Essential role of rho kinase in the ca2+ Sensitization of Prostaglandin F2α-Induced Contraction of Rabbit Aortae

Although the prostate gland is a rich source of α1-adreno- (α1-AR) and m1-cholino receptors (m1-AChR), the membrane processes associated with their activation in glandular epithelial cells is poorly understood. We used the whole-cell patch-clamp technique to show that the agonists of the respective receptors, phenylephrine (PHE) and carbachol (CCh), activate cationic membrane currents in lymph node carcinoma of the prostate (LNCaP) human prostate cancer epithelial cells, which are not dependent on the filling status of intracellular IP₃-sensitive Ca²⁺ stores, but directly gated by diacylglycerol (DAG), as evidenced by the ability of its membrane permeable analogue, OAG, to mimic the effects of the agonists. The underlying cationic channels are characterized by the weak field-strength Eisenman IV permeability sequence for monovalent cations ( P _K(25) > P _Cs(4.6) > P _Li(1.4) > P _Na(1.0)), and the following permeability sequence for divalent cations: P _Ca(1.0) > P _Mg(0.74) > P _Ba(0.6) > P _Sr(0.36) > P _Mn(0.3). They are 4.3 times more permeable to Ca²⁺ than Na⁺ and more sensitive to the inhibitor 2-APB than SK&F 96365. RT-PCR analysis shows that DAG-gated members of the transient receptor potential (TRP) channel family, including TRPC1 and TRPC3, are present in LNCaP cells. We conclude that, in prostate cancer epithelial cells, α1-ARs and m1-AChRs are functionally coupled to Ca²⁺-permeable DAG-gated cationic channels, for which TRPC1 and TRPC3 are the most likely candidates.     

Protection of hepatocytes from apoptosis by a novel substance from actinomycetes culture medium

A novel substance, #675, found from an Streptomyces sp. SM675 culture medium, dose-dependently stimulates the proliferation of human functional liver cell 4 (FLC4). When FLC4 cells were incubated under conditions without fetal bovine serum (FBS), typical features of apoptotic cell death such as shrinkage and nuclear condensation appeared; high molecular weight (HMW) DNA fragments were found; and caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were cleaved. When FLC4 cells were incubated with #675 and without FBS, the cells grew healthy, no HMW DNA fragments were found, and caspase-3 and PARP cleavage weakened, suggesting that #675 protects FLC4 cells from apoptosis induced by FBS-deprivation. The quantitative reverse-transcribed polymerase chain reaction did not show differences in PARP or Bcl-2 mRNA expression in FLC4 cells incubated with or without #675, indicating other genes may be involved in this anti-apoptosis effect. These results show that #675 enhances FLC4 proliferation via an apoptosis-inhibition pathway, implying potential pharmacological and clinical applications.     

Functional evaluation of lung by Xe-133 lung ventilation scintigraphy before and after lung volume reduction surgery (LVRS) in patients with pulmonary emphysema

We evaluated the respiratory functions of patients with pulmonary emphysema who underwent lung volume reduction surgery (LVRS) by the mean transit time (MTT) with Xe-133 lung ventilation scintigraphy, forced expiration volume in 1 sec (FEV1.0), residual volume (RV), distance walked in 6 min (6-min walk), and the Hugh-Jones classification (H-J classification) before and after LVRS. In 69 patients with pulmonary emphysema (62 men, 7 women; age range, 47-75 years; mean age, 65.4 years +/- 6.1, preoperative H-J classification, III (two were II)-V) who underwent LVRS, all preoperative and postoperative parameters (MTT 3 weeks after LVRS and the others 3 months after LVRS) were judged statistically by the Wilcoxon signed-ranks test and Odds ratio. Every postoperative parameter was improved with a significant difference (P < 0.05) compared to preoperative parameters. MTT at 3 weeks after LVRS was not associated with %FEV1.0 and the H-J classification at 3 months after LVRS, but was associated with RV and a 6-min walk at 3 months after LVRS. MTT was useful for the clinical evalution of aerobic capability after LVRS.     

Immune responses in newly developed short-lived SAM mice. IV. Chromosomal location of a gene controlling defective helper T-cell activity.

Short-lived SAMP-P/1 mice are low responders in in vitro antibody responses because of a selectively impaired helper T(Th)-cell activity. After crossing with high responders (B10.BR mice), about 12% of (B10.BR x SAM-P/1) (BRP)F2 mice showed low responsiveness, as did SAM-P/1 mice, against two T-dependent antigens, sheep and horse red blood cells (RBC), both of which were not cross-reactive to each other at helper T- and B-cell levels. The immune activities against the two antigens in individual BRPF2 mice showed a good correlation (r = 0.81), thereby suggesting that SAM-P/1 mice have an antigen non-specific Th cell dysfunction. Based on the incidence of the low responders in F2 generation and statistical analyses, the hypo-responsiveness was postulated to be controlled by two genes. To survey the location of these genes, linkage analyses were performed in the F2 mice using a large set of genetic markers. Low responders in the F2 generation showed a significantly higher incidence of SAM-P/1 genotype at the Gpi-1 as well as c locus on chromosome 7 (Chr.7). However, no linkage of low responsiveness to the Hbb locus was evident, an area present at a more distal site to the centromere on the same chromosome. These results suggest that one of the genes controlling the hypo-responsiveness of SAM-P/1 mice is linked to both Gpi-1 and c loci and that it locates at a more proximal site on Chr.7.     

