The role of cytokines TGF-beta1 and FGF-1 in the expression of characteristic markers of rat liver myofibroblasts cultured in three-dimensional collagen gel.

Rat liver myofibroblasts (MFB) are the key cells involved in the deposition of extracellular matrix in fibrotic liver. They were isolated by repeated passaging of non-parenchymal cell fraction and cultured in 3-dimensional (3D) collagen gel mimicking tissue. The transfer of MFB from plastic dishes to collagen resulted in the change in their shape from large and spread to slender with long extensions. The expression of transforming growth factor-beta1 (TGF-beta1) and of MFB markers, alpha-smooth muscle actin (alpha-SMA) and cellular fibronectin (EDA-FN), on protein level was significantly decreased in collagen gel. The gel did not change the expression of metalloproteinase MMP-2 but activated the proenzyme. The experiments with inhibitors of metabolic pathways showed that EDA-FN and alpha-SMA were differently regulated. The expression of EDA-FN required functional TGF-beta1 receptors and was also dependent on the activity of protein kinases MEK1 and MEK2. alpha-SMA expression was primarily determined by the 3D environment. Fibroblast growth factor-1 (FGF-1) in combination with heparin decreased the expression of alpha-SMA and increased the expression of EDA-FN in the cells on plastic. The cellular environment may influence the cells per se and may modify the action of other agents.

Histiocytic necrotizing lymphadenitis (Kikuchi-Fujimoto disease) with CNS involvement in a child.

We describe the case of a 9-year-old boy with encephalitis associated with histiocytic necrotizing lymphadenitis (HNL), also known as Kikuchi-Fujimoto disease. The child presented with unilateral cervical lymphadenopathy and fever that evolved to encephalitis in 3 weeks. Brain MRI showed bilateral temporal lobe hyperintense signal on T2 and FLAIR, hyperintense FLAIR signal in the periaqueductal gray matter, medial walls of the third ventricle, and mammillary bodies, multiple diffusion restriction foci in a central perivascular distribution and central perivascular enhancement. The perivascular distribution and nodularity of the diffusion restriction seen in this case has not been previously reported in HNL encephalitis.

Collagenolytic potential of rat liver myofibroblasts.

Rat liver myofibroblasts (MFB) were isolated by repeated passaging of nonparenchymal liver cell fraction. They were cultured on polystyrene Petri dishes, on fibrin or on type I collagen gels for 5 days. Quantitative RT-PCR, Western blotting, zymography and immunocytochemistry were used to study differences in cell morphology and protein expression. MFB were large and spread on plastic substrate, with prominent alpha-smooth muscle (alpha-SMA) fibres. They turned much smaller and elongated on collagen which was accompanied by the rearrangement of the cytoskeleton and a decrease in alpha-SMA and beta-actin content. Collagen gel induced the expression of a group of metalloproteinases (MMP-2, -3, -9, -13), on mRNA and protein level which resulted in the degradation of the gel. This response was accompanied by changes in the mRNA expression of cytokines of TGF-beta family, CTGF and interleukin-6, as well as of osteopontin and thrombospondin-2 that are involved in metalloproteinases (MMPs) regulation. The expression of MMPs substrates, collagen types I, IV and XII did not change or decreased. The effects of fibrin gels on MFB were milder than those of collagen. MFB assumed to deposit collagen and other ECM components in fibrotic liver, besides hepatic stellate cells, also possess a great collagenolytic potential.

Selective IgM deficiency in CD30+ cutaneous lymphoproliferative disorder.

Primary cutaneous anaplastic large cell lymphoma with local lymph node involvement was diagnosed in a 13-year-old boy with an ulcerative facial lesion and a history of skin lesions of lymphomatoid papulosis. The tumor regressed with chemotherapy. He continued to develop recurrent self-limited lesions of lymphomatoid papulosis , with a halo surrounding these lesions during the healing phase. He developed selective immunoglobulin M deficiency with decline in levels even 4 years after the chemotherapy with no recurrent infections noted and adequate IgG response to immunizations. Both peripheral blood IgM+ and memory B cells were low, suggesting a possible cause-effect relationship between selective immunoglobulin M deficiency and chronic CD30+ cutaneous lymphoproliferative disorders.

