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The electromagnetically spinning method for measurement of fluid viscosity was improved to obtain accurate values of low viscosity in the low shear rate region. Harmful effects derived from the mechanical friction to the smooth rotation of the rotor are thoroughly eliminated by employing a viscosity probe suspended by a thin metal wire. In our previous study, the motion of the meniscus of the sample surface also acted as a troublesome resistant torque for probe rotation, which was addressed by employing a sandwiched structure of the disk probe between the bottom and top plates. The measurements were carried out in two procedures. In the freely oscillating operation, we could measure the viscosity of the atmosphere with a viscosity of approximately 1/100 of that of water. The second type of quasi-steady measurement enabled a measurement of pure water in the range of shear deformation rates smaller than 1 s−1.

The electromagnetically spinning method for viscosity measurement was enhanced for applications in a confined reaction chamber. In the reaction chamber, the immersion of the rotor into the sample might trigger the precipitation and aggregation of ingredients to the pivot of the probe rotor, which leads to harmful frictional torque of the rotation. To address this problem, an upper-point-type auto-standing probe rotor was developed and applied for the long-term measurement of the solution process of rock sugar in water. In addition, we propose the adoption of a punched board, instead of a solid board, as the lower substrate of the plate-plate-type rotational viscometer, which allows rapid exchange of the sample fluid between the interior and exterior of the narrow measurement area through molecular diffusion. We investigated the effect of punches from the perspective of the diffusion process of momentum and determined the geometrical conditions.

We evaluated the usefulness of the total myocardial uptake ratio (TMUR) of 15-(p-123Iiodophenyl)-3(R,S)-methyl-pentadecanoic acid (123I-BMIPP) for predicting cardiac complications in patients with mitochondrial encephalomyopathy or myotonic dystrophy. Six patients with mitochondrial encephalomyopathy, four with myotonic dystrophy, and 10 control subjects were studied. Quantitative assessment of 123I-BMIPP dynamic myocardial imaging was performed, and the TMUR of 123I-BMIPP was calculated according to the Ishii-MacIntyre method. Then, the TMUR was compared in the 10 patients and 10 healthy controls, and all patients were followed for 56.1±22.1 months to evaluate cardiac complications. TMUR in patients (2.69±0.64) was significantly (P0.01) lower than that in controls (3.28±0.25). Three patients in whom the TMUR value was above 3.00 had no cardiac complications. On the other hand, all patients in whom TMUR was below 3.00 had some kind of cardiac complication during the follow-up period. Two patients showed progressive conduction abnormality and underwent pacemaker implantation, one patient had sick sinus syndrome and underwent pacemaker implantation, another patient showed non-sustained ventricular tachycardia and paroxysmal atrial fibrillation, and four of seven patients, including one with a pacemaker, showed an increased cardiothoracic ratio value over 50. In conclusion, measurement of the TMUR by the Ishii-MacIntyre method is useful for evaluating the development of cardiac complications in patients with mitochondrial encephalomyopathy or myotonic dystrophy.

Abstract: PE-35 monoclonal antibody, detecting a cell-surface antigen of various types of carcinoma and normal epithelium, reacts exclusively with the medullary epithelium in the thymus; therefore, the antigen has been considered as a marker of medullary differentiation in thymomas. Using the catalyzed signal amplification method, which made it possible to apply PE-35 to routinely processed, archival tissues, we examined expression of this antigen, together with CD1a reactivity of lymphocytes, in 40 thymic epithelial tumors subclassified using the Mü1ler-Hermelink system. Medullary thymomas infiltrated with a small number of CD1a-negative lymphocytes were PE-35 positive, although many of the long spindle tumor cells were PE-35 negative. Mixed thymomas and predominantly cortical thymomas, both with prominent CD1a-positive lymphocytes, were also PE-35 positive, although some areas of the latter type were PE-35 negative. Cortical thymomas with decreased numbers of CD1a-positive lymphocytes were largely PE-35 negative. In well-differentiated thymic carcinomas with a few CD1a-positive lymphocytes, two cases were negative, but four cases were at least focally positive with PE-35. All high-grade thymic carcinomas infiltrated with some CD1a-negative lymphocytes were PE-35 positive. These results suggested that medullary thymoma generally possesses the medullary nature, although the latter tends to be lost in the long spindle tumor cells. Mixed and predominantly cortical thymomas may have mixed medullary phenotype and cortical function. Cortical thymoma and many well-differentiated thymic carcinomas may possess the cortical nature, while the large polygonal tumor cells tend to lose immature T-lymphocyte-retaining function.

