Research in similarities between EEP34 and lipocortin

EDTA-extractable protein (EEP) is a major extrinsic protein of lens fiber membrane. The EEP of calf lens separated three major bands by SDS-PAGE, and the molecular weights of these major bands were 34 kilodalton (kd.), 33 kd. and 32 kd. Combination of EEP with F-actin and some kinds of phospholipid containing liposomes depends on Ca2+. In two-dimensional gel electrophoresis, some spots of the calf-EEP partially overlapped with spots of lipocortin, which is a substrate for epidermal growth factor receptor/kinase and an inhibitor of phospholipase A2. The 34 kd. protein of the EEP cross-reacted to the antibody prepared against lipocortin of pig's aorta: These results suggest the important function of EEP in lens.     

An immunohistochemical study on the ontogeny of cells immunoreactive for spot 35 protein, a novel Purkinje cell-specific protein, in the rat cerebellum

A time course study on the appearance and distribution of cells immunoreactive for spot 35 protein, a novel cerebellar Purkinje cell-specific protein, was conducted in the developing cerebella of fetal and early postnatal rats by PAP immunohistochemistry. Spot 35-immunoreactive cells were first noted in the cerebellar anlage on the 17th embryonic day, appearing as large cellular aggregations in the mantle layer and a small number of elongated cells dispersed between the cell aggregations and the ependymal layer. As the development proceeded, the spot 35-immunoreactive cells gradually accumulated beneath the external granular layer. At birth, they were arranged compactly in 4-5 irregular rows to form a primitive Purkinje cell layer. During their subsequent development, immunostaining for spot 35 protein demonstrated the rearrangement of the Purkinje cells into a single row and the maturation of their somata, axons and dendrites. All these findings indicate that spot 35 protein is a specific marker for the cerebellar Purkinje cells, from their migrating stage throughout the course of their maturation. The present study further describes the transitory lamellar arrangement of spot 35-immunoreactive Purkinje cells located horizontally at a short distance away from the ependymal layer of the cerebellum on the 17th embryonic day.     

Novel responses of peripheral lymphocytes of cancer patients to chemical induction of sister chromatid exchanges

Sensitivities to sister chromatid exchange (SCE) induction by chemicals of peripheral lymphocytes from 26 cancer patients were estimated under conditions identical to those for healthy humans which had been reported (Cancer Res., 43: 439-442, 1983). The sensitive individual was defined as one whose cells give a mean induced SCE frequency more than 2 standard deviation units above the population mean of induced SCEs in cells from the healthy humans. When cells were treated with 3-amino-1-methyl-5H-pyrido[4, 3-b]indole in the presence of rat liver S9 mix, 8 in 10 stomach cancer patients, 4 in 4 colon cancer patients, 3 in 9 lung cancer patients, 0 in 3 patients bearing other cancers, and 0 in 9 non-cancerous individuals were sensitive. The corresponding frequency of individuals in the healthy population, reported previously, was 1 in 33 persons. Thus, the frequency of sensitive individuals in the combined group of stomach and colon cancer patients was very significantly higher than were frequencies in control groups. Three in 10 patients with stomach cancer and 4 in 16 patients with other cancers were sensitive to induction of SCE by methyl methanesulfonate. Six in these 7 methyl methanesulfonate-sensitive patients were also 3-amino-1-methyl-5H-pyrido[4,3-b]indole sensitive. The frequency of methyl methanesulfonate-sensitive individuals in the healthy populations was 2 in 50. There was no patient who was sensitive to SCE induction by 4-nitroquinoline 1-oxide. The frequency was not significantly different from the healthy population, in which 3 in 50 persons were sensitive. These results suggest that a particular cancer correlates with the sensitivity of peripheral lymphocytes to SCE induction by particular chemicals.     

A prostacyclin analogue reduces free radical generation in heart-lung transplantation.

The mechanism by which prostacyclin acts to prevent in vivo reperfusion injury is still uncertain. This study was therefore undertaken to assess the effect of a stable prostacyclin analogue (OP 41483-alpha-CD [OP]) on oxygen-derived free radicals after heart-lung transplantation. OP was administered to the heart-lung graft through the pulmonary artery for 25 minutes encompassing the reperfusion process. Free radicals were directly measured by electron spin resonance spectroscopy. The radical intensities of pulmonary venous blood were significantly lower in the OP group than in the control group, suggesting that fewer free radicals were generated in the lungs of the OP group. The cardiac and respiratory function were better in the OP group than in the control group. The lung is the primary source of oxygen free radical attack, and the beneficial action of OP on free radical generation is almost exclusively restricted to the lung and does not apply to the heart. This result suggested that OP probably is effective in inhibiting free radical generation from the endothelium.     

Bronchial hyperresponsiveness to acetylcholine during acute pulmonary congestion in guinea pigs]

To understand the precise mechanism of bronchial hyperresponsiveness in patients with congestive heart failure, we studied the effect of mild pulmonary congestion on bronchial responsiveness to inhaled acetylcholine (ACh) in guinea pigs. We induced mild pulmonary congestion by inflation of a balloon placed in the left atrium, and maintained the left atrial pressure (Pla) at 10 mmHg for 30 minutes with continuous monitoring of lung resistance (RL) and dynamic compliance (Cdyn). Furthermore, we determined the provocative concentration of ACh producing 100% increase in RL (PC100-ACh), before and during balloon inflation. In animals with propranolol pretreatment, but not in animals without propranolol pretreatment, mild pulmonary congestion caused slight increase in RL (N.S.) and significant decrease in Cdyn (p less than 0.01) and PC 100-ACh (p less than 0.01). Cutting of bilateral vagal nerves partially inhibited the decrease of PC100-ACh, but pretreatment with either phenoxybenzamine, indomethacin, AA-861 or OKY-046 had not effect. These results suggest that blockade of beta-adrenergic receptors and the vagal reflex, but not of alpha-adrenergic receptors or arachidonates, contributes to bronchial hyperresponsiveness during acute pulmonary congestion.     

