正常椎间盘胶原的研究

腰椎间盘突出症是引起腰腿痛常见的原因。胶原作为椎间盘结构的主要成分,构成椎间盘的纤维框架,其类型与分布直接决定着椎间盘结构的强度和功能的稳定。本文利用溴化氰消化椎间盘胶原产生多肽,借助于梯度层析。SDS-PAGE及光密度定量扫描等对正常人椎间盘胶原进行了研究。结果表明:正常人椎间盘含Ⅰ型及Ⅱ型两种胶原,它们的分布呈明显而特征性的移行性变化:纤维环外层边缘以Ⅰ型胶原为主(83％),由外向内Ⅰ型胶原逐渐移行为Ⅱ型胶原,靠近髓核处以Ⅱ型胶原为主(72％);髓核中心含有Ⅱ型胶原。此为椎间盘的一个结构特性,以满足椎间盘的特殊功能的需要。     

Participation of the adhesive disc during karyokinesis in Giardia lamblia

Evidence is presented for a potential involvement of the adhesive disc on the nucleus division in Giardia lamblia. The trophozoite mitotic nucleus was studied by transmission electron microscopy, freeze-fracture, freeze-substitution and also by immunofluorescence microscopy using anti-tubulin antibodies specific to spindle microtubules and Panotic staining. Prior to cell division the nucleus elongated and a displaced disc fragment, established contact with the nucleus. A progressive nucleus indentation was coincident with the concomitant presence of a disc fragment at the constricted region. One nucleus each time progressively divided until the karyokinesis was finished and two daughter-nuclei were observed. After the first karyokinesis a second karyokinesis takes place following the same procedure. When Giardia gets the four nuclei, cytokinesis occurs. Duplicated basal bodies were seen in between the first and the second karyokinesis. Immunofluorescence microscopy, using a panel of anti-tubulin antibodies, and electron microscopy of cells processed using microtubule stabilizer buffers, or cells fast-frozen and freeze-substituted, did not reveal the presence of a typical spindle. We propose that Giardia lamblia presents an uncommon mitotic behavior where the adhesive disc, a microtubular structure, seems to participate in the karyokinesis process.     

Correlation between MRI evidence of degenerative condylar surface changes, induction of articular disc displacement and pathological joint sounds in the temporomandibular joint

Objectives: The relationship of bony changes in the condylar surfaces in articular disc displacement without reduction in temporomandibular joint (TMJ) was investigated using diagnostic imaging. The study also evaluated whether the bony changes in the condylar surfaces limit disc and condyle motion, and produce pathological joint sounds. Materials and methods: Thirty‐seven joints in 28 patients diagnosed with degenerative bony changes in the condylar surfaces radiographically and anterior disc displacement without reduction using magnetic resonance imaging (MRI) were studied. The bony changes were assessed by radiographic examination and classified into two types: pathological bone changes (PBCs) including erosion, osteophyte formation and deformity, and adaptive bone changes (ABCs) including flattening and concavity. MRI was performed on the TMJ to examine the configuration and position of the discs. Joint sounds in the TMJ were determined using electrovibratograghy with a joint vibration analysis. Results: The articular disc motion to the condyle in the PBC group was smaller than in the ABC group irrespective of the configuration of the disc, even though there were no significant differences between the two types of bony changes in the disc position during jaw closing. The joint vibration analysis of the TMJ showed that joint sounds with a higher frequency were observed in the PBC group than in the ABC group. High energy levels needed to produce the higher frequencies (over 300 Hz) were observed only in the PBC group.     

Changes in the interfacial shear resistance of disc annulus fibrosus from genipin crosslinking

Crosslinking soft tissue has become more common in tissue engineering applications, and recent studies have demonstrated that soft tissue mechanical behavior can be directly altered through crosslinking. Using a recently reported test method that shears adjacent disc lamella, the effect of genipin crosslinking on interlamellar shear resistance was studied using in vitro bovine disc annulus.     

Use of the disc fermenter to examine production of citric acid by Aspergillus niger

Summary The effects of mode of growth on citric acid production by A. niger were studied using surface culture, shake culture and the disc fermenter. The disc fermenter enabled the effects of medium exchange with biomass retention to be examined.     

