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Biomedical subjects

S Usami

Publications and source records attributed to S Usami.

At least 127 records · Page 7Linked to original sources

Deformability measurements on individual sickle cells using a new system with pO2 and temperature control.

Although the rheologic behavior of sickle erythrocytes (SS cells) is highly dependent on oxygen tension (pO2) and temperature, very little data exist regarding the effects of deoxygenation and reoxygenation on the rheology of "individual" SS cells at body temperature. We have devised and assessed a new experiment system, in which micropipette aspiration can be performed on individual cells in a constant-temperature chamber that has ports for changing media with different pO2 (effected in 30 to 120 seconds) and sensing probes for monitoring pO2 and temperature. This system enabled us to simultaneously alter and monitor pO2 at 37 +/- 0.5 degrees C, and to monitor and study a single cell under microscopic observation. The static rigidity (E) and dynamic rigidity (eta) of individual SS cells were determined by repeated aspirations of the same cell under various pO2. With stepwise reductions in pO2, E and eta showed no significant changes before sickling, but once sickled, their values markedly increased by 10(2)- to 10(3)-fold concomitantly with morphologic alteration of the cell. Thus, the deformability of a single SS cell behaves in an "all or none" manner at a critical pO2, and earlier studies on the effect of deoxygenation on the rheology of SS cell suspensions probably reflect the overall behavior of SS cells with widely distributed critical pO2.

Anemia, Sickle Cell↗

Prostaglandin E1 and dibutyryl cyclic AMP enhance platelet resistance to deformation.

The effect of prostaglandin E1 (PGE1) on platelets is mediated through the PGE1 receptor and the consequent maintenance of the platelet's discoid shape. The effects of PGE1 and dibutyryl cAMP (dbcAMP) on the deformability of human platelets were studied. Deformability tests based upon the micropipette aspiration on the platelets were performed by using pipettes with radii (Rp) of 0.26-0.36 microns. The time course of the extension length (Dp, in microns) of the platelets in response to aspiration with a negative pressure (delta P) of 5 cm H2 O (delta P x Rp = 0.15 dynes/cm) was analyzed. PGE1 treatment (0.1 microM) resulted in a decrease of platelet deformability as compared with results obtained for apparently non-activated, control platelets. The deformation index, i.e., Dp/Rp (PGE1-treated)/Dp/Rp (control), was significantly reduced to 0.90 +/- 0.04. DbcAMP treatment also significantly decreased the deformability of platelets and this decrease was dbcAMP dose dependent. In contrast, colchicine- or cytochalasin D-treated platelets increased deformability. PGE1-treated platelets had a higher [cAMP]i than controls. Platelets treated with PGE1 or dbcAMP showed a reduced [Ca2+]i increment induced by thrombin as compared to non-treated controls. These results indicate that PGE1 and dbcAMP treatment of platelets is accompanied by an enhancement of platelet resistance to deformation. The increased [cAMP]i and low [Ca2+]i after PGE1 treatment may limit the rearrangement of cytoskeleton and thus enhance platelet resistance to deformation.

Alprostadil↗

Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate.

The light- and electron microscopic localization of glutamate and glutamine in the rat organ of Corti was studied by means of antisera raised against the respective amino acids coupled to carrier proteins. The light microscopic analysis was performed in semithin sections treated according to the peroxidase-antiperoxidase procedure. The two amino acids were visualized in the same ultrathin sections by use of postembedding immunocytochemistry with two different gold particle sizes. The distribution of aspartate-like immunoreactivity was also recorded, but only at the light microscopic level. In the hair cells, the level of glutamate-like immunoreactivity was higher than that in supporting cells but lower than that in the presumed glutamatergic terminals of cerebellar parallel and mossy fibres. The latter types of terminal were sampled from ultrathin sections that had been incubated under the same conditions as the cochlear sections. Within the hair cells, gold particles signalling glutamate were enriched on mitochondria but not on clusters of synaptic vesicles. Glutamine-like immunoreactivity was present in hair cells as well as supporting cells. The glutamate/glutamine ratio, expressed as the ratio between the respective gold particle densities, was considerably lower for hair cells compared with the cerebellar excitatory terminals. No consistent difference was found between outer and inner hair cells in relation to the levels and subcellular distribution of glutamate and glutamine immunoreactivities. Aspartate-like immunoreactivity was accumulated in outer hair cells, with some labelling also of border cells and Böttcher cells. While the present study confirmed the presence of glutamate in hair cells and demonstrated that these cells are also endowed with the important glutamate precursor glutamine, it revealed notable differences between hair cells and presumed glutamatergic terminals in the CNS. These could reflect differences in the synthesis and compartmentation of transmitter glutamate. Methodological factors could also contribute. Alternatively, the differences could be interpreted to suggest that the hair cell transmitter is not glutamate, but a similar compound. Aspartate could be a candidate in the case of the outer hair cells.

