Expression of FKBP12 and ryanodine receptors (RyRs) in the spinal cord of MND patients.

We investigated the FKBP12 and ryanodine receptor (RyR) immunoreactivity (IR) in the spinal cords of neurological controls and patients with motor neuron disease (MND). In the neurological controls, the cytoplasm of the spinal anterior horn neurons was stained with anti-FKBP12 antibodies and anti-RyR (type 1 and type 2) antibodies. In the MND cases, the residual neurons in the anterior horn of the spinal cord showed IR for RyR (type 1 and 2) antibodies, while weak IR for anti-FKBP12 antibodies was comparable to that of controls. The numbers of neurons recognized with the anti-FKBP 12 or anti-RyR (type 1 and 2) antibodies were counted in the anterior horn of spinal cords from the MND cases and neurological controls. Frequency of neurons stained with anti-FKBP 12 antibody was significantly decreased in the MND cases compared to that in controls (48.7+/-23.2%, 71.0+/-18.5%, respectively, mean+/-SD, p<0.0005). In the MND cases, numbers of normal-appearing, chromatolytic neurons showing IR to anti-FKBP12 (N19) antibody were significantly decreased compared to those in the controls. Immunoreactivities to anti-RyR antibodies (type 1and 2) in MND cases were present and there was no difference compared to those of the controls. Neurons in the spinal cord anterior horn of Kii-ALS cases with prolonged clinical duration were immunostained with both anti-FKBP12 and anti-RyR (type 1 and 2) antibodies similar to that in the controls. The anterior horn neurons of MND cases of short clinical duration showed absent IR to FKBP 12 antibody but present IR to RyR (type 1 and 2) antibodies. The present result suggests that FKBP12 IR was decreased in the MND cases with short clinical duration. RyR (type 1 and 2) is a major component of the intracellular calcium channel, which mediates calcium-induced calcium release. FKBP12, which is an endogenous ligand for RyR, stabilizes the calcium channels preventing calcium leakage in the absence of receptor activation. Imbalance between FKBP12 and RyR IR may play an important role in degeneration due to MND. Further study of the correlation between RyR and FKBP12 should contribute to clarifying the mechanisms of neurodegeneration in MND, including calcium-induced neuronal loss.     

Aerosolized methacholine-induced bronchoconstriction and pulmonary hyperinflation in rats

The body plethysmography (BPG) is a useful tool for analyzing pulmonary function in small animals because it simultaneously measures airway resistance (R _aw) and functional residual capacity (FRC). We previously described a BPG with the enclosed environment maintained at body temperature and water vapor-saturated. We found dose-dependent increases in R _aw in response to inhaled methacholine (Mch) with no apparent increase in FRC in intratracheally intubated rats without paralysis. To resolve this apparent inconsistency in clinical observations, we repeated the study using a newly developed BPG that allowed us to shorten the interval between Mch-inhalation and measurements by about two-thirds. Using Mch concentrations of 0, 0.5, 1.0, 2.0 and 4.0 mg/ml each for 2 min, both parameters increased in a dose-dependent fashion with FRC (mean ± SE) values of 3.77 ± 0.16, 4.43 ± 0.26, 4.75 ± 0.34, 5.02 ± 0.49 and 5.34 ± 0.38 ml and R _aw values of 18.6 ± 3.9, 21.6 ± 4.9, 35.0 ± 6.9, 49.0 ± 8.8 and 65.7 ± 8.8 ml/s/Pa, respectively. Immediate measurement after Mch-inhalation demonstrated profound bronchoconstriction associated with dose-dependent increases in FRC.     

A clinical study on 34 cases of urothelial cancer of upper urinary tract

Thirty-four cases of tumor of the renal pelvis or ureter or both have been treated in our department during the past decade. The primary tumor was in the renal pelvis in 11 cases, in the ureter in 21 cases and in the ureter and renal pelvis in 2 cases, a co-existent tumor in the bladder was found in 4 cases. Seventeen patients had a tumor on the right side and 17 on the left side. The most frequent symptom was gross hematuria (70.6%) and flank pain was the presenting symptom in 7 cases (20.6%). On the intravenous pyelography, a filling defect in the renal pelvis or ureter (41.2%) and nonvisualization (53.0%) were frequent findings. Twenty-nine cases had undergone total nephroureterectomy with resection of a bladder cuff, 3 had simple nephrectomy and 2 had open biopsy alone. Postoperative radiation therapy was done in 1 case, chemotherapy in 10 cases, and 6 cases of them were treated by CAP therapy (cis-dichlorodiamine platinum, doxorubicin and cyclophosphamide). Actual and relative 5-year survival rates were 53.8% and 63.5%, and no significant difference was found in survival rate between the patients with renal pelvic tumors and those with ureteral tumors.     

