Lymphokine-independent, leukemia cell-mediated induction of tumor necrosis factor in human monocytes.

Unactivated human blood monocytes and monocytic THP-1 cells were found to respond to some leukemia cells by tumor necrosis factor (TNF) production. The TNF production by THP-1 cells in response to K562 cells was preceded by a rapid rise in [Ca2+]i, initiated within 1 h and terminated within 4 h as a refractory state took over. Neither the amount nor the duration of TNF production was enhanced by gamma-interferon. The P32/ISH cells did not induce a significant [Ca2+]i change of TNF production, while MOLT-4 cells failed to induce TNF despite their capacity to mobilize Ca2+ in THP-1 cells. The failure of P32/ISH or MOLT-4 to induce TNF was attributed primarily to a lack of stimulatory membrane molecules rather than to suppression by an inhibitory component, since liposomes carrying membrane components of K562 and MOLT-4 or P32/ISH in varying proportions elicited TNF production that precisely reflected the K562 proportion. The ability of K562 to induce TNF was selectively impaired by trypsin, whereas the ability to mobilize [Ca2+]i was more sensitive to glutaraldehyde, although once the latter activity was extinguished, the K562 cell could no longer induce TNF. These results suggest that some leukemia cells are equipped with two or more signaling membrane moieties which together stimulate monocytes for transient tumoricidal expression in the preimmune stage.

Significance of myocytes with positive DNA in situ nick end-labeling (TUNEL) in hearts with dilated cardiomyopathy: not apoptosis but DNA repair.

BACKGROUND: The presence of apoptotic myocytes has been reported in human hearts with dilated cardiomyopathy (DCM) on the basis of a positive finding of DNA in situ nick end-labeling (TUNEL). However, ultrastructural evidence of myocyte apoptosis has not been obtained. METHODS AND RESULTS: A total of 80 endomyocardial biopsies were obtained from right and left ventricles of 20 patients with DCM and 20 normal control subjects. TUNEL-positive myocytes were found by light microscope in 15% of DCM specimens (controls, 0%, P<0.05), and the percentage of TUNEL-positive myocytes per section in DCM was 1. 0+/-2.7% (mean+/-SD). According to TUNEL at the electron microscopic level (EM-TUNEL), immunogold particles, which label DNA breaks with 3'-OH terminals, were markedly accumulated in the bizarre-shaped nuclei, with widespread clumping of chromatin (so-called "hypertrophied nuclei") of the myocytes obtained from DCM. Their ultrastructure was neither apoptotic nor necrotic but rather that of living cells. Taq polymerase-based DNA in situ ligation assay, which detects double-stranded DNA fragments more specifically than TUNEL, did not detect a positive reaction in any case. In mirror sections, all of the TUNEL-positive myocytes in DCM simultaneously expressed proliferating cell nuclear antigen, which is required for both DNA replication and repair, but Ki-67, a replication-associated antigen, was completely negative in all cases, which appeared to rule out cell proliferation activity. CONCLUSIONS: Most of the TUNEL-positive myocytes in hearts with DCM are not apoptotic but rather living cells with increasing activity of DNA repair.

Monoclonal antibodies recognizing different epitopes of the 27-kDa gap junction protein from rat liver.

Monoclonal antibodies (2-3E2, 6-3G11, and 7-3H6) against gap junction plaques purified from rat liver were prepared and characterized. Immunoblot analysis of liver gap junctions revealed that all three antibodies reacted with the 27-kDa protein, but not with the 22-kDa one. The 2-3E2 and 6-3G11 antibodies both reacted with the 27-kDa protein in gap junctions purified from livers of the rat, mouse, rabbit, and guinea pig; the 7-3H6 antibody, however, failed to react with the 27-kDa protein from guinea pig liver. The 7-3H6 antibody reacted strongly with the 24- to 26-kDa degradation products of the 27-kDa protein. Indirect immunofluorescence showed that the 6-3G11 and 7-3H6 antibodies both gave the same specific fluorescence labeling on rat liver cryosections, suggesting that these two antibodies recognized the cytoplasmic sites of the 27-kDa protein. Immunoblot analysis of protease-digested fragments from the 27-kDa protein revealed that the 7-3H6 antibody reacted with the 24- and 17-kDa fragments (including portions of the carboxyl-terminal domain of the 27-kDa protein) produced with endoproteinases Arg-C and Lys-C, respectively. Immunoblot analysis of CNBr fragments of the 27-kDa protein revealed that all three antibodies reacted with the 10-kDa fragment, which is thought to be the carboxyl-terminal domain of the 27-kDa protein. These results demonstrate that three monoclonal antibodies recognize different epitopes of the cytoplasmic sites (probably the carboxyl-terminal domain) of the 27-kDa liver gap junction protein.

