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PubMed1989

Stability of the transformants obtained by phage particle-mediated gene transfer.

Recombinant lambda phage DNA, encapsulated in phage particles and coprecipitated with calcium phosphate, efficiently transforms cultured mammalian cells without a requirement for carrier DNA. The present paper analyzes the stability of the transformants obtained by the phage transfer method. lambda phage particles containing recombinant DNA that includes the thymidine kinase (TK) gene of herpes simplex virus type 1 as a selective marker were introduced into Ltk- cells deficient in TK activity, and TK+ transformants were selected in HAT medium. To test the stability of the TK+ phenotype of the transformants, seven individual transformant clones were isolated, cultured in HAT selective medium and then in non-selective medium for various lengths of time. After such culture, transformants were allowed to develop colonies in both selective and non-selective medium. For all seven transformant clones, the numbers of colonies obtained in the two types of medium were almost identical, irrespective of whether or not each transformant clone had been previously cultured for 15 to 50 days in non-selective medium. This result suggests that most transformants obtained by the phage transfer method maintain the TK+ phenotype stably, for at least 50 days, when grown in non-selective medium.

Animals↗
PubMed2005

Simple and noninvasive breath test using 13C-acetic acid to evaluate gastric emptying in conscious rats and its validation by metoclopramide.

The (13)C-breath test has been used to clinically evaluate gastric emptying. However, this method has not been sufficiently validated in experimental animals. The present study aimed to establish a simple and noninvasive (13)C-breath-test system in Sprague-Dawley male rats. After fasting, rats were orally administered Racol containing (13)C-acetic acid and housed in a desiccator. The expired air in the chamber was collected in a breath-sampling bag using a tube and aspiration pump. The level of (13)CO2 in the expired air was measured using an infrared spectrometer at appropriate intervals for 120 min. During this period, the rate of (13)CO2 excretion increased, peaked, and decreased thereafter. The maximum concentration (Cmax) and area under the curve (AUC120 min) of (13)CO2 excretion increased in volume- and dose-dependent manners. The time taken to reach the maximum concentration (Tmax) of (13)CO2 excretion increased as the volume increased, but was not affected by the dose of (13)C-labeled acetic acid. Metoclopramide dose-dependently increased the Cmax and shortened Tmax of (13)CO2 excretion compared with those of the control rats, whereas the AUC120 min was not affected. These results confirm that this simple method can successfully evaluate gastric emptying. Moreover, this system is suitable for investigating additional physiological functions using other labeled compounds.

Acetic Acid↗
PubMed1998

Invasion beyond interlobar pleura in non-small cell lung cancer.

STUDY OBJECTIVE: To assess the outcome of lung cancer with invasion beyond interlobar pleura and to clarify whether it should be treated in the same way as invasion to the parietal pleura or to other visceral pleura. DESIGN: Retrospective analysis. SETTING: Tokyo Medical College Hospital. PATIENTS: Eighteen resected non-small cell lung cancers with invasion beyond interlobar pleura were studied. The outcomes of those patients, those with parietal pleural invasion, and those with other visceral pleural invasion were compared. Patients with rib invasion, mediastinal organ invasion, or distant metastasis were excluded. RESULTS: The 5-year survival rate for patients with invasion beyond interlobar pleura was 34.2% and the median survival time was 56.5 months. The outcome was significantly better than that of patients with parietal pleural invasion. There was no significant difference between the outcome of invasion beyond interlobar pleura and that of other visceral pleural invasion. In patients without lymph node metastasis, similar results were obtained. There was no difference between the outcome of patients with invasion beyond interlobar pleura, who undergo lobectomy with a parietal resection of the invaded lobe, and that of patients with visceral pleural invasion, who undergo lobectomy. CONCLUSIONS: The behavior of patients with invasion beyond interlobar pleura is different from that of patients with parietal pleural invasion and should be categorized as T2. The optimum operative method was lobectomy with only parietal resection of the invaded lobe to preserve the pulmonary function.