Visibility of epidermoid tumors on steady-state free precession images.

PURPOSETo determine whether steady-state free precession sequences improve the MR visibility of epidermoid tumors in comparison with spin-echo images.METHODSPatients were four women and three men with epidermoid tumors in the subarachnoid spaces. MR was performed with a 1.5-T superconductive unit. For steady-state free precession imaging, three-dimensional Fourier transform fast imaging with steady-state free precession (FISP) images were used (20-40/7 [repetition time/echo time], flip angle of 25 degrees). The visualization and contrast-to-noise ratio were compared in FISP images and spin-echo images. In one case, the images of FISP and fast low-angle shot were obtained with variable repetition times and flip angles to evaluate the best pulse sequences for the visualization of epidermoid tumors.RESULTSThe contrast-to-noise ratios between tumors and cerebrospinal fluid ranged from 7.9 to 17.5 (average was 12.9) on FISP images. The average of contrast-to-noise ratios on T1, T2, and proton density-weighted spin-echo images were 1.6, 2.0, and 4.2, respectively. Three-dimensional Fourier transform FISP images best showed central nervous system and demonstrated epidermoid tumors excellently.CONCLUSIONSEpidermoid tumors in the subarachnoid spaces were better demonstrated on steady-state free precession (three-dimensional Fourier transform FISP) than on conventional spin-echo images.     

A mechanism for reducing intraocular pressure in normal volunteers using UF-021, a prostaglandin-related compound]

The mechanism of reduction of intraocular pressure (IOP) and other ocular effects were studies after topical application of prostaglandin (PG) F2 alpha and UF-021, a new PG related compound, in eight normal volunteers. IOP, aqueous humor flow rate and outflow rate were evaluated during a period of four hours after the application. Both PGF2 alpha and UF-021 caused significant and similar IOP reduction for four hours. Neither compound produced any significant change in the aqueous humor flow rate or outflow rate, suggesting the increase of unconventional outflow rate as being the possible mechanism of IOP reduction in normal human eyes.     

Germinoma originating in the basal ganglia and thalamus: MR and CT evaluation.

PURPOSETo describe MR and CT features of germinoma originating in the basal ganglia and thalamus and to discuss the roles of each modality for its diagnosis.METHODSMR and CT studies of six cases of germinomas, five of which were histologically proved, were retrospectively reviewed. T1-weighted, T2-weighted, and contrast-enhanced T1-weighted conventional spin-echo images, and unenhanced and contrast-enhanced CT images were evaluated.RESULTSTypically, the tumor consisted of an irregular solid area with contrast enhancement and various-size cysts. Cystic components were found in five cases and calcification in four. Intratumoral hemorrhage was noted in one. Ipsilateral cerebral hemiatrophy and brain stem hemiatrophy were noted in three cases each. MR was superior to CT in evaluating precise tumor extension, cystic components, and intratumoral hemorrhage, although in one case, extension of the tumor was better defined on CT in its early stage. Calcification was difficult to identify by MR alone. The solid components of the tumors generally showed slightly high density on CT, which seemed to be characteristic compared with nonspecific intensity pattern on MR.CONCLUSIONThe combination of CT and MR findings allows early detection and appropriate diagnosis of the mass in the basal ganglia and/or thalamus.     

Role of Ca2+ on vasoactive intestinal peptide-induced glucose and adenosine 3',5'-monophosphate production in the isolated perfused rat liver.

Livers from fed rats (180-240 g) were perfused noncyclically with a hemoglobin-free medium in vitro to determine whether vasoactive intestinal peptide (VIP) increases hepatic glucose production through a cAMP- or a Ca(2+)-dependent mechanism. Glucose output did not increase, but cAMP increased maximally during 10(-9) M VIP infusion. When VIP was perfused at 10(-8) M or more, glucose output increased dose dependently, whereas cAMP increased only a little during the VIP infusion, but increased greatly after the infusion. When Ca2+ was excluded from the perfusate, glucose output produced by 10(-8)-10(-7) M VIP was only 40% of that observed in the Ca(2+)-containing perfusion, and the increase in cAMP was abolished almost completely. By adding 10(-7) M A23187 for 10 min during the infusion of 10(-9) M VIP, cAMP, which increased with VIP alone, decreased during the A23187 infusion and increased again after the cessation of the A23187 infusion, whereas glucose output increased during the A23187 infusion. These results were similar to those observed with higher concentrations of VIP. When 10(-4) M isobutylmethylxanthine and 10(-8) M VIP were infused concurrently, cAMP increased rapidly during the infusion and decreased after the infusion. In conclusion, 1) glycogenolysis is produced by VIP through a Ca(2+)-dependent mechanism, rather than a cAMP-dependent one; and 2) the restriction of cAMP accumulation during the infusion of high concentrations of VIP is caused by Ca(2+)-induced phosphodiesterase activation.