Thin film growth of CaAgAs by molecular beam epitaxy.

We have grown thin films of CaAgAs by molecular beam epitaxy, which was theoretically proposed to be a topological insulator. The temperature dependence of resistivity and the carrier concentration at 4 K were similar to the reported results of bulk samples. However, the magnetoresistance exhibited a steep increase at low magnetic fields, a behavior not observed for bulk samples. This steep increase of resistivity is ascribable to the weak antilocalization effect and provides clues to the nature of the topological surface state of CaAgAs.

Reconstruction of band structure induced by electronic nematicity in an FeSe superconductor.

We have performed high-resolution angle-resolved photoemission spectroscopy on an FeSe superconductor (T_{c}∼8 K), which exhibits a tetragonal-to-orthorhombic structural transition at T_{s}∼90 K. At low temperature, we found splitting of the energy bands as large as 50 meV at the M point in the Brillouin zone, likely caused by the formation of electronically driven nematic states. This band splitting persists up to T∼110 K, slightly above T_{s}, suggesting that the structural transition is triggered by the electronic nematicity. We have also revealed that at low temperature the band splitting gives rise to a van Hove singularity within 5 meV of the Fermi energy. The present result strongly suggests that this unusual electronic state is responsible for the unconventional superconductivity in FeSe.

Groin hernias in adults: value of color Doppler sonography in their classification.

PURPOSE: We assessed the abilities of color Doppler and power Doppler sonography to distinguish among types of groin hernias by demonstrating the inferior epigastric artery (IEA) and its relationship with the hernia sac. METHODS: Nineteen consecutive patients (14 men and 5 women), clinically diagnosed as having groin hernias and scheduled to undergo herniorrhaphy, were prospectively enrolled in this study. Ultrasound examinations were performed preoperatively with a 6-12-MHz linear-array transducer. The IEA was identified, if possible, and its relationship to the hernia sac assessed. The sonographic diagnoses were compared with the operative findings. RESULTS: There were 15 indirect inguinal hernias, 4 direct inguinal hernias, and 1 femoral hernia; 1 patient had bilateral inguinal hernias (indirect and direct). In 18 (90%) of 20 hernia cases, the trunk segment of the IEA could be visualized. In 11 (55%) of 20 hernia cases, the origin segment of the IEA could be visualized and its relationship with the hernia sac assessed. In 9 (82%) of the 11 hernia cases, hernia types were correctly diagnosed by sonography. The overall accuracy of sonography for diagnosing the type of hernia was 45% (9 of 20 hernias). CONCLUSIONS: Color Doppler sonography can accurately differentiate types of groin hernias if the origin segment of the IEA and the hernia sac can be visualized simultaneously. However, color Doppler sonography sometimes failed to visualize this segment.

Nonenzymatic elimination of ascorbic acid in clinical samples.

OBJECTIVES: Ascorbic acid interferes significantly in the oxidative reaction of chromogenic reagents by peroxidase and hydrogen peroxide. Currently, ascorbate oxidase is commonly utilized for eliminating the interference of ascorbic acid in the oxidative colorimetric reaction. This enzyme, however, displays several disadvantages, such as high cost, variation from lot to lot, and low stability. We applied a series of commercially available and stable radicals (ascorbic acid quenchers [AAQs]) for nonenzymatic quenching of ascorbic acid in the uricase-based uric acid determination in serum and urine. DESIGN AND METHODS: In order to evaluate the quenching activity of AAQs, a commercially available uric acid detection kit was used. TBA-80FR.NEO biochemical analyzer was utilized for the assay. RESULTS: 4-Hydroxy-2,2,6,6-tetramethyl-1-piperidinyloxy free radical (AAQ-2) was the most effective ascorbic acid quencher among the four stable radicals, and the uric acid assay suffered no interference by AAQ-2. The ascorbic acid quenching ability of 2 mmol/L of AAQ-2 in reagent solution (reagent-I) was > or = 2 U/ml ascorbate oxidase in reagent solution. CONCLUSIONS: AAQ-2 was proven to be a suitable quencher of ascorbic acid in clinical samples.

Modification of release rates of cyclosporin A from polyl(L-lactic acid) microspheres by fatty acid esters and in-vivo evaluation of the microspheres.

