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Biomedical subjects

H Ohuchi

Publications and source records attributed to H Ohuchi.

At least 37 records · Page 2Linked to original sources

[Aortic regurgitation caused by the proximal dissecting flap invagination to the left ventricle].

A 68-year-old male with sudden back pain and cardiogenic shock status transferred to our ward. Transthoracic echocardiography revealed that the abnormal round shape string was in the left ventricular outflow tract. The continuity from the staring to the aortic valve was unclear. Intimal flap could not be detected at the level of the ascending aorta. Color Doppler flow imaging showed that the severe AR jet extended into the round string. TEE showed that the intimal tear and flap was seen just above the left subclavian artery. Preoperative diagnosis was acute Stanford type A dissection and acute severe AR due to the inversion of the proximal intimal flap to the left ventricular outflow tract through the aortic valve. At operation, the proximal intimal flap was dissected circumferentially and was cut all the way around 8 cm above the aortic valve ring and was inverted to the left ventricular outflow tract. The aortic valve was preserved because of its normal character after exclusion of the proximal intimal flap. Ascending and arch replacement was carried out. Postoperative TEE and TTE slowed no findings of AR. The patient's postoperative course was uneventful. To our knowledge, this is the first reported case that severe AR caused by the proximal intimal invagination to the left ventricle.

Acute Disease

[Aprotinin and recombinant human erythropoietin reduce the need for homologous blood transfusion in cardiac surgery].

The effects of low dose aprotinin (Trasylol) and preoperative administration of recombinant human erythropoietin (EPO) were evaluated in 144 patients undergoing cardiopulmonary bypass divided into four groups. Group I (n = 43) received a subcutaneous administration of EPO (18,000 U) one week before operation and intraoperative administration of low-dose aprotinin (mean; 1.38 +/- 0.26 x 10(6) kallikrein inactivator units; KIU) from extracorporeal circulation, group II (n = 39) received only preoperative administration of EPO, group III (n = 28) received only intraoperative administration of low-dose aprotinin (mean; 1.46 +/- 0.25 x 10(6) KIU), and group IV (n = 34) were not administered either drug. Compared with group IV, the intraoperative blood loss was significantly lower in group I (p < 0.01), and in group II or III (p < 0.05). The postoperative drainage in 24 hours was significantly lower in groups I and III receiving aprotinin than in the other groups. The mean volume of total homologous blood transfusion and the percentage of cases not requiring a homologous blood transfusion in each group was, respectively, 74 +/- 235 ml and 88.4% in group I, 282 +/- 1289 ml and 87.2% in group II, 414 +/- 584 ml and 60.7% in group III, and 976 +/- 1931 ml and 44.1% in group IV. Significant differences were recognized between group I and group IV (p < 0.05). These findings indicate that when used in combination, both drugs reduce blood loss and the need for a homologous blood transfusion more effectively than either drug alone.

Aged

Correlation of wing-leg identity in ectopic FGF-induced chimeric limbs with the differential expression of chick Tbx5 and Tbx4.

It has been reported that members of the fibroblast growth factor (FGF) family can induce additional limb formation in the flank of chick embryos. The phenotype of the ectopic limb depends on the somite level at which it forms: limbs in the anterior flank resemble wings, whereas those in the posterior flank resemble legs. Ectopic limbs located in the mid-flank appear chimeric, possessing characteristics of both wings and legs; feather buds are present in the anterior halves with scales and claws in the posterior halves. To study the mechanisms underlying the chimerism of these additional limbs, we cloned chick Tbx5 and Tbx4 to use as forelimb and hindlimb markers and examined their expression patterns in FGF-induced limb buds. We found that Tbx5 and Tbx4 were differentially expressed in the anterior and posterior halves of additional limb buds in the mid-flank, respectively, consistent with the chimeric patterns of the integument. A boundary of Tbx5/Tbx4 exists in all ectopic limbs, indicating that the additional limbs are essentially chimeric, although the degree of chimerism is dependent on the position. The boundary of Tbx5/Tbx4 expression is not fixed at a specific position within the interlimb region, but dependent upon where FGF was applied. Since the ectopic expression patterns of Tbx5/Tbx4 in the additional limbs are closely correlated with the patterns of their chimeric phenotypes, it is likely that Tbx5 and Tbx4 expression in the limb bud is involved in determination of the forelimb and hindlimb identities, respectively, in vertebrates.

Amino Acid Sequence

Sympathetic reinnervation demonstrated on serial iodine-123-metaiodobenzylguanidine SPECT images after cardiac transplantation.

