Connective Tissue Ground Substance Research Articles

PATHOANATOMICAL FEATURES OF SHAR-PEI’S HEREDITARY AMYLOIDOSIS

Shar-Pei's amyloidosis is a hereditary disease which is characterized by amyloid deposition in stromal-vascular organs. The contributory cause of amyloidosis is а genetic disease Shar Pei's fever. Amyloidosis is based on enzymatic processes abnormality in connective-tissue cells. Enzymopathy is followed by synthesis of the abnormal fibrous protein entering strong compound with proteins of plasma and glycosaminoglycans of ground substance of connective tissue.

Comparative analysis of gingival crevicular fluid β-glucuronidase levels in health, chronic gingivitis and chronic periodontitis.

Background and Objectives:Current methods available for periodontal disease diagnosis are seriously deficient in terms of accuracy, in the ability to predict ongoing or future disease activity and indeed in determining whether previously diseased sites are in an arrested phase or still active. One area that is receiving a great deal of attention is the biochemical investigation of gingival crevicular fluid (GCF). β-glucuronidase (βG) is one of the enzymes found in GCF that is involved in degradation of the ground substance and fibrillar components of host connective tissue. GCF βG activity might be a good indicator or predictor of periodontal disease activity. This study was conducted to estimate and compare the GCF βG levels in patients with healthy periodontium, chronic gingivitis, and chronic periodontitis.Methodology:Subjects were classified into three groups of 20 patients each; healthy individuals, chronic gingivitis, and chronic periodontitis. After recording the plaque index, gingival index and probing pocket depth, 1 μL GCF was collected by placing a calibrated microcapillary pipette extracrevicularly and transferred to sterile plastic vials containing 350 μL of normal saline with 1% bovine serum albumin. Analysis of βG was done by spectrophotometryResults:βG levels in GCF were significantly higher in chronic periodontitis group (mean value - 2.04743), followed by chronic gingivitis group (mean - 1.11510) and healthy group (0.53643).Conclusion:Increased βG levels were observed in patients with increased periodontal destruction, hence GCF βG levels can be used as biochemical marker for periodontal disease activity.

ACCUMULATION OF CONNECTIVE TISSUE GROUND SUBSTANCE IN THE HEART OF THE DWARF MOUSE.

ACCUMULATION OF CONNECTIVE TISSUE GROUND SUBSTANCE IN THE HEART OF THE DWARF MOUSE.

Scleromyxoedema (lichen myxoedematosus) associated with a paraprotein, IgG 1 of type kappa.

SUMMARY A case of scleromyxoedema (lichen myxoedematosus) associated with an IgG1, type kappa paraproteinaemia is reported. Culture of bone marrow material demonstrated that this tissue synthesized monoclonal IgG (subclass γ I) with a light chain of type kappa. The culture of a biopsy specimen of the diseased skin demonstrated low-speed IgG synthesis. It is concluded that the bone marrow and pathological skin synthesize a paraprotein with the same characteristics as that in the serum. Microchemical analysis of pathological skin demonstrated an increase in the content of neutral glycoproteins, hyaluronic acid and chondroitin sulphates as well as the presence of heparin and keratan sulphates. It could not be demonstrated that the serum of this patient contained antibody against connective tissue ground substance or other skin constituents.

