ABSTRACT
The immunolocalization of phospholipids has not yet been clearly demonstrated in temporomandibular joints (TMJs). We have examined the distribution of one of phospholipids, phosphatidyl-choline (PC), in the rat mandibular condylar surface and lower joint cavity. Some fresh resected TMJs with their disks attached were immediately plunged into isopentane-propane cryogen (-193 degrees C). Cryostat sections were cut, mounted on NH3+-coated slides, and fixed with paraformaldehyde (PF). Cryosections were first immunostained with anti-mouse PC antibody (JE-1). Subsequently, they were labeled with immunogold particles following silver enhancing for light microscopic analyses. Some cryosections were subjected to double immunofluoresecence labeling with anti-fibronectin antibody or hyaluronic acid-binding protein in combination with the anti-PC antibody. As an immunocontrol, other cryosections were pretreated with phospholipase A2 before such immunofluorescence labeling. We have confirmed the presence of PC in the lower joint cavity of rat TMJs as well as on the mandibular condylar surface layer, which was colocalized with hyaluronic acid and fibronectin respectively. However, by treatment with phospholipase A2, such immunolabeling for PC was clearly decreased, showing that the PC is a component in the rat in vivo TMJ. These findings suggest that PC, hyaluronic acid and fibronectin may interact each other in the TMJ articular surface areas to play a functional role for lubrication in TMJ.
Subject(s)
Mandibular Condyle/metabolism , Phosphatidylcholines/metabolism , Temporomandibular Joint/metabolism , Animals , Fibronectins/metabolism , Fluorescent Antibody Technique , Frozen Sections , Histological Techniques , Hyaluronic Acid/metabolism , Immunohistochemistry , Mandibular Condyle/anatomy & histology , Rats , Rats, Wistar , Temporomandibular Joint/anatomy & histologyABSTRACT
The purpose of the present study is to clarify native ultrastructures of upper surface layers of the rat mandibular condylar cartilage in vivo by a quick-freezing method. The mandibular cartilaginous tissues were removed with their articular discs attached without opening the lower joint cavity. The specimens were processed for light microscopy, transmission or scanning electron microscopy. Deep-etching replica membranes were also prepared after the routine quick-freezing method. The upper surface layer was well preserved by the quick-freezing method. The cartilaginous tissues, which were fixed without opening their articular discs, appeared to keep better morphology than those after opening them. The upper surface layer was thicker than the corresponding layer as reported before. It consisted of atypical extracellular matrices with lots of apparently amorphous components, which were distributed over typical collagen fibrils, by conventional electron microscopy. As revealed with the replica membranes, it also consisted of variously sized filaments and tiny granular components localized on the typical collagen fibrils. A pair of stereo-replica electron micrographs three-dimensionally showed compact filaments within the upper surface layer. The quick-freezing method was useful for keeping native ultrastructures of the fragile upper surface layer in the mandibular condylar cartilage, which may be functionally important to facilitate smooth movement of the temporomandibular joint.
Subject(s)
Cartilage, Articular/ultrastructure , Cryoelectron Microscopy/methods , Mandibular Condyle/ultrastructure , Animals , Freeze Etching , Frozen Sections , Male , Microscopy, Electron, Scanning , Rats , Rats, WistarABSTRACT
The purpose of this study was to examine time-dependent topographical changes of leaking proteins from blood vessels in the mouse cerebellum to assess the effect of normal blood circulation on the blood-brain barrier (BBB). The distribution of leaking serum proteins was immunohistochemically examined by various cryotechniques including our "in vivo cryotechnique". The cryofixed cerebellar tissues were processed for the freeze-substitution method, and finally embedded in the common paraffin wax. Serial de-paraffinized sections were immunostained by anti-mouse immunoglobulin-G (IgG) or albumin antibody. By combination of the "in vivo cryotechnique", in which normal blood flow into the cerebellum was always kept in vivo, with the freeze-substitution method, serum IgG and albumin were clearly localized inside of cerebellar blood vessels. To examine abnormal leakage of blood vessels as a model of anoxia, some cerebellar tissues were partially removed from brains in the mouse skull and quickly frozen in the isopentane-propane within a minute. In such resected cerebellar tissues, serum IgG and albumin were diffusely immunostained in large areas around the blood capillaries, probably because of easy leakage of the serum components through the immediately changed BBB. To the contrary, no serum protein could be identified outside blood capillaries under living conditions of the anesthetized mice. The present combination method, both "in vivo cryotechnique" and freeze-substitution, for immunohistochemistry enabled us to examine the in vivo localization of serum components in mouse brains due to alteration of the BBB.