Studies on heterogeneity of Taka-amylase A: isolation of an amylase having one N-acetylglucosamine residue as the sugar chain.

Crystalline Taka-amylase A, prepared from Takadiastase, was fractionated into four fractions by DEAE-Sephacel and Concanavalin A-Sepharose column chromatography. The relative weight ratio of the fractions was 90 : 4 : 4 : 2. They had similar molecular weights (51,000), amino acid compositions, and hydrolytic activity against soluble starch, but different phenyl maltosidase activities and electrophoretic mobilities on polyacrylamide gel electrophoresis. Three of the fractions mainly had the high mannose type sugar chain with the sugar composition of Man5-GlcNAc2, but the other fraction had only one N-acetylglucosamine residue as the sugar chain. These results suggested that Taka-amylase A was heterogeneous both in the sugar portions and in the polypeptide portions.

Down regulation of myelin-specific mRNAs in the mechanism of hypomyelination in the undernourished developing brain.

The expression of myelin-specific protein genes, i.e. myelin proteolipid (PLP), basic (BP), and myelin associated glycoproteins (MAG) was studied in normal and severely undernourished 20-day-old rats. The undernutrition paradigm resulted in reductions of approximately 50, 25 and 65% in body weight, brain weight and brain myelin yield, respectively. The amount of total brain RNA was not significantly altered, although the amount of cyclophilin (CYC) mRNA was increased. In contrast, the steady-state levels of myelin specific mRNAs were significantly decreased by approximately 40, 20 and 40% for PLP, BP and MAG, respectively. In addition, polyadenylation of the PLP transcript was altered, producing an abnormal ratio of the 1.6 kb to the 3.2 kb PLP mRNAs. The results indicate that down-regulation of myelin-specific gene expression is involved in the mechanisms of hypomyelination in hunger disease, although the individual genes are differently altered. Furthermore, undernutrition may have additional effects on the posttranscriptional processing of the transcripts as indicated by the abnormal size distribution of PLP messages.

Chemotherapy of brain metastases from lung carcinoma: a controlled randomized study.

A controlled randomized study was carried out to evaluate the effects of chemotherapy in patients with brain metastases from lung carcinoma. One hundred patients were randomly divided into three groups at the time of diagnosis or after surgery for metastases. Group A received radiotherapy alone; Group B received radiotherapy and chloroethylnitrosoureas (methyl-CCNU, 100-120 mg/m2, or ACNU 80-100 mg/m2, every 6-8 weeks), and Group C received radiotherapy and a combination of chloroethylnitrosoureas and tegafur (300 mg/m2, daily). Of the 100 patients, 88 could be evaluated. The reduction rates of the tumors of the patients in whom tumor was not surgically removed or not totally removed were compared. Complete resolution of the tumor was noted in 29, 69, and 63% of the patients in Groups A, B, and C, respectively. Tumor regression of greater than or equal to 50% was seen in 36, 69, and 74% of the patients in Groups A, B, and C, respectively. The difference in the response rates of Groups A and C was statistically significant (P less than 0.05). Median survival after the start of treatment for brain metastasis was 27, 30.5, and 29 weeks in Groups A, B and C, respectively. There was 1 long-term survivor (more than 5 years) in Group A, 3 in Group B, and 1 in Group C. The main cause of death was deterioration attributable to the primary lesion or systemic metastasis, and no statistical difference was noted in survival time among the groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytidylate cyclase activity is stimulated via activation of a guanine nucleotide-binding protein.