Pharmacology Review

Extravasation or inadvertent infiltration of fluids into subcutaneous tissue from peripheral intravenous (IV) devices is a common adverse event in newborns. (1)(2) Although fluids occasionally extravasate from central venous lines, the complication is much more common from peripheral catheters, which are used widely in sick neonates. (3) This article focuses on extravasations from peripheral IV devices. Injury to the skin, even in a very immature neonate, results in an inflammatory response and heals by scar formation. (4) Although the scars from many of these injuries appear to improve with time, (5) tissue necrosis from extravasation injury could result in partial or complete skin loss, infection, and nerve and tendon damage, with the potential risk of permanent cosmetic and functional impairment. (6)(7)(8) The incidence of extravasations from Teflon® catheters has been reported to vary from 23% to 63%. (1) A recent survey of regional neonatal intensive care units in the United Kingdom recorded the prevalence of extravasation injury resulting in skin necrosis as 38 per 1,000 neonates, with 70% of these injuries occurring in infants of 26 weeks’ gestation or less. (2) The incidence of extravasation is related closely to the type of device used, insertion site, duration of therapy, infusate administered, patient activity, and gestational age. (1)(3)(9)(10) The fragility of the skin, particularly in the first 2 weeks after birth, and the lack of subcutaneous tissue in preterm neonates makes them uniquely susceptible to injury and skin loss. Extravasation may occur from the tip of the cannula or needle piercing the vessel wall. Alternatively, distal obstruction of the vein due to thrombosis or venoconstriction from irritation of the vessel wall may lead to increased back pressure and leakage from the entry point of the needle or …

An electrochemical illustration of the mathematical modelling of chlorine impact and acidification in electrochemical tumour treatment and its application on an agar–agar gel system

Electrochemical treatment (EchT) of tumours, which is also known as electrochemical therapy, implies that tumour tissue is treated with a direct current through the use of electrodes. This was shown earlier when the major electrochemical reactions at the anode were chlorine and oxygen evolution and the impact of chlorine in EchT was investigated by mathematical modelling. In this work the electrochemical illustration of a mathematical model of electrochemical treatment of tumours is presented using a Pt electrode placed in agar–agar gel containing 1% phenolphthalein and NaCl. The appearance of the bright red color around the Pt wire due to the pH change caused by hydrogen evolution is used for the simulation of the shape and the boundaries of the tumour tissue. After the polarity was changed, oxygen evolution takes place on the Pt wire electrode and the red color disappears simulating the destructive influence of the electrochemical process on tumour tissue. A better understanding of the mathematical model presented earlier is achieved and it can be used for educational purposes in oncological praxis because agar–agar is the closest approximation to the ground substance of connective human tissue and successfully simulates the processes which occur in vivo. There is also some quantitative agreement between the results calculated earlier and those obtained in this work. The previous model was based on tissue, which was treated as an aqueous solution of sodium chloride containing bicarbonate and proteins and the results presented a detailed and useful quantitative analysis of the system. In this work, the quantitative analysis presented is based on an agar–agar system and the experimental conditions give the possibility of more detailed qualitative analysis. The combination of these two models offers a better simulation of the processes in vivo.

Neuromuscular Techniques in Orthopedics

Summary Alternative medicine offers approaches to health care that are complementary to or integrative with conventional medicine. Many possibilities have emerged from this background, of methods that can enhance medical care, such as acupuncture and lymphatic drainage techniques. Another method is neuromuscular therapy, which considers both local or global causes of pain and dysfunction, as well as perpetuating factors that impact the musculoskeletal system. Neuromuscular therapy aims specifically to alter the ground substance of connective tissue, to deactivate myofascial trigger points and reduce ischemia, and to assess and beneficially modify postural alignment, musculoskeletal dysfunction, and neural entrapment. Additionally, nutritional awareness and emotional distress are considered in an effort to alter chronic pain and myofascial dysfunction.

The morphology of autotomy structures in the sea cucumber Eupentacta quinquesemita before and during evisceration.

