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Biomedical subjects

Y Yajima

Publications and source records attributed to Y Yajima.

At least 91 records · Page 5Linked to original sources
PubMed1994

Overproduction of a Ca(2+)-independent protein kinase C isozyme, nPKC epsilon, increases the secretion of prolactin from thyrotropin-releasing hormone-stimulated rat pituitary GH4C1 cells.

Rat pituitary GH4C1 cells express protein kinase C (PKC) transcripts for cPKC alpha, cPKC beta II, nPKC delta, nPKC epsilon, nPKC eta, and aPKC zeta, but not for cPKC gamma or nPKC theta. Of the transcripts produced, the nPKC epsilon isoform is the most abundant. Transfection of GH4C1 cells with an expression plasmid containing nPKC epsilon cDNA leads to the transient overexpression of cellular nPKC epsilon and confers enhanced phorbol ester binding activity. Transient expression of an inactive point mutant (nPKC epsilon K-->R) of nPKC epsilon, where Lys436 at the putative ATP-binding site is replaced with Arg, also confers elevated binding activity. However, only overproduction of the wild type in transfected cells increases the basal levels and stimulates the secretion of prolactin (PRL) by 12-O-tetradecanoylphorbol-13-acetate or thyrotropin-releasing hormone (TRH). In stable clones overexpressing nPKC epsilon, immunocytofluorescence and immunoblot experiments indicated that TRH causes the rapid translocation and down-regulation of an appreciable fraction of nPKC epsilon. Both the basal and TRH-stimulated levels of PRL secretion are clearly correlated with the expression level of nPKC epsilon but not with the TRH receptor densities in these clones. The dose dependence of TRH-stimulated secretion were similar in all cells overexpressing cPKC alpha, cPKC beta II, nPKC epsilon, and nPKC delta, but the enhancement of PRL secretion was specific for the overproduction of nPKC epsilon, no effect was found when other isozymes were overproduced. These findings clearly demonstrate that the expression level of nPKC epsilon in GH4C1 cells is rate-limiting for basal and TRH-stimulated PRL secretion, and they provide the first direct evidence that nPKC epsilon plays a key role in hormonal secretory processes.

Animals↗
PubMed1994

Neuronal activity in nucleus ambiguous during deglutition and vocalization in conscious monkeys.

Extracellular recordings were made from the nucleus ambiguous in three conscious Macaca nemestrina monkeys during spontaneous vocalizations and swallows. The temporal relationship of neuronal activity to swallowing was inferred through correlation with the thyroarytenoid electromyographic (EMG) activity. Videofluoroscopic analysis of a fourth monkey during swallows of barium-impregnated fruit juice established the temporal relationship between swallowing and thyroarytenoid EMG activity. Of 691 cells recorded from the nucleus ambiguous and its adjacent area, the neuronal activity of 80 cells showed modulation during swallowing. Sixty-two cells were classified as "active" cells, with increased activity in relation to swallowing, while 18 cells were classified as "suppressed" cells, with tonic activity that reduced with swallowing. A continuum of latency was seen between the onset of modulation of these cells and the onset of swallowing, from "early" before the swallow to "late" after the swallow onset with most of the cells (44 cells) showing modulation near the onset of the swallow. A majority (37) of the 62 active swallowing-related cells also discharged with vocalization, but they demonstrated a lower discharge frequency and a longer burst duration during swallowing. Of the 18 suppressed swallowing-related cells, 11 cells discharged phasically with the respiratory cycle and may be a part of the ventral medullary respiratory center. This chronic awake primate model would preserve the many sensory inputs that may modify the oropharyngeal swallow and may better approximate human physiology.

Animals↗
PubMed1994

Effects of sarpogrelate on serotonin-induced increases in cytosolic Ca2+ in cultured rat mesangial cells.

The effects of the new 5-HT2A receptor antagonist sarpogrelate on the cellular action of serotonin were examined in cultured rat mesangial cells by measuring cytosolic free calcium concentration ([Ca2+]i). Sarpogrelate inhibited serotonin-induced increases in [Ca2+]i in a concentration-dependent manner. M1, a major metabolite of sarpogrelate, also exhibited an inhibitory effect exceeding that of sarpogrelate. The inhibitory effects of sarpogrelate and M1 were abolished by washing out these compounds. In contrast, the increase in [Ca2+]i induced by angiotensin II or arginine vasopressin was not affected by pretreatment of the cells with sarpogrelate or M1. These results suggest that sarpogrelate and its major metabolite (M1) act as reversible and specific 5-HT2A receptor antagonists against the contractile action of platelet-derived serotonin in mesangial cells.