Effects of hyaluronan and iodine on wound contraction and granulation tissue formation in rat skin wounds.

BACKGROUND: Hyaluronan (HA) plays an important role in the repair of damaged skin and has been used for the treatment of wounds. Iodine is a mild topical antiseptic. AIM: A mixture of high molecular weight HA with the iodine complex KI(3) (hyiodine) was reported to accelerate wound healing in patients with diabetes and patients after surgery. We investigated how this mixture affects wound contraction, granulation tissue (GT) and wound edges in excision skin wounds in rats. METHODS: Hyiodine was applied to full-thickness wounds made on the back of rats. The areas of the contracting wounds were calculated from digital photographs. The moving edges of the wound were studied by histological methods. The properties of GT were studied in wounds in which contraction was prevented by the insertion of plastic rings. The effects of hyiodine were compared with those of high molecular weight (1200 kDa) HA, low molecular weight (11 kDa) HA and KI(3) solution. RESULTS: Hyiodine accelerated wound contraction significantly in the first 5 days of healing. On day 3, hyiodine-treated wounds had reduced to 63% of the original area, whereas the wound area in saline-treated animals was 75% of the original size. The proliferating epidermis was thicker in hyiodine-treated animals on day 7. In the wounds with inserted rings, hyiodine caused little change in GT, but the weight of the crust/exudate formed on the top of the wound was increased by 351% compared with only minor changes caused by the hyiodine components alone. CONCLUSIONS: Hyiodine supports wound healing by stimulating wound contraction and epidermal proliferation and by keeping the wound moist through increased exudation.

Selective inhibition of hepatic collagen accumulation in experimental liver fibrosis in rats by a new prolyl 4-hydroxylase inhibitor.

Fibrosis and cirrhosis of the liver are often the result of chronic liver damage by a variety of different agents. Pathological accumulation of collagen, disruption of the lobular structure, and impaired hepatocellular function frequently lead to systemic involvement and fatal complications. Drugs inhibiting collagen hydroxylation and accumulation are expected to improve this situation, making prolyl 4-hydroxylase (P4H), the key enzyme of intracellular collagen processing, a rational target for pharmacological intervention. S 4682, a novel inhibitor of purified P4H (Ki = 155 nmol/L), reduced hydroxyproline (Hyp) synthesis in chicken embryo calvaria (IC50 = 8.2 micromol/L) and in cultured hepatic stellate cells (HSC) (IC50 = 39 micromol/L). S 4682 inhibited hepatic collagen hydroxylation in vivo after metabolic labeling with [14C]proline. In the CCl4 model of chronic hepatic injury, characterized by histologically and biochemically evident fibrosis and highly elevated levels of serum procollagen type III N-peptide, S 4682 reduced hepatic collagen accumulation, decreased prevalence of ascites, and lowered serum procollagen type III N-peptide (PIIINP) levels. The hepatic Hyp content of drug-treated animals was closely correlated with serum levels of PIIINP S 4682 had no influence on Hyp content of heart, lung, and kidney.

Hepatocyte proliferation in silicotic rat liver after partial hepatectomy.

Liver silicosis was induced in rats by an intravenous injection of silica particles. The presence of silicotic granulomas in the liver lowered the initial response of hepatocytes to partial (37%) hepatectomy and slowed down the decline in the number of dividing cells 6 days after the stimulus, but did not affect significantly the total number of mitotic cells.

Prolonged reduction of hepatocyte proliferative ability in rats after a single treatment with carbon tetrachloride.

