[Utility of the uric acid clearance method as a marker of antitumor effect of chemotherapy in cases of malignancies].
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Adult↗
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[An operated case of dissecting aortic aneurysm of Marfan's syndrome after Cabrol's operation].
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Adult↗
Fluorescent staining of human chromosomes: identification of some common aberrations.
With the development of fluorescent staining techniques and their application to human chromosomes, identification of the entire human karyotype is now possible. Of considerable value are the elimination of the guesswork in the pairing of the C group and sex chromosomes and the recognition and characterization of the translocation chromosomes in mongolism.
Chromosome Aberrations↗
Manifestations of carcinoma of the uncinate process by means of superior mesenteric arteriography.
Identification of carcinoma arising in the uncinate region of the pancreas is difficult because of the peculiar topographic relationship of this lesion with adjacent structures. Among the various diagnostic procedures performed upon ten patients with this malignant lesion, superior mesenteric arteriography provided the most helpful information, including signs of invasion in the main trunk of the superior mesenteric artery and vein and in the proximal portion of the jejunal arteries. A clearer visualization of these vascular involvements was attained in the right posterior oblique projection rather than in the anteroposterior projection. On the contrary, the arterial and portal vessels belonging to the celiac territory, such as superior pancreaticoduodenal arteries, dorsal pancreatic artery, gastroduodenal artery, splenic vein and portal vein, were almost free of involvement. At laparotomy, almost all of the patients had direct extension of the tumor to the superior mesenteric vessels, forming a cancerous core in the root of mesentery. All died of cachexia within six months after a palliative surgical procedure. Serial roentgenograms of superior mesenteric artery, especially taken in the oblique projection, are the best means of confirming carcinoma of the uncinate process, for which only a few available methods have been established to evaluate the clinical aspects.
Adenocarcinoma↗
[Erythroid colony formation in vitro by human marrow cells--fundamental studies by a modified plasma clot culture system and its clinical application (author's transl)].
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Cells, Cultured↗
Inactivation of glucose-6-phosphate dehydrogenase by 4-hydroxy-2-nonenal. Selective modification of an active-site lysine.
Incubation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides with 4-hydroxy-2-nonenal (HNE) results in a pseudo first-order loss of enzyme activity. The pH dependence of the inactivation rate exhibits an inflection around pH 10, and the enzyme is protected from inactivation by glucose 6-phosphate. Loss of enzyme activity corresponds with the formation of one carbonyl function per enzyme subunit and the appearance of a lysine-HNE adduct. The data presented in this paper are consistent with the view that the epsilon-amino group of a lysine residue in the glucose 6-phosphate-binding site reacts with the double bond (C3) of HNE, resulting in the formation of a stable secondary amine derivative and loss of enzyme activity. We have described a mechanism by which HNE may, in part, mediate free radical damage. In addition, a method for the detection of the lysine-HNE adduct is introduced.
Aldehydes↗
New methods to detect donor-type DNA in HLA-DRB1-matched living-related liver transplant recipients.
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Adult↗
Prophylactic effect of neuroleptics in symptom-free schizophrenics.
Prophylactic effects of psychotropic drugs on 55 schizophrenics in remission were evaluated for 3 years in a double-blind controlled study employing a cross-over design. Patients were randomly assigned to the following drugs orally administered twice a day: placebo; diazepam 15 mg; imipramine 50 mg; chlorpromazine 75 mg; and haloperidol 3 mg. The number of days of remission for each patient was recorded. Since only two patients received all five drug treatments, the data were analyzed using the number of days allocated to the "first assigned drugs" only and the cross-over aspect of the experimental design was disregarded. All patients treated with either the placebo, diazepam or imipramine relapsed within a year. On the other hand, four patients treated with chlorpromazine, or with haloperidol, were in remission for more than 1 year. Fifty percent of the patients relapsed within 16 days with placebo; 88 days with diazepam; 30 days with imipramine; 165 days with chlorpromazine; and 74 days with haloperidol. Within a year, only chlorpromazine significantly prolonged the remission state as compared to placebo and imipramine. At the end of the 3-year trial, both chlorpromazine and haloperidol significantly prolonged the remission state as compared to the other three drugs. These data suggest that neuroleptic treatment for a longer period is vitally important to prevent relapse even in schizophrenics in remission and that such a trial seems an efficient method for investigating the prophylactic effects of neuroleptics.
Adult↗
Usefulness of a colorimetric method for testing antifungal drug susceptibilities of Aspergillus species to voriconazole.