Pattern Regulation of the Leg Disc of the Fleshfly, Sarcophaga peregeina

A fate map of the hind leg disc of Sarcophaga peregrina was constructed by examining the adult structures of implanted disc fragments. The locations of presumptive adult structures in the disc were similar to those of fore leg disc of Drosophila and Sarcophaga ruficornis . However, the concentric borderlines of the segments could not be ascertained in the present case.
Pattern regulation of disc fragments was studied by culturing them either in adult females for several days or for 3 days in mature larvae placed on wet condition. Cultured disc fragments regenerated or duplicated as in Drosophila , with some exceptions. For instance, the region with a high density of positional values, the upper medial quarter, of the fore leg disc of Drosophila was not found. A characteristic difference in the rate of regeneration or duplication was observed in the implanted fragments, when cultured in larvae or adult hosts. This variable pattern regulation in larval and adult hosts could be due to different compositions of the hemolymph in which would healing of the implanted disc fragments takes place.     

The elongation of the human cervical vertebral column

Cervical vertebral elongation has been studied using serial cephalometric radiographs of 32 children examined regularly from 0.25 to 17 years. Mean vertebral body heights increased rapidly to about 2.5 years and then decelerated except for a spurt at about the age of peak height velocity. There were only small sex differences in vertebral body elongation to 12 years. From then to 15 years, the vertebral body heights in the girls exceeded those in the boys; later this sex difference was reversed. There was no pubertal spurt in disc elongation. The correlation coefficients were negative between vertebral body heights and the heights of adjoining intervertebral discs, e.g., body C3 and disc C3–4, but those between body heights or between disc heights were positive. The heights of adjacent cervical vertebral bodies were correlated more highly than the heights of non-adjacent bodies. There was a similar pattern of differences between correlation coefficients for the heights of adjacent and non-adjacent intervertebral discs.     

Development of a method for the quantitative assessment of the microorganism sensitivity to antibiotics by using discs. A study of the patterns of doxycycline diffusion from discs into the nutrient agar]

Doxycycline concentrations provided by the antibiotic diffusion from paper discs into sterile agarized medium and the medium plated with staphylococci and Coli bacteria were studied at different distances from the disc center. Infection of the nutrient medium with the test cultures had no effect on the antibiotic diffusion rate. A linear relation between the logarithm of the antibiotic concentration in the agar and distance from the disc centre were found. Probably it is possible to determine the MIC of the antibiotic with respect to various microorganisms by the value of the radius of the growth inhibition zone around the disc using diagrams expressing such a relation.     

Comparison of disc diffusion and epsilometer (E-test) testing techniques to determine antimicrobial susceptibiliy of Campylobacter isolates of food and human clinical origin

The antibiotic resistance profiles of 75 Campylobacter isolates of food and human clinical origin was determined by two agar diffusion susceptibility methods; disc diffusion and epsilometer-test (E-test). The most common therapeutic antimicrobials, erythromycin, ciprofloxacin and tetracycline were studied, along with chloramphenicol, ampicillin and naladixic acid. The resistance observed for each antimicrobial, as determined by both of methods, were statistically compared using Fisher two-tailed analysis.Of the six antimicrobials studied only two were shown to have statistically different patterns when resistance was compared by disc diffusion and E-test. The percentage of isolates resistant to clinically relevant antimicrobials using both techniques ranged from 6.6 to 21.3% for erythromycin, 25.3–26.6% for tetracycline and 33.3–36.0% for ciprofloxacin. The prevalence of multi-drug resistant (MDR) campylobacters (isolates resistant to 2 or more antimicrobials) for both disc diffusion and E-test was 44%. It can be concluded that, for four of the six antimicrobials assessed, antimicrobial resistance prevalences could be equally determined by either of the methods studied.     

The effect of collagen reinforcement in the behaviour of the temporomandibular joint disc

In this paper, the influence of collagen fibres in the behaviour of the temporomandibular joint disc is studied. A three-dimensional finite element model of the joint is developed from a set of medical images. The model comprises the mandible, part of the cranium and both temporomandibular joints. Joints have been considered to be composed of the articular discs and the temporomandibular ligaments. A fibre-reinforced porohyperelastic model was used to study the response under clenching of the fibrocartilage that composes the articular disc. This was divided in an intermediate zone, and two bands, an anterior and other posterior, in order to define the orientation of collagen fibres. The study demonstrates that the introduction of collagen fibres in the biphasic behaviour of the articular disc implies for a prescribed displacement not only an increase of the pressurization in the tissue, but also higher stresses in the anterior and posterior bands, as well as in the lateral zone of the disc. Thus, modelling the disc as an isotropic solid matrix leads in this case to an overestimation of the stresses in the intermediate zone, an underestimation of the pore pressure in this area, and an underestimation of the stresses in the rest of the disc.     

