The human prohibitin gene located on chromosome 17q21 is mutated in sporadic breast cancer.

A gene called "prohibitin" was isolated as a candidate antiproliferating gene in rat liver cells. We have isolated the human homologue of the rat prohibitin gene and mapped it to chromosome 17q12-21 where a gene responsible for hereditary breast cancer was localized. DNA sequence analysis of 2 exons in this gene in 23 sporadic breast cancers, which showed loss of heterozygosity on the long arm of chromosome 17 or developed in patients 35 years old or younger, identified 4 cases of somatic mutation; 2 of these were missense mutations; 1 showed a 2-base deletion resulting in truncation of the gene product due to a frame shift; the other had a C to T transition in an intron adjacent to an intron-exon boundary. These results suggest that this gene may be a tumor suppressor gene and is associated with tumor development and/or progression of at least some breast cancers.

Correlation of loss of alleles on the short arms of chromosomes 11 and 17 with metastasis of primary breast cancer to lymph nodes.

To examine the role of loss of heterozygosity (LOH) during tumor development and/or progression, we looked for correlations between metastasis of breast cancer to a regional lymph node(s) and LOH of chromosomal arms 11p, 13q, 16q, 17p, and 17q, where frequent losses in primary tumors have been detected. No correlation between lymph node metastasis and LOH of chromosomes 13q, 16q, or 17q was observed. However, tumors showing LOH of chromosomes 11p (chi 2 = 10.82, P less than 0.01) and 17p (chi 2 = 6.78, P less than 0.01) revealed a significantly higher incidence of metastasis to a regional lymph node(s) than tumors without LOH on these chromosomal arms. Furthermore, only four of 30 (13%) patients with tumors that retained both 11p and 17p had metastasis to a regional lymph node(s), compared with 24 of 32 (75%) patients with tumors that had lost both 11p and 17p. Analysis of LOH with markers on chromosomes 11p and 17p in a large number of tumors indicated that the peritelomeric region of each of these chromosomal arms contains a tumor suppressor gene that may be associated with tumor progression, particularly metastasis to a regional lymph node(s).

Actions of opioids on respiratory activity via activation of brainstem mu-, delta- and kappa-receptors; an in vitro study.

Opioid-induced respiratory depression is well documented. However, exact sites of action and mechanisms for opioid-induced effects on respiration have not yet been elucidated. The present study was carried out on isolated brainstem-spinal cord preparations from newborn rats in order to explore the opioid activity on brainstem mu-, delta- and kappa-receptors. The brainstem-spinal cord was isolated from 0- to 4-day-old Sprague-Dawley rats. The preparation was perfused with artificial cerebrospinal fluid (28.5 degrees C) equilibrated with 95% O2 and 5% CO2 at a pH of 7.4. Neuronal respiratory activity was recorded from the ventrolateral part of the medulla oblongata and efferent impulses from C4 or C5 ventral roots. Effects of the mu-receptor agonist DAGO, the delta-receptor agonist DPDPE and the kappa-receptor agonist U50,488 on respiratory frequency (fR), inspiratory time (Ti) and peak integrated C4 amplitude (Int[C4]) were measured. In addition, the effect of pre-treatment with the mu1 receptor antagonist naloxanazine (35 mg/kg, subcutaneous injection) was evaluated. DAGO reduced fR and Ti in a concentration-dependent manner and caused a reduction of Int(C4) at high concentrations (10 microM). The mu1 receptor antagonist naloxanazine shifted the fR concentration-response curve for DAGO to the right (P < 0.05). DPDPE had no effect on respiratory activities whereas U50,488, like DAGO, reduced fR and Int(C4) in a concentration-dependent manner. It was concluded that mu-opioid receptors, including the mu1 were involved in fR reduction whereas kappa-opioid receptors were involved in reduction of both fR and respiratory amplitude. Delta-opioid receptors do not seem to participate in respiratory modulation at this age.

Age- and temperature-dependent effects of opioids on medulla oblongata respiratory activity: an in vitro study in newborn rat.

Influences of post-natal age (P0-P4) and temperature (22.5 degrees-31.5 degrees C) on the action(s) of opioids on respiratory activities from neonatal rat brainstem-spinal cord preparations were examined in this study. A temperature-dependent mu-opioid receptor effect on respiration was found. In addition, the effect of morphine increased with postnatal age (P0-P4). Hence, age and temperature must be taken into account when performing studies on medullary respiration-related structures using the neonatal rat brainstem-spinal cord preparation.

Effects of neuroactive substances on the morphine-induced respiratory depression; an in vitro study.