The usefulness of a colorimetric method using Alamar Blue (Alamar Blue method) for susceptibility testing of Aspergillus species to voriconazole and three existing antifungal agents, itraconazole (ITCZ), flucytosine (5-FC), and amphotericin B (AMPH-B), was studied using four ATCC strains of three species, including two strains of A. fumigatus and one each of A. flavus and A. niger. For comparison, the broth microdilution method for antifungal susceptibility testing of filamentous fungi proposed by the National Committee for Clinical Laboratory Standards (NCCLS method M38-P) was also used. Minimum inhibitory concentration (MIC) endpoints were read spectrophotometrically for the Alamar Blue method and visually for the NCCLS method after 46-50 h. Like the NCCLS method, Alamar Blue produced highly reproducible results for all the drugs and strains tested; most MIC values obtained by nine tests were within the range of 2 twofold dilutions for each strain. Voriconazole and ITCZ susceptibility testing with the Alamar Blue method and the NCCLS method yielded comparable results in 94% of the tests, meaning that the endpoints obtained were identical or differed by no more than 2 twofold dilutions. On the other hand, susceptibility testing for 5-FC and AMPH-B yielded scores of 25% and 64%, respectively. Our study suggests the potential value of the Alamar Blue method as a convenient alternative to the NCCLS M38-P method for routine testing of Aspergillus species susceptibility to at least voriconazole and ITCZ.
Antifungal Agents↗
Nephrotoxicity assessment by measuring cellular ATP content. I. Substrate specificities in the maintenance of ATP content in isolated rat nephron segments.
To clarify the characteristics of cellular ATP synthesis in individual nephron segments for assessing nephrotoxicity of chemicals, cellular ATP content was measured by the luciferin/luciferase system under various conditions using intact nephron segments isolated from male Sprague-Dawley rats. Increasing the duration of collagenase treatment of kidney slices significantly lowered the cellular levels of ATP newly synthesized from 2 mM glutamine in PST at 37 degrees C over 30 min (p less than 0.01). The tubular incubation time significantly affected the cellular ATP content in the early and middle portions (S2) of the proximal tubule (p less than 0.05 and p less than 0.01, respectively) over 20 min and in the late proximal tubule over 10 min. Among numerous substrates tested, such as D-glucose, glutamine, pyruvate, DL-lactate, and beta-hydroxybutyrate, the substrate utilization for maintaining cellular ATP content was entirely variable according to each nephron segment. Pyruvate and glutamine were the best substrates in the proximal tubule. On the other hand, ATP production from glutamine was less than that from the other substrates in the distally located nephron segments: medullary and cortical thick ascending limbs of Henle's loop (MAL and CAL, respectively), distal tubule, cortical and medullary collecting tubules (CCT and MCT, respectively). In general, glucose, pyruvate, and lactate appear to be equivalent in maintaining ATP content in the distal segments of renal tubules. A monovalent cation ionophore, monensin, at 10 micrograms/ml decreased the cellular ATP content in MAL, CAL, and MCT significantly. Mercuric chloride (HgCl2) was used as a model compound to study nephrotoxicity by investigating its effects on cellular ATP metabolism in microdissected nephron segments. HgCl2 at 1 x 10(-6) M significantly decreased ATP content only in S2 (p less than 0.05), clearly demonstrating S2 to be the most sensitive segment within the nephron. These results indicate that measurement of cellular ATP content would be a useful method forecasting the intrarenal toxic site and potency of possible nephrotoxic chemical compounds.
Adenosine Triphosphate↗
Detection of replication-competent adenoviruses spiked into recombinant adenovirus vector products by infectivity PCR.
The presence of replication-competent adenovirus (RCA) in clinical lots of adenovirus vectors raises a variety of safety concerns. To detect RCA in adenovirus vector products, the cell culture/cytopathic effect (CPE) method has generally been preferred. However, it is difficult to evaluate the amount of RCA clearly and quantitatively by this method. In addition, the cell culture/CPE method requires large-scale cell culturing and a substantial amount of time. For the purpose of establishing a method to detect RCA more sensitively and rapidly, we developed the infectivity PCR, a hybrid method that combines the infectivity assay and quantitative PCR. This method allows RCA to be quantified by real-time quantitative PCR using primers and a probe designed for E1 DNA. By infectivity PCR, 1 pfu of RCA spiked into 10(9) particles of adenovirus vectors could be detected. In contrast, CPE was observed in the cells infected with 10(4) pfu of RCA spiked into 10(9) particles of adenovirus vectors. The glass-beads method was suitable for extracting DNA rapidly from the RCA-infected cells. These results showed that infectivity PCR combined with the glass-beads-based DNA extraction method was useful for the detection of RCA in adenovirus vector products.