Immunocytochemical localization of epidermal growth factor receptor in early embryos of the Japanese medaka fish (Oryzias latipes)

This study was undertaken to localize epidermal growth factor receptor (EGFR) during early development of Japanese medaka embryos using immunocytochemistry. Specific staining was observed in all stages studied. All of the cells of the embryonic disc from the germinal disc (1 cell) through the late high blastula stages stained moderately for EGFR. Beginning with the flat blastula stage, the surface and lateral cells of the embryonic disc and the cells migrating around the yolk stained intensely for EGFR, and this continued throughout the study period. The presence of the keel at the late gastrula stage did not affect the moderate staining of the majority of the embryonic disc cells. When somites first appeared, the keel region stained less intensely than before, but scattered individual cells stained intensely for EGFR. Embryos with 12 somites had a neural tube that was lightly stained except for a few intensely stained individual cells. The neural tube, notochord and somites in 24-somite embryos lacked immunostaining. However, the surface epithelium, aorta, intestinal epithelium and pronephric duct demonstrated EGFR immunostaining. This study demonstrates that EGFR is present during medaka development and supports the hypothesis that EGFR ligands are important during cleavage, gastrulation and early organogenesis.     

A fluorescence histochemical study of the monoamine-containing cell in the developing frog taste organ

Summary Sequential changes in the monoamine-contianing cell (MC cell) of the developing frog tongue has been studied by fluorescence histochemistry using uptake of 5,6-dihydroxytryptamine. At st. 16, a few yellow fluorescent cells, here called MC cells, appeared in random order at the uppermost layer of the dorsal epithelium. They were round or elliptical in shape. At st. 18 the MC cells, greatly transformed, were found at the periphery of the sensory disc primordium which first appears during this stage. The MC cell was made up of three parts: perikaryon, process and terminal portion. The perikaryon was located at the upper half of the epithelium and from it a single process stretched vertically toward the basal lamina, above which the dilated terminal portion was found. Thereafter the perikaryon gradually moved toward the basal layer while remaining at the periphery of the disc primordium. Meanwhile the terminal portion moved over the basal lamina toward the center of the disc primordium. At st. 22, the whole of the MC cell lay flat above the basal lamina. The perikaryon was localized at the periphery of the sensory disc and from there the process stretched toward the center. Thus, the morphology of MC cells resembled the adult state, except for smaller size. MC cells were never observed in the subepithelial connective tissue in the present study. This seems to suggest that the MC cell of the frog fungiform papilla is of epithelial origin.     

Occurrence of beta-lactamase type ESBL and IBL in Pseudomonas aeruginosa rods]

The aim of the study was to determine extended spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) among Pseudomonas aeruginosa strains. A total of 43 strains isolated from humans (6), hospital sink (1), fish (15), cattle (5), swine (5), dog (1), redder (1) fur animals (9) were studied. ESBL-producing strains were detected with double disc diffusion test according to Jarlier et al. (8). Clavulonate and tazobactam were used as the inhibitors of ESBL. Inducible beta-lactamases were determined using double disc method according to Sanders (15). Cefoxitin was the inductor of these beta-lactamases. The susceptibility study was carried out using the disc diffusion method according to NCCLS standards. A total of 8 ESBL (18.6% of all strains) and 31 (72%) IBL producing strains were detected. The obtained results indicate the necessity of monitoring of ESBL- and IBL-producing strains of Pseudomonas aeruginosa.     

Modeling The Nutrientsbehavior in Intervertebral Discs: A Boundary Integral Simulation

It is a well-known fact that computational biomechanics and mechanobiology have deserved great attention by the numerical-methods community. Many efforts and works can be found in technical literature. This work deals with the modeling of nutrients and their effects on the behavior of intervertebral discs. The numerical modeling was carried out using the Boundary ELement Method (BEM) and an axisymmetric model of the disc. Concentration and production of lactate and oxygen are modeled with the BEM. Results agree well enough with those obtained using finite elements. The numerical efforts in the domain and boundary discretizations are minimized using the BEM. Also, the effect of the calcification of the disc that causes the vascularization loss has been studied. The glucose, oxygen and lactate components behavior has been analyzed applying a mixed loading-unloading process, then allowing the study of the disc-height variations due to the degradation of the disc.     

Nonlinear numerical analysis of the structural response of the intervertebral disc to impact loading

The analysis of intervertebral disc dynamics under impact loading, using computational simulation, is scarcely reported. In this study, the contribution of the characteristic structure of the disc to its dynamic response has been evaluated. The influence of several model features on the dynamic response was investigated. A hyperelastic large deformation formulation was used to describe the nonlinear behaviour of the soft tissues. The material parameters were determined by the fitting of experimental data from the literature. The model demonstrated pressure wave propagation and reflection through the disc, with a periodic oscillation of the system in response to a single impulse load, and highlighted a potential primary role played by the collagen fibre reinforcement. Their tensioning contributes to changing the stress propagation and oscillation, with a faster reduction in the internal pressure peak. The natural frequency of the disc was predicted to be approximately 9.8 Hz for the vertical oscillation.     