Dendritic polyamides having a high degree of branching and a narrow polydispersity were synthesized by a “one-pot” procedure which consists of successive activation of carboxyl groups followed by condensation with an aminodicarboxylic acid, as the AB2 monomer. 5-[3-(4-Aminophenyl)propionylamino]isophthalic acid hydrochloride (4) was used, as the AB2 monomer, without a protecting group. The carboxyl group of trimesic acid (8) was activated with an equimolar condensing agent diphenyl (2,3-dihydro-2-thioxo-3-benzoxazolyl)phosphonate (DBOP) to produce the corresponding active amides 1,3,5-benzenetris(3-carbonylbenzoxazoline-2-thione). Then, 3 equiv of 4 was added to 8 and condensed. The dendritic polyamides were synthesized divergently by repeating these activations and condensations. The terminal carboxyl groups were end-capped by diethylamine and the products were isolated. Dendritic polyamides obtained were characterized by IR and NMR spectroscopies and GPC. Number-average molecular weights (Mn) of dendritic polyamides estimated by the end group analysis were in good agreement with the calculated values. The degrees of branching and the polydispersities Mw/Mn were 0.8−0.9 and 1.2−1.5, respectively.

Abstract: A patient with stage III vulvar squamous cell carcinoma who was successfully treated with intra-arterial and intravenous chemotherapy and subsequent local excision is presented. The tumor was located on the left side of the vulva. Chemotherapy initially was administered into the left internal iliac artery. After two courses of the intra-arterial infusions, the tumor was no longer visible macroscopically. Two additional courses of chemotherapy were given intravenously to treat possible metastases to the lymph nodes. After the chemotherapy, the tumor site in the vulva was excised. A few small nests of tumor cells were identified in the excised specimen. The appearance of the vulva after surgery was quite natural and similar to that of a woman without disease. The patient is alive and disease-free 36 months after surgery. Conservative therapy, which includes intra-arterial and intravenous chemotherapy, should be considered for selected women with advanced vulvar squamous cell carcinoma.

The findings on computed tomography (CT) of a huge cystic craniopharyngioma in a 3-year-old girl are described. The cyst occupied both anterior cranial fossae and a part of it extended to the region of the third ventricle which was displaced posteriorly. The tumor showed no contrast enhancement after the intravenous administration of contrast medium.

Papillomaviral E2 genes encode proteins that regulate viral transcription. While the full-length bovine papillomavirus type 1 (BPV-1) E2 peptide is a strong trans activator, the homologous full-length E2 product of human papillomavirus type 16 (HPV-16) appeared to vary in function in previous studies. Here we show that when expressed from comparable constructs, the full-length E2 products of HPV-16 and BPV-1 trans activate a simple E2- and Sp1-dependent promoter up to approximately 100-fold in human keratinocytes and other epithelial cells as well as human and animal fibroblasts. Vaccinia virus-expressed, purified full-length HPV-16 and BPV-1 E2 proteins bound a consensus E2 site with high specific affinities (Kd = approximately 10(-9) M) and stimulated in vitro transcription up to six- to eightfold. In vivo and in vitro trans activation by either E2 protein required cooperation with another activator, such as Sp1, or other factors that interact with papillomavirus promoters, such as AP-1, Oct-1, nuclear factor 1/CTF, transcriptional enhancer factor 1, or USF. The glutamine-rich domain B of Sp1 or the mutually unrelated activation domains of other transcription factors were necessary and sufficient for cooperation with either E2 factor. We conclude that like BPV-1 E2, the HPV-16 E2 protein has the potential to function as a strong activator of viral gene expression in cooperation with cellular transcription factors.

A form of systemic tetanus with atypical symptoms was observed in mice injected in the left thigh with a mixture of tetanus toxin and antibody produced in guinea pigs against a fragment of toxin obtained from a subtilisin digest of the crystallized toxin. The mice did not show typical symptoms of the local tetanus such as convulsions or spastic paralysis of the injected limb.

The covalent structure of bovine brain calreticulin, a major Ca(2+)-binding protein in the lumen of the endoplasmic reticulum, was determined by analysis of the purified protein. The protein consisted of 400 amino acids, with an N-linked oligosaccharide attached to the polypeptide chain. The polypeptide sequence determined was compatible with the sequence of calreticulin deduced from cDNA of different sources, with a number of differences presumably due to species-specific amino acid substitutions. The protein retained the C-terminal tetrapeptide, KDEL, involved in retention of proteins resident in the endoplasmic reticulum, whereas the N-terminal signal peptide predicted from the cDNA sequence had been removed in the purified protein. The bovine brain protein contained a high-mannose type of oligosaccharide attached to Asn162, which is typical of resident endoplasmic reticulum proteins. The carbohydrate moiety was heterogeneous and had the composition GlcNAc2Man4-9, of which GlcNAc2Man5 was the most abundant in the bovine brain preparation. Glycosylation of calreticulin, however, appeared to be a species-specific modification, as Asn162 is replaced by Asp in the sequences already determined for a number of species. Analysis of the purified protein also identified an intramolecular disulphide bridge between Cys120 and Cys146.