Angiotensin II↗
PubMed1994

Anaerobic threshold can provoke microalbuminuria in non-insulin-dependent diabetics.

We examined two groups of non-insulin-dependent diabetic men (group A, 13 patients without microalbuminuria; group B, 9 patients with intermittent microalbuminuria) to ascertain whether the anaerobic threshold (AT) can provoke microalbuminuria, comparing them with 12 healthy subjects matched for age and sex (group C). All subjects exercised on a bicycle ergometer until the AT was reached. In intermittent microalbuminuria, the albumin:creatinine ratio (ACR) was over 0.25 mg/mmol.Cr 1-3-fold in 5 measurements. The ACR after exercise was increased to over 0.25 mg/mmol.Cr in 4/9 cases in group B (P < 0.05), in 2/13 cases in group A, but not at all in group C. We also studied the mechanism of exercise-induced microalbuminuria. In group B, ACR before exercise correlated positively with the baseline plasma glucose. Furthermore, positive correlation was found between ACR after exercise and HbA1c in group B. The AT did not affect the urinary beta 2-microglobulin in any groups. The plasma atrial natriuretic factor (ANF) after exercise was elevated most prominently in group B (P < 0.05). Positive correlation was found between increments of ACR and increments of plasma ANF after exercise in group B. We conclude that the AT can provoke microalbuminuria in some non-insulin-dependent diabetics. The plasma ANF and metabolic control may play an important role in the pathophysiology of exercise-induced microalbuminuria.

Adult↗
PubMed1994

[A case of NIDDM with non-clostridial gas-producing infection in the lower limb--the effects of hyperbaric oxygen therapy].

A 36-year-old male was admitted because of swelling and pain in the lower left limb. He had been diagnosed as having non-insulin dependent diabetes mellitus 4 years previously and had been treated unsuccessfully with insulin. Since gas formation was radiographically demonstrated in the soft tissue of the left foot, we treated him with antibiotics and hyperbaric oxygen under the diagnosis of gas gangrene. Despite this therapy, the gangrenous lesions progressed. Amputation of the left foot was necessary. Peptstreptococcus was isolated from the gangrenous area and identified as such on the seven hospital day. Nine cases of diabetic patients with non-clostridial gas gangrene who were treated with hyperbaric oxygen therapy have been reported in Japan. The results indicated that hyperbaric oxygen therapy is ineffective in the treatment of non-clostridial gas gangrene in diabetic patients.

Adult↗
PubMed1994

Modification in activity of medullary respiratory-related neurons for vocalization and swallowing.

1. The medullary ventral respiratory group (VRG) in and near the nucleus ambiguus contains neurons related to respiration. Also found here are neurons related to vocalization and swallowing as well as motoneurons of laryngeal, pharyngeal, palatal, and esophageal muscles. Previous reports in anesthetized animals have characterized discharge properties of neurons as they relate to a single behavior, e.g., respiration. Relatively few studies have documented discharge properties during more than one behavior, e.g., respiration and swallowing. Neurons were recorded extracellularly from awake Macaca nemestrina monkeys engaged in a vocalization task. The present paper describes how respiratory-related neurons (RRNs) modify their discharge during vocalization and swallowing. 2. The temporal relation between vocalization, subglottal pressure (SP), and diaphragm electromyogram (EMG) was established from recordings in anesthetized monkeys. Vocalization was elicited by electrical stimulation of the midbrain periaqueductal gray (PAG). Vocalization is preceded by deep inspiration and a brief pause in diaphragm EMG and begins with a rapid increase in positive SP. 3. Extracellularly recorded neural potential from the VRG in three awake monkeys were related to respiration by correlating their discharge with EMGs from the posterior cricoarytenoid or intercostal muscles during quiet respiration. Neurons were classified as inspiratory (INSP; N = 27), phase spanning (PS; N = 20), or expiratory (N = 6) on this basis. 4. A fourth category of cells was defined as a subgroup of INSP cells on the basis of their discharge during vocalization. This group, inspiratory-pause (INSP-PS; N = 10), paused for approximately 100 ms just before vocalization and resumed their activity during vocalization. 5. Of 63 fully analyzed RRNs, 40 (63%) also modulated their activity with vocalization and 3 (5%) with swallowing. Thirteen (21%) RRNs modulated with vocalization, respiration, and swallowing. Seven (11%) cells were modulated only with respiration. 6. Most cells demonstrated a shorter period of activity and a higher discharge rate associated with vocalization in comparison with quiet respiration. Six (30%) of the PS cells demonstrated an augmenting discharge pattern before vocalization, which was different from the other PS cells and different from their pattern during quiet respiration. Thirteen RRNs showed a pause in activity during vocal fold closure associated with swallowing, whereas three cells gave a burst at this time. 7. The higher discharge rate and shorter burst duration preceding vocalization, compared with quiet respiration, may be related to the greater positive SP necessary to support vocalization.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Ultrastructural study of glomerular basement membrane in diabetic rats by quick-freezing and deep-etching method.