Evisceration in the dendrochirotid sea cucumber Eupentacta quinquesemita is a whole-body response involving a predictable series of events including muscle contraction and failure of three autotomy structures: (i) the introvert, the dexterous anterior extensible portion of the body wall, (ii) the tendon linking the pharyngeal retractor muscle to the longitudinal body wall muscle and (iii) the intestine-cloacal junction. The autotomy structures are histologically complex, consisting of muscle, nervous and connective tissue. Autotomy resulted from complete loss in the tensility of the connective tissue ground substance. Separation of the autotomy structures was facilitated by muscle contraction. The cell and tissue changes involved with autotomy were documented by microscopic examination of autotomising tissue. Change in the autotomy structures appears to initiate from the peritoneal side with delamination of the peritoneum followed by a wave of disruption as the connective tissue is infiltrated by coelomic fluid. Evisceration and autotomy in E. quinquesemita are neurally controlled, so particular attention was paid to the fate of neuronal elements. Neurosecretory-like processes containing large dense vesicles and axons were present in the connective tissue layers of the autotomy structures in association with extracellular matrix, muscles and neurons. These neuronal elements remained largely intact during autotomy and did not appear to be a source of factors that effect connective tissue change. They may, however, be involved in muscle activity. Holothuroid autotomy structures are completely or partially bathed in coelomic fluid, so there is potential for hormonal or neurosecretory activity using the coelomic fluid as a conduit. Connective tissue change during evisceration appears to be effected or mediated by an evisceration factor present in coelomic fluid that has a direct transmitter-like or neurosecretory-like mode of operation. The final outcome, expulsion of the viscera, is likely to result from a suite of factors that interact in a manner yet to be determined.

Ultrastructure, Distribution, and Density of Lamellar Bodies in Human Peritoneum

To determine the ultrastructure, relative density, and location of lamellar bodies in the various regions, structures, cells, and intercellular matrix in normal human peritoneum; to carry out engineering analysis of the role of lamellar structures in serosal lubricancy and deduce what effect this system may have on the process of peritoneal dialysis. Five samples of normal human parietal peritoneum obtained at elective operation were fixed in a tannic acid-glutaraldehyde mixture and submitted to examination by transmission electron microscopy. Detailed analysis using reconstruction of serial electron micrographs and tracings of montages were employed in determining location, and disposition, density, and geometric patterns of lamellar bodies in all levels of the peritoneal membrane. Lamellar profiles were found in greatest density enmeshed in surface microvilli and in mesothelial cytoplasm. Lamellar bodies were frequently observed capping the external portion of mesothelial junctional complexes, and within intercellular junctions. Lamellar bodies were also encountered in macrophages, both in the peritoneal cavity and submesothelial tissue, and also in fibroblasts. Lamellar bodies were present in low density in the matrix ground substance of submesothelial connective tissue, in blood vessel walls between smooth muscle, in endothelial cell cytoplasm, and in vascular lumina. Three-dimensional analysis of lamellae on mesothelial surfaces indicates that an arrangement of constantly changing microscopic spheres and cylinders would act at "ball and roller bearings" among the microvilli for the lubrication of opposing surfaces. The entrapment of fluid in lamellar bubbles, which in normal peritoneum fill the microvillous layer, would, if maintained in peritoneal dialysis, constitute a stagnant layer of considerable stability and inertia.

An extracellular enzyme with hyaluronidase and chondroitinase activities from some oral anaerobic spirochaetes.

Treponema denticola, Treponema vincentii and Treponema socranskii produce an enzyme that hydrolyses hyaluronic acid (HA) and chondroitin sulphate (CS). The secreted enzyme is specifically inhibited by gold sodium thiomalate and anti-bee-venom antibodies. The use of saturated substrate (HA or CS) transblots allowed the visualization of active enzyme directly from culture supernatants and is a useful tool in clarification of complex polysaccharide-degrading enzyme specificities. The affinity-purified extracellular enzyme of T. denticola contains a single molecular species with a molecular mass of 59 kDa. Since it hydrolyses both HA and CS, it can more appropriately be termed a hyaluronoglucosaminidase (HGase). The HGase has been localized at the cell surface by electron microscopy and may play an active role in the degradation of connective tissue ground substance in the initiation and progression of periodontal disease.