The transplanted heart is without autonomic nervous control in the early postsurgical period. We present here a case of cardiac transplantation in which 123I-metaiodobenzylguanidine (MIBG) SPECT and an exercise-loading test were used to monitor the sympathetic reinnervation. The distribution of myocardial 123I-MIBG uptake extended with time from 1 to 2 yr after surgery. However, functional improvement, estimated by the heart rate response to exercise, was not discernable during this period. The findings in this case suggest the feasibility of 123I-MIBG SPECT imaging in the serial monitoring of sympathetic reinnervation after transplantation and that scintigraphic evidence of reinnervation precedes functional recovery.

3-Iodobenzylguanidine

Differential expression of the two closely related LIM-class homeobox genes LH-2A and LH-2B during limb development.

We have isolated the chicken homeobox genes LH-2A and LH-2B encoding two related LIM domain-containing homeodomain proteins and examined the expression pattern during chick limb development. LH-2A is most closely related to human and rat LH-2, while LH-2B is less conserved. Although both LH-2A and LH-2B are expressed in the limb mesenchyme throughout stage 16 to stage 32, LH-2A transcripts are detectable in the distal limb bud and LH-2B transcripts are detectable in the anterior limb bud. Signals from the apical ectodermal ridge positively regulate LH-2A expression, since removal of the apical ectoderm resulted in the rapid reduction of LH-2A expression in the distal limb mesenchyme. Ectopic expression of the sonic hedgehog gene in the anterior margin of the limb bud resulted in the rapid reduction of LH-2B expression accompanying respecification of the positional value to the posterior phenotype. These results suggest that LH-2A and LH-2B play important roles in the determination and specification of the proximal-distal and anterior-posterior positional values, respectively.

Amino Acid Sequence

Fibroblasts expressing Sonic hedgehog induce osteoblast differentiation and ectopic bone formation.

We investigated the role of Sonic hedgehog (SHH) in osteoblast differentiation and bone formation. The numbers of ALP-positive cells in the mouse fibroblastic cell line C3H10T1/2 and the mouse osteoblastic cell line MC3T3-E1 were increased by co-culture with chicken fibroblasts transfected with chicken Shh cDNA encoding amino-terminal peptide (Shh-N). The conditioned medium of Shh-N-RCAS-transfected chicken fibroblast cultures also significantly increased ALP activity in both C3H10T1/2 and MC3T3-E1 cells. Intramuscular transplantation of Shh-N-RCAS-transfected chicken fibroblasts into athymic mice induced ectopic bone formation. These results indicate that SHH induces osteoblast differentiation and ectopic bone formation.

3T3 Cells

Cardiorespiratory responses to exercise after repair of the univentricular heart.

The purpose of this study is to evaluate cardiorespiratory responses to exercise in patients with univentricular heart according to the type of repair used. Forty-three patients with univentricular heart were divided into three groups: 15 preoperative patients (group A), 18 who had Fontan repair (group B) and 10 who had ventricular septation (group C). Group C was further divided into two subgroups, 7 with normal atrioventricular valve function (group C1) and 3 with atrioventricular valve regurgitation (group C2). Cardiorespiratory variables were determined after performance of cardiopulmonary exercise testing. One-hundred-and-twenty-five healthy subjects, age 5-26 years, served as controls. Oxygen uptake in group C1 at both ventilatory threshold and peak exercise was highest in all groups of univentricular heart (P < 0.05), while peak oxygen uptake in group C1 was significantly lower vs controls (P < 0.001), and that in group B was significantly higher than that for group A. Although chronotropic incompetence was noted in all groups of univentricular heart, marked improvements in both the relationship between heart rate and oxygen uptake and in the ventilatory efficiency were observed after definitive repair. While ventilatory efficiency was still impaired in group B, there was no significant difference between that in group C1 and the control group. When patients with univentricular heart of the left ventricular type (Van Praagh's type A single ventricle) were analyzed separately, superior cardiorespiratory response after ventricular septation was also found. In view of these findings, the ventricular septation procedure is preferred to the Fontan method in patients with univentricular heart when morphological conditions are suitable for this procedure so as not to make residual complications, such as significant atrioventricular valve regurgitation.

Adolescent

Lay-open pulmonary arterioplasty for postoperative hilar pulmonary artery stenosis.

