normal pre-B-I cells, do not express surrogate light chain on their surface. Pre-B-I cells from fetal liver and bone marrow of lambda 5 T/lambda 5 T mice differentiate with normal kinetics and in normal numbers to sIg+, mitogen-reactive B cells. These results suggest that the delayed generation of sIg+ B cells in the peripheral, mature compartments of CD5+ and CD5- cells could be accounted for by the daily production of approximately 5 x 10(5) sIg+ B cells from the pre-B-I cell pool in the absence of a normal pool of pre-B-II cells.
Animals↗
Isolation of rat GnRH receptor cDNA having different 5'-noncoding sequence.
A rat GnRH receptor cDNA was cloned with 1317 base pairs (bp) from a pituitary cDNA library. This clone had a full amino acid coding region, with 172 and 161 bp noncoding sequences in 5'- and 3'-flanking regions of the coding region, respectively. The nucleotide sequence of amino acid coding region was completely identical to that of previous report by Kaiser et al. (1992), but 5'-noncoding region was much longer than the previously reported one and its sequence was different. This 5'-noncoding region could be divided into two subregions on the basis of similarity between the two sequences. The 32 bp sequence flanking the coding region was completely identical to the previous report, but remaining 5'-end sequence almost totally lacked similarity. In the 3'-flanking region of coding region, 167 bp-long sequence was also newly identified. The new species of GnRH receptor mRNA found in the present study suggests that the presence of various mechanisms of GnRH receptor expression in the rat pituitary.
Amino Acid Sequence↗
A critical role of lambda 5 protein in B cell development.
The lambda 5 gene is a homolog of immunoglobulin J lambda-C lambda genes, expressed specifically in immature B-lineage cells. Lambda 5-encoded molecules form membrane complexes with mu or D mu proteins in association with an additional protein specifically expressed in immature B cells that is encoded by the Vpre-B gene. We have generated mice in which the lambda 5 gene is inactivated by targeted gene disruption in embryonic stem cells. In these mice, B cell development in the bone marrow is blocked at the pre-B cell stage. However, the blockade is leaky, allowing B cells to populate the peripheral immune system at a low rate. These cells are allelically excluded and able to respond to antigen.
Animals↗
Pre-B lymphocyte-specific transcriptional control of the mouse VpreB gene.
The VpreB genes, which encode surrogate immunoglobulin light chain molecules, are expressed as RNA almost exclusively in pre-B cells. We have investigated the transcriptional control mechanisms which are responsible for the pre-B cell-specific RNA expression of the mouse VpreB1 and VpreB2 genes. Nuclear run-on analyses demonstrate that the pre-B cell-specific expression of both VpreB genes is controlled primarily at the level of initiation of transcription. S1 nuclease protection-mapping defined two or three major start sites of transcription for the VpreB genes. To find a promoter and other potential cis-acting regulatory elements, a 700-bp fragment 5' of the transcription start sites of the VpreB1 gene was used in gene transfer experiments and found to act as a promoter in pre-B lymphocytes. Deletion experiments showed that 191 bp upstream of the most 5' transcription start site is required for the pre-B cell promoter activity. DNA sequence analysis of the 5' region of the mouse VpreB1, VpreB2 and human VpreB genes reveal that this region of approximately 200 bp is strongly conserved. This 200-bp promoter region contains several conserved nucleotide sequence motifs which may act to mediate the pre-B cell-specific transcription of the VpreB genes.
Animals↗
A pre-B- and B cell-specific DNA-binding protein, EBB-1, which binds to the promoter of the VpreB1 gene.
The VpreB1 protein is thought to be expressed on the surface of pre-B cells in association with lambda 5 and mu heavy chain, and to play an important role on B cell differentiation. The expression of VpreB1 and lambda 5 is pre-B cell specific, and regulated at the initiation of transcription. We have identified at least two sequence-specific DNA-binding proteins which bind to the region -191 to -74 of the promoter of the mouse VpreB1 gene. These DNA-binding proteins also bind to the promoter of the mouse lambda 5 gene. One of the two DNA-binding proteins, called EBB-1, is restricted to pre-B and B cells, but not detected in plasma cells, T cells and cells of other lineages. Transient transfection analysis of reporter constructs revealed that the binding sites of these proteins play a significant role in the activity of the promoter, especially the binding site of EBB-1. Taken together these results suggest that EBB-1 might be one of the crucial factors which regulates a series of intracellular events in B cell differentiation.
Animals↗
Time-dependent changes of collagen cross-links and their precursors in the culture of osteogenic cells.