Ultramicroassay of red cell group using Terasaki-plate

In order to simplify the testing-method of blood groups (ABO, Rh, MN, Ss, P, I, Lewis, Duffy, Kidd and Diego) and the assay of glycosyl transferases activity, we have developed a new method using Terasaki plate (T-plate method), and examined its usefulness in the routine examination. As results followings were obtained: 1) The sensitivity of T-plate method was equal or superior than that of the common method using test-tube. 2) Samples used for T-plate method were smaller in volume compared with those for the common tube test method. 3) The percent of coincidence between T-plate method and tube test method was 100% for ABO, Rh, MN, Ss, P, I, Duffy, Kidd and Diego grouping. They were 98% for Lewis(a) and 96% for Lewis(b) grouping, respectively. These results indicate the superiority of T-plate method than the common method using test-tubes in respects of smaller volume of samples, lower price and handling of a large amount of samples.     

Disposition of amino acid synaptic transmitters, acetylcholine and substance P in the LM-suprageniculate nuclear complex of the cat's thalamus.

Immunocytochemical analysis with antibodies raised against aspartate, glutamate, gamma-aminobutyrate (GABA), choline acetyltransferase (ChAT), and substance P (SP) have allowed the transmitter characterisation and distribution of cells of the lateralis medialis-nucleus suprageniculatus (LM-SG) complex to be made at the level of the light microscope. We have found that the intranuclear distributions of aspartate and glutamate differed substantially from that of GABA, as well as there being specific and, in some cases, major differences in the respective populations of cells labelled with all three amino-acid-sensitive antibodies. ChAT-labelled elements were disposed very similarly to acetylcholinesterase (AChE)-positive subregions of the nuclear complex, while SP labelling was comparatively weak, albeit present, throughout the region. These data provide an important first step towards the further understanding of the details of the neurochemical and functional identity of the LM-SG complex.     

Depolarization thresholds for hippocampal damage, ischemic preconditioning, and changes in gene expression after global ischemia in the rat

Induced ischemic tolerance in rat hippocampus was investigated in a forebrain ischemia model of repeated 4-vessel occlusion (4-VO). Ischemic insult variability was reduced by the use of dc potential measurements to determine the duration of ischemic depolarization in hippocampus. The results demonstrate a depolarization threshold for ischemic injury to CA1 neurons of 4-6 min and a window for optimal preconditioning of 2.5-3.5 min. Levels of induced mRNAs encoding hsp72 and several immediate-early genes were also shown to vary with depolarization interval. Immediate-early genes were maximally induced after depolarization periods inducing optimal preconditioning, while hsp72 expression increased with insult severity over the range leading to neuron loss. These results are similar to those obtained in gerbil studies indicating that preconditioning does not require large increases in hsp72 expression, and demonstrate the fundamental comparability of rodent global ischemia models when monitored by this approach.     

"True" posterior communicating artery aneurysm presenting the abducens nerve palsy

Most of "so-called" posterior communicating artery aneurysms previously reported, originated from the internal carotid-posterior communicating junction. Aneurysms arising from the posterior communicating artery itself are very rare. The abducens nerve palsy caused by cerebral aneurysm is also very rare. We are reporting a case with the saccular aneurysm arising directly from the distal half of the posterior communicating artery presenting the abducens nerve palsy. This 73-year-old woman who had no treatment with hypertension for several years was admitted for sudden onset of severe headache, vomitting and unconsciousness on March 1, 1984. She opened her eyes when addressed and had disorientation, urinary incontinence, right-hemiparesis and left-abducens nerve palsy. A 4-vessel angiography revealed the saccular aneurysm originating directly from the distal half of the posterior communicating artery. The patient underwent left-frontotemporal craniotomy on the 27th day after subarachnoid hemorrhage under Hunt & Kosnic Grade 3. The aneurysm originated directly from the distal half of the posterior communicating artery and directed inferior-posterior-laterally below the oculomotor nerve. The neck was successfully clipped. Immediate post-operative course was uneventful until the 7th day after surgery. On the 8th day she had hypertensive intraventricular hemorrhage and expired. The autopsy could not be obtained. The saccular "true" posterior communicating artery aneurysm with isolated unilateral abducens nerve palsy as seen in our case has not been reported. Considering the operative findings, we thought the aneurysmal dome contacted directly with the abducens nerve.     

Treatment of advanced bladder cancer with intra-arterial infusion of cisplatinum (CDDP) and aclacinomycin (ACR), combined with angiotensin II

Ten patients with advanced bladder cancer were treated with intra-arterial infusion therapy. The patients consisted of nine males and one female between 55 and 82 years old (median: 70 years). In all patients, cisplatinum (CDDP) (2 mg/kg), aclacinomycin (ACR) (0.5 mg/kg) and Angiotensin II (25 mg) were infused via the internal iliac artery for a period of about 30 minutes. Seven patients also received X-ray therapy with a linac. The efficacy of this therapy was assessed by computed tomographic scanning, sonography and cystoscopy. As a result of this assessment, 2 patients were rated complete response "(CR)", 6 partial response (PR) (showing 50% or more reduction in the lesion) and 2 no change "(NC)". To compare the efficacy of this therapy for two histopathologically defined groups of patients (patients with grades 2 and 3 cancer), one patient was rated "CR", four "PR" and two "NC" in the grade 3 group (total 7 patients), while one was rated "CR" and two "PR", in the grade 2 group (total 3 patients). In effective cases, pollakiuria and miction pain disappeared shortly following intra-arterial infusion therapy. As for side effects of the therapy, mild nausea or vomiting was observed in all patients, while leukopenia was noted in one patient.