Evaluation methods of speech perception ability through cochlear implant.
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Cochlear Implants↗
Biomedical subjects
J Kawano
Publications and source records attributed to J Kawano.
Genetic analysis of equine methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis.
Pulsed-field gel electrophoresis (PFGE) was used to determine genetic relationships among 15 methicillin-resistant Staphylococcus aureus (MRSA) isolates from mares with metritis and from a stallion with dermatitis in Hokkaido. All the 15 isolates showed phage pattern 6/47/54/75, coagulase type IV, and enterotoxin type A. The restriction endonuclease SmaI cut their genomic DNAs into 15 or 16 fragments ranging in size from 8 to 630 kb. Fourteen of the 15 isolates showed the same PFGE pattern, whereas the remaining one appeared to be closely related. The 9 human MRSA isolates showing the same phenotypic characteristics as the horse isolates gave different PFGE patterns from those of the horse isolates.
Animals↗
Comparison of pulsed-field gel electrophoresis and phage typing for discriminating poultry strains of Staphylococcus aureus.
OBJECTIVE: To compare pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus aureus from chickens in England, Belgium, Bulgaria, Argentina, and Japan, to assess the value of PFGE for discriminating strains, and to compare results obtained by PFGE with those obtained by biotyping and phage typing. SAMPLE POPULATION: 78 S aureus isolates from diseased and healthy chickens. PROCEDURE: Chromosomal DNA of S aureus was digested with restriction endonuclease Sma I, and fragments were separated by PFGE in 1% agarose gel. RESULTS: All 78 strains from 5 countries were classified as poultry ecovar according to a previously established biotyping system. Chromosomal DNA was cut by Sma I into 18 to 23 fragments ranging from about 3 to 685 kb. Seventy-eight strains produced 15 types, arbitrarily designated A to O, and 45 subtypes. Some differences were observed in PFGE patterns among countries. However, 10 fragments (333, 190, 110, 63, 55, 42, 34, 19, 10, and 3 kb) were highly conserved and were shared by almost all (> 78%) of the strains examined. The PFGE patterns were compared with those obtained by phage typing. All 29 strains belonging to avian phage-group II produced type A and 19 subtypes. Of the 15 strains belonging to phage-group I, 11 produced 8 types (B to H, O) and 5 subtypes that were different from those of type A. CONCLUSIONS: Genomic DNA fingerprinting by PFGE is an effective technique for discriminating poultry S aureus strains and appears to be a useful method for subtyping strains of avian phage groups or the poultry-specific ecovar.
Animals↗
Crossed projection neurons are generated prior to uncrossed projection neurons in the lateral superior olive of the rat.
The present study examined in the lateral superior olive (LSO) of the rat whether LSO neurons projecting to the ipsilateral inferior colliculus (IC) might be generated later than those projecting to the contralateral IC. Rat fetuses were exposed in utero to 5-bromodeoxyuridine (BrdU), a thymidine analogue, to label neurons proliferating at different embryonic stages from day E11 through to E20. Upon reaching adulthood, the rats were given unilateral injections of fluoro-gold (FG), a retrograde fluorescent tracer, into the IC. Subsequently, the tissue sections of the brains obtained from the rats were immunostained for BrdU to simultaneously detect neurons that were BrdU-positive and/or FG-positive. BrdU-positive LSO neurons were found in the rats which had been exposed to BrdU during E12-E16. In E12 and E13 BrdU-exposure cases, the vast majority of doubled-labeled (BrdU-positive and FG-positive) neurons were seen on the contralateral side to the FG injection. In E14, E15 and E16 BrdU-exposure cases, in contrast, all double-labeled neurons were found on the ipsilateral side to the FG injection. The distribution of these double-labeled neurons within the nucleus was diffuse in all the BrdU-exposure cases. Thus, the results indicate that LSO neurons are generated during E12-E16, that the crossed projection neurons are generated 1-4 days earlier than the uncrossed projection neurons, and that no topographical relationships exist between the early- and the late-generated populations of the LSO neurons.
Animals↗
Isolation of methicillin-resistant coagulase-negative staphylococci from chickens.