Summary Eighteen cases of intracranial meningioma that received preoperative or post-operative irradiation with60Cobalt or Linac have been reviewed. Eight patients received preoperative irradiation because their tumours were very large, hypervascular, and surgically not readily accessible. Two angioblastic meningiomas of the haemangiopericytoma type responded well to irradiation and were easily extirpated. One of these developed a local recurrence and metastases to the lungs and liver nine years after surgery. The recurrent intracranial tumour and nodules in the lungs responded well to irradiation. The other six cases of meningiomas of meningothelial, transitional, or haemangioblastoma types, did not improve clinically or radiologically after irradiation.

Thermotolerance under chronic exposure to moderate hyperthermia at 41°C was hardly induced in the mouse temperature-sensitive mutant ts85 cells, in contrast to the parental wild-type FM3A cells. Thermotolerance was induced at a reduced level in the mutant cells compared with the wild-type cells by incubation at 33°C (permissive temperature), but not at 39°C (non-permissive temperature), after a brief exposure at 44°C. Under conditions where protein synthesis was inhibited by cycloheximide at 41°C, significant amounts of thermotolerance developed in FM3A cells. FM3A cells depleted of cellular ATP by treatment with 2,4-dinitrophenol and 2-deoxyglucose were not sensitized to thermal cell killing at 44°C, when drug-treated cells were washed and exposed to hyperthermia in drug-free growth medium, where cellular ATP rapidly recovered. However, the cells deprived of ATP under the treatment at 41°C failed to develop thermotolerance, indicating a requirement of ATP for thermotolerance development. The decay of thermotolerance was not affected by ATP levels after it was developed. The degradation of abnormal cellular proteins which contained amino acid analogues was promoted at 33°C relative to normal protein degradation in FM3A and ts85 cells. Both normal and abnormal proteins were degraded at a reduced rate at 43°C. Pretreatment of cells at 41°C decreased the rate of degradation of abnormal proteins at 33°C by 20% in FM3A cells and by about 100% in ts85 cells. Pretreatment of cells at 41°C increased significantly the conjugation of 125I-labeled ubiquitin to cellular endogenous proteins in extracts of FM3A cells, but decreased the conjugation in extracts of ts85 cells. The data presented here, in conjuntion with the observations by others that the ts85 cell is a mutant defective in the ubiquitination of cellular proteins at non-permissive temperatures, suggest that the ATP-dependent ubiquitination may be crucial for the development of thermotolerance.

Objective and Design:The role of cyclooxygenase (COX)-2 was examined using a rat endotoxin shock model and the potency and selectivity of NS-398, a COX-2 selective inhibitor in vitro, for COX-2 activity was examined in vivo.¶Material:Male Wistar rats (weighing 140–180 g) were used.¶Methods:Lipopolysaccharide (LPS, 1 mg/kg, i.v.) was administered to rats (LPS-treated rats) and expression of COX-1 mRNA and COX-2 mRNA in the aorta and peripheral blood leukocytes was examined by RT-PCR. COX activity was assessed by measuring the plasma 6-keto prostaglandin (PG) F1α, PGE2and thromboxane (TX) B230 s after administration of arachidonic acid (AA, 3 mg/kg, i.v.). NS-398 (0.3–100 mg/kg, p.o.) or indomethacin (0.3–3 mg/kg, p.o.) was administered 1 h before the AA injection.¶Results:COX-2 mRNA was detectable in the aorta and peripheral blood leukocytes at least from 3 to 9 h after the LPS injection but not in non-LPS-treated rats. Plasma 6-keto PGF1α, PGE2and TXB2levels after AA injection into LPS-treated rats were significantly enhanced compared to findings in non-LPS-treated rats. NS-398 showed significant inhibition of the increase in PGs in LPS-treated rats, the ED50values being 0.35 mg/kg for 6-keto PGF1α, 1.5 mg/kg for PGE2and < 0.3 mg/kg for TXB2. NS-398 even at 100 mg/kg did not significantly suppress the increased PGs levels in non-LPS-treated rats. In contrast, indomethacin significantly inhibited plasma PGs levels after AA injection into LPS-treated rats and non-LPS-treated rats. The ED50values in LPS-treated rats, determined by 6-keto PGF1α, PGE2and TXB2production, were 1.0, 1.3 and 2.3 mg/kg and those in non-LPS-treated rats were 0.42, 0.24 and 0.93 mg/kg, respectively.¶Conclusions:In a rat endotoxin shock model, expression of COX-2 plays a role in an increase in COX activity. NS-398 showed preferential inhibitory effects on COX-2 activity in vivo. This approach is useful to directly analyze the inhibitory activity of NSAIDs for COX-1 and COX-2 in vivo.