Subject(s)
Blood-Brain Barrier/physiopathology , Cerebellum/physiopathology , Frozen Sections/methods , Hypoxia/physiopathology , Immunoglobulin G/blood , Serum Albumin/metabolism , Animals , Blood Vessels/metabolism , Blood-Brain Barrier/metabolism , Cerebellum/metabolism , Cerebellum/pathology , Hypoxia/blood , Hypoxia/pathology , Immunohistochemistry/methods , Mice , Mice, Inbred C57BLABSTRACT
Both hyaluronic acid and fibronectin localizations were examined in the upper surface layer of rat mandibular condylar cartilages by immunohistochemical techniques. Their delicate structure was successfully preserved by preparation procedures of joint condyles with disks. Paraformaldehyde-fixed cartilaginous tissues were cut in a cryostat, and cryosections were analyzed using streptavidin-peroxidase and indirect immunofluorescence methods. Another immunogold method with conventional preparation procedures and a quick-freezing method was performed for their ultrastructural analyses. Both hyaluronic acid-binding protein and anti-fibronectin antibody were used to localize hyaluronic acid and fibronectin in the mandibular condylar cartilage, respectively. Some cryosections were pre-treated with hyaluronidase and chondroitinase before such labeling. The upper surface layer was composed of double laminar structures. One bordered with the cartilage matriceal surface, which was positive for fibronectin. The hyaluronic acid was localized over the fibronectin layer. Therefore, the hyaluronic acid in vivo was bound with fibronectin in the cartilaginous matrix, performing lubrication for the mandibular joint movement.
Subject(s)
Cartilage, Articular/cytology , Mandibular Condyle/cytology , Animals , Cartilage, Articular/chemistry , Cartilage, Articular/ultrastructure , Fibronectins/metabolism , Fibronectins/ultrastructure , Freeze Drying , Hyaluronic Acid/metabolism , Hyaluronic Acid/ultrastructure , Immunohistochemistry , Mandibular Condyle/chemistry , Mandibular Condyle/ultrastructure , Microscopy, Confocal , Microscopy, Immunoelectron , Rats , Rats, WistarABSTRACT
This report explains a rapid procedure (approximately 50 min) for the isolation of highly purified total RNA from Leishmania promastigotes based on extraction with acidic phenol. The simplicity of the manipulations required make this method ideal for processing multiple samples; the quality of the RNA obtained is suitable for reverse transcription polymerase chain reaction analysis.
Subject(s)
Leishmania , RNA, Protozoan/isolation & purification , Animals , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
In this work we demonstrate that the PCR-ELISA technique is sufficiently sensitive and specific for use as a diagnostic test in cases of mucocutaneous leishmaniasis. DNA was extracted from cultures of Leishmania braziliensis, Leishmania infantum, Leishmania tropica, Leishmania mexicana, Trypanosoma cruzi, and blood samples from individuals who presented a clinical diagnosis of leishmaniasis as well as from healthy individuals. The DNA was PCR amplified and the product obtained was hybridised with a biotin-labelled probe, the sequence of which was designed in our laboratory. The result of the hybridisation was visualised by means of an ELISA technique using antifluorescein antibody labelled with alkaline phosphatase and p-nitrophenylphosphate (pNFF) as chromogen. The optical density of the products of the pNFF hydrolysis was quantified in a spectrophotometer at a wavelength of 405 nm. Using this technique the percentage of detection was 83.3% in blood samples from patients clinically diagnosed as having mucocutaneous leishmaniasis. No false positive results were obtained.
Subject(s)
Enzyme-Linked Immunosorbent Assay , Leishmania braziliensis/isolation & purification , Leishmaniasis, Mucocutaneous/diagnosis , Polymerase Chain Reaction , Animals , Colorimetry , DNA Probes , DNA, Kinetoplast/analysis , Humans , Leishmania braziliensis/genetics , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
La presente publicación describe una visión de las expresiones de la medicina tradicional de acrácter social y mide los cambios que se producen en el campo de la cultura de salud
Subject(s)
Family Practice , Public Health , PeruABSTRACT
La presente publicación describe una visión de las expresiones de la medicina tradicional de acrácter social y mide los cambios que se producen en el campo de la cultura de salud
Subject(s)
Family Practice , Public Health , PeruABSTRACT
Proyecto de investigación sobre medicina tradicional realizada en las comunidades de Saylla y Huasas (Cuzco) con la finalidad de modificar la dependencia en materia de salud promoviendo la utilización de los recursos propios y difundir las tecnologías médicas andinas
Subject(s)
Medicine , Medicine, Traditional , Plants, Medicinal , Community Health Services , PeruABSTRACT
El presente volumen recoge el producto de 115 entrevistas a personas integrantes de 23 familias de las comunidades de Saylla y Huasao (Cusco), se pudo conocer y controlar los efectos del cambio en los hábitos, usos y costumbres en salud en estas dos comunidades; una con fuerte influencia de la medicina formal y la otra de la medicina tradicional
Subject(s)
Attitude/ethnology , Medicine , Medicine, Traditional , Population Characteristics , PeruABSTRACT
Se hizo un estudio sobre la existencia y trayectoria de la arteria dorsal del pie, mediante palpacion clinica del dorso del pie en 2000 pacientes, y por diseccion anatomica bilateral en 40 cadaveres. La palpacion fue negativa solo en 3.6% de los pacientes examinados y en el estudio anatomico 3.48%. Se encontro reemplazo de la arteria por una arteria perforante de la peronea o fibular en 1.6% de los casos y un trayecto aberrante en 0.8%. La presencia de la arteria dorsal del pie tiene importancia clinica, en el sentido de valorar facilmente la circulacion distal del miembro inferior. Debido a la rareza de su falta en el medio nacional (3.6%), dato que va en contra de estadisticas tradicionales, la cateterizacion de esta arteria ofrece un via facil, elegante y segura cuando se requiera dicho procedimiento. Ademas su acceso es mucho mas sencillo y rapido que el de la arteria tibial posterior