Effects of different neuroactive substances on morphine-induced respiratory depression were studied in medullary respiration-related structures using in vitro brainstem-spinal cord preparation from 1 to 4-day-old rats. Application of morphine (10 microM) reduced respiratory rhythm (fR) as measured by C4 ventral root activity. The depressant effects of morphine were reversed by acetylcholine (10 microM), substance P (50 nM), thyrotropin releasing hormone (TRH) (100 nM) and forskolin (10 microM). The adenosine receptor antagonist, theophylline (100 microM), the dopamine receptors antagonist, haloperidol (10 microM), the cyclooxygenase inhibitor, indomethacin (10 microM) and the phospholipase A(2) inhibitor, quinacrine (10 microM) had no effect on morphine-induced respiratory depression.

Immunofluorescence localization of type I and type III collagen and fibronectin in mouse dental tissues in late development and during molar eruption.

Affinity-purified antibodies produced intense staining for type I collagen in alveolar bone matrix and predentine, and moderate staining in the dentine matrix, lamina propria, connective tissue invaginating into papillary layer of the enamel organ, dental sac and periodontal ligament. No staining occurred in oral epithelium, stellate reticulum, stratum intermedium, ameloblasts and odontoblasts. Fibronectin was distributed similarly except at the interface between the epithelial diaphragm and pre-odontoblasts where type I collagen was absent but fibronectin was present. In contrast, type III collagen showed strong staining in the periodontal ligament and lamina propria but no staining in bone matrix, predentine, dentine and at the interface between the epithelial diaphragm and pre-odontoblasts. The staining pattern for type III collagen was similar to that of type I and fibronectin in other tissues including endosteal reticular tissue, the connective tissue invaginating into papillary layer and the extracellular matrix of the pulp.

Mutational analysis of TGF-beta type II receptor, Smad2, Smad3, Smad4, Smad6 and Smad7 genes in colorectal cancer.

Transforming growth factor-beta(TGF-beta) is known to play an important role in controlling embryonal development, cell proliferation and homeostasis. The purpose of this study is to elucidate the involvement of the TGF-beta pathway in colorectal carcinogenesis. DNA was extracted from 100 patients with colorectal cancer. Then, all coding regions of the TGF-beta type II receptor (TRII) and the genes for Smad2, Smad3, Smad4, Smad6, and Smad7 were analyzed by PCR-SSCP and direct sequencing. Also, a LOH analysis of 18q21, where the Smad2 and Smad4 genes are located, was performed. We detected 11 cases of frameshift mutation in the TRII gene (11%) and 5 cases of point mutations in the Smad4 gene (5.0%); LOH at 18q21 was detected with 33% frequency. No abnormalities were found in the genes for Smad2, Smad3, Smad6, and Smad7. These results suggest that the abnormalities of TRII and Smad4 play an important role inhibiting TGF-beta signaling in colorectal carcinogenesis.

Increased sensitivity to vecuronium and prolonged duration of its action in patients with end-stage renal failure.

STUDY OBJECTIVES: To determine whether the duration of action of vecuronium is influenced by chronic renal failure. DESIGN: Prospective clinical comparison. SETTING: Operating room, Hokkaido University Hospital. PATIENTS: 7 adult ASA physical status II and III patients with impaired renal function undergoing living-related renal transplantation (Group A), 5 adult ASA physical status II and III patients with impaired renal function undergoing elective surgery other than renal transplantation (Group B), and 13 adult ASA physical status I and II patients with normal hepatorenal function undergoing elective surgery (Group C). INTERVENTIONS: Anesthesia was induced with thiamylal 5 mg x kg(-1) and fentanyl 100 microg IV and maintained with 60% nitrous oxide and 1% isoflurane. An initial dose of vecuronium was administered and patients' tracheas were intubated. MEASUREMENTS: Neuromuscular function was monitored by acceleration of thumb adduction with train-of-four stimulation. Vecuronium 60 microg x kg(-1) was administered as the initial dose via a central catheter, and if the first twitch was more than 3% of the control, another dose of vecuronium 20 microg x kg(-1) was given as necessary. Both onset time and duration of action until 25% recovery were measured. Plasma vecuronium and its metabolite, 3-desacetyl-vecuronium, levels were measured at onset and at 25% recovery in Groups A and C. MAIN RESULTS: The total dose of vecuronium and initial concentration of vecuronium showed no significant difference between Group A and Group C. Duration of action was significantly prolonged in Group A and Group B compared with Group C. CONCLUSIONS: Duration of action of vecuronium is prolonged in patients with end-stage renal failure mainly due to higher sensitivity to vecuronium.

Serum and urine inorganic fluoride levels following prolonged low-dose sevoflurane anesthesia combined with epidural block.