OBJECTIVE: Lay-open pulmonary arterioplasty, a novel surgical technique to enlarge postoperative stenosis at the hilar pulmonary artery, was evaluated. METHODS: Lay-open arterioplasty, in which the enlarged hilar stenotic pulmonary artery is partially made up of previous surgical scar tissue instead of being covered by a patch, was performed on 10 patients whose ages ranged from 2.2 to 15.7 years. Surgical results were assessed by angiography. RESULTS: All patients tolerated the procedure without bleeding or embolic complications associated with pulmonary arterioplasty. Nine patients underwent concomitant procedures including total repair (n = 5), central interposing shunt (n = 3), and right ventricular outflow tract reconstruction (n = 1). No deaths or life-threatening events occurred during the total follow-up period of 18 patient-years. The stenotic segment was significantly enlarged from the preoperative diameter of 0.9 +/- 1.1 mm (mean +/- standard deviation) to the postoperative diameter of 8.0 +/- 1.3 mm, values which correspond to 7.0% +/- 8.8% and 68.4% +/- 11.5% of the normative values, respectively. A follow-up angiogram (n = 5) revealed an increase in the pulmonary artery diameter balanced with somatic growth (initial value, 65.2% +/- 9.0% of normal; second value, 69.1% +/- 7.7% of normal). No aneurysms or clinically significant restenoses were seen on the angiograms. CONCLUSIONS: Our initial midterm results with this method were promising. The pulmonary arteries subjected to this procedure grew in proportion to somatic growth.

Angioplasty

A chick wingless mutation causes abnormality in maintenance of Fgf8 expression in the wing apical ridge, resulting in loss of the dorsoventral boundary.

We analyzed a Japanese chick wingless mutant (Jwg) to know a molecular mechanism underlying wing development. We observed expression patterns of eleven marker genes to characterize the mutant. Expressions of dorsoventral (DV) and mesenchymal marker genes were intact in nascent Jwg limb buds. However, expression of Fgf8, a marker gene for the apical ectodermal ridge (AER), was delayed and shortly disappeared in the wing regressing AER. Later on, ventral expression of dorsal marker genes of Wnt7a and Lmx1 indicated that the wing bud without the AER became bi-dorsal. In addition, the posterior mesoderm became defective, as deduced from the impaired expression patterns of Sonic hedgehog (Shh), Msx1, and Prx1. We attempted to rescue a wing by implanting Fgf8-expressing cells into the Jwg wing bud. We found that FGF8 can rescue outgrowth of the wing bud by maintaining Shh expression. Thus, the Jwg gene seems to be involved in maintenance of the Fgf8 expression in the wing bud. Further, it is suggested that the AER is required for maintenance of the DV boundary and the polarizing activity of the established wing bud.

Animals

Theory and computer programs for calculating solution pH, buffer formula, and buffer capacity for multiple component system at a given ionic strength and temperature.

PURPOSE: A theory and computer programs running on Microsoft Excel for Windows for calculation of solution pH, buffer formula, and buffer capacity at a given ionic strength and temperature were developed. The theory does not limit the category of buffer components, the number of buffer components, or the number of ionizations for each buffer component. The usefulness of the programs was examined. METHODS: The formulas for 7 single component buffer solutions and 2 multiple component buffer solutions composed of citrate, phosphate, Tris, borate, and glycine were calculated. The solution pH values were measured at 25, 40, 55 and 70 degrees C for comparison with the calculated pH values. RESULTS: Of the 108 predictions made, 96 were of pH values with +/- 0.1 pH unit of the measured values, at temperatures ranging from 25 degrees C to 70 degrees C and at ionic strengths ranging from 0.1 M to 0.5 M. CONCLUSIONS: These programs will be useful for identifying appropriate buffer solutions at various temperatures and/or ionic strengths.

Buffers

Effect of ionic strength on solution stability of PNU-67590A, a micellar prodrug of methylprednisolone.

PURPOSE: PNU-67590A is a water-soluble micellar prodrug of methylprednisolone (MP). The major products of degradation of PNU-67590A are MP by hydrolysis and methylprednisolone 17-suleptanate (17-E) by 21-->17 acyl migration. The effect of ionic strength on micelle formation and stability of PNU-67590A in aqueous solution was examined. METHODS: PNU-67590A solutions at pH 2 and 8 and ionic strength of 0.05, 0.1, 0.2, and 0.4 M were maintained at 25 degrees C in the dark to measure MP and 17-E levels over time. RESULTS: The rate of degradation of micellar PNU-67590A at pH 8 was less than that of monomeric PNU-67590A, and vice versa at pH 2. Increase in ionic strength decreased both the critical micelle concentration of PNU-67590A and the degradation of micelle PNU-67590A at both pHs, resulting in improved overall stability of PNU-67590A. CONCLUSIONS: Formulation of PNU-67590A in a concentrated solution with high ionic strength will maximize stability and shelf-life.