The early stage of cross-link formation in bone collagen was studied in a cell culture system. An osteogenic cell line that produces and accumulates a remarkably high amount of collagen, and that eventually forms bone-like structures, was used in this study for its time-dependent development of reducible cross-links. It was found that precursors of the cross-link, dehydro-dihydroxynorleucine and dehydro-hydroxynorleucine became detectable as soon as the cells attained a confluent state. They showed maximal amounts at days 3-5 after confluence, but substantially disappeared at day 10 after confluence. In contrast, two characteristic cross-links of bone collagen, dehydro-dihydroxylysinorleucine (dehydro-DHLNL) and dehydro-hydroxylysinorleucine (dehydro-HLNL), which were present in trace amounts at the stage of cell confluence, gradually increased in amount and reached a plateau at day 10, just when their precursors disappeared. Thus, it was found that there was a time lag of about a week between the maximal formations of precursors and cross-links of bone collagen in this system. The significance of this time lag was interpreted in terms of the minimum essential accumulation of collagen for the precursor-product transition. The ratio of dehydro-DHLNL to dehydro-HLNL was as low as 0.7 at day 3 after confluency, increased to 4.2 at day 20, the period just before mineralization began, and decreased thereafter, suggesting a qualitative change in bone collagen associated with mineralization.
Animals↗
The expression of the mouse VpreB/lambda 5 locus in transformed cell lines and tumors of the B lineage differentiation pathway.
The expression of RNA transcripts from two pre B lymphocyte related genes, VpreB and lambda 5, has been studied in a series of transformed cell lines which appear frozen at different states of B lineage differentiation, from early progenitors to surface Ig positive B cells. In the HAFTL-1 cell line, which arose from fetal liver by transformation with a retrovirus containing the Hras oncogene, Northern analysis of poly A+ mRNA as well as in situ hybridization of RNA in single cells revealed that lambda 5 and VpreB are already expressed at the progenitor stage and increase in expression as the progenitors differentiate to precursor (preB) cells, or are turned off as the progenitors differentiate to myeloid cells. Continued rearrangements of Ig genes in pre B cell lines leading to Ig expression on the surface of NFS-5 pre B cells do not influence the continued expression of VpreB and lambda 5. Surface Ig-positive B lineage cell lines also express the pre B-related genes. Both Ly1+ as well as Ly1- pre B cells are VpreB- and lambda 5-positive. Lipopolysaccharide (LPS) stimulation of 70Z/3 pre B cells does not turn off lambda 5 expression. It therefore appears that, at least in transformed cell lines, the expression of VpreB and lambda 5 is not directly regulated by the expression of microH, kappa L, or lambda L chains, LPS reactivity, or the Ly1 surface antigen. Fusion of plasmacytoma cells with normal pre B cells to generate pre B hybridomas leads to down-regulation of VpreB/lambda 5 expression.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
[Clinical study on total intravenous anesthesia with droperidol, fentanyl and ketamine. 14. Effect on epidural pressure].
We studied effect of total intravenous anesthesia using ketamine, fentanyl and droperidol (DFK) on epidural pressure as an index for cerebrospinal fluid pressure in six surgical patients who underwent gastrectomy. The epidural catheter was inserted on the previous day. The epidural puncture was made at Th7-12 and the tip of the catheter was located 5 cm cephalad. The epidural pressure was measured before, just after and 30 minutes after the induction. The induction dose of fentanyl was 5 micrograms.kg-1 and that of ketamine was 1 mg.kg-1. The epidural pressure at the induction decreased in significantly by 19% as compared with that before the induction. The result suggested that DFK would not increase cerebrospinal fluid pressure when the doses of ketamine and fentanyl were changed.
Adult↗
Long-term proliferating early pre B cell lines and clones with the potential to develop to surface Ig-positive, mitogen reactive B cells in vitro and in vivo.
Cell lines and clones were established from PB76-positive mouse fetal liver at day 13 and 14 of gestation, which proliferated with division times of a day in serum-substituted cultures under the stimulatory influence of adherent stromal cells and the cytokine IL-7 for periods longer than half a year. These lines expressed varying levels of the B lymphocyte lineage related markers PB76, B220, BP-1, VpreB and lambda 5, but no surface Ig or MHC class II molecules. All clones expressed PB76, VpreB and lambda 5 in a high percentage of cells, while B220 and/or BP-1 expression was low or undetectable in some. A cell line, and several clones established from it, all had kappa and lambda light chain genes in germ-line configuration. Either one or both of their H-chain-gene containing chromosomes carried a DH to JH. These pre B cell lines and clones could be induced to VH to DH and VL to JL rearrangements. This resulted in the development of varying percentages of sIg-positive surface, MHC class II negative, LPS-reactive B cells within 2-3 days, in the absence of contacts with stromal cells and/or IL-7. When injected into SCID mice, the cultured pre B cells populated the spleen of these mice to 5% with surface Ig-, MHC class II-positive LPS-reactive cells for greater than 25 weeks. The long-term in vitro proliferative capacity of these DH-JH rearranged pre B cell clones makes them major candidates for committed stem cells of the B lineage.