Female Wistar rats were pretreated with I ml of carbon tetrachloride/kg of body weight or with olive oil. All the rats were given this dose of CCl4 20 or 40 days later. Liver regeneration as evaluated by 3H-thymidine incorporation into liver DNA and by the number of mitotic hepatocytes was markedly impaired in CCl4-pretreated rats when compared with olive oil-pretreated controls. DNA labelling reached only 83 and 59% and mitotic index 35 and 58% of control values, respectively, at 20-day and 40-day time intervals. The variables characteristic of liver damage did not parallel the changes in cell division. About 20% of hepatocytes were necrotic both in the CCl4-pretreated and in the control rats. The activity of serum alanine aminotransferase was higher in the CCl4-pretreated rats. Only serum aspartate aminotransferase activities were somewhat lower when compared to controls. Similarly, serum aminotransferases were much less affected by the pretreatment than the markers of regeneration when two low doses of CCl4 (0.125 ml/kg) were given to rats 20 days apart. The activities of microsomal enzymes aniline hydroxylase and pethidine demethylase were equal in control and in experimental rats 20 days after CCl4 pretreatment which indicated that the effects of CCl4 were not mediated by an overall decrease in cytochrome P-450 enzymes. In summary, a single pretreatment of rats with CCl4 induced changes in liver that lasted for 40 days and impaired liver regeneration when another dose of CCl4 was applied.

Ultrastructure of rabbit tracheal epithelium after the administration of Hexabrix--a hexaiodated contrast agent for tracheobronchography.

The ultrastructure of the tracheal epithelium after the application of 1 ml of Hexabrix (a hexaiodated contrast agent for tracheobronchography) into the airways was studied. Severely altered pseudostratified columnar epithelium was found in the trachea. Eighty-one +/- 5% of the goblet cells had been stimulated to discharged mucus. Forty-three +/- 3% of them were completely exhausted and had degenerated. The ciliated cells displayed marked signs of pathological alteration up to vacuolar degeneration. The ciliary border above the epithelium was severely damaged. The mean number of cilia per micron 2 fell to 3.3 +/- 0.5, but the proportion of intact cilia did not drop below 95%. As morphological signs of impaired self-cleaning ability of the airway epithelium, a large amount of inspissated mucus and numerous bacteria were found in the area of the impaired ciliary border. Coarse grained contrast substance of supreme electron density was intermingled with the condensed mucus. The most pronounced damage to the cells was observed at the sites of direct contact of the contrast agent with the cell membranes.

[Epiglottitis and subglottic laryngitis]. / Epiglotitis a subglotická laryngitis.

Account of the course and conclusions of a working session of the Paediatric Society devoted to laryngeal obstruction in children. The participants discussed in particular the differential diagnosis of subglottic laryngitis and epiglottitis in children.

Regenerative ability of hepatocytes is inhibited in early stages of liver fibrosis.

Female Wistar rats were given three doses of carbon tetrachloride, 10.4 mmol/kg of body weight. The doses were administered within 16 days and another 16 days were allowed to pass before partial (37%) hepatectomy was done. The liver showed very mild fibrosis at that time. DNA synthesis (measured by 3H-thymidine incorporation) was decreased by 53% and mitotic activity of hepatocytes was decreased by 56% when compared to olive oil-pretreated partially hepatectomized controls. The results show that the mitotic potential of hepatocytes in early stages of liver fibrosis is impaired which may influence the course of the disease.

Relationship of liver damage and liver regeneration after carbon tetrachloride treatment in rats.

Various doses of carbon tetrachloride (0.625 mmol to 10 mmol CCl4/kg body weight) were administered to female Wistar rats. Liver damage after a single treatment was evaluated by serum aminotransferase levels and by the extent of necrotic areas in parenchyma. Liver regeneration was evaluated by 3H-thymidine incorporation into liver DNA and by the number of dividing hepatocytes. Mitotic index of hepatocytes rose in parallel with the specific activity of DNA and with the extent of necrosis. However, the activities of serum aminotransferase AST and ALT increased much more rapidly and did not correlate either with necrosis or with regeneration rate. Increased membrane permeability in morphologically intact cells, increased synthesis of the enzymes by the liver as well as the leakage from necrotic cells are discussed as possible causes of the high aminotransferase activities in serum.