Histochemical localization of oxidized glutathione-catalysing enzymes in human term placenta

This paper reports a new cytochemical affinity technique for detecting oxidized-glutathione-binding enzymes by light microscopy and for carrying out fine structural analyses by means of oxidized glutathione labelled with colloidal gold. Albumin-colloidal gold particles were prepared. Oxidized glutathione, added to the solution, replaced albumin on the surface of the gold, thus forming a new histochemical reagent. In human placenta, oxidized-glutathione-catalysing activity was detected on the brush border of the placental villi, over the foldings of endothelial membranes, and in the ground substance of connective tissue in the villous core. Every process of the cell membrane demonstrated enhanced oxidized-glutathione-catalysing activity. This new reagent is a unique example of a low-molecular-weight compound labelled with colloidal gold, and it permits direct measurement of oxidized-glutathione-binding activity in tissue sections.

Hyaluronic acid metabolism and its clinical significance in patients treated by continuous ambulatory peritoneal dialysis

Musculoskeletal syndromes are common in patients treated by dialysis for end-stage renal failure and abnormal connective tissue metabolism has been implicated. Hyaluronic acid is a major component of connective tissue ground substance. Serum, dialysate, and 24-h urine hyaluronic acid was therefore measured in 43 patients treated by CAPD to determine hyaluronic acid metabolism and to relate these variables to morbidity and mortality over an 18-month period. Serum hyaluronic acid was elevated in 71% patients, being correlated with patient age, length of time on dialysis, and weight loss over the preceding 6 months. Small quantities of predominantly low-molecular-weight hyaluronic acid were lost in the urine, whereas much larger amounts of mixed-molecular-weight hyaluronic acid were excreted in peritoneal dialysate. Dialysate hyaluronic acid exceeded serum hyaluronic acid. Baseline serum hyaluronic acid was closely correlated with morbidity and mortality over the following 18 months. Serum hyaluronic acid is an accurate predictor of mortality and morbidity over an 18-month period in patients treated by CAPD. Large quantities of hyaluronic acid are excreted in peritoneal dialysate, which in part represents local hyaluronic acid production.

Cervical distensibility changes in pregnancy, term, and preterm labor.

T he changes in the mechanical behavior of cervical tissue observed at the end of gestation are in fact due to histological and biochemical changes. The inner circumference of the cervix in the nongravid state, having a well-ordered collagenous network, cannot reach the inner circumference value necessary to accommodate the fetal head. Therefore, the rearrangement of the collagen fibers into a more disorganized, looser network during pregnancy seems to be an important process in cervical ripening. In support of this idea are the numerous histological studies showing disorganization of collagen fibers at term in animals1,2 and women. 3,4 Biochemical changes of the cervical connective tissue ground substance components (eg, proteoglycans, glycoproteins, water content) are likely to play a major role in the disorganization of the collagen network. 5~6 In the human cervix the fibroelasticity seems to dominate, since the test results observed by several authors’.8 with the uterine cervix closely resemble the results for fibroelastic materials. In obstetrics, it is of great interest to quantitatively determine mechanical characteristics of the uterine cervix, particularly the ease with which the cervix can be dilated, in order to describe and understand the mechanism of human cervical dilatation. Digital internal pelvic examination is a fast and simple technique to study physical characteristics of the uterine cervix, and a number of clinical systems of quantifying cervical ripeness have been devised, most notable that of Bishop.g With these systems, a low score indicates an unripe cervix, and a high score, a ripe cervix. Unfortunately, these methods are subjective, and amalgamate several parameters, eg, that cervical consistency is presumably related to biochemical connective tissue changes whereas the other elements of the pelvic score (dilation, effacement, position, level of the presentation) are most likely related to myometrial activity. This is why several instruments were designed to quantitate the mechanical properties of the uterine cervix related to connective tissue changes (measurement of cervical compliance and cervical consistency),7,810 but the clinical application of such methods is very difficult and/or not accurate enough. We have developed a cervicotonometer to measure a distensibility index in the clinical practice that is based on the values of the force used to open the cervix and the inner diameter of the cervix obtained with each force used. The use of this instrument and results obtained with it will be described in this article.