STUDY OBJECTIVES: To determine whether serum and urine inorganic fluoride levels with prolonged (more than 7 hours) low-dose (0.8 to 2.0 vol %) sevoflurane anesthesia plus epidural anesthesia were increased as compared with isoflurane anesthesia plus epidural anesthesia. To measure the urine tubular enzymes N-acetyl-beta-glucosaminidase (NAG), alpha 1-microglobulin (alpha 1-M), and beta 2-microglobulin (beta 2-M) for renal tubular injury in both groups. DESIGN: Randomized, prospective study. SETTING: University hospital. PATIENTS: 15 ASA physical status I and II adults (7 males, 8 females) who were scheduled for prolonged laparotomy (lasting 9.5 to 10.2 hours) with general anesthesia. MEASUREMENTS AND MAIN RESULTS: Epidural anesthesia was administered before induction of general anesthesia. General anesthesia was induced with thiamylal administered intravenously (IV), and the trachea was intubated following administration of vecuronium IV. It was maintained with either sevoflurane or isoflurane in nitrous oxide and oxygen. Standard monitoring was used in all patients. Serum and urine inorganic fluoride and urine tubular enzymes were measured periodically. Serum inorganic fluoride was 54 mumol/L at 4.3 minimum alveolar concentration (MAC) hours of sevoflurane; the peak level for isoflurane was 8 mumol/L at the same MAC hours. Sevoflurane also increased urine inorganic fluoride excretion to 96 mumol/hr 8 hours. NAG excretion started to increase after inhalation of either sevoflurane or isoflurane. alpha 1-M and beta 2-M excretion increased markedly postoperatively. Even though fluoride levels and tubular enzymes were high, there was no evidence of postoperative renal dysfunction. CONCLUSIONS: There was no increase in urinary enzymes, which are indicators of tubular injury, specific to sevoflurane. There was no postoperative renal dysfunction, as indicated by unchanged serum creatinine and blood urea nitrogen levels.

[Fundamental and clinical studies on T-1982 (cefbuperazone) in the field of obstetrics and gynecology].

Fundamental and clinical studies on T-1982 (cefbuperazone) in the field of obstetrics and gynecology were carried out. Transfer of T-1982 to various location in uterus tissue was more than 10 micrograms/g over 2 hours after T-1982 1 g intravenous injection. T-1982 was distributed in cervix uteri at the highest concentration followed by ovarium, oviduct, portio vaginalis, endometrium and myometrium. Mean transfer ratio of cervix uteri to uterus arterial blood was 67.6%. Ten cases of gynecological infections receiving T-1982 demonstrated "good" results in 9 cases, except 1 case excluded from the evaluation of efficacy. Neither side effect nor clinical test abnormality was observed. Based on the results of basic and clinical studies, T-1982 is considered to have efficacy in the treatment of gynecological infections.

[Clinical studies on cefoperazone in the pediatric field (author's transl)].

Cefoperazone (CPZ) at dose levels of 80 approximately 100 mg/kg/day, divided 3 approximately 4 times, was drip-infused or intravenously injected for a period of 2 approximately 6 days to 10 patients. All 10 cases, 4 cases of bronchopneumonia from which H. influenzae was detected (Group A S. pyogenes and S. pneumoniae were also detected in each 1 case). 2 cases of coli urinary tract infection, 1 case of acute colitis from which pathogenic E. coli was detected, 1 case of E. coli carrier, 1 case of acute bronchitis (bacteria were not detected), and 1 case of urinary tract infection (bacteria were not detected) showed rapid improvement of clinical symptoms with rapid eradication of the pathogenic bacteria. In one case of urinary tract infection where S. epidermidis and S. faecalis were simultaneously detected, S. epidermidis was removed but S. faecalis was merely decreased. The effective antibacterial concentration after intravenous injection of CPZ in the feces was determined and found to be present in sufficient concentrations to prevent colon infection. No particular side effects were observed during CPZ therapy.

[Do all patients with mandibular hypoplasia present difficulty in endotracheal intubation?].

On 7 patients with Pierre-Robin syndrome (PR), 8 patients with first and second brachial arch syndrome (BA) and 7 patients with Treacher Collins syndrome (TC), we examined retrospectively the number of patients whom anesthesiologists found difficult to perform laryngoscopy and intubation, the number of attempt necessary for proper insertion of the tracheal tube, the number of patients in whom the endotracheal intubation with laryngoscopy was given up and the time necessary for endotracheal intubation. We compared these with those from 22 control patients. Our results suggest that the difficult laryngoscopy and intubation are more frequent in TC. On the other hand, most of PR and BA may not present difficulty in intubation.

[The duration of action of vecuronium in five patients after kidney transplantation--comparison with that during kidney transplantation].