Biodegradable microspheres containing cyclosporin A (CyA, cyclosporine) were prepared using poly(L-lactic acid) (PLA) and poly(lactide-co-glycolide)(PLGA) by a solvent evaporation method. CyA was efficiently entrapped in PLA, PLGA(50/50) and PLGA(75/25) microspheres in a range of 81-85%. CyA released constantly from PLA microspheres without any lag time, whereas the drug from PLGA(50/50) and PLGA(75/25) microspheres was released after lag time of about 1 and 3 weeks, respectively. Addition of fatty acid esters enhanced the release rates of CyA from PLA microspheres. In-vivo study was performed using rats with adjuvant-induced arthritis. PLA microspheres with ethyl myristate sustained high blood levels of CyA compared with the microspheres with no additives over 4 weeks. In addition, the PLA microspheres improved the symptoms such as the decrease in body weight and the increase in paw swelling occurred by adjuvant-induced arthritis in rats. Consequently, the release rate of CyA from PLA microspheres can be improved by adding fatty acid esters and PLA microspheres with fatty acid esters seem to be a useful dosage form for autoimmune disease therapy.

Determination of glycated albumin by enzyme-linked boronate immunoassay (ELBIA).

A new affinity method for quantification of glycated albumin by an enzyme-linked boronate-immunoassay (ELBIA) has been established, based on the interaction between boronic acids and the cis-diols of glycated human serum albumin (HSA) trapped by anti-HSA antibody. To evaluate the ELBIA, we first examined the accuracy of the conventional boronate affinity chromatographic (BAC) method. In the BAC method, 8.1-18.9% of nonglycated albumin calibrator nonspecifically bound to the boronate affinity column, values that were regarded as the column blank. In the modified BAC method, therefore, we substracted the column blank value from the measured glycated albumin value to obtain the true value. Because glycated albumin values measured by ELBIA were exactly the same as reported by the modified BAC method, we suggest that the ELBIA results reflect the real status of albumin glycation. We have also developed a fully automated ELBIA system, allowing multiple, rapid, and precise measurements of glycated albumin.

Hydrolysis of xylan by Aspergillus niger immobilized on non-woven fabrics.

Aspergillus niger, which produces xylan-degrading enzymes, was immobilized on non-woven fabrics. The maximum xylan hydrolysis activity (15 U/cm3-support) and the highest stability were obtained when the fungus was immobilized on non-woven fabric made of silk. The enzymatic properties of the immobilized preparation were similar to those of the free enzyme. Ten times repeated batch hydrolysis of birch-wood xylan was done over a period of 450 h. Hydrolysis of different xylan substrates such as oat spelts and rice bran by the immobilized mycelia was also investigated.

Lipoprotein abnormalities in type 2 diabetic patients. Increased distribution of triglycerides (TG) in very low density lipoprotein (VLDL) and decreased apolipoprotein E/VLDL-TG ratio are risk for ischemic heart disease.

Plasma lipoproteins and clinical characteristics of type 2 diabetic patients with (n = 50) and without (n = 108) ischemic heart disease (IHD) were compared. The patients with IHD were older (64 +/- 9 vs 59 +/- 9 years, mean +/- SD, p < 0.01) and had a longer duration of diabetes (14 +/- 9 vs 11 +/- 8 years, p < 0.05). Lipids, lipoproteins and apolipoproteins were comparable in the two groups. The percent distribution of triglycerides (TG) in the very low density lipoprotein (VLDL) fraction was significantly higher (55 +/- 16 vs 50 +/- 17%, p < 0.05). The level of low density lipoprotein (LDL) (33 +/- 14 vs 39 +/- 15%, p < 0.05) and the apo E/VLDL-TG ratio (0.085 +/- 0.045 vs 0.120 +/- 0.097, p < 0.01), however, was significantly lower in patients with IHD than in those without IHD. Multiple regression analysis also indicated that age, duration of diabetes, percent distribution of TG in VLDL, percent distribution of TG in LDL and the apo E/VLDL-TG ratio were significantly related to the presence of IHD. Hypertriglyceridemia, particularly when characterized by abnormalities of TG distribution, may play an important role in the development of IHD in type 2 diabetic patients.

Polymerase chain reaction amplification of Asp f I and alkaline protease genes from fungus balls: clinical application in pulmonary aspergillosis.