Drug Stability

The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor.

Vertebrate limb formation has been known to be initiated by a factor(s) secreted from the lateral plate mesoderm. In this report, we provide evidence that a member of the fibroblast growth factor (FGF) family, FGF10, emanates from the prospective limb mesoderm to serve as an endogenous initiator for limb bud formation. Fgf10 expression in the prospective limb mesenchyme precedes Fgf8 expression in the nascent apical ectoderm. Ectopic application of FGF10 to the chick embryonic flank can induce Fgf8 expression in the adjacent ectoderm, resulting in the formation of an additional complete limb. Expression of Fgf10 persists in the mesenchyme of the established limb bud and appears to interact with Fgf8 in the apical ectoderm and Sonic hedgehog in the zone of polarizing activity. These results suggest that FGF10 is a key mesenchymal factor involved in the initial budding as well as the continuous outgrowth of vertebrate limbs.

Amino Acid Sequence

Pax-6 is involved in the specification of hindbrain motor neuron subtype.

Pax-6 is a member of the vertebrate Pax gene family, which is structurally related to the Drosophila pair-rule gene, paired. In mammals, Pax-6 is expressed in several discrete domains of the developing CNS and has been implicated in neural development, although its precise role remains elusive. We found a novel Small eye rat strain (rSey2) with phenotypes similar to mouse and rat Small eye. Analyses of the Pax-6 gene revealed one base (C) insertion in an exon encoding the region downstream of the paired box of the Pax-6 gene, resulting in generation of truncated protein due to the frame shift. To explore the roles of Pax-6 in neural development, we searched for abnormalities in the nervous system in rSey2 homozygous embryos. rSey2/rSey2 exhibited abnormal development of motor neurons in the hindbrain. The Islet-1-positive motor neurons were generated just ventral to the Pax-6-expressing domain both in the wild-type and mutant embryos. However, two somatic motor (SM) nerves, the abducent and hypoglossal nerves, were missing in homozygous embryos. By retrograde and anterograde labeling, we found no SM-type axonogenesis (ventrally growing) in the mutant postotic hindbrain, though branchiomotor and visceral motor (BM/VM)-type axons (dorsally growing) were observed within the neural tube. To discover whether the identity of these motor neuron subtypes was changed in the mutant, we examined expression of LIM homeobox genes, Islet-1, Islet-2 and Lim-3. At the postotic levels of the hindbrain, SM neurons expressed all the three LIM genes, whereas BM/VM-type neurons were marked by Islet-1 only. In the Pax-6 mutant hindbrain, Islet-2 expression was specifically missing, which resulted in the loss of the cells harboring the postotic hindbrain SM-type LIM code (Islet-1 + Islet-2 + Lim-3). Furthermore, we found that expression of Wnt-7b, which overlapped with Pax-6 in the ventrolateral domain of the neural tube, was also specifically missing in the mutant hindbrain, while it remained intact in the dorsal non-overlapping domain. These results strongly suggest that Pax-6 is involved in the specification of subtypes of hindbrain motor neurons, presumably through the regulation of Islet-2 and Wnt-7b expression.

Animals

Essential roles of the winged helix transcription factor MFH-1 in aortic arch patterning and skeletogenesis.

Mesenchyme Fork Head-1 (MFH-1) is a forkhead (also called winged helix) transcription factor defined by a common 100-amino acid DNA-binding domain. MFH-1 is expressed in non-notochordal mesoderm in the prospective trunk region and in cephalic neural-crest and cephalic mesoderm-derived mesenchymal cells in the prechordal region of early embryos. Subsequently, strong expression is localized in developing cartilaginous tissues, kidney and dorsal aortas. To investigate the developmental roles of MFH-1 during embryogenesis, mice lacking the MFH-1 locus were generated by targeted mutagenesis. MFH-1-deficient mice died embryonically and perinatally, and exhibited interrupted aortic arch and skeletal defects in the neurocranium and the vertebral column. Interruption of the aortic arch seen in the mutant mice was the same as in human congenital anomalies. These results suggest that MFH-1 has indispensable roles during the extensive remodeling of the aortic arch in neural-crest-derived cells and in skeletogenesis in cells derived from the neural crest and the mesoderm.

Animals

Alveolar-arterial gas tension differences during progressive exercise in patients after the Fontan operation.