Aged↗
PubMed1991

[Pars plana lensectomy preserving a clear anterior capsule in vitreous surgery].

The anterior capsule was preserved intact in pars plana lensectomy in 14 cases of pars plana vitrectomy carried out for proliferative diabetic retinopathy in 9 cases, sarcoid uveitis in one case, proliferative vitreo-retinopathy in 3 cases and retinal detachment with a giant break in one case. Follow-up periods ranged from 6 to 17 months, with a mean of 11 months. Simultaneous posterior chamber intraocular lens implantation anterior to the anterior capsule was performed in 7 cases, and in one case secondary implantation was performed 4 months later. In 11 cases anterior capsules remained clear and in 9 of them lens epithelial cells had been thoroughly removed up to the equator using a Terry squeegee. Posterior synechia occurred in 4 cases in which air or gas tamponade had been done. This method allows complete anterior vitrectomy and is adaptable not only to simultaneous but also secondary posterior chamber intraocular lens implantation.

Adolescent↗
PubMed1989

[A case of gall bladder cancer with high level alpha-fetoprotein].

A case of primary gall bladder tumor with high level AFP were demonstrated, which showed hepatocellular carcinoma like pattern with trabecular proliferation histologically, positive AFP stain of cancer cells by a peroxidase conjugated antibody method immunohistochemically. This case was no metastasis with liver and lymph nodes.

Aged↗
PubMed1981

[Significance of non-invasive parameter (PSP/ESVI) in assessing left ventricular contractility (author's transl)].

Since myocardial pump function is affected by preload or afterload, it is difficult to assess left ventricular (LV) contractility independently from these in clinical cardiology. In this study, we performed inotropic intervention by isoproterenol or propranolol, and changed afterload by angiotensin or nitroprusside, to estimate the usefulness of peak systolic pressure/end-systolic volume index (PSP/ESVI) as a parameter of LV contractility. The data were obtained by cuff method or catheter-tip manometer and echocardiography. We calculated PSP/ESVI in various heart diseases, and results were analyzed for a significant change using the t test. Ejection fraction, mean VCF (mVCF) were calculated simultaneously, and compared with PSP/ESVI. The results were as follows: 1) PSP/ESVI ratio significantly reflected inotropic intervention with isoproterenol and propranolol. 2) PSP/ESVI ratio was less affected by the change of peak systolic pressure than EF or mVCF. 3) There was a significant difference in PSP/ESVI among congestive cardiomyopathy, valvular heart diseases and normal controls. 4) There was also a significant difference in PSP/ESVI between patients of NYHA I and those of NYHA II--IV. 5) PSP/ESVI ratio, measured by non-invasive method, was thought to be a useful parameter to assess LV contractility.

Adolescent↗
PubMed1993

[Prophylactic combination therapy after TUR of superficial bladder cancer].

We evaluated 63 patients with superficial bladder cancer (pTa, pTl) who were treated with instillation of bleomycin +/- bacillus Calmette-Guerin (BCG) and administration of uraciltfutraful (UFT) for prophylaxis of tumor recurrence after transurethral resection (TUR). The patients were randomly assigned to groups A and B after transurethral resection by the closed envelope method. Group A (34 cases) was designed as continuous diluted intravesical instillation of bleomycin (120 mg/2,000 ml saline solution/day repeated for 3 days), instillation BCG (40 mg/40 ml saline solution/weekly 6 times) and UFT (400 mg orally/day for 2 years maximum). Group B (29 cases) was designed as the aforementioned minus BCG instillation. Cumulative non-recurrence rates in group A and group B were 80.1% and 72.1% at the time of three years, which revealed no significant difference between the two groups (p = 0.265, generalized Wilcoxon test). The high recurrent incidence of superficial bladder cancer is primarily due to the multifocal nature of the cancer or implantation of tumor cells at the time of the subsequent transurethral resection. The procedure was performed safely with no severe side effects. Our method might be useful to reduce the recurrences of superficial bladder cancer after transurethral resection of bladder (TUR).