Animals↗

Analgesia inhibitory system involvement in nonacupuncture point-stimulation-produced analgesia.

Acupuncture analgesia (AA), caused by low-frequency stimulation of an acupuncture point (AP)--in this case the tibial muscle--was augmented. Nonacupuncture analgesia (NAA), caused under certain circumstances by stimulation of a nonacupuncture point (NAP)--in this case the abdominal muscle--was unmasked by lesion in the lateral centromedian nucleus of the thalamus (L-CM) or part of the posterior hypothalamus (I-PH). Stimulation in these regions suppressed the augmented part of the AA and blocked the NAA. These regions were, collectively, given the name analgesia inhibitory system. NAA was abolished, the same as AA, by hypophysectomy. The pathways from the AP and NAP to the pituitary gland were different. AA was naloxone reversible, and NAA was dexamethasone reversible. The analgesia inhibitory system is activated nonspecifically by stimulation of either an AP or NAP. It ascends to the I-PH, thence to the L-CM, and ultimately inhibits the pathway nonspecifically connected to the NAP and AP in the lateral part of the periaqueductal central gray (PAG), without affecting the pathway specifically connected to the AP. Thus, only stimulation of an AP will produce analgesia, whereas stimulation of an NAP will not normally produce analgesia. Stress-induced analgesia (SIA) is produced in a different way than AA or NAA.

Afferent Pathways↗

Preprotachykinin A mRNA detected by in situ hybridization in guinea pig vestibular ganglion.

In situ hybridization histochemistry was conducted to examine the expression of mRNA encoding preprotachykinin A (PPTA), the precursor of the neuropeptide substance P (SP) and neurokinin A (NKA), in guinea pig vestibular ganglion. Signals for PPTA mRNA moderately accumulated on most ganglion cells. SP and NKA are thus confirmed to be present in vestibular ganglion cells of the guinea pig.

Animals↗

Locomotion forces generated by a polymorphonuclear leukocyte.

There have been very few studies which have measured the physical forces generated by cells during active movements. A special micropipette system has been designed to make it possible to observe cell motion within the pipette and to apply a pressure to counter the chemotactic migration of the cell. This provides a direct measure of the locomotion force generated by the cell. The average velocity of forward motion is 0.33 microns/s in the absence of counter-pressure. The application of a positive counter-pressure (C-P) causes a decrease in the velocity of the forward motion of the cell. At 17 cm H2O of C-P, the cell velocity drops to zero and even moves backward with a higher C-P. The results show that the decrement of velocity is linearly related to the magnitude of the C-P with a complete stoppage at a pressure of 17 cm H2O which corresponds to a force of 0.003 dyn. The maximum work rate of the cell is approximately 2.5 x 10(-8) erg/s.

Biomechanical Phenomena↗

[Lymphoid infiltration of colon cancer and clinical significance of the skin test].