Animals↗
B cell development in fetal liver.
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Animals↗
Long-term proliferating early pre-B-cell lines and clones with the potential to develop to surface immunoglobulin-positive, mitogen-reactive B-cells in vitro and in vivo.
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Animals↗
Potassium ion channels operated by receptor stimulation can be activated simply by raising temperature.
Application of either dopamine (DA), acetylcholine (ACh), or histamine (HA) to the identified ganglion cells of Aplysia elicits a K(+)-dependent slow hyperpolarization. When temperature of the bathing solution was raised from 22 to 32 degrees C, these cells were also hyperpolarized with a marked increase in K+ conductance. The warm- and transmitter-induced current responses recorded under voltage clamp were not blocked by either 1 mM Ba2+ or 10 mM TEA. Intracellularly injected guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) depressed both warm- and transmitter-induced K+ responses immediately after the injection. Intracellular application of guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) caused a gradual, irreversible increase in K+ conductance of the plasma membrane and occluded both responses. Transmitter-induced response markedly decreased when the temperature was raised from 22 to 32 degrees C, suggesting that the response to transmitter was occluded during the warm-induced response. These results suggested that the G-protein regulating the receptor-operated K+ channels could be activated simply by raising temperature.
Acetylcholine↗
A slow voltage-dependent Na(+)-current induced by 5-hydroxytryptamine and the G-protein-coupled activation mechanism in the ganglion cells of Aplysia.
Application of 5-hydroxytryptamine (5HT) induces a slowly depolarizing response in the neurons of Aplysia abdominal ganglion. In voltage-clamped cells, 5HT induced a slow inward current that increased steeply with membrane depolarization from -85 mV showing a negative slope conductance, but never reversed into outward when hyperpolarized beyond the equilibrium potential for K+. The 5HT-induced response was markedly augmented in Ca(2+)-free media, but depressed in Na(+)-free media, and unaffected by a change in external potassium. Intracellular injection of guanosine 5'-O-(2-thiodiphosphate) (GDP beta S) significantly depressed the 5HT response in a dose-dependent way. Injection of cholera toxin (CTX) selectively blocked the 5HT-induced response, the effect being irreversible. Neither 3'-deoxyadenosine, an inhibitor of adenylate cyclase, nor H-8, an inhibitor of protein kinase A, depressed the 5HT response. 3-Isobutyl-1-methylxanthine (IBMX) did not augment the 5HT response appreciably. The 5HT responses were not depressed at all during a saturated response to Br-cyclic AMP injected intracellularly. It was concluded that the 5HT response is produced by opening of the voltage-dependent Na(+)-channels with activation of CTX-sensitive G-protein but not necessarily with an increase in intracellular cyclic AMP.
1-Methyl-3-isobutylxanthine↗
[Clinical study on total intravenous anesthesia with droperidol, fentanyl and ketamine--11. Effect on epidural pressure].
We have developed a new method of total intravenous anesthesia using ketamine, fentanyl and droperidol (NLA-FK). There are many reports describing a significant increase in cerebrospinal fluid pressure (CSFP) during ketamine administration, but little is known about changes in CSFP during NLA-FK. As epidural pressure (EP) is considered as a good index for CSFP, we measured it in 12 patients under either NLA-FK or isoflurane anesthesia, who underwent gastrectomy. In the NLA-FK group the EP increased significantly by 26% at the induction of anesthesia as compared with the preinduction level, and it decreased to the preinduction level 30 minutes after the induction. Thereafter the pressure was not appreciably changed. In the isoflurane group EP significantly decreased by 16% at induction, but the pressure significantly increased by 29% 30 minutes after the induction as compared with the preinduction level. The change was similar to that of the NLA-FK group. Although the increase in the EP during the induction of NLA-FK is significant, it is considered to be within normal ranges. However, further detailed clinical study is needed to attenuate the significant increase in the EP during the induction of NLA-FK.
Anesthesia, Intravenous↗