We compared the duration of vecuronium action in five patients after the kidney transplantation with that during kidney transplantation. After the transplantation, three patients required no hemodialysis therapy but two patients underwent hemodialysis therapy again. In all these five patients, including patients who were back to hemodialysis therapy, the durations of vecuronium action after receiving transplanted kidney were shorter than those during kidney transplantation. These shortened durations are speculated to be mainly due to excretion of vecuronium by the transplanted kidney and the effect of long term steroid therapy. However in this study the durations of vecuronium action in patients who required further hemodialysis therapy were also shorter than those during kidney transplantation. To determine whether this is a common or exceptional phenomenon, further evaluation should be needed.

[Pharmacodynamics of vecuronium in the kidney transplant recipient and the patient with normal renal function].

Pharmacodynamics of vecuronium were evaluated in seven kidney transplant recipients and compared with seven patients with normal renal function. Vecuronium 0.12 mg.kg-1 was used for the initial dose and 0.03 mg.kg-1 for the second dose for each patient under general anesthesia with either isoflurane, sevoflurane or halothane plus nitrous oxide after induction by thiamylal. The effect of vecuronium was evaluated by a muscle relaxation monitor. The time to the maximum blockade (onset time) and the time of 25% recovery of the first twitch height (duration time) were measured after each administration of vecuronium in patients of both groups. The onset times after the initial and second doses were similar in both groups (180.0 +/- 15.5 sec for recipients versus 185.7 +/- 14.9 sec for patients of normal renal function). However, duration was significantly longer in recipients than in patients of normal renal function. Durations after the initial and second doses were 130 +/- 19 min for the initial dose and 86 +/- 12 min for the second dose in recipients, whereas they were 51 +/- 5 min and 37 +/- 5 min in patients of normal renal function. The prolonged durations of vecuronium in the kidney transplant recipient were speculated to be mainly due to delayed elimination of the drug and effect of cyclosporin, an immunosuppressive agent in the recipients. These results suggest that the titrated administration of vecuronium by keen monitoring of muscle relaxation is needed in the kidney transplant recipient to avoid unnecessary prolongation in duration of action.

[Perioperative management for patients with hypertrophic cardiomyopathy undergoing noncardiac surgery].

We had two patients with hypertrophic cardiomyopathy for noncardiac surgeries. Case 1: A 74-year-old man for right nephrectomy received epidural lidocaine and nitrous oxide combined with 0.2-0.6% isoflurane. During the operation, heart rate and blood pressure were relatively unstable, but he woke up promptly after the operation. Early on the morning of the 2nd post-operative day, he was found dead on his bed. Case 2: A 52-year-old man for gastrectomy was anesthetized with nitrous oxide and halothane with continuous propranolol infusion. Through the operative period, heart rate and blood pressure were stable and postoperative course was uneventful. In these two patients, preoperative Holter ECG showed ventricular tachycardia, which may increase the risk of a sudden death. These cases demonstrate that general anesthesia with nitrous oxide combined with halothane, can be administered with a low risk in patients with HCM for noncardiac surgery and that postoperative intensive care unit monitoring is necessary for these patients for several days to prevent a sudden death.

[Analysis of microsatellite alteration in colorectal cancer].

We investigated the possible correlation between the microsatellite alterations (replication error: RER, and loss of heterozygosity: LOH) and clinicopathologic factors and survival in colorectal cancer. A total of 78 colorectal cancers was examined for microsatellite alteration at three microsatellite loci containing D2S123, D18S58 and C117-703. RER is considered positive when at least one microsatellite locus is detected. RER was positive in 28.2%, and the respective positivity was 12.8%, 15.3% and 11.5%. The positivity of LOH was 6.4%, 10.3% and 19.2%, in that order. RER-positive cancers were more significantly found in the proximal colon than the distal colorectum. Node-negative colorectal cancers were more noted in RER (+)-positive cancers. Multivariate analysis showed that LOH in D18S58 locus and RER in CI17-703 locus were independent prognostic factors.

[K-ras gene mutations in adenomas from familial adenomatous polyposis].

A 18-year-old woman underwent total colectomy for familial adenomatous polyposis. In order to clarify the significance of K-ras mutations in early colorectal carcinogenesis, K-ras mutations were analyzed in multiple adenomas by PCR-SSCP method. A total of 256 adenomas were found throughout the entire colon and rectum, and the distribution was a sparse type. The correlation between K-ras gene and clinicopathological factors was examined in 90 adenomas. There was no correlation among K-ras mutations and anatomical distribution, or morphological classification, but K-ras mutation was more frequent in severe compared with slight atypia. We investigated the correlation between the size of adenoma in the horizontal and vertical directions and K-ras mutation. K-ras mutation was more frequent in the horizontal size greater than 6 mm in diameter, and also more frequent in vertical size greater than 20 mm in height. It was concluded that the adenomas detecting K-ras mutations might have proliferating potential, and would be applied to determine polypectomy.