Administration, Intravesical↗
PubMed1995

[Phosphoglyceric acid mutase].

Human phosphoglyceric acid mutase comprises M-, B- and MB-type isozymes composed of the combination of the muscle-specific (M) and nonmuscle-specific (B) subunits. Human DNAs encoding M and B subunits were respectively reconstructed at their 5' regions without changing the amino acid sequences, and expressed directly in Escherichia coli under controls of the trp promoter. M- and B-type isozymes were over-produced in the bacterial cytoplasm as soluble, active forms, which have been purified and characterized. MB-type was synthesized in vitro by recombining M- and B-type. All the three recombinant isozymes thus obtained were the same in properties tested as the naturally-occurring ones. Polyclonal IgGs specific to the M-type, B-type and MB-type isozymes were prepared from rabbits immunized with the respective isozymes mainly by treating with the columns bound with the M- or B-type isozyme. A method for the immunoassay of the MB-type isozyme which exists specifically in cardiac muscle, is now under development.

Animals↗
PubMed2003

Long-, intermediate- and short-acting benzodiazepine effects on human sleep EEG spectra.

The effects of 10 mg haloxazolam (HAX), 4 mg flunitrazepam (FNZ), and 0.5 mg triazolam (TRI), each administered for seven consecutive nights were studied in the sleep electroencephalograms (EEG) of 17 (six HAX, five FNZ and six TRI) healthy male student volunteers. Recordings of C3-A1 EEG data from one baseline night, three drug nights (first, fourth and seventh) and two withdrawal nights (second and fourth) were analyzed using a fast Fourier transformation method. All three drugs induced similar changes in the 0.5 Hz to 40 Hz power spectrum; namely (i) higher frequency (including the sigma and beta bands) activity increased and lower frequency activity reduced on the drug nights; (ii) the enhancement of sigma activity peaked during non-rapid eye movement sleep following the first administration and was maintained at high levels on all drug nights; (iii) beta activity increased through the night after administration of HAX and FNZ, but not TRI, which suggests a blood concentration level dependent increase of beta activity; and (iv) only HAX showed a residual effect on the fourth withdrawal night. These results indicate that (i) chronic administration of these three benzodiazepine derivatives produce similar profiles in sleep EEG spectral changes, with some differences depending on their half-lives and doses; (ii) the mechanism of sigma enhancement is sensitive to even the initial administration night of BDZ; and (iii) frequencies below and above the sigma band are less sensitive to BDZ and also show an increase through the night after administration, suggesting differences in the mechanisms reflected by these EEG frequency bands.

Adult↗
PubMedDate not supplied

Endovascular placement of an extraluminal femoropopliteal bypass graft in human cadavers.

PURPOSE: A method to create an extraluminal femoropopliteal bypass graft using endovascular techniques was evaluated in situ on cadaver extremities in an attempt to develop a minimally invasive alternative technique for the management of infrainguinal occlusive arterial disease. METHODS: The endovascular placement of an extraluminal femoropopliteal bypass graft was undertaken in 5 cadaver legs. Following percutaneous access to the popliteal artery (PA) or common femoral artery (CFA), a Rosch-Uchida needle was used to perforate the vascular wall, followed by the creation of an extraluminal tract using a looped wire and catheter. Once the desired level was reached the needle was again used to perforate the vascular wall of the proximal superficial femoral artery (SFA) or PA depending on the access used. Self-expanding expanded polytetrafluoroethylene (ePTFE) stent-grafts were then deployed to establish the extraluminal femoropopliteal bypass connecting the two arterial puncture sites. Following dilatation of the stent-graft, angiography was performed to assess the endoprostheses and to look for contrast leaks. RESULTS: Technical success was achieved in all 5 legs. Procedure time varied from 15 to 30 min. The angiographic studies performed immediately after completion of the bypass procedure showed patency of the grafts with no evidence of kinking or leakage in any of the cases. CONCLUSION: This study has proved that the endovascular placement of an extraluminal femoropopliteal bypass graft in human cadaver legs using endovascular techniques under fluoroscopic control is technically feasible.