[Hemangiomas of the larynx in children--results of surgical treatment]. / Hemangiomy hrtanu u detí--výsledky chirurgické lécby.

Surgical treatment of six cases of haemangiomas of the larynx. Surgery was used only after all conservative means were exhausted. After complete removal of the tumour after a two-year interval no relapse was recorded. The function of the larynx was not impaired.

[Tracheobronchography in neonates and infants with obstruction of the upper respiratory tract]. / Tracheobronchografie novorozencu a kojencu s obstrukcí horních dýchacích cest.

In the period of 1980 to 1989 the authors performed contrast examinations of tracheobronchial tree in 38 children up to one year of age, three of them being newborns. In 29 examined children (74.4%) there was an anatomical defect causing the obstruction of upper respiratory pathways. In 15 of them the trachea compression was caused by pressure of hyperplastic thymus on the brachiocephalic arterial truncus crossing trachea. For visualization of upper respiratory pathways Hexabrix 320 (Byk Gulden) was used in 30 children, administered in 10 cases by jet ventilation. The filling was recorded by radiocinematographic method.

[Occlusion of a pseudoaneurysm of the internal carotid using a catheterization technic]. / Uzáver pseudoaneuryzmatu vnitrní krkavice katetrizacní technikou.

The authors describe a successful therapeutic occlusion of the left internal carotid artery in a five years old boy by the catheterization technique. The intervention treated a pseudoaneurysm originating after incision of a peritonsillar abscess.

Elastin content in human fibrotic and cirrhotic liver.

Human liver tissue obtained at autopsy was extracted with hot 0.1 M NaOH. Determination of amino acids desmosine and isodesmosine was used to quantify elastin in the insoluble residue. Hydroxyproline content was used as an index of collagen content in the liver. Hydroxyproline content was twofold in fibrotic liver and threefold in cirrhotic liver when compared to normal liver, while the sum of desmosine and isodesmosine increased threefold and sixfold, respectively. Elastin content is low in normal liver when compared to collagen but appears to grow much faster in diseased liver.

Effect of bronchoalveolar lavage on the ultrastructure of the tracheal epithelium in rabbits.

Serious damage to the tracheal epithelium due to bronchoalveolar lavage (BAL) was recorded. Immediately after BAL 99 +/- 2% of goblet cells were exhausted and degenerated. Their regeneration began 24 h after BAL resulting in hyperplasia of goblet cells with the formation of endoepithelial mucous glands. The most pronounced injury to the ciliated cells was apparent 2 h after BAL. BAL markedly impaired the ciliary border. The mean number of cilia/microns 2 fell to 1.5 +/- 0.3/microns 2, then gradually rose to 7.5 +/- 0.5/microns 2 (controls 9.7 +/- 0.5/microns 2). The morphological signs of impairment of the self-cleaning ability of the epithelium were the most pronounced after 24 h and were still present at the end of the experiment.

The effect of Hexabrix tracheobronchography on the ultrastructure of the airway epithelium.

The ultrastructure of the airway epithelium was studied after experimental Hexabrix tracheobronchography. Serious damage to the tracheal epithelium due to Hexabrix was recorded. 81 +/- 5% of goblet cells were stimulated to discharge mucus. 43 +/- 3% were completely exhausted and degenerated. The ciliated cells displayed vacuolar degeneration or severe pathological alteration. The average number of cilia/micron 2 fell to 3.3 +/- 0.5. As morphological signs of impaired self-cleaning ability the inspissated mucus and numerous bacteria appeared in the area of the disorganized ciliary border.

Regeneration of changes produced in rabbit tracheal epithelium by saline lavage of the airways.

The authors studied the course of the repