Sepharose derivatives containing citric acid as affinity ligand. Purification of fumarase.

Six Sepharose derivatives, in which citrate was immobilized via methylene carbons, were prepared by coupling of the alpha- and beta-isomers of citrylpolymethylenediamine to Sepharose. The purification of fumarase from pig heart was dependent on the length of the spacer arm, but not on the isomeric configuration of the immobilized citrate. Gels having six methylene carbons had the largest adsorption capacity for the enzyme and therefore were the most suitable for use in affinity columns for its purification.

In vitro IgE-secreting cells in man. I. Mitogen-independent and -dependent subpopulations.

In vitro IgE secretion by atopic and normal peripheral-blood lymphocytes was examined in culture with pokeweed mitogen or Staphylococcus aureus strain Cowan-I (StaCw) or without mitogen. IgE secreted in culture supernatants was measured with double antibody radioimmunoassay. Enumeration of IgE-secreting cells was made by a protein-A plaque assay.

Pharmacokinetics of ceftazidime in patients with renal insufficiency and in those undergoing hemodialysis.

The pharmacokinetics of ceftazidime, a new cephalosporin antibiotic, were studied in 7 healthy volunteers and 32 patients with renal insufficiency after a single 500-mg intravenous injection. The mean plasma half-life during beta-phase was 1.67 h in normal subjects and was prolonged to 15.

Inhibition of a membrane-bound enkephalin-degrading aminopeptidase by bestatin analogs.

A variety of bestatin analogs were examined as potent inhibitors of a membrane-bound enkephalin-degrading aminopeptidase that was purified from monkey brain. Bestatinyl amino acid derivatives showed strong inhibition of this enzyme. The most effective was bestatin-L-Arg X AcOH, with a Ki value of 0.

[Seminal vesicle cyst associated with contralateral renal agenesis: a case report].

A 37-year-old man was hospitalized for a more extensive examination of azoospermia. A seminal vesiculogram demonstrated a cystic lesion of the right seminal vesicle, but failed to show the left seminal vesicle because of intrascrotal adhesion caused by previous orchidopexy. A transrectal ultrasonogram and CT-scan revealed a large cystic mass in the retrovesical region.

Reductive cleavage of Xaa-proline peptide bonds by mild alkaline borohydride treatment employed to release O-glycosidically linked carbohydrate units of glycoproteins.

During the deglycosylation reaction of fish egg polysialoglycoproteins under the conditions of 1 M NaBH4 in 0.1 M NaOH at 37 degrees C for 48 h, a marked loss of the glycine content has been encountered, besides the serine and threonine residues to which the carbohydrate units are linked. The chemical basis behind this phenomenon has been elucidated by amino acid analysis first of the major glycopeptides (carbohydrate-(O)Thr-Gly-Pro-Ser) derived from desialylated polysialoglycoproteins and subsequently six proline-containing peptides before and after treatment under similar conditions.

[Studies on an oral calcium loading test in upper urinary tract stone formers].

An oral calcium loading test was performed on 55 patients with upper urinary tract stones to find absorptive or renal hypercalciuria. The mean urinary calcium excretion level of this patient group was significantly greater than that of a control group of 10 patients without abnormal upper urinary tract. Idiopathic hypercalciuria defined as more than 250 mg per day in males and more than 200 mg per day in females was observed in 13 patients; 4 patients with absorptive hypercalciuria, 6 patients with renal hypercalciuria and 3 patients of origin-unknown hypercalciuria.

Purification and characterization of two distinct dipeptidyl aminopeptidases in soluble fraction from monkey brain and their action on enkephalins.

Two distinct dipeptidyl aminopeptidases, which were designated DPP-A and DPP-B, were purified from soluble fraction of monkey brain using Leu-enkephalin as the substrate. The enzymes were purified 187 and 136 fold, respectively. Both enzymes showed the optimum pH in neutral range.

Role of joint afferents in relation to the initiation of forelimb stepping in thalamic cats.

To analyze the roles of joint afferents in relation to initiation of forelimb stepping in thalamic cats, we recorded the unit spikes of the cervical dorsal roots, stimulated the joint afferents, and applied local anesthesia to the joint capsule. Almost all of the joint afferents of the shoulder, elbow, wrist and finger adapted slowly and exhibited alternating firing during forelimb stepping. About 45% of the afferents of each joint showed firings as the limb moved from forward to backward.

Purification and characterization of an enkephalin-degrading aminopeptidase from guinea pig serum.

An enkephalin-degrading aminopeptidase using Leu-enkephalin as a substrate was purified about 4100-fold from guinea pig serum. The purified preparation was apparently homogenous, showing one band on polyacrylamide gel electrophoresis. The molecular weight of the enzyme was approx.

Fish egg polysialoglycoproteins: structures of new sialooligosaccharide chains isolated from the eggs of Oncorhynchus keta (Walbaum). Fucose-containing units with oligosialyl groups.

A series of acidic oligosaccharide alditols having different neutral core oligosaccharides were isolated from salmon egg polysialoglycoproteins by alkali-borohydride treatment followed by anion-exchange chromatography and Iatrobead chromatography. Their structures were determined by methylation analysis, molecular secondary ion mass spectrometry of underivatized oligosaccharides, and enzymatic desialylation. The molecular secondary ion mass spectra of intact sialooligosaccharides exhibit pronounced quasi-molecular-ion peaks, (M + H)+, (M + Na)+, (M + 2Na - H)+, and/or (M + K)+, as well as some diagnostic sequence ion peaks.

[Eosinophilic cystitis: a report of 2 cases].

Two patients with eosinophilic cystitis are presented in this report. The first case was a 39-year-old man who was admitted to our clinic with complaints of pollakisuria, pain on urination and hematuria for a month, which had not been relieved by antibiotic therapy. He had both a family and a personal past history of allergy.

Enkephalin degrading enzymes in cerebrospinal fluid.

Enkephalins were rapidly degraded by specific enzyme systems in vivo. In cerebrospinal fluid (CSF), however, it has been undefined whether these enzyme systems existed. Our experiments showed enkephalins were hydrolyzed by the enzymatic activity in both CSF of human and monkey.

[Two cases of renal angiomyolipoma].

Two cases of renal angiomyolipoma without tuberous sclerosis are presented. The 1st case was a 33-year-old woman who had left flank pain and macroscopic hematuria on August 26, 1980. Physical examinations revealed a large left renal tumor, and selective renal angiograms showed the presence of a hypovascular mass located in the central to median lower part of the left kidney.

A membrane-bound aminopeptidase isolated from monkey brain and its action on enkephalin.

A membrane-bound aminopeptidase which cleaves the tyrosine-glycine bond of enkephalin was purified about 1600-fold from monkey brain. This aminopeptidase hydrolyzed Leu-enkephalin with a Km value of 35 microM and also hydrolyzed basic, neutral and aromatic amino acid beta-naphthylamides. An apparently homogeneous enzyme consisted of a single polypeptide chain with a molecular weight of approx.

A novel neutral oligosaccharide chain found in polysialoglycoproteins isolated from Pacific salmon eggs. Structural studies by secondary ion mass spectrometry, proton nuclear magnetic resonance spectroscopy, and chemical methods.

A novel carbohydrate chain possessing a hitherto unknown disaccharide unit, alpha-L-Fuc leads to D-GalNAc, has been isolated from salmon egg polysialoglycoproteins on alkali-borohydride treatment. Salmon egg polysialoglycoproteins contain O-glycosidically linked neutral pentasaccharide chains in addition to a number of oligosialosyl group containing sugar chains. Composition analysis of the neutral pentasaccharide gave fucose, galactose, 2-acetamido-2-deoxygalactose, and 2-acetamido-2-deoxygalactitol in a molar ratio of 1:2:1:1.

Discharge patterns of reticulospinal neurons corresponding with quadrupedal leg movements in thalamic cats.

To analyze the discharge patterns of the reticulospinal (R-S) neurons associated with four-limb movement, we recorded the unit spikes of 108 R-S neurons in 18 thalamic cats. (1) Unit spikes of R-S neurons exhibited alternating firings during leg movements, not only stepping on the treadmill but also upon passive flexion and extension movement by the experimenter's hand. (2) R-S neurons manifested firing patterns associated with diagonal, reciprocal and quadrupedal leg movements.

Pharmacokinetics of cefotetan in normal subjects and patients with impaired renal function.

The elimination kinetics of cefotetan (YM09330), a new parenteral semisynthetic cephamycin derivative, were studied in eight healthy volunteers and 41 patients with renal insufficiency after the administration of a single 500-mg dose intravenously. Concentrations of cefotetan in serum and urine were determined by both bioassay and high-pressure liquid chromatography. The pharmacokinetic parameters for cefotetan were calculated on the basis of a two-compartment open model.

Elimination kinetics of cefotaxime and desacetyl cefotaxime in patients with renal insufficiency and during hemodialysis.

The pharmacokinetics of cefotaxime, a new semi-synthetic cephalosporin for injection, was studied in 30 subjects with various degrees of renal function after a single 1-gram intramuscular injection. Serum and urinary concentrations of cefotaxime and desacetyl cefotaxime were determined by high pressure liquid chromatography. The pharmacokinetic parameters of cefotaxime were obtained using a one-compartment open model.

Reticular neuron activities associated with locomotion in thalamic cats.

Unit discharges of the reticular neurons and EMGs of extensors and flexors were recorded in the 4 limbs of thalamic cats during stepping on a motor-driven treadmill to study the functional role of the spino-bulbo-spinal (SBS) reflex in locomotion. (1) Reticular neurons showed alterations of increased or decreased discharge rate depending on the phase of the step cycle; they were divided into input (ascending), output (reticulospinal, R-S) and internuncial neurons underlying the SBS reflex. (2) R-S neurons showed increased spikes from the end o the stance to the beginning of the swing phase; in this period, flexor muscle EMGS were obtained.

Sensory evoked potentials and their changes with respiration in man and cat.

Cutaneously evoked cortical potentials were recorded in man and chloralosed cats, in order to analyse the neural mechanisms responsible for their fluctuation with respiration. From the Cz region of the human scalp, 2 components of potentials, early and late, were evoked following mechanical stimulation of a finger contralateral to the recording side. These potentials varied with the phase of respiration, the late component being larger in amplitude and longer in duration when the stimulus was applied during expiration.