It was reported that lymphoid infiltration (LI) of colon cancer tended to be found more commonly in patients with colonic cancer in an early stage than in an advanced stage. This time, the correlation among LI of colon cancer, cell-mediated immunity and the cumulative 5-year survival rate was studied in 124 patients with resected colon cancer. The results were as follows: 1) The cumulative 5-year survival rate in patients with positive LI was higher than that in the patients with negative LI. 2) There was correlation between the degree of LI of the cancer and Su-PS skin reaction. 3) Postoperative cell-mediated immunity was reduced in the cases of poor prognosis. Therefore we insist on paying attention to immunochemotherapy for 24 months after surgery.

Colonic Neoplasms↗

[Early diagnosis and management against perioperative fungus infection].

Candida spp. is frequently detected from perioperative patients, especially from those requiring central venous catheter (CVC) management. CAND-TEC could detect the blood fungus infection easily and quickly. We analyzed 438 specimens from 57 patients requiring perioperative CVC management for more than three weeks, and investigated usefulness of CAND-TEC comparing with other serological methods. The dilution rate of CAND-TEC showed a positive correlation with amount of beta-D-glucan which was measured by Toxicolor test and Endotoxin-specific assay. It also showed a positive correlation with amount of D-arabinitol. When blood samples diluted more than four times were positive with CAND-TEC, the patients were considered to be infected, and accumulative infection rate was up to 84.2% three weeks after introduction of CVC management. CAND-TEC would be useful in detecting Candida infection compared with conventional methods, and the fungus infection was much more frequent incidence than ever expected. It is important to detect fungus infection early and respond immediately to avoid nosocomial infection.

Candidiasis↗

Localization of substance P-like immunoreactivity in guinea pig vestibular endorgans and the vestibular ganglion.

The immunocytochemical distribution of substance P (SP) in guinea pig vestibular endorgans and the vestibular ganglion was investigated. Two kinds of SP-immunoreactive fibers were distinguished. Most were thick, and found around or beneath sensory hair cells. These SP-immunoreactive fibers were distributed predominantly on the slope of the crista and the peripheral region of the macula. By electron microscopy, we confirmed this type of SP-like immunoreactivity to be restricted within primary afferent neurons. Some vestibular ganglion cells also showed SP-like immunoreactivity, suggesting that SP is present in some primary afferent neurons, and is involved in afferent neurotransmission. The characteristic distribution of SP may indicate functional differences within each endorgan. The other group of immunoreactive nerve fibers, varicous thin fibers, could be found in the stroma of vestibular endorgans, nerve trunk, vestibular ganglion, and along blood vessels of the vestibular ganglion. These fibers may have a different origin, and have an influence on blood flow and certain other functions.

Animals↗

The mechanism of irritative nystagmus and paralytic nystagmus. A histochemical study of the guinea pig's vestibular organ and an autoradiographic study of the vestibular nuclei.

To establish the difference of mechanism between irritative and paralytic nystagmus, alterations of Na-K-ATPase and succinic dehydrogenase activity in the vestibular sensorineural elements were investigated for 20 guinea pigs, and glucose uptake of the vestibular nuclei for 13 guinea pigs were measured by the [14C]-2-deoxy-D-glucose method. Irritative and paralytic nystagmus were experimentally provoked by introducing K+ into the perilymphatic space. From the results it was concluded that irritative nystagmus is provoked by increased excitability of vestibular sensory cells, while paralytic nystagmus is provoked by decreased excitability. However, the direction of nystagmus was eventually decided by the tonus imbalance between the bilateral vestibular nuclei. The ipsilateral vestibular nucleus was predominant during irritative nystagmus, while the contralateral vestibular nucleus was predominant during paralytic nystagmus.

Animals↗

Coexistence of substance P and calcitonin gene-related peptide-like immunoreactivities in the rat vestibular endorgans.

The immunocytochemical distribution and coexistence of substance P (SP) and calcitonin gene-related peptide (CGRP) in the rat vestibular endorgans were investigated. SP-like immunoreactivity was found in nervous elements beneath and around hair cells. CGRP-like immunoreactivity was also abundantly distributed beneath and within the sensory epithelia. In the present study, double-staining immunocytochemistry revealed that three different types of immunoreactivities: SP-positive/CGRP-negative, SP-negative/CGRP-positive, and SP/CGRP-positive immunostaining can be distinguished. SP/CGRP-immunoreactive fibers were localized within as well as beneath the sensory epithelia. These fibers often penetrated the epithelia and nearly reached the surface. The present immunocytochemical evidence suggests that different types of peripheral nervous systems may exist in the vestibular periphery.