Significance of the hippocampus in the development of convulsive syndrome by kindling electrostimulation of the caudate nucleus

The experiments were conducted on rabbits with chronically implanted electrodes (neocortex, hippocampus, amygdala, caudate nucleus). The rostral part of caudate nucleus was stimulated once daily at a current maximum 300 microA, using 60 Hz square pulses (1.0 Ms) 12.0 s in duration. The severity of motor seizures and its developmental velocity were studied in the relation with a degree of the epileptization of the brain. The data obtained illustrated the next relation: the more stable and intensive were discharges in hippocampus the harder motor seizures.     

Quantitative analysis and comparative regional investigation of the extracellular matrix of the porcine temporomandibular joint disc.

Characterization of the extracellular matrix of the temporomandibular joint (TMJ) disc is crucial to advancing efforts in tissue engineering the disc. However, the current literature is incomplete and often contradictory in its attempts to describe the nature of the TMJ disc matrix. The aim of this study was to identify the variation of key matrix components along the three axes of the porcine disc using ELISAs to quantify these matrix components, immunohistochemistry to identify their regional distribution, and SEM to examine collagen fiber diameter and orientation. The overall GAG content of the TMJ disc (including the dermatan sulfate proteoglycans) was 5.3+/-1.2% of the dry weight. Chondroitin sulfate, which comprised 74% of this total GAG content, was 4.4, 8.2, and 164 times more abundant than dermatan sulfate proteoglycan, keratan sulfate, and hyaluronic acid, respectively. In general, these GAGs were most concentrated in the intermediate zone of the TMJ disc, appearing in dense clusters, and least concentrated in the posterior band. Additionally, chondroitin sulfate was more abundant medially than laterally. Collagen II was discovered in trace amounts, with higher relative amounts in the intermediate zone. Collagen fibers were observed to run primarily in a ring-like fashion around the periphery of the disc and anteroposteriorly through the intermediate zone, with a mean fiber diameter of 18+/-9 mum. Characterization studies of the TMJ disc, including prior biomechanical and cell studies along with the current study of the extracellular matrix, collectively reveal a distinct character of the intermediate zone of the disc compared to its anterior and posterior bands.     

Effect of Extracts of the Bark of Saccoglottis gabonensis on the Microflora of Palm Wine

Direct addition of the bark of Saccoglottis gabonensis to fresh palm juice followed by microbial enumeration every 6 hr showed that the bark inhibited bacterial growth appreciably. The effect of various extracts of the bark on two bacterial species isolated from palm wine (Leuconostoc mesenteroides and Lactobacillus plantarum) was studied by using the paper disc method. It was observed that three of the five constituents of the bark showed inhibitory effect on the bacteria.     

Expression of lumican related to CD34 and VEGF in the articular disc of the human temporomandibular joint

Lumican belongs to the small leucine-rich repeat proteoglycan (SLRP) gene family and has been reported to exist in the cornea, intervertebral disc and tendon. Lumican plays a significant role in the assembly and regulation of collagen fibres. The human temporomandibular joint (TMJ) disc is made up of fibrocartilage with an extracellular matrix (ECM) composed of collagen and proteoglycans. The existence and behaviour of lumican have not been studied in the human TMJ disc. Therefore, we used immunohistochemical methods to detect lumican, CD34 and vascular endothelial growth factor (VEGF) and histochemical staining with toluidine blue in 13 human TMJ specimens (10 surgically removed and 3 obtained from autopsy). In both normal and deformed discs we observed staining with toluidine blue. We found that the area of metachromasia inside the deformed disc was uneven and expression of lumican was strong in the areas negative for metachromasia. Staining of VEGF and CD34 inside the deformed disc was seen. We confirmed the expression of lumican in the human TMJ disc and showed that a large number of fibroblast-like cells existed in the area of strong lumican expression. These new findings about the behaviour of lumican suggest that it may play a key role in the generation of a new collagen network by fibroblast-like cells.Key words: TMJ disc, lumican, CD34, VEGF, immunohistochemistry, metachromasia.     

Simulation of Electrical Interaction of Cardiac Cells

A model of the electrical activity of excitable membrane was used to simulate action potential propagation in cardiac cells. Using an implicit method for solving finite difference equations, propagation through the intercalated disc region between two abutting cells was studied. A model of interaction was constructed and parameters of the cellular junction determined. Estimates of the intercalated disc resistance were then made from these junction parameters using a field analysis of the junction. Values of approximately 4 Ω-cm² were found and correlate well with experimentally measured values.