Influence of duration of complete global brain ischemia on neurologic outcome in dogs

The author studied the influence of duration of complete global brain ischemia (CGBI) on neurologic outcome (NO) in dogs. The CGBI of various durations, from 10 to 20 minutes, were induced by clamping of the ascending aorta combined with bypass formation between the aorta and the right atrium. The dogs were divided into four groups according to the duration of CGBI: dogs in group A, B, C or D suffered 10, 15, 18 or 20 minutes of CGBI respectively. In four groups, NOs were evaluated for 7 days after the ischemic insult. In group A, no significant damages were observed and all survived for 7 days. In group B, marked differences in the degree of neurologic deficits were observed, and nearly a half were almost normal and the remainder were severely damaged. In groups C and D, all were so severely damaged as to become vegetative and any difference in the degree of neurologic deficits was not observed among the groups. The author concludes that dogs which have suffered CGBI for 15 minutes or less are not suitable for use in order to evaluate the efficacy of treatments for ischemic brain damage with NOs, and that dogs which suffered CGBI for 18 minutes or more are appropriate for this purpose.     

Effects of the yeast extract components pyrroloquinoline quinone and aspartic acid on vitamin B12 production in Klebsiella pneumoniae IFO 13541

The production of vitamin B12 from carbohydrates, peptone, casamino acid, etc., by intestinal bacteria was investigated. Klebsiella pneumoniae IFO 13541 was the most efficient strain for vitamin B12 production, which depended exclusively on the concentration of yeast extract added to the medium. A concentrated solution of yeast extract (1 ml) was chromatographed on a Sephadex G-25 column (1 x 180 cm) and eluted with H2O (eighty fractions of 3 ml each were collected). It was found that fractions in which bacterial growth was most prevalent also exhibited the highest amount of vitamin production. The effectiveness of yeast extract was shown by the participation of pyrroloquinoline quinone and aspartic acid in the growth stimulation and in the vitamin B12 production in this strain.     

Extracellular adenosine triphosphate completely reverses the thyrotropin-induced morphological change in FRTL-5 cells

We quantified the TSH-induced morphological change in FRTL-5 thyroid cells according to a morphological index corresponding to the mean cell area measured from microscopic photographs. Within 15 min, TSH induced, at 10 pM and higher concentrations, a decrease in morphological index together with a rise in cAMP levels in a TSH dose-dependent manner. Forskolin, 3-isobutyl-1-methylxanthine, and RO 20-1724, the latter two being phosphodiesterase inhibitors, mimicked these TSH effects, indicating that the rise in cAMP levels is responsible for the TSH effect. Extracellular ATP and its derivatives, known as purinergic receptor agonists, decreased cAMP levels and caused a complete reversal of the TSH morphological effect. Prior exposure of the cells to islet-activating protein (pertussis toxin), the depletion of extracellular Ca2+, or the addition of low doses of protein kinase-C inhibitors completely abolished the inhibitory action of ATP on the TSH effect, whereas phorbol 12-myristate 13-acetate, which activates protein kinase-C, mimicked the ATP action to some extent. Thus, although the TSH-induced change in cell morphology seems to be dependent on cAMP levels, the inhibition of TSH action by ATP seems to be mediated by at least two signal transduction pathways involving islet-activating protein substrate G-proteins: one inhibiting adenylate cyclase and the other involving Ca2+ and protein kinase-C.     

The prognostic significance of cell cycle markers in esophageal cancer after neoadjuvant chemotherapy

Proliferating cell nuclear antigen (PCNA), p27 and cyclin A were analyzed by immunohistochemistry in 89 patients (untreated control n = 40, neoadjuvant chemotherapy n = 49) with esophageal cancer invading the submucosal lesion. The mitotic index (MI) was calculated as the percentage of mitotic cells. In control subjects, the mean PCNA, p27, cyclin A and MI were, respectively, 60.4%, 18.0%, 19.9% and 1.7%; in the chemotherapy group, these values were 46.8%, 15.1%, 18.0% and 1.2% respectively. Neoadjuvant chemotherapy decreased PCNA and the MI significantly. As prognostic indicators, PCNA and the MI were significant in control subjects and p27 and cyclin A were significant in the chemotherapy group. Using multivariate analysis, p27 was a prognostic factor in both groups and cyclin A was prognostic only in the chemotherapy group. Although PCNA and the MI were useful growth and prognostic markers in untreated control subjects, their significance was lost after neoadjuvant chemotherapy. p27 and cyclin A were determined to be significant markers in the neoadjuvant chemotherapy group, especially p27, which was independent in both groups.     

Evaluation of surgical procedures for cyanotic congenital heart disease by using MR imaging

ECG-gated MR imaging has been shown to be effective for the diagnosis of congenital heart disease. In this study, we assessed its role in the postoperative evaluation of surgical procedures in patients with complex congenital heart disease. MR images of 26 patients with Rastelli (five), Fontan (three), Senning (three), Damus (one), Jatene (eight), Waterston (four), and Potts (two) procedures were evaluated retrospectively. The accuracy of MR imaging was compared with that of angiography in 20 patients. The surgical anastomoses were identified in all patients. Patency, atresia, or hypoplasia of central pulmonary arteries and postoperative complications (focal stenoses of pulmonary arteries, thrombosed conduit, peri-conduit abscess) were shown. Narrowing of the right ventricular outflow tract and focal compression of the proximal pulmonary arteries were recognized as specific complications of the Jatene procedure. MR imaging appears to be effective in the postoperative evaluation of surgical procedures used for congenital heart disease. It should be considered as an alternative to repeated catheterization and angiography for the postoperative examination of children with complex congenital heart disease.