The three-dimensional ultrastructure of glomerular basement membrane (GBM) in streptozotocin (STZ)-induced diabetic rats was examined by quick-freezing and deep-etching method. In three layers of the GBM of control rats, the outer and inner layers were formed by files of perpendicular fibrils, which connected the epithelial or endothelial cell surfaces with meshwork structures of the middle layer. In the diabetic rats, the inner layer was diffusely enlarged and the meshwork structure of the middle layer became markedly irregular due to the rupture of fine fibrils and thickening of material adherent to the fibrils. These ultrastructural changes correspond to those of subendothelial oedema, lamellation of lamina densa and fluffy material in the GBM, as revealed on conventional ultra-thin sections. It is suggested that the initial morphological change of STZ-induced diabetic nephropathy is disruption of matrix fibrils in the GBM, seemingly indicating a disturbance of size and/or charge barriers.

Animals↗
PubMed1993

Somatostatin induces release of the alpha subunits of pertussis toxin-sensitive G proteins in native membranes and in intact GH4C1 rat pituitary cells.

Incubation of GH4C1 rat pituitary cell membranes with the poorly hydrolyzable GTP analogue, GTP gamma S, produces a decrease in the pertussis toxin-catalyzed ADP-ribosylation of 40-kDa protein in the membrane pellet and the release of an alpha-like substrate from the membrane into the supernatant fraction; these effects do not occur with the inactive GDP analogue, GDP beta S. The resolved supernatant fraction from GTP gamma S-stimulated membranes is significantly activated to pertussis toxin-catalyzed [32P]ADP-ribosylation by the addition of purified beta gamma complex. Immunoblot analysis identifies the released pertussis toxin substrate as alpha subunits of Gi2, Gi3, and G(o) in the resolved supernatant. The physiological agonist, somatostatin, also stimulates the release of Gi2 and G(o) alpha subunits but not Gi3 from GH4C1 cell membranes in the presence of a low concentration of GTP gamma S (20 nM). The effects of somatostatin are inhibited by pretreatment of GH4C1 cells with pertussis toxin. Furthermore, the addition of somatostatin to intact GH4C1 cells decreases the level of Gi2 alpha subunits in the crude membrane whereas immunoblot analysis of the 274,000 x g supernatant (cytosolic fraction) clearly shows the presence of Gi2 alpha subunits. These data indicate that pertussis toxin-sensitive G proteins in GH4C1 cells dissociate into alpha subunits and beta gamma complex with the release of the alpha subunits from the membranes upon somatostatin activation.

Adenosine Diphosphate Ribose↗
PubMed1993

Multifunctional properties of ambiguous neurons identified electrophysiologically during vocalization in the awake monkey.

1. The nucleus ambiguus (NA) of the medulla contains motoneurons (MNs) for muscles of the larynx, palate, pharynx, and esophagus. Previous studies in anesthetized animals have demonstrated neural discharge correlated closely with respiration, swallowing, and electrical stimulation-elicited vocalization. A preliminary study confirming the above findings was done by recording NA motoneuron extracellular potentials from awake vocalizing monkeys. The present study was undertaken to quantitatively describe discharge properties of a large number of NA neurons recorded from awake, vocalizing monkeys (Macaca nemestrina). 2. In monkeys trained to vocalize for a food reward, extracellular recordings of neurons in and around the nucleus ambiguus were correlated with laryngeal electromyographic activity during vocalization. A nerve cuff electrode on the ipsilateral recurrent laryngeal nerve allowed identification of laryngeal MNs by antidromic activation of laryngeal MNs and the collision test. 3. Most laryngeal MNs became active 100-200 ms before vocalization. They ceased discharging during or immediately after vocalization. Some MNs discharged in close synchrony with bursts of EMG associated with variations in the vocalization. The mean discharge rate of MNs during vocalization was 18 Hz, and the maximum rate in many cells was over 100 Hz. MNs were also active during swallowing. One MN was related only to respiration and one exclusively to swallowing. 4. Some non-motoneurons (Non-MNs) and cells that may possibly be MNs (PossMNs), recorded in and near the NA, showed properties similar to MNs. Many (147) were active only with vocalization, whereas others were active with swallowing (23); respiration (9); vocalization and swallowing (32); vocalization and respiration (40); or vocalization, swallowing, and respiration (17). 5. The present study demonstrates the importance of studying laryngeal MNs in the chronic preparation. Namely, it is shown that both MNs and Non-MNs of the NA are active with more than one activity. Moreover, some Non-MNs are active for only one activity, e.g., vocalization or swallowing. These findings imply the existence of subsets of medullary neurons involved in multiple behaviors for control of generalized laryngeal functions and other subsets related to specific behaviors.