Cytidylate cyclase is a membrane-bound enzyme which catalyzes the biosynthesis of cytidine 3',5'-cyclic monophosphate (cCMP) from CTP. By using a sensitive and specific enzyme immunoassay method, we evaluated the participation of guanine nucleotide-binding protein (G-protein) in the regulation of cytidylate cyclase activity in rat brain and other tissues. AlF4-, an activator of G-proteins, effectively elevated the cyclase activity. The stimulation by GTP gamma S, a nonhydrolyzable GTP analogue, was also observed in time- and concentration-dependent manner during the preincubation and this effect was competitively inhibited by the addition of GDP beta S. However, islet-activating protein and cholera toxin which affected adenylate cyclase activity had no effect on cytidylate cyclase activity. These results indicate that cytidylate cyclase is associated with a certain G-protein and its activity is regulated by the mode different from that of adenylate cyclase.

Visual evoked potentials associated with the verbal and non-verbal problem-solving processes.

VEPs to six repetitive checkerboard and letter or number stimuli (presented in two triplets) were measured. The task solution process was separated into the store, retention, solve and post-solve stages. Three stimuli in intratrial stimulus positions 1, 2 and 3 were identical and another three in positions 4, 5 and 6 were also identical. The subject's task was either naming the letter-number or matching the checkerboard stimuli in two triplets. For both stimuli, the occipital P270 was almost the same in amplitude as positions 1 and 4. During the memory retention (at position 2) the P270 enhanced for the letter-number stimulus, but not for the checkerboard. During the post-solve stage (at positions 5 and 6) the P270 enhanced for both stimuli. The occipital P270 enhancement seems to correlate with the decrease in the degree of significance of the task relevant stimuli.

Task redundancy and an enhancement of the late positive components in human visual evoked potentials.

VEPs to four or six repetitive letter and checkerboard stimuli were measured. These stimuli were presented in a first triplet and a single stimulus or in first and second triplets. Three stimuli in a triplet were identical. The subject's task was either naming the letter stimulus or pattern matching the checkerboard. A degree of task-redundancy of the stimuli at intratrial stimulus positions 2 and 3 was varied by means of stimulus duration and indication of the task stimulus prior to the triplet. The stimulus duration was 300 msec and the prior information of the task stimulus was delivered by a letter or checkerboard warning stimulus (WS) in session 1. The stimulus duration was shorter (10 msec in session 2, 20 microseconds in session 3) and the WS of random pattern did not deliver the information of the task stimulus in session 2 and 3. At stimulus positions 2 and 3, the P260 of the occipital VEP enhanced in amplitude for the letter and checkerboard stimuli in session 1, whereas the P260 decreased in amplitude for both stimuli at the same positions in session 3.

Evaluation of mitogen-induced responses in marine mammal and human lymphocytes by in-vitro exposure of butyltins and non-ortho coplanar PCBs.