We investigated alveolar-arterial gas tension differences as a measurement of ventilatory impairment during exercise in patients who had undergone the Fontan operation. The ventilatory response to exercise in 13 operated patients was compared with that of 11 control subjects. The difference between end-tidal and arterial oxygen tension (P(ET-a)DO2) and between arterial and end-tidal carbon dioxide tension (P(a-ET)DCO2) as well as the physiologic dead space-tidal volume ratio were calculated during progressive treadmill exercise testing. In the Fontan group, P(ET-a)DO2 and P(a-ET)DCO2 were significantly higher than in control subjects and increased in parallel during the study period. The physiologic dead space-tidal volume ratio was higher in the Fontan group than in the control group, and the difference in these ratios between the values obtained using end-tidal carbon dioxide tension and those using arterial carbon dioxide tension correlated well with P(a-ET)DCO2 (r = -0.79 to -0.98, p < 0.001). The physiologic dead space-tidal volume ratio during exercise was significantly higher in patients with low exercise capacities than in those with high exercise capacities (p < 0.05). The alveolar-arterial gas tension differences during exercise were greater in Fontan patients than in control subjects. We conclude that the size of the physiologic dead space can be evaluated from measurements of arterial carbon dioxide tension and is correlated with impaired exercise capacity after the Fontan operation.

Adolescent

[Constrictive pericarditis complicated with hepatic coma--a case report].

A 39-year-old man with pedal edema and icterus was admitted to our hospital. Laboratory findings revealed hyperbilirubinemia. Echocardiography and chest CT revealed the calcified mass front of the right ventricle. As cardiac catheterization showed dip and plateau in right ventricle, the diagnosis of constrictive pericarditis was established. Considering his condition and bleeding tendency with hepatic failure, plasma exchange was done for three days before operation. As the patient's condition deteriorated with hepatic coma, pericardiectomy was performed. The median sternotomy approach without cardiopulmonary bypass was selected. The calcified mass was removed, and the thickened pericardium was excised. Postoperative hemodynamic findings were improved, and the serum bilirubin returned to normal. In conclusion even in this case with severe hepatic failure due to constrictive pericarditis, surgical treatment should have be considered.

Adult

[Surgical outcomes of transverse aortic arch replacement].

The surgical outcomes of transverse aortic arch replacement were analyzed for the cases of 33 consecutive patients who underwent operations during the seven years between July 1989 and March 1996. Sixteen patients were atherosclerotic aneurysm (group T), 17 patients were acute or chronic aortic dissection involving aortic arch (group D). As for brain protection during aortic arch reconstruction, selective cerebral perfusion was employed in 16 patients of group T and 11 patients of group D. In 6 patients of group D with acute type A aortic dissection or chronic type B dissection, deep hypothermic circulatory arrest at 18 approximately 20 degrees C of rectal temperature was employed. In 14 patients of group T, selective cerebral perfusion was simultaneously started when we begun cardiopulmonary bypass. There were one operative death in group T patients (operative mortality 6.2%) and two in group D patients (operative mortality 11.8%). Perioperative stroke was occurred in two patients of group T with complete recovery in one and with partial recovery in the other. The latter patient died of medistinitis and graft infection. There was no hospital death nor stroke in group D patients. Duration of selective cerebral perfusion was 157.8 +/- 54.4 minutes in group T patients, 140.1 +/- 66.5 minutes in group D patients (n = 11), duration of circulatory arrest was 49.5 +/- 11.4 minutes in 6 patients of group D patients, respectively. In 27 patients who underwent operation with selective cerebral perfusion, morbidity of stroke was 7.4% (two patients). These were caused by technical failure during arch vessel reconstruction and seemed to be avoidable. In conclusion, it is reasonable to employ selective cerebral perfusion for aortic arch reconstruction in atherosclerotic aneurysm of transverse aortic arch. In aortic dissection, either selective cerebral perfusion or deep hypothermic circulatory arrest is justified as cerebral protection during operation.

Adult

Sonic hedgehog is expressed in epithelial cells during development of whisker, hair, and tooth.

Sonic hedgehog (Shh) is a vertebrate gene homologous to a Drosophila segment polarity gene, hedgehog, and functions as a secreted signaling molecule in limb pattern formation, differentiation of motor neurons, and sclerotome induction. We found that Shh is also expressed in epithelia of the developing whisker, hair, tooth, rugae, and thyroid primordium of mouse embryos. In whisker and hair development, Shh is expressed in epithelial cells before condensation of the underlying mesenchymal cells and then in the placode. The expression of Shh continues in the hair bulb surrounding the dermal papilla. Shh is also expressed in epithelial cells of the tooth bud, then localized to the enamel knot. The Shh expression continues in developing ameloblasts. These results suggest that SHH is an essential epithelial signaling molecule in epithelio-mesenchymal interactions for the terminal differentiation.

Animals