Angiography↗
PubMed1997

[A new surgical management of esophageal stenosis without esophagectomy].

In many cases, reflux esophagitis following surgical treatment for esophageal stenosis is caused by the recurrence of that after esophagectomy and esophogogastrostomy. We performed a new management without esophagectomy for a 66-year-old man with sliding hiatal hernia and esophageal stenosis induced by reflux esophagitis. A Expanding Metalic Stent (MES) was inserted to the stenotic portion of the esophagus, and then Collis-Nissen's procedure was done through left thoracotomy and phrenotomy. The postoperative course was satisfactory, and no gastroes-ophageal reflux was detected with the use of 24h pH-monitoring of the esophagus after surgery.

Aged↗
PubMed1999

Safety and efficacy of autologous platelet transfusion in cardiac surgery: comparison of cryopreservation, blood collection on the day before surgery, and blood collection during surgery.

In a group of 39 patients with ischemic heart and valvular disease (January 1997 to May 1998), three platelet collection methods were compared in terms of safety and effectiveness. The methods were: (i) collection of autologous platelets over several weeks and freezing them for storage until surgery (frozen group, 12 patients); (ii) collection of autologous platelets on the day before surgery and preserving them without freezing (fresh group, 8 patients); and (iii) collection of autologous platelets intraoperatively (intraoperative group, 9 patients). Ten patients served as controls (control group). Blood pressure was not significantly affected by platelet collection in the frozen and fresh groups, but both systolic (P < 0.01) and diastolic blood pressure (P < 0.05) decreased significantly after collecting platelets in the intraoperative group. Similarly, heart rate was unaffected by platelet collection in the frozen and fresh groups, while it increased significantly in the intraoperative group (P < 0.05). Blood loss after 24 h was significantly smaller in the fresh group than in the frozen group (P < 0.05). Total blood transfusion volume was significantly smaller in the frozen and fresh groups than in the intraoperative and control groups (P < 0.05). Bleeding time 2 h postoperatively, when administration of autologous platelets had been completed, was reduced compared with immediately postoperative values in all three groups receiving autologous platelets (P < 0.05). However, only the frozen and fresh groups showed a significantly shorter bleeding time than the control group (P < 0.05). In all three groups receiving autologous platelets, the platelet count was significantly increased after administration of autologous platelets, but only the fresh group had a platelet count that was significantly greater than the control group (P < 0.05). From these results we conclude that the frozen and fresh groups received safer treatment than the intraoperative group. Although hemostasis improved after all three regimes of autologous platelet transfusion, only the frozen and fresh groups had a reduced need for allogeneic blood transfusion compared with the control group. For this reason we conclude that the frozen and fresh groups were also superior to the intraoperative group in terms of effectiveness. However, the recovery of platelets after frozen storage was low, and to obtain a good effect with the freezing method it is necessary to collect and store large volumes of platelets. In terms of simplicity, safety, and efficacy, the fresh method seems to be the preferred technique.

Aged↗
PubMed1994

Molecular basis for subtypic differences of the "a" subunit of coagulation factor XIII with description of the genesis of the subtypes.

The "a" subunit of human coagulation factor XIII (F13A) exhibits genetic polymorphism defined by four common alleles, F13A*1A, *1B, *2A, and *2B. We have previously suggested on the basis of the isoelectric focusing patterns of the four allele products that point mutations at two separate sites and one intragenic crossing over might be involved in the genes of F13A polymorphism. Here, we report nucleotide substitutions associated with F13A polymorphism. A C/T transition of the second nucleotide of codon 564 in exon 12 is responsible for the difference between F13A*1A and *1B and that between F13A*2A and *2B, and a set of two base changes in codons 650 and 651 in exon 14 leads to the differences between F13A*1A and *2A and those between F13A*1B and *2B. The four combinations of the point mutations at the two exons thus correspond to the four alleles, two of which were generated by the point mutations from ancestral monomorphic gene. The results suggest strongly that intragenic crossing over must be involved in the genesis of the fourth allele. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods discriminating these base changes in exons 12 and 14 are also presented.