A series of balloon-borne experiments to search for particles of cosmic origin is being planned. The experimental apparatus to be borne on the balloon includes a drift chamber as one of its main components. Flash analog-to-digital convertors digitize signals from the chamber at the rate of 30 megacycles/s. The apparatus on the balloon is linked by radio to a ground station. The apparatus is remotely controlled through a terminal placed at the ground station. A description is given of the apparatus on the balloon, data transmission from the balloon to the ground station, the software on the balloon, command transmission, and the computer complex at the ground station.<>

Twelve cases of tracheobronchomalacia (TBM) cases were reviewed: five were pediatric, and seven were adult, two of which were due to relapsing polychondritis (RPC). In pediatric TBM, the malacic segments were short. Resection of the malacic segment in one case and laryngotracheoplasty with autologous costal cartilage in one case were unsuccessful. However, aortopexy gained good results. Two cases managed conservatively experienced gradual improvement of their symptoms. In adult TBM, plication of pars membranacea was not effective in one case. The insertion of a stent was minimally effective in one case, and distinctly in one polychondritic case. The other four cases managed conservatively have deteriorated gradually. From these findings, a new classification system is proposed.

Abstract1. To improve the production of chimeras, the distribution of donor blastodermal cells after transferring into recipient embryos was examined morphologically.2. Donor blastodermal cells were distributed near the site of injection in the epiblast and in the subgerminal cavity and yolk. Some filled the hole made by the micropipette and were distributed outside the epiblast. Many were buried in yolk. In some cases, more donor blastodermal cells were located in the yolk than in the subgerminal cavity and some were located 800 μm below the under‐surface of the epiblast.3. It is recommended that injection should be as shallow as possible to increase the proportion of chimeras produced, and that some means is needed to prevent blastodermal cells from escaping from the hole produced by injection.

Three infants with congenital tracheal stenosis and three adults with various diseases of the central airway underwent esophageal tracheobronchoplasty to repair long-segment stenoses and defects. The primary operative goal was enlargement of the stenosis ( n = 4), repair of the defect ( n = 1), or both ( n = 1). Cardiopulmonary support was required in two cases. All three infants were operated on for generalized congenital tracheal stenoses. There was one postoperative death on the fifth day. Another infant died of pneumonia 3 months after operation. Tracheal patency was excellent in two infants. One infant is well without symptoms 6 years after the operation, although balloon dilation was required three times during the first postoperative year. In the three adult patients, the primary diseases were congenital tracheal stenosis, iatrogenic injury associated with relapsing polychondritis, and malignant mediastinal tumor involving the trachea. All lesions involved both the trachea and main stem bronchi. Postoperative airway patency was excellent in all three adults, although expandable metallic stents had to be inserted in one patient. Postoperative pulmonary function was improved, particularly forced expiratory volume in 1 second and peak expiratory flow rate. Although the postoperative mortality rate was still high, especially among the infants, and prolonged postoperative ventilatory support was required for five of the six patients, long-term patency and postoperative pulmonary functional improvement are encouraging. (J Thorac Cardiovasc Surg 1996;112:124-9)

There are few specific pathologic findings that can be relied on to distinguish primary thymic carcinomas from lung carcinomas with mediastinal extension or showing metastasis to the anterior mediastinum. The immunohistochemical reactivity on frozen sections of thymic carcinomas and lung carcinomas, which are histologically similar to each other, was examined with the use of monoclonal antibodies to cytokeratins 7 and 13. Among keratinizing squamous cell carcinomas, all thymic carcinomas reacted with antibody specific for cytokeratin 7 (9/9, 100%), whereas no staining reaction was seen in lung carcinomas (0/5, 0%) ( p < 0.01). This finding can be used as a diagnostic aid in primary thymic keratinizing squamous cell carcinomas to expedite treatment and prognosis. Cytokeratin 7 and cytokeratin 13 monoclonal antibodies reacted with almost all cases of thymic carcinoma. Applications of monoclonal antibodies specific for certain cytokeratins, especially 7 and 13, may be helpful in the diagnosis of other subtypes of thymic carcinomas. (J THORAC CARDIOVASC SURG 1995;110:1670-5)

Prothymosin-alpha, the precursor of thymosin-alpha1, may play a role in cell proliferation, and the plasma level of thymosin-alpha1 may reflect the degree of proliferation of the tumor cells.