Asp fI(18 kDa) and alkaline protease (33 kDa) are the 2 major antigens which are derived from Aspergillus (A.) fumigatus and have been implicated as possible virulence factors in the pathogenesis of Aspergillus-induced diseases. We attempted to detect fragments of genes encoding both proteins from fungus balls obtained at surgery or autopsy by polymerase chain reaction (PCR) amplification and then used PCR to test clinical samples. Frozen-stored fungus ball samples from a patient with acute myeloid leukemia complicated by Aspergillus pneumonia and from a patient with pulmonary aspergilloma were studied. We successfully amplified a 315 bp PCR product, the target sequence for Asp f I, and a 747 bp PCR product as a target sequence for alkaline protease (ALP) in both cases. In addition, 13 clinical samples including sputum specimens from patients with pulmonary aspergillosis were also examined. PCR analysis for the Asp f I (ALP) gene in clinical samples showed positive results in 5/10 (6/10) patients with pulmonary aspergilloma and in 3/3 (1/ 3) patients with invasive pulmonary aspergillosis. Culture data on A. fumigatus revealed positive results in 3/9 patients with pulmonary aspergilloma and in 2/3 patients with invasive pulmonary aspergillosis. This method can be used to recognize the involvement of A. fumigatus in various clinical settings where conventional culture results are not readily available.

Enzymatic method for assaying uric acid in serum with a new tetrazolium salt produces water-soluble formazan dye.

OBJECTIVES: To apply an enzymatic method for assaying uric acid in serum based on the uricase (EC 1.7.3.3)-catalase (EC 1.11.1.6)-formaldehyde dehydrogenase (FADH, EC 1.2.1.46) coupled with the new tetrazolium salt producing a water-soluble formazan dye as an indicator system. Unlike the traditional tetrazolium salts, e.g., iodonitrotetrazolium (INT) and nitrotetrazolium blue (NTB), the corresponding formazan dye produced did not absorb to the test tube and the reaction cells of the analyzer. Moreover, this formazan dye had a higher water solubility and more sensitivity. DESIGN AND METHODS: A two electron reduction of tetrazolium salt with NADH, produced by the uricase-catalase-FADH reaction, was mediated by an electron carrier, 1-methoxy PMS. The generation of formazan, monitored at 440 nm, was proportional to the concentration of uric acid in serum. The sensitivity of this method was about 1.5 times that of the peroxidase-TOOS [N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine] method. The assay was evaluated with TBA-80FR.NEO biochemical analyzer. The average within-run and between-day imprecision (CV) were 0.75-2.44% and 3.20-3.33%, respectively. RESULTS: Results of the proposed method (y) correlated well with those determined by the conventional chromogen method (x): y = 0.970 x - 0.018 mmol/L (Sy l x = 0.019 mmol/L, r = 0.993, n = 67). CONCLUSIONS: We also present data showing that the method is rapid, relatively free of interference, and amenable to automation.

Peripheral alpha-linked N-acetylglucosamine on the carbohydrate moiety of mucin derived from mammalian gastric gland mucous cells: epitope recognized by a newly characterized monoclonal antibody.

To obtain a tool to study the structural characterization and the detection of mucin derived from the gastric gland mucous cells, we developed a monoclonal antibody, designated HIK1083, against mucin purified from rat gastric mucosa. In an ELISA, HIK1083 reacted strongly with the mucin purified from a deep layer of the corpus and antrum but only slightly reacted with that obtained from the surface mucosal layer. The reaction of mucin and HIK1083 was inhibited by the oligosaccharides obtained by the alkaline borohydride reduction of antigenic mucin. Two purified oligosaccharide alditols reacting with the monoclonal antibody obtained from the antigenic mucin had one and two peripheral alpha-linked N-acetylglucosamine residues, respectively, according to the evidence from NMR spectroscopy. Moreover, among the commercially available p-nitrophenyl derivatives of monosaccharides, only p-nitrophenyl-N-acetyl-alpha-D-glucosaminide inhibited the reaction of this monoclonal antibody and the antigenic mucin in a concentration-dependent manner. These results, as well as the immunohistochemical observations, indicate that alpha-linked N-acetylglucosamine residues are specifically attached to the peripheral region of the carbohydrate moiety of the mucin synthesized in and secreted from the gastric-gland-type cells, and indicate that the monoclonal antibody HIK 1083 recognizes this structure.

Simple detection of the 5S ribosomal RNA of Pneumocystis carinii using in situ hybridisation.