Scanning electron microscope observations on collagen fibers in human dentin and pulp

Human permanent teeth were examined in the scanning electron microscope after demineralization and exposure to preparative procedures based on hydrogen peroxide, trypsin, and EDTA. These substances removed the inorganic material, the cellular structures, the homogeneous connective tissue ground substance, and interfibrillar matrix. The remaining tissue components comprised a network of distinct collagen fibers whose organization was related to the type of tissue in which these were incorporated. A similar or identical method has not been developed or applied to teeth previously. Dentin and predentin comprised a compact mass of fibers which basically were parallel to the continuously growing interior surface of the predentin, or arranged at an acute angle to this plane. Collagen fibers in the pulp were numerous, but lacked any particular orientation in most areas. Interodontoblastic fibers crossed the odontoblastic zone at a right angle to the pulp chamber wall and mingled with collagen fibers in predentin. When previously published findings of ours are taken into account, it is possible to conclude that other factors than the organization of the collagen fibers are responsible for the stainability of these fibers in predentin and in interglobular dentin with silver methenamine, and that aldehyde groups on collagen fibers in predentin may be actively and directly involved in the mineralization of the dentin.

Cytochemical visualization of anions in collagenous and elastic fiber-associated connective tissue matrix in neonatal and adult rat lungs using iron-containing stains

The cytochemical reactivity of pulmonary connective tissue matrix component in neonatal and adult rat was evaluated using high iron diamine (HID) to detect sulfate ester end groups and dialyzed iron (DI) to detect sulfated and carboxylated end groups of complex carbohydrates, including glycoproteins and glycosaminoglycans at the ultrastructural level. The HID reaction product, in the form of discrete 5-12 nm silver particles following appropriate intensification with thiocarbohydrazide-silver proteinate, was found associated with cell surfaces, the elastin component of elastic fibers, and at regular intervals along the length of collagen fibers in large airways and deep lung interstitium. Staining was similar in adult and neonatal rats, except in areas where connective tissues were presumably still rapidly developing in the neonatal animals. Here large gaps or spaces containing filamentous structures were observed between collagen and elastic fibers. The distribution of DI-reactive sites was similar to that seen with HID with the exception of elastic fibers in which only the microfibrillar portion stained. The collagen-associated reaction was not regularly disposed like that stained with HID, but rather it formed a tight continuous density around the fiber. These results indicated the presence and location of glycoproteins and glycosaminoglycans in connective tissue ground substance regions prior to the full development of elastic and collagenous elements in neonatal pulmonary airways and parenchyma. They also demonstrate cytochemically the presence of a sulfate ester-containing complex sugar found associated with the elastin component of elastic fibers in the lung.

Uptake of 67Ga in the lung of mice during bleomycin treatment.

Changes of 67Ga uptake in the lungs and changes of components of the so-called ground substance of the lung connective tissues of mice were followed for 7 weeks after the start of bleomycin (BLM) administration (20 mg/kg body weight IP, twice weekly for 5 weeks; this treatment induced fibrosis of the lung). 67Ga uptake of the lung was elevated at 1 week, and reached a maximum at 5 weeks (3.00 +/- 0.11% dose/g lung), and then decreased slightly at 7 weeks. The uronic acid content in the 1.2 M NaCl-soluble fraction, which contained predominantly heparan sulfate (HS), was increased at 1 week, peaked at 3 weeks, and then remained unchanged up to 7 weeks. This pattern was similar to that of 67Ga accumulation in the lungs. The uronic acid content of the 0.4 M NaCl-fraction, which contained predominantly hyaluronic acid (HA), was decreased at 1 week, but increased to a maximum at 3 weeks, then decreased to about the initial level at 5 weeks and decreased further at 7 weeks. Lung hydroxyproline content, an index of collagen content, was increased at 3 weeks and continued to increased rapidly thereafter, reaching approximately 1.5 times the control value at 7 weeks. Serum iron, measured as an indicator of iron metabolism, was slightly increased at 3 weeks and there was a corresponding decrease of unsaturated iron-binding capacity (UIBC). No corresponding change of 67Ga uptake was apparent. These results indicate that HS increased before the collagen accumulation at an early stage of pulmonary fibrosis of the lung during BLM treatment of mice, and support our earlier proposal that HS is a major acceptor for 67Ga accumulation.