Methicillin-resistant coagulase-negative staphylococci were isolated from the nares and skin of 1- to 8-week-old healthy chickens in three flocks from a farm. Isolation of methicillin-resistant coagulase-negative staphylococci was positive for 72 (25.7%) of the 280 chickens tested, with the frequency varying from 2.2 to 100% according to flock. A total of 45 appropriate isolates were selected and subjected to identification. Of the 45 methicillin-resistant coagulase-negative staphylococcal isolates selected, 37 were identified as Staphylococcus sciuri, 5 were identified as Staphylococcus epidermidis, and 3 were identified as Staphylococcus saprophyticus. The distribution of the species was different among the flocks. Comparative analysis of the SmaI-digested chromosomal DNA by pulsed-field gel electrophoresis revealed that the isolates could have originated from a single clone of each of S. sciuri and S. saprophyticus and three clones of S. epidermidis. By two methods based on the PCR technique, the mecA gene was detected in all five representative isolates of each methicillin-resistant coagulase-negative staphylococcal clone. The nucleotide sequence of a PCR fragment obtained from an isolate of S. sciuri was completely identical to the corresponding region of mecA genes reported in human methicillin-resistant Staphylococcus aureus isolates and Staphylococcus epidermidis isolates. The representative methicillin-resistant coagulase-negative staphylococcal isolates were resistant to many beta-lactam antibiotics, and some isolates were also resistant to macrolide and aminoglycoside antibiotics. This is the first evidence of the existence of methicillin-resistant coagulase-negative staphylococci from animals possessing the mecA gene.
Animals↗
Desulfation of tyrosine-O-sulfated peptides by some eukaryotic sulfatases.
Three mammalian and eight non-mammalian arylsulfatases were investigated for their activities toward tyrosine-O-sulfate (TyrS) in peptides. None of the mammalian arylsulfatases exhibited detectable activities toward TyrS-containing peptides. Of the non-mammalian arylsulfatases tested, Types VII, VIII, and H-1, 2, and 5 displayed strong activity on endo-TyrS residues. The prokaryotic sulfatase, Type VI, was active only on free TyrS and N-terminal TyrS of Leu-enkephalin. All the sulfatases were active on p-nitrophenyl sulfate and p-nitrocatechol sulfate.
Peptides↗
[Intraoperative diagnosis. The reason of restricted indication and suggestion].
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Diagnosis↗
Electron microscopic study of synaptogenesis and myelination of the olfactory centers in developing rats.
Development of the central olfactory system was studied in the rat with an electron microscope at three main structures: the olfactory bulb, the lateral olfactory tract, and the primary olfactory cortex (the piriform cortex). As a parameter of development, the synaptic density was examined quantitatively in the bulbar glomerulus and layer Ia (termination of bulbofugal fibers) of the piriform cortex. which are the key stations of the olfactory pathway. The synaptic densities in the glomerulus and those in layer Ia were 5.7% and 4.6% on embryonic day 19, 15.8% and 12.5% on postnatal day (P) 0, and 57.3% and 37.2% on P10, as compared with the adult (100%). As another parameter of development, the density of myelinated axons in the lateral olfactory tract was examined quantitatively. The densities of myelinated axons in the tract were 0% on P5, 15.1% on P10, and 73.5% on P21 of the adult density. Maturation in the tract was still progressing, even at P21, in terms of bundle formation and the thickness of myelin sheaths. The results show that synaptogenesis in the bulbar glomerulus is followed by synaptogenesis in layer Ia of the piriform cortex, and that myelination in the lateral olfactory tract occurs over a prolonged period, even in the stages after P21.
Aging↗
Postnatal development of the projection from the medial superior olive to the inferior colliculus in the rat.
Normal projection development from the medial superior olive (MSO) to the inferior colliculus (IC) was examined by injecting Fluoro-Gold (FG), a retrograde tracer, into the IC unilaterally at postnatal days 0 (P0), P3, P7 and maturity. The rats were killed 1 day after FG injection. At all ages, labeled neurons in the MSO appeared on the ipsilateral side only, as in adult controls. The total number of MSO neurons counted in Nissl-stained sections was constant throughout the postnatal periods. The labeled frequency index of MSO neurons was increased stepwise (from 35% to 90%) with increasing postnatal stages (from P0 to adulthood), suggesting differential growth of early- and late-developing axons.
Age Factors↗
Anatomical plasticity in the medial superior olive following ablation of the inferior colliculus in neonatal and adult rats.