The effects of exposure to butyltin compounds (BTs: tributyltin; TBT, dibutyltin; DBT and monobutyltin; MBT) and non-ortho coplanar PCBs (IUPAC 77, 126 and 169) on marine mammals and human lymphocyte were evaluated. Peripheral blood mononuclear cells (PBMCs) isolated from Dall's porpoises (Phocoenoides dalli), bottlenose dolphins (Tursiops truncatus), a California sealion (Zalophus californianus), a larga seal (Phocoa largha) and humans (Homo sapiens) were exposed at varying concentrations of BTs and coplanar PCBs. Concanavalin A (Con A)-stimulated mitogenesis found significantly suppressed (P<0.01) when the cells were exposed at 300 nM (89 ng/ml) of TBT and 330 nM of DBT (77 ng/ml), while MBT showed little cytotoxicity at treatment levels of up to 3,600 nM (620 ng/ml). BTs concentrations in the liver of Dall's porpoises from Japanese coastal waters ranged between 81-450 ng/g for TBT and 200-1,100 ng/g (wet wt.) for DBTs, which is greater than the cytotoxic levels registered in this study. In contrast, non-ortho coplanar PCBs did not suppress cell proliferation at concentrations of up to 30 nM (10 ng/ml). The residue levels of coplanar PCBs in the blubber of Dall's porpoises were 0.12-1.3 ng/g, which were one order of lower than those levels that do cell proliferation. When cells were exposed to a mixture of TBT/DBTand coplanar PCBs, the proliferation was significantly reduced to 33 nM DBT plus 34 nM CB-77 and 33 nM DBT plus 28 nM CB-169 mixtures, respectively. The investigations relating the contaminant-induced immunosuppression in marine mammals have been focused on persistent organochlorines such as PCBs. pesticides and dioxin compounds. However, this study suggested the possibility of BTs could also pose a serious threat to the immune functions in free-ranging marine mammals and humans.

Fifteen-year follow-up of a heterozygous Fabry's disease patient associated with pre-excitation syndrome.

A 47-year-old woman with heterozygous Fabry's disease with pre-excitation syndrome has been followed up for 15 years. Diagnosis was confirmed by the typical electron microscopic feature of the endomyocardial specimen and a decreased plasma alpha-galactosidase activity. As the disease progressed, the interventricular septum thickened from 11 to 17 mm as measured by echocardiography, while the AH interval was prolonged from 80 to 140 msec. In Fabry's disease, the PR interval has been reported to be variable from short PR to AV block. Therefore, this case may be helpful to understand the time course in the AV conduction abnormalities with the progression of Fabry's disease.

Relationship between thallium-201 myocardial SPECT and findings of endomyocardial biopsy specimens in dilated cardiomyopathy.

The purpose of this study was to clarify which myocardial histological findings associated with dilated cardiomyopathy (DCM) are reflected in quantitative 201Tl myocardial SPECT. We obtained studied SPECT images from 21 patients with DCM 10 minutes and 2 hours after they received an injection of 111 MBq 201Tl at rest. We calculated the percent coefficient of variation of myocardial 201Tl counts [%CV(TI)], the washout rate (WR), standard deviation of WR [SD(WR)], extent score (ES) and severity score (SS). We used image analysis to measure % fibrosis, % myocytes, the ratio of fibrous tissue to myocyte tissue (F/My), myocyte size and standard deviation of myocyte size [SD(My)] in left ventricular endomyocardial biopsy specimens. The %CV(Tl) was correlated with % fibrosis and F/My. The ES and SS also correlated with F/My. The correlation between SD(WR) and SD(My) was significant. The present findings suggest that %CV(Tl), ES and SS of rest 201Tl SPECT reflect myocardial fibrosis and that the standard deviation of washout reflects the distribution of myocyte size.

[Recurrent cerebral aneurysm suggestive of misplaced clipping. Case report].

A case of recurrent cerebral aneurysm after complete neck clipping is described. A 47-year-old male who presented with headache and nausea underwent neck clipping of a berry aneurysm of the left middle cerebral artery. Ten days later, angiographic findings suggested the presence of a second, large aneurysm adjacent to the first, which suggested misplaced clipping. Reoperation confirmed that a new aneurysm had formed next to the original aneurysm. A possible explanation of the recurrence is as follows. The M1 flowed into the M2 at a right angle. The aneurysmal neck was situated on the distal end of the M1 and the dome protruded antero-inferiorly at an angle of nearly 90 degrees to the long axis of the M1 opposite the origin of the M2. The parent artery bulged slightly, and its wall was thin and reddish, just distal to the aneurysmal neck. Proximal to the neck there was another small bulge, but the wall here was normal. These bulges were coated with Oxycel and Biobond at the time of aneurysmal neck clipping. After clipping, blood flow into the dome was interrupted, and the consequent hemodynamic stress caused the bulges to expand dramatically and form a new aneurysm. The authors conclude that there is a likelihood of early recurrence after neck clipping if the parent artery exhibits such morphological features as observed in this case.