Structure and properties of tendon collagen complex during disorganization of the ground substance of connective tissue

Structure and properties of tendon collagen complex during disorganization of the ground substance of connective tissue

Local immune system of nasal nucosa in inflammation. IgA distribution and secretory activity.

The local immune system of human mucosa (inferior turbinates and nasal polyps) from 53 humans with chronic inflammation was studied by examining the distribution and secretory activity of IgA using an immunofluorescent technique. Diffuse fluorescence for IgA was observed throughout the connective tissue ground substance in every nasal turbinate and nasal polyp. In mucosa with mild inflammatory changes, most IgA seemed to be produced locally by IgA-producing plasma cells. However, in severely inflamed nasal mucosa, including nasal polyps, plasma cells were relatively fewer in number and the distribution of IgA generally corresponded to the pattern of fibrin deposition. Secretory activity of IgA in the glandular cells of nasal turbinates was well retained. Only in dilated glandular ductules in nasal polyps was the secretory activity impaired.

Effect of mast cell chymase of rat skin on intercellular matrix: a histochemical study

The effects of purified mast cell chymase of rat skin on intercellular matrix were studied histochemically using cartilage as a model. The enzyme effectively releases proteoglycans from neonatal human costal cartilage. An analysis of the primary site of proteolytic attack by chymase in the matrix was attempted. The results suggest that components containing the more acidic radicals (e.g. sulphates) are hydrolyzed first. After a prolonged attack, the matrix loses most of its staining with Toluidine Blue and Alcian Blue, and staining of collagen is increased. The hydrolyzed regions of cartilage in Alcian Blue-stained sections exhibited a purple interference colour in polarized light. A similar interference colour appeared after incubating sections of rat skin with chymase. The mast cell chymase presumably promotes diffusion of plasma to the site of injury by the direct breakdown of connective tissue ground substance.

Mucopolysaccharidosis III A (Sanfilippo Disease Type A). Histochemical, Electron Microscopical and Biochemical Findings

This is a report of two brothers iwth mucopolysaccharidosis. The 8- and 10-year-old boys presented the characteristic clinical symptoms of the syndrome in their entirety. Both had a highly increased excretion of heparan sulfate in urine. The elder boy died and was autopsied and diagnosed as having Sanfilippo disease Typ A by a drastic reduction of heparan sulfate sulfamidase activity in organ extracts. Histochemically, highly water-soluble, sulfate acid mucopolysaccharides were demonstrated in liver, spleen, and cerebrum of the deceased child. Chemical analyses revealed a 12-fold increase of sulfated mucopolysaccharide in the patients liver and a 4.5-fold increase in the cerebrum when compared with normal controls. The sulfated polysaccharide consisted mainly of heparan sulfate, which was of low molecular size, heterogeneous in charge, and rich in its sulfamino hexose content. In addition, the gangliocytes of cerebrum and cerebellum had accumulated glycolipids. Electron microscopically the storage cells were overloaded with lysosomal residual bodies. The mitral valve was also involved in the storage process, which is a rare manifestation of the Sanfilippo syndrome. Acid mucopolysaccharides were deposited intracellularly as well as extracellularly in the mitral valve tissue. Polarisation microscopically there was found a change from normally positive to negative birefringence in the connective tissue ground substance of the mitral valve when containing stored mucopolysaccharides.