AIMS: To investigate the effectiveness of digoxigenin incorporated double stranded DNA probes produced by the polymerase chain reaction (PCR), for the detection of Pneumocystis carinii, using in situ hybridisation (ISH). METHODS: Formalin fixed, paraffin wax embedded sections of 26 human lung samples from 11 patients with P carinii pneumonia (PCP), and 15 with various types of fungal and viral pneumonia, were obtained during necropsy or transbronchial lung biopsy. Three additional PCP induced rat lung samples were also tested. PCR probes were prepared using the digoxigenin labelling mixture, and they were amplified from the DNA of a PCP induced rat lung after administration of dexamethasone, on the basis that 5S ribosomal RNA sequences are identical in human and rat P carinii. ISH was performed using this probe, and visualised using the digoxigenin nucleic acid detection kit. An immunohistochemical study using anti-human Pneumocystis monoclonal antibody was also carried out in parallel. RESULTS: ISH positively stained eight (of eight) lung necropsy specimens from patients with PCP, three (of three) transbronchial lung biopsy specimens from patients with PCP, and none of the three PCP induced rat lung specimens. In contrast, none of the specimens from patients with pneumonia caused by Aspergillus sp (n = 5), Candida sp (n = 4), Cryptococcus sp (n = 2), mucormycosis (n = 2), or cytomegalovirus (n = 2) were positive on ISH or immunohistochemistry. CONCLUSIONS: Using a digoxigenin labelled PCR probe for the entire 5S rRNA sequence in conjunction with conventional staining, ISH is highly reactive and specific for the diagnosis of PCP.

[Unclassified chronic myeloproliferative disorder (Ph negative/bcr rearrangement negative CML) with late appearing Philadelphia like chromosome].

A 48-year-old male was diagnosed to be unclassified chronic myeloproliferative disorder (UCMPD)/Ph negative bcr rearrangement negative (Ph-/bcr-) CML by hematological, cytogenetical and DNA analyses (Jpn. J. Clin. Hematol. 33(4): 525-531, 1992). Three years and a half after the diagnosis of UCMPD/Ph-bcr- CML, Ph chromosome was observed in 17 of 20 examined cells. Hematological findings showed a transformation into blast crisis. The late appearing of Ph in a case of UCMPD/Ph1-bcr- CML described here is rare. Southern blot analysis using 3' and 5' bcr probe showed no bcr rearrangement. Analyses of BCR/ABL chimeric RNA by RT-PCR method were negative in both of Major- and Minor BCR/ABL chimeric RNA. In the present case it is speculated that Ph is developed as the result of multistep progression and also speculated that the breakpoint at BCR gene differs from Major- and Minor-bcr in usual Ph+CML and de nove Ph+ ALL. Therefore, it may be reasonable that the present case is understood to be a case with late appearing Ph-like chromosome.

[Genetic analysis by fluorescence in situ hybridization of lung cancer cells obtained by bronchial brushing].

Fluorescence in situ hybridization was done with specimens obtained by bronchial brushing from 25 patients with abnormal lung shadows. A satellite DNA probe, specific for chromosome 11, was used to detect numerical chromosome aberrations in tumor cell nuclei. Normal diploid human lymphocyte nuclei, which served as control, had two signal spots in 99.6% of the nuclei in response to the chromosome 11 probe. The most frequent signal spots in class V cells (Case 1-7) ranged from 3 to 5, followed by 6 to 8, regardless of histopathological findings of lung cancer. In class I cells (cases 8-11) the signal appearance was similar to that in class V cells. The disease in patients from whom class I cells were obtained was found to be malignant by other diagnostic procedures performed afterward. The abnormalities in cases 13-25 were diagnosed as non-malignant by brush cytology, and clinical course showed a little more than 3 spots. These data indicate that fluorescence in situ hybridization with specimens obtained by bronchial brushing can be useful for detecting numerical chromosome abnormalities and can aid in the rapid diagnosis of lung cancer.

In vitro and ex vivo free radical scavenging activities of carazostatin, carbazomycin B and their derivatives.

Free radical scavenging activities of various carbazole compounds, carazostatin, carbazomycin B and their chemically modified derivatives were studied in vitro and ex vivo. Among these compounds, carazostatin, which was isolated as a free radical scavenger from the culture of Streptomyces chromofuscus, showed the most potent inhibitory activity against lipid peroxidation of rat brain homogenate in vitro. Carbazomycin B, a known antimicrobial antibiotic, also exhibited strong activity in this system. Although O-modified derivatives of carazostatin and carbazomycin B retained considerable activity, N,O-dimethyl derivatives did not suppress the peroxidation. On the other hand, the results from the ex vivo evaluation of these carbazoles in the lipid peroxidation system of mouse blood plasma showed that the original compounds as well as their O-modified derivatives had a strong inhibitory activity upon oral administration to mice. These findings suggest that these natural carbazoles and their effective derivatives can protect tissues from the peroxidative damage due to generation of free radicals.

[Blood leukotriene B4 response to endotoxin in fish oil fed rats].