We evaluated the consequences of unilateral ablation of the inferior colliculus (IC) upon the ascending projection from the medial superior olive (MSO) to the IC. Ablation of the IC was performed in rats aged between postnatal day 1 (P1) and maturity. All the rats were given injections of Fluoro-Gold (FG) into the ipsilateral IC at birth (P0) (before the ipsilateral IC was ablated in any case) so that growth of early-developing axons to the ipsilateral IC could be examined for any labeled neurons in the ipsilateral MSO. Upon reaching adulthood, the rats received injections of Fluoro-Ruby (FR) into the contralateral (intact) IC so that aberrant crossed projections to the intact IC could be examined for any labeled neurons in the ipsilateral MSO. These rats were killed 2 days after FR injections. The number of surviving cells in the ipsilateral MSO were counted in Nissl-stained sections for quantitative analysis of retrograde degeneration. The results show that: (1) the total number of neurons was reduced to 64-34% in the ipsilateral MSO as a result of IC ablation; (2) cell reduction by retrograde degeneration followed a U-shaped curve with a maximal effect in rats operated at P7 (reduced to 34%); (3) adult ablation of the IC led to retrograde degeneration that was less severe than that in late neonatal (P7) ablation; (4) an aberrant projection from the MSO to the contralateral IC occurred in rats operated at P1 and P3 but not in rats operated at P7 or maturity. Thus, our findings suggest that growth of late-developing axons is a major factor in the plasticity of this system of projection.
Age Factors↗
Differentiation of the olfactory epithelium during development.
Maturation of the olfactory epithelium was studied in the rat. As a developmental index of the olfactory epithelium, quantitation was based on the thickness and the number of cell layers of the epithelium. Mean epithelial thicknesses were 84 microns on embryonic day (E) 19, 94 microns on postnatal day (P) 0, 98 microns on P10 and 57 microns in adult. Average numbers of cell layers were 7 (E19), 8 (P0), 10 (P10) and 7 (adult). Thus the olfactory epithelium progressively increased in thickness between P0-P10, and later decreased in thickness as well as in number of cell layers toward adult. To clarify the number of mature receptor cells sending their axons to the olfactory bulb, the percentages of receptor cell labeling (number of labeled cells/total number of receptor cells) were estimated with an aid of retrograde fluorescent tracer injected into the olfactory bulb. As a result, their percentages were 34% (P1-P3), 50% (P8-P10) and 78% (adult). The ablation experiment of the bulb on P1 pups and adult rats revealed that the percentages of receptor cell degeneration were 31% in neonates (P1) and 76% in adults, confirming the results obtained from the retrograde tracing. Thus the present study shows the substantial increase of mature receptor cells during development and may provide a morphological basis in understanding of the functional maturity of olfaction.
Animals↗
Glycoconjugates in rat small intestinal mucosa during infection with the intestinal nematode Nippostrongylus brasiliensis.
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Animals↗
Purification and immunohistochemistry of Griffonia simplicifolia agglutinin-II-binding mucus glycoprotein in rat stomach.
Gastric mucus is thought to protect the gastric wall from mechanical trauma, desiccation, pathogenic microorganisms, acid and proteases. We purified Griffonia simplicifolia agglutinin-II (GSA-II)-binding mucus glycoprotein (GMG) from rat gastric mucosa by solubilization in a guanidine-containing buffer, gel permeation chromatography, Ricinus communis agglutinin-I (RCA-I)-affinity chromatography and GSA-II-affinity chromatography. Rat GMG showed high molecular weight on a Sephacryl S-1000 column, and a single band in 0.5% agarose-2% polyacrylamide composite gels and blots. A protein of approximately 60 kDa was contained in the GMG preparation. GMG was deglycosylated with trifluoromethanesulphonic acid treatment. An antibody was raised against deglycosylated GMG (deGMG). The antibody recognized deGMG, GMG, periodic acid-treated deGMG and O-glycanase-digested deGMG, but did not react to trypsin-digested deGMG. These results suggest that the antibody recognizes proteinase-sensitive region or peptide backbone of GMG. In immunohistochemistry, the mucous gel layer of the stomach luminal surface was stained with antibody. The antibody recognized not only gastric mucous neck cells and pyloric gland cells, but also gastric surface mucous cells, mucous cells in the duodenal gland, and goblet cells in the small intestine and colon. These results indicate that GMG is a component of rat gastric mucus, and that the antibody recognizes mucous-secreting cells in rat stomach and intestine.
Animals↗
A new type of staphylococcal exfoliative toxin from a Staphylococcus aureus strain isolated from a horse with phlegmon.