Animals↗

[A case of graft-versus-host disease following irradiated fresh blood transfusion].

We reported a case of a fatal graft-versus-host disease (GVHD) which developed in a 65-year-old, male patient which was considered to have been induced by irradiated fresh blood donated by his son after a coronary bypass surgery. Fresh blood was obtained from his relatives, and a 15 Gy irradiation was performed before transfusion. The diagnosis of acute GVHD was made by clinical symptoms and histological examinations of the skin and the bone marrow. He died of sepsis on the 19th post-operative day. The HLA typing of the lymphocytes, revealed that the patient had A 2, A 24, Bw 52, Bw 62, Cw 4, DR 2, and his son had A 24, Bw 52, DR 2. A 24 and Bw 52 were homogeneous making his son histocompatible with one of the patient's haplotype. This might well be attributable to the occurrence of GVHD in this case, meaning that 15 Gy irradiation was not sufficient for the prevention of this disease.

Aged↗

Control of expression of Agrobacterium vir genes by synergistic actions of phenolic signal molecules and monosaccharides.

Most virulence (vir) genes of Agrobacterium tumefaciens that are required for the formation of crown gall tumors are expressed in response to such plant signal molecules as acetosyringone and lignin precursors. The phenolic signals are transduced through a receptor VirA protein in the inner membrane of the bacterial cell. The expression of these genes triggers the transfer of a specific DNA segment, called transferred DNA (T-DNA), from the Ti plasmid to plant cells, and its integration into their nuclear DNA. We show here that a group of aldoses (L-arabinose, D-xylose, D-lyxose, D-glucose, D-mannose, D-idose, D-galactose, and D-talose) can markedly enhance acetosyringone-dependent expression of vir genes when the concentration of acetosyringone is limited (10 microM) but does not enhance the expression of noninducible genes. Likewise, a 2-deoxy-D-glucose, a nonmetabolized sugar, is also effective. When a deletion was introduced into the virA gene in the region encoding the periplasmic portion of the VirA protein, enhancement by glucose disappeared, but vir expression was induced by acetosyringone in this mutant. These results suggest that these sugars directly enhance a signaling process initiated by phenolic inducers that results in an increase in expression of the vir genes.

Journal Article↗

The dynamics of shear disaggregation of red blood cells in a flow channel.

Red blood cell (RBC) rouleaux were formed in a flow channel in the presence of 2 g/dl dextran (molecular weight 76,000). The partial separation of RBC rouleau doublets adhering to the floor of the flow channel in response to small oscillatory shear stresses was observed experimentally. Theoretical analyses on displacement and drag force were performed to determine whether the motion of the cell involves membrane rotation (i.e., rolling) or sliding. From the experimental data and the results of theoretical analyses, it is concluded that, under the conditions of the experiments, the RBCs in a doublet separate from each other by rolling, rather than sliding of the sheared cell.

Biomechanical Phenomena↗

[Effects of ulinastatin on plasma polymorphonuclear leukocyte elastase activity and respiratory function in patients undergoing cardiopulmonary bypass].

Plasma polymorphonuclear leukocyte elastase (PMNE) activity is known to increase during cardiopulmonary bypass (CPB), and is considered to mediate pulmonary tissue damage causing postoperative respiratory failure. We made a clinical study in order to clarify the effects of ulinastatin (Miraclid), a proteolytic enzyme inhibitor, on plasma PMNE activity and respiratory function. Twenty adult patients undergoing coronary artery bypass grafting were divided into 2 groups; 10 to group U and 10 to group C. The patients in group U received 500 U/kg body weight of ulinastatin intravenously before and after CPB, and the patients in group C not receiving the dose served as controls. Arterial blood samples were obtained before the operation, 1 hour after CPB, 3 hours, 1 day and 4 days after the operation. The leukocyte count was significantly lower in group U 1 hour after CPB and 3 hours after the operation compared to group C (p less than 0.01). The plasma PMNE activity rose rapidly after starting CPB and the peak level appeared 1 hour after CPB; 1165 +/- 560 micrograms/l in group U and 1981 +/- 562 micrograms/l in group C (p less than 0.02). There was a significant correlation between the leukocyte reduction and the rate of increase of beta-glucuronidase activity were diminished in group U compared to preoperative values (p less than 0.05). The recovery of the oxygenation index (PaO2/F1O2), which was used for evaluation of the respiratory function, was worse in group C (p less than 0.01). These results suggest that the administration of ulinastatin in patients undergoing CPB is useful for the prevention of the deleterious effects of PMNE on the lung.

Aged↗

[Histology and immunopathology of colonic carcinoma].

The correlation among lymphoid cell infiltration (LI), cell-mediated immunity and immunoglobulin-containing cells distribution in the colon cancer was studied in resected materials of 136 patients with colonic cancer. The results were as follows: 1) LI of the colon cancer tended to be found more commonly in patients with colonic cancer in clinical early stage than advanced stage. 2) The cumulative 5-year survival rate in patients with positive LI was longer than that in the patients with negative or normal LI. 3) In colonic cancer patients with the better prognosis, IgA-containing cells were distributed around the tumor tissues more abundantly than that in the poor ones. 4) These results suggest that the IgA system somewhat participates in cancer immunity.

Colonic Neoplasms↗

Immunocytochemical study of the GABA system in chicken vestibular endorgans and the vestibular ganglion.

The immunocytochemical distribution of gamma-aminobutyric acid (GABA), GABA synthesizing enzyme; L-glutamate decarboxylase (GAD) and degradative enzyme; GABA transaminase (GABA-T) in the chicken vestibular endorgans and the vestibular ganglion was investigated. GABA and GAD-like immunoreactivity were confined to the sensory hair cell cytoplasm, suggesting that GAD synthesizes GABA in the hair cell. GABA-T-like immunoreactivity, indicative of GABA degradation, was found around hair cells, along nerve fibers running through the stroma and within the ganglion cell. These immunocytochemical findings indicate that the GABAergic system exists in the chicken vestibular endorgans and that GABA may function as an afferent neurotransmitter at the level of hair cells.

4-Aminobutyrate Transaminase↗

Shear stress-induced detachment of human polymorphonuclear leukocytes from endothelial cell monolayers.

We employed a static-incubation assay to determine the intensity of wall shear stress (tau) needed to detach human polymorphonuclear leukocytes (HPMNs) from human umbilical vein endothelial cell (HUVE) monolayers. Confluent monolayers of HUVE were placed in a parallel-plate flow chamber which was mounted on the stage of an inverted tissue culture microscope, attached to a perfusion system and maintained at 37 degrees C. All events in the selected fields were recorded using videomicroscopy. HPMNs were co-incubated for 15 minutes with the HUVE monolayers under control conditions or in the presence of 10(-7) M formyl-methionyl-leucyl-phenylalanine (FMLP). Following this static incubation, a series of five individual flows, each 1 minute in duration, were driven through the flow channel, exposing the cells to 1.0, 2.0, 3.8, 7.6 and 14.8 dyn/cm2 wall shear stresses. Under control conditions, the percentage of HPMNs remaining attached to the HUVE monolayers following exposure to each shear stress was 61, 38, 25, 12 and 5, respectively. In the FMLP-treated condition, the percentage of HPMNs remaining attached to the monolayers was significantly greater than control at all five levels of tau. Thus, under control conditions, adherent HPMNs can be detached from endothelial cell monolayers in vitro with levels of shear stress normally found in the microcirculation (18). In the presence of FMLP, the level of shear stress needed to overcome the adhesions is increased significantly.

Cell Adhesion↗