Adenoviridae↗
[Rapid and direct detection of Mycobacterium tuberculosis complex and mycobacteria in sputum by advanced method, PCR].
We developed two PCR methods, which amplify bovine tuberculous MPB70 gene and mycobacterial 16S rRNA gene, for detection of tubercle bacilli and mycobacteria in sputum, respectively. Among 27 Mycobacterium species and 57 species of 30 genuses other than Mycobacterium, only M. tuberculosis (TB) complex, i.e., M. tuberculosis, M. bovis, M. africanum, M. microti showed DNA amplification by PCR for MPB70, and amplification of 16S rRNA gene were observed specific in Mycobacterium species. A combination of these PCR abilities were available to differentiate the TB complex and nontuberculous mycobacteria (NTM). We investigated the correlation between these methods and conventional methods with 311 sputa that were suspected mycobacteriosis. The PCR method could detect 12 cases of TB complex and 4 cases of NTM in 17 specimens, which were positive by conventional methods, but could not for one specimen. Among 294 specimens that were negative with conventional methods, the PCR method detected 13 and 8 cases of TB complex and NTM, respectively. These results were confirmed by commercial tuberculous specific DNA probe or investigation of the clinical background of the patients. On the other hand, 273 specimens showed negative result either PCR nor conventional methods. The PCR method did not detect tuberculous DNA in normal 197 sputa, which were not suspected mycobacteriosis. These results indicate that each one of these PCR methods is highly specific to TB complex or Mycobacterium species. We concluded that these PCR methods are useful and advanced methods for rapid and direct detection of tuberculosis and mycobacteriosis.
DNA, Bacterial↗
Quantitative introduction of a given macromolecule into cells by fusion with erythrocyte ghosts using a fluorescence activated cell sorter.
FITC-conjugated bovine serum albumin (FITC-BSA) molecules were quantitatively introduced into human erythrocyte ghosts by gradual hemolysis. When the ghosts and L cells were fused with UV-inactivated HVJ (Sendai virus), FITC-BSA was introduced into the cytoplasm of the L cells and fluorescence could be observed inthe cells with a fluorescence microscope. A mixture of L cells and ghosts was introduced into a fluorescence activated cell sorter (FACS), which could separate the mononuclear cells on the basis of their light-scattering profile. Four distinct populations of mononuclear cells were found by fluorescence analysis. These populations were separated from the cell mixture and found to correspond to cells fused with one, two and three ghosts and unfused cells. After separation, the cells from each population could form colonies in culture. As a given macromolecule can be quantitatively introduced into erythrocyte ghosts with the FITC-BSA, after fusion of these ghosts with cells, this sorting method is useful for separating cells containing a definite number of macromolecules.
Cell Fusion↗
A new method for evaluating the bitterness of medicines by semi-continuous measurement of adsorption using a taste sensor.
We describe a new method for the evaluation of the bitterness of medicines by semi-continuous measurement of adsorption using a multichannel taste sensor or 'electric tongue'. The bitterness of 10 basic medicines was evaluated by both the taste sensor and in human gustatory sensation tests with 11 volunteers. The sensor part of the taste sensor consists of eight electrodes made of lipid/polymer membranes. Three variables were obtained from the taste sensor data: sensor output (S), the change of membrane potential caused by adsorption, corresponding to aftertaste (C), and the ratio C/S. These variables were used to predict an estimated bitterness score in multiple regression analysis. Semi-continuous measurement of C (every 30 s up to 150 s) was adopted as an additional explanatory variable, and the attenuation rate of C was defined as C'. These data were also subjected to multiple regression analysis. The correlation coefficient (r) estimated for the bitterness score predicted by the taste sensor, using C' for channel 2 and C/S for channel 4, and the score obtained by human gustatory sensation, was 0.824. This value was greater than that obtained using C/S for both channels 2 and 4 (0.734). The method described in the present study seems to offer good predictability for the evaluation of bitterness.
Adsorption↗
[Effect of anti-ulcer drugs and PGE2 on the proliferation of rat gastric cultured cells].