Alleles↗
PubMed1983

Determination of atrial size by esophageal echocardiography.

The sizes of both left atrial (LA) and right atrial (RA) cavities were assessed in 16 patients by esophageal echocardiography and biplane cineangiography. The changes in echocardiographic dimension and cineangiographic volume during 1 cardiac cycle showed excellent correlations in both atria. In the left atrium, the relation between the echocardiographic dimension and the cineangiographic volume was significant (r = 0.83) and was fitted by the following power function: LA volume (ml) = 0.94 X LA dimension (mm) 1.24. In the right atrium, the relation between the dimension and the volume was significant; RA volume (ml) = 0.015 X RA dimension (mm) 2.34 (r = 0.95). Thus, esophageal echocardiography is a useful method for evaluating LA and RA size and simultaneously observing of both atria.

Adult↗
PubMed1994

Molecular analysis of T cell receptor V beta chain to detect leukemia cell clonality in patients by adaptor ligation-mediated polymerase chain reaction.

We have developed a simple and rapid method to analyze the clonality of leukemia cells. After three rounds of amplification by adaptor-ligation polymerase chain reaction (PCR), the cDNA is cut with AluI, HaeIII, RsaI, and Sau3AI, and analyzed by polyacrylamide gel electrophoresis. The size of the restriction fragments is compared to that of the published restriction fragments size each TCR-beta subfamily V region. The sensitivity of adaptor-ligation PCR restriction enzyme analysis (AL-PCR-REA) was 10(-4) MOLT-4 T-ALL cell population in the normal peripheral blood lymphocytes (PBL). Application of AL-PCR-REA to PBL and bone marrow (BM) cells from eight clinical leukemia samples indicated that a detection sensitivity was rather low, but revealed the clonality of all eight clinical samples. This AL-PCR-REA method can detect clonality without the need for either radioisotopes or sequencing procedures.

Base Sequence↗
PubMed2006

Midterm results of extensive primary repair of the thoracic aorta by means of total arch replacement with open stent graft placement for an acute type A aortic dissection.

OBJECTIVES: We sought to describe the midterm results of extensive primary repair of the thoracic aorta by means of the modified elephant trunk technique with a stent graft for acute type A aortic dissection, particularly the changes of the false lumen shown by enhanced computed tomographic scanning. METHODS: The subjects were 35 consecutive patients who received arch replacement with open stent grafting for type A acute aortic dissection between December 1997 and April 2002. The mean follow-up period was 55 months (range, 30-83 months). Computed tomographic scanning was performed at 1, 3, 12, and 36 months postoperatively to detect thrombosis and obliteration of the false lumen after its exclusion by the stent graft. The diameter of the aorta was measured at 3 levels: the distal edge of the stent graft, the diaphragm, and the origin of the superior mesenteric artery. RESULTS: Two patients died in the initial operation, but no patients required additional surgical treatment of the thoracic aorta. The mean diameter of the stent grafts was 26.2 mm, and the mean length was 8.9 cm. Thrombus formation in the false lumen was recognized at the distal edge of the graft in all patients, at the diaphragmatic level in 26 patients, and at the superior mesenteric artery level in 15 patients. Obliteration of the false lumen was recognized at the distal edge of the graft in all patients, at the diaphragmatic level in 20 patients, and at the superior mesenteric artery level in 15 patients. The aorta distal to the stent graft showed minimal changes. CONCLUSIONS: In patients with acute type A aortic dissections, it is possible to perform extensive primary repair of the thoracic aorta with relative safety by using a synthetic graft with a self-expanding stent, and this method might reduce the necessity of further operations not only for the distal descending aorta but also for the thoracoabdominal aorta.

Aged↗