Six-week-old male Wistar rats were given a low fat diet containing 15% cod liver oil (FO; n = 30) or 15% safflower oil (SO; n = 30) for 6 weeks. In the polymorphonuclear leukocytes, arachidonic acid (AA) was significantly lower in the FO group than in the SO group (p < 0.001), and eicosapentaenoic acid (EPA) in the FO group was present in large amounts showing the EPA/AA ratio of 1.76. When lipopolysaccharide (LPS; 10mg/kg) was intraperitoneally injected, white blood cell counts in the blood significantly decreased in both groups (p < 0.001) compared to the controls 2 hours later, and at 4 hours, the counts in the SO group (2,033 +/- 151/mm3) were lower than in the FO group (3,189 +/- 624/mm3), (p < 0.01). In both groups, leukotriene B4 (LTB4) in the whole blood increased at 4 hours following LPS-administration compared to the controls (1,219 +/- 167.3pg/ml in the SO group and 600.0 +/- 109.0pg/ml in the FO group, p < 0.001). It should be noted that the level of LTB4 in the SO group was significantly higher than in the FO group (p < 0.001). Changes of the metabolites including decreased LTB4 production in the arachidonic cascade related to the increase of EPA may play a role in the inhibition of the decrease in white blood cell counts.

Adenomyomatosis of the gallbladder: a clinical survey of 30 surgically treated patients.

Thirty patients with adenomyomatosis of the gallbladder (AMG) were operated on between January 1983 and June 1990. They were made up 3.3% of patients who underwent cholecystectomy during the same interval. Of the 30 patients, ages ranged from 22 to 77 years (mean 52.3 years) and the male-to-female ratio was 8:7. Among the macroscopic types, 10 cases of generalized, 12 of segmental (S) and 8 of fundal (F) were noted, and the size of the affected portion in type S (0.8 +/- 0.2 cm, mean +/- SD) was significantly thinner than in other two types (p less than 0.05). Although the main symptom was abdominal pain, the majority of patients with type F had no complaints. Twenty patients (27%) were accompanied by gallstones including cholesterol stones in 60% of cases, and all six cases showing microbes in the bile had gallstones. Only six patients were diagnosed as AMG by preoperative imaging techniques. Other diagnoses comprised 15 of chronic cholecystitis and 3 of suspected gallbladder carcinoma. To identify the expanded Rokitansky-Aschoff sinuses, endoscopic retrograde cholangiography and/or ultrasonography of the abdomen were most useful. No preponderant coexistent lesion other than gallstones was noted. Levels of carcinoembryonic antigen in gallbladder bile in cases of AMG (2.5 +/- 1.5 ng/ml, mean +/- SD) were significantly lower than in gallbladder carcinoma (p less than 0.01). All the patients were easily treated with cholecystectomy, and 24 patients who have been followed up after surgery are doing well.(ABSTRACT TRUNCATED AT 250 WORDS)

[Progress in glycated albumin assay and its clinical significance].

Glycated albumin (G-Alb) measured as one of the glycated proteins is recognized to be the clinical significant index of blood glucose control at earlier stage rather than HbA1c. The borate affinity column (BAC) method is regarded as a reference assay method, but the assay of fructosamine (FRA) is widely used at present as the easier method of G-Alb. In this paper, the characteristics, problems and usefulness of BAC method and FRA assay were described.

Extraction method for preparing pyridylamino sugar derivatives and application to porcine gastric mucus glycoprotein analysis.

For the sensitive detection of free sugars and oligosaccharides, the use of their pyridylamino derivatives has now found general acceptance. To remove excess 2-aminopyridine from this derivative in a reaction mixture, gel filtration and ion-exchange chromatography were conducted. It was found in the present study that contaminated 2-aminopyridine could be selectively removed from the reaction mixture by adjusting the pH with saturated sodium bicarbonate at above 8.5 followed by extraction with benzene. By using this method, fewer purification steps and less time are required, with minimum loss of pyridylamino sugar derivatives.

[Care of cardiac insufficiency and laboratory test informations].

Measurement of serum myoglobin, CK-MM isoforms and CK-MB isoenzyme protein are useful for diagnosis and therapy of acute myocardial infarction (AMI). As latex agglutination turbidimetry of serum myoglobin is an easy and rapid assay method, it can be applied for early diagnosis of AMI and its observation of progress after break out of AMI. CK-MM isoforms also can be applicable for early diagnosis of AMI. The assay method of CK-MB isoenzyme protein by immunochemiluminescence is useful for anticipating the infarct size of AMI since this format is not influenced by CK-BB, myokinase, or mitochondria CK.