The effects of anti-ulcer drugs and PGE2 on the proliferation of rat gastric cultured cells were investigated. To study the cell proliferation, the rate of incorporation of 3H-thymidine into the cultured cells was measured. ULTROSER G, corresponding to fetal bovine serum, was found to dose-dependently increase the incorporation of 3H-thymidine into the cultured cells at a dose range of 1-4%. The incorporation of 3H-thymidine was highest at 36 hr after the inoculation of cells. Therefore, the effect of the drugs on the proliferation was performed under the condition of 2% ULTROSER G and evaluated at 36 hr. Aldioxa, cetraxate HCl, cimetidine (10(-7)-10(-5) M) and PGE2 (10(-8)-10(-6) M) had no effect on the cell proliferation, while indomethacin (0.35-1.41 x 10(-3) M) inhibited the proliferation in a dose-dependent manner. Aldioxa, cetraxate HCl (10(-5) M) and PGE2 (10(-6), 10(-8) M) antagonized the inhibitory effect by indomethacin at a concentration of 0.5 x 10(-3) M, but cimetidine (10(-7)-10(-5) M) did not. From the above results, it was concluded that this method was useful for investigating the effect of drugs on the proliferation of gastric cultured cells.
Animals↗
Ileal schwannoma in which blood loss scintigraphy was useful for diagnosis.
An 85-year-old woman was hospitalized with severe melena of unknown origin. Upper gastrointestinal (GI) endoscopy and lower GI endoscopy did not detect the origin and we could not establish any diagnosis. To explore the bleeding site, 99mTc-HSA blood loss scintigraphy was performed and a tumor of the small intestine was suspected. Fluoroscopic examination of the small intestine and abdominal CT scan confirmed an ileal tumor measuring 4x3 cm. The mass was a well-demarcated tumor about 80 cm proximal to Bauhin's valve. Partial resection of the ileum was carried out and the tumor was histologically diagnosed as schwannoma. Thereafter, there has been no recurrence of melena nor metastasis of the tumor. It is thought that blood loss scintigraphy is a useful method for unexplained exacerbation of melena.
Aged↗
Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential.
AIM: To analyze the relationship between sperm mitochondrial membrane potential and sperm motility parameters by means of a computer-assisted sperm analyzer (CASA) and in-vitro fertilization rate(%FR). METHODS: Semen samples were obtained from 26 men undergoing in vitro fertilization-embryo transfer (IVF-ET). Informed consent was obtained from all men prior to the study. Samples were prepared using wash and swim-up method in HEPES-HTF medium. The sperm motility (%MOT), progressive motility (%PMOT), average path velocity (VAP) microm/s), straight line velocity (VSL) (micro m/s), curvilinear velocity (VCL) (microm/s) and %hyperactivated sperm (%HA), and the %FR were assessed. The samples were incubated in the presence of 2.0 mciromol/L of 5,5',6,6'-tetra-chloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) for 30 min at 37 degrees C in air and washed in PBS before flow cytometry (FACSCalibur: Becton Dickinson) analysis. The mitochondrial probe JC-1 was used to identify the mitochondrial membrane potential. The sperm was divided into three populations according to the fluorescence pattern as follows: the high mitochondrial membrane potential group (n=8), the moderate group (n=5), and the low group (n=13). Statistical analysis was performed using unpaired t-test. RESULTS: Significant differences were found between the high and the low groups in %MOT (91.1+/-8.5 vs 63.0+/-32.7, mean+/-SD), VAP (73.0+/-14.2 vs 52.1+/-12.5), VCL (127.0+/-28.1 vs 87.0+/-22.6), %HA (27.3+/-23.6 vs 7.2+/-9.0) and %FR [73.2 (48/56) vs 59.0 (69/117)]. No significant differences were found in other CASA parameters. CONCLUSION: When the sperm mitochondrial membrane potential increases, sperm motility parameters and fertility potential will also increase. The JC-1 dye method is useful to predict sperm fertility potential.
Embryo Transfer↗
[Single-stage correction of coarctation of the aorta with aortic regurgitation].
A successful single-stage correction of coarctation of the aorta with aortic regurgitation is reported. A 39-year-old man was referred for angina, ankle swelling, and numbness of lower extremities. Aortogram revealed severe aortic regurgitation and coarctation of the aorta with 2.8 mm minimum diameter. Aortic valve replacement was performed under separate perfusion of upper and lower body, after which the coarctectomy and graft interposition were done under the left heart bypass. The patient was discharged in good condition with minor medication of digitalis and anticoagulant agents. As far as the case is concerned, the left heart bypass during graft interposition has possible risk of cardiac arrest and usual cardiopulmonary bypass is hazardous when complicated by severe coarctation of the aorta. Surgical procedure and assist methods used in this case were quite useful for coarctation of the aorta with aortic regurgitation.
Adult↗