A new type of staphylococcal exfoliative toxin (sET) was isolated from the culture filtrate of a Staphylococcus aureus strain isolated from a horse with skin infection including phlegmon. The new sET was purified by precipitation with 80% saturated ammonium sulfate, column chromatography on DEAE-cellulofine A-500, gel filtration on a Sephadex G-75 column, and polyacrylamide gel electrophoresis (7.5% polyacrylamide). The new sET elicited general exfoliation of the epidermis with the so-called Nikolsky sign when inoculated into both 3-day-old mice and 1-day-old chicks, whereas sETA and sETB from human strains of S. aureus caused exfoliation in a 3-day-old mouse alone and shET from a porcine strain of Staphylococcus hyicus caused exfoliation in 1-day-old chicks alone. Intraepidermal splitting was observed at the granular layer of the epidermis of mice inoculated with the new sET as well as those inoculated with sETA. Exfoliation at the germinative layer of the epidermis was also observed in the chicks inoculated with the new sET as well as those inoculated with shET. The new sET was serologically different from sETA, sETB, and shET and showed the same molecular weight on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was thermolabile and lost its toxicity after being heated at 60 degrees C for 15 min. We propose that the new sET be designated as sETC.
Animals↗
A protein-specific monoclonal antibody to rat liver beta 1-->4 galactosyltransferase and its application to immunohistochemistry.
We have produced a new protein-specific monoclonal antibody (MAb) to rat liver beta 1-->4 galactosyltransferase. This MAb, GTL2, was selected as the most reactive IgG to a periodate-treated antigen. Antigen and protein specificities of GTL2 were verified by immunoblotting of a non-glycosylated recombinant protein of human galactosyltransferase and enzymatically deglycosylated rat galactosyltransferase. Using GTL2, an immunohistochemical study was done in rat liver, epididymis, and salivary glands. Intense staining was observed in Golgi areas of epididymal duct epithelial cells, and submandibular and sublingual acinar cells. Hepatocytes showed weaker staining. Immunoelectron microscopic observation revealed that the staining was exclusively localized in trans-Golgi membranes of these cells.
Animals↗
Human colorectal carcinoma-specific glycoconjugates detected by pokeweed mitogen lectin.
Pokeweed mitogen (PWM) lectin, known to bind branched poly-N-acetyllactosamines, has a highly selective affinity for human colorectal carcinomas. We performed light microscopic (LM) histochemistry with PWM lectin on paraffin sections of human colorectal tissues. In histological sections, normal mucosae and adenomas with mild dysplasia exhibited negative reaction (0/10, 0/13, respectively) with or without neuraminidase pre-digestion, whereas adenomas with moderate dysplasia showed a small increase in PWM lectin reactivity after neuraminidase digestion (4/23). In contrast, we observed a high incidence of positive reactivity in colorectal carcinoma without neuraminidase pre-digestion (38/44). After digestion with neuraminidase, there was increased reactivity of colorectal carcinomas in situ (7/12) and invasive carcinomas (13/32). These results imply that human colorectal carcinomas consistently contain substantial amounts of PWM-reactive branched poly-N-acetyllactosamine glycoconjugates structures. We also compared the staining patterns of PWM lectin and monoclonal antibodies (MAb) directed to Lewis X (LeX) or Lewis Y (LeY) antigen. PWM lectin reactivity was largely confined to the apical membranes of carcinoma tissues. MAb-LeX or MAb-LeY immunoreactivity was seen on the apical membranes and in the cytoplasm of both adenomas and carcinomas. Therefore, histochemical studies with this lectin should be useful for identification of carcinoma tissues and analysis of glycoconjugates associated with colorectal carcinoma.
Adenoma↗
Differential projections to the superior collicular layers from the perihypoglossal nuclei in the cat.
The primary objective of the present study is to demonstrate the presence of a projection to the superficial layers of the superior colliculus (SC) from the perihypoglossal nuclei, specifically from the nucleus intercalatus (INT) in the cat. Iontophoretic application of WGA-HRP into the perihypoglossal complex produced orthogradely labeled terminals in the SC contralaterally forming two bands: one is in the superficial gray layer, and the other in the intermediate gray layer. The superficial band was evenly distributed in the upper portion of the superficial gray layers (layers II1-2) and the deeper band existed in the intermediate gray layer (layer IV) being arranged in a discontinuous manner. Injections of the tracer into the superficial layers of the SC yielded retrogradely labeled cells only in the rostral part of the contralateral INT; by contrast, the injection confined to the deep layers produced labeling of cells exclusively in the nucleus prepositus hypoglossi (PH). Thus, the INT and the PH each project separately to the functionally different superficial and intermediate layers of the SC, respectively. On the basis of the present anatomical findings, it is suggested that the perihypoglossal nuclei as a whole contribute not only to the oculomotor but also to the visuosensory regulatory function in the SC.
Afferent Pathways↗