Epidermoid of the quadrigeminal cistern--case report.

The authors report a case of an epidermoid in the quadrigeminal cistern in a 40-year-old female who presented with headache and nausea. According to the literature, epidermoids are rarely found in the quadrigeminal cistern. Additional unusual features in this case were the absence of a capsule and T1-weighted magnetic resonance images suggestive of both solid (less intense) and fluid (more intense) components.

[Postoperative infections related to pacing wires, pulmonary arterial catheters, and drainage tubes temporarily inserted during open-heart surgery].

Bacterial examinations of temporary pacing wires (P-wires), pulmonary arterial (P-A) catheters, and drainage tubes temporarily inserted during open-heart surgery were performed in 213 patients. Bacteria were detected in 19 (2.8%) of 672 specimens gathered from the subject patients, with coagulase-negative Staphylococcus (CNS) being most frequently observed. P-wires accounted for 17 out of 19 of the culture-positive specimens, and 7 of the P-wires remained in place for more than two weeks. The frequency of infection with the P-wires was significantly higher than with the P-A catheters or drainage tubes. The period of time that the P-wire was left in place significantly longer than for P-A catheter or drainage tube. There was, however, no statistically significant difference between the culture-positive and negative groups in respect to age, detention periods, operation times, CPB times, or length of ICU stay. As a result of these findings, we have concluded that P-wires should be removed as soon as possible following surgery, and in any case, a meticulous care should be taken to prevent transcutaneous infection.

[Presence of human epidermal growth factor (hEGF), basic fibroblast growth factor (bFGF) in human aqueous].

Thirty-eight cataract eyes and 15 artificial aphakic or pseudophakic eyes were enrolled in this study to determine levels of human epidermal growth factor (hEGF) and basic fibroblast growth factor (bFGF) in human aqueous, using a radioimmunoassay. Neither hEGF nor bFGF were detected in either congenital cataract eyes or senile cataract eyes without any complication. In cases of senile cataract complicated with glaucoma and traumatic cataract, bFGF range from 0.4 to 0.8 ng/ml whereas hEGF was not detected. In cases of senile cataract with myocardial or brain infarction and anterior subcapsular cataract, bFGF was detected ranging from 0.4 to 1.0 ng/ml and in some of the cases hEGF was detected at a level of 1.0 ng/ml. hEGF was detected at 1.0 ng/ml in some cases of secondary cataract after uveitis, cataract complicated with retinal detachment, cataract after scleral buckling or vitrectomy, and aphakic or pseudophakic eyes after extracapsular cataract extraction or phacoemulsification. These results showed that hEGF and bFGF exist at a level of about 1 ng/ml in human aqueous in some pathological states and it seems possible that proliferation of lens epithelial cells is promoted by the growth factors, with the result that after cataract, anterior capsular opacities and shrinkage become severe.

[Intraventricular arachnoid cyst--on the origin of intraventricular arachnoid cysts].

Arachnoid cysts very rarely occur within the ventricular system, where no arachnoid tissue exists. We present three cases of intraventricular arachnoid cyst with special reference to its origin. The first patient was a 5-year-old boy who complained of headaches and enlargement of his head. A CT scan revealed obstructive hydrocephalus and a large cystic lesion in the right lateral ventricle. The symptoms resolved after fenestration of the cyst and cystoperitoneal shunt. The second patient was a 49-year-old woman who complained of headache and numbness in her left upper extremity. A CT scan and MRI revealed a large cyst in the trigone of the right lateral ventricle. Fenestration of the cyst wall and cystoperitoneal shunt were performed relieving her complaints. The third patient was a 42-year-old man who complained of frequent seizures and dizziness. A CT scan and MRI demonstrated a moderate size cystic mass in the inferior horn of the right lateral ventricle. The symptoms were improved by partial resection of the cyst wall. Immunohistochemical studies and light microscopy confirmed that the cyst walls were composed of arachnoid membrane, implying that the cysts were arachnoid cysts. CT and MRI in these three cases showed widening of the choroidal fissure bordering the cyst wall, occasionally involving a part of the protruding cyst wall. On enhanced CT and MRI, the choroid plexus in the trigone of ipsilateral lateral ventricle was displaced anterolaterally, implying that the cysts had grown from outside the choroid plexus. Postoperative MRI demonstrated the shrunken cyst wall attached to the choroidal fissure. The surgical findings in case 3 also showed that the cyst wall was attached firmly to the choroid plexus. These findings appeared to indicate that the intraventricular arachnoid cyst originated from the arachnoid layer drawn into the choroidal fissure with choroidal vascular mesenchyme.