Animals↗
PubMed1993

Loss of glomerular anionic sites and the development of albuminuria in rats with streptozotocin-induced diabetes.

Examination was made of changes in the anionic sites of the glomerular basement membrane (GBM) in rats with streptozotocin (STZ)-induced diabetes by the immersion method of polyethyleneimine (PEI). PEI particles in GBM of diabetic rats significantly decreased from the 1st through the 8th week. Urinary albumin excretion in diabetic rats significantly increased at the 2nd but not earlier week. Insulin treatment effectively prevented decrease in PEI particles in STZ-injected rats. In rats with STZ-induced diabetes, initial renal alteration was disturbance of the charge barrier, followed by the development of albuminuria. Continued deterioration of anionic sites and possibly additional disturbance of size barrier were considered responsible for the development of albuminuria. Insulin treatment appears to prevent the loss of anionic sites of GBM.

Albuminuria↗
PubMed1992

Increase in cytoplasmic Ca2+ and stimulation of calcitonin secretion from human medullary thyroid carcinoma cells by the gastrin-releasing peptide.

Examination was made of the effects of gastrin-releasing peptide (GRP) on human medullary thyroid carcinoma cells (TT cells). GRP stimulated calcitonin(CT) release in a concentration-dependent manner at 0.1-1000 nmol/l. On adding forskolin along with GRP, CT release was greater than by GRP alone. The stimulatory effect of A23187 was not additive. Intracellular free calcium concentration ([Ca2+]i) was measured for individual TT cells loaded with fura-2. The addition of GRP caused a rapid and transient rise in [Ca2+]i in a concentration-dependent manner followed by a sustained increase in [Ca2+]i. In the medium without Ca2+, this sustained increase did not occur and the concentration of CT release from TT cells by GRP was reduced by approximately a half. GRP would thus appear to be importantly involved in the regulation of thyroid C cell function through modulation of [Ca2+]i.

Calcimycin↗
PubMed1992

Catabolism of hemoglobin-haptoglobin complex in microsome subfractions.

After internalization of hemoglobin-haptoglobin complex (Hb-Hp) via receptor-mediated endocytosis (RME) into liver parenchymal cells, organelles containing the complex distribute in the microsome fraction (Ms). Prior to the catabolism, Hb-Hp dissociates symmetrically into two 82,000-dalton (82 kDa) subunits. In the present investigation, the first event of Hb-Hp metabolism in Ms were further examined after [3H-heme, 14C-glogin]Hb-Ho or [125I-Hb]Hp injection to rats. Shortly after the internalization of Hb-Hp, this complex in Ms was intact. At 60 min after injection, radioactive materials of Ms extracted by freezing and thawing (F&T) with yield of 15% were composed of Hb-Hp, 82 kilodaltons (kDa) subunits and Hb metabolites with a ratio of 1:6:13. The heme metabolites were identified as [3H]bilirubin by high performance liquid chromatography (HPLC). The ratio of Hb-Hp/82 kDa subunits/Hb metabolites in microsome residue of the F&T was 40:8:1. The radioactivity in Ms at 60 min localized microsomes subfraction except Golgi light fraction. In electron microscope radioautography of microsome subfraction using [125I]Hb-Hp, silver grains were observed over or within morphologically heterogenous vesicles, e.g. vesicles containing very low density lipoprotein (VLDL) particles with appendage like multi-vesicular body (MVB) or compartment of uncoupling of receptor and ligand (CURL) in Goligi light and intermediate fractions. These studies suggest that Hb-Hp internalized by RME is dissociated symmetrically into two 82 kDa subunits in organelles of Ms, and that organelles with MVB or CURL-like structures are associated with Hb-Hp metabolism.

Animals↗