A chemotactic protein for polymorphonuclear leukocytes (lung carcinoma-derived chemotaxin [LUCT]) was purified from culture fluid of the human lung giant cell carcinoma LU65C cells to electrophoretically homogeneous form through five sequential purification steps: DEAE-Sepharose, CM-Sepharose, HPLC on carboxyl-methylated-polyvinylalcohol resin, hydrophobic, and reversed-phase. The molecular mass was determined as approximately 10 kD by SDS-PAGE and isoelectric point was 10.7. The chemotactic activity (ED50 0.75 x 10(-9) M) was sevenfold more potent than that of FMLP (5 X 10(-9) M) and comparable with that of C5a (10(-9) M). NH2-terminal amino acid sequence and amino acid composition of LUCT strongly suggest that it may be closely related to the putative protein encoded by the cDNA clone (3-10C) and almost identical with a part of sequence of the chemotactic factor derived from stimulated human leukocytes in the 6th to 32nd, but not the NH2-terminal 5 amino acids. These results indicate that the carcinoma cells produce LUCT without any added stimulant and suggest that the previously isolated chemotactic monokines may correspond to des(1-5) of LUCT in the NH2-terminal region.

Clostridium perfringens produced at least three distinct proteases in a synthetic medium containing calcium. Two of them, thiol and ethylenediaminetetraacetic acid disodium salt-sensitive proteases, appeared at an early stage of growth, but the other one, perhaps being identical to the one produced in a calcium-deficient medium, appeared at a late stage. The production of these proteases depended on Ca2+ but not on Zn2+ in the medium. Alpha-toxin, perhaps being a zinc-containing metalloenzyme, was rather resistant to the proteases, but toxin, produced in a zinc-deficient medium or deprived of zinc with ethylenediaminetetraacetic acid disodium salt, was very sensitive. By adding Zn2+, the toxin lacking zinc may have been converted to the zinc-containing metalloprotein that is resistant to proteases. This may explain why alpha-toxin activity increased progressively in a zinc-containing medium in spite of simultaneous production of potent proteases and why it disappeared rapidly in a zinc-deficient medium.

Hepatitis E virus (HEV) is a pathogenic agent that causes fecally-orally transmitted acute hepatitis. The genome, a single-stranded positive-sense RNA, encodes three forward open reading frames (ORFs), in which an approximately 2-kb structural protein is located in the 3' end. To produce HEV-like particles the structural protein, with its N terminus truncated (amino acid residues 112 to 660 of ORF2), was expressed in insect Tn5 cells by a recombinant baculovirus. In addition to the primary translation product with a molecular mass of 58 kDa, a large amount of a further-processed molecule with a molecular mass of 50 kDa was generated and efficiently released into the culture medium. Electron microscopic observation of the culture medium revealed that the 50-kDa protein self-assembled to form empty virus-like particles (VLPs). The buoyant density of the VLPs in CsCl was 1.285 g/cm3 and their diameter was 23.7 nm, a little smaller than the 27 nm of native HEV particles secreted into the bile or stools of experimentally infected monkeys. The yield of the VLPs was 1 mg per 10(7) cells as a purified form. The particles possess antigenicity similar to that of authentic HEV particles and, consequently, they appear to be a good antigen for the sensitive detection of HEV-specific immunoglobulin G (IgG) and IgM antibodies. Furthermore, the VLP may be the most promising candidate yet for an HEV vaccine, owing to its potent immunogenicity.

An extremely rare case of a sellar mature teratoma with a melanotic progonoma is reported. The patient was a 14-year-old boy who showed diabetes insipidus and panhypopituitarism. The tumor was composed of mature teratoma and clusters of pigmented and non-pigmented polyhedral cells which were identical to those seen in a melanotic progonoma in other sites. Premelanosomes were observed in the cytoplasm of the polyhedral cells under an electron microscope. Melanotic progonoma was considered to be of teratoid origin in the present case.

The carboxylic acids (I) are coupled with the alkynes (II) in the presence of bis(η5‐cyclooctadienyl)ruthenium to form the enol esters (III).

The secondary amines (I) are coupled with carbon dioxide (II) and 1‐hexyne (III) in the presence of Ru(0) and phosphines to form the O‐hexenyl carbamates (IV).