[The prevalence and the clinical characteristics of silent myocardial ischemia detected by stress thallium scintigraphy].

The prevalence of silent myocardial ischemia was retrospectively assessed in a group of 100 consecutive patients with angiographically proved coronary artery disease, and diagnostic ECG, by symptom-limited exercise thallium-201 scintigraphy. Twenty-four patients had no evidence of ischemia despite adequate exercise level. So among 76 patients with exercise induced ischemia, only 33 patients (43%) stopped exercise due to anginal pain (symptomatic ischemia: Group 3). And 43 patients with asymptomatic ischemia composed of 23 patients (30%) with ECG change (Group 2B) and 20 patients (26%) without ECG change (Group 2A). Patients background including the history of old myocardial infarction and diabetes mellitus, were similar among Group 2A, 2B, and Group 3. And our major observation was that the extent and severity of quantified SPECT perfusion defects was nearly identical between 3 groups Thus in this study group, there was a rather high prevalence rate of silent ischemia (57%) by exercise thallium-201 criteria. Patients with silent ischemia, associated with positive and negative exercise ECG findings, and those with exercise angina had similar background and comparable amount of jeopardized myocardium.

[Two cases of stent graft repair for thoracic aortic dissection: usefulness of three-dimensional computed tomogram].

We carried out stent graft repair in two patients with Stanford type B thoracic aortic dissection. A 51-year-old male was admitted to our hospital because of thoracic aortic dissection. Chest CT revealed an aneurysm of the distal aortic arch. The entry was pointed out 1 cm distal from the take off of the left subclavian artery in three-dimensional CT (3 D-CT). He was treated with a Gianturco stent which was anchored into the 30 mm Hemashield graft under selective cerebral perfusion. Another case was a 72-year-old male with a descending aortic aneurysm. 3 D-CT showed that the entry existed 4 cm proximal to the celiac artery. We performed transluminal implantation of the spiral Z-stent covered with the woven Dacron graft. 3 D-CT was useful for the preoperative management and the surgical treatment of thoracic aortic dissection.

[A difficult case of antiphospholipid syndrome with repeated restenosis of coronary artery and grafts].

A 65-year-old man complained of exertional angina. Coronary angiography revealed 99% stenosis in the left anterior descending artery (LAD). He underwent percutaneous transluminal coronary angioplasty (PTCA) and stenting. Repeat angiography demonstrated restenosis of the previous PTCA and stenting site. He underwent coronary artery bypass grafting (CABG) with placement of the left internal thoracic artery (LITA) to LAD. Chest discomfort and V1-V3 ST elevation appeared on the first post operative day. Coronary angiography revealed occlusion of the LITA graft. He underwent re-operation. Because the radial artery was severely sclerotic, the great saphenous vein was used for the graft. Two weeks later, he began to show edema in the left lower extremity. Echoangiogram showed occlusion of the left deep vein with thrombus. He tested positive for anticardiolipin IgG and IgM antibodies. Eighteen months after re-operation, he had recurrent chest discomfort on exertion. Coronary angiography revealed 90% stenosis of the anastomosis (SVG-LAD). A second re-operation was performed. We used the right internal thoracic artery (RITA) for the graft. The postoperative angiography showed patent graft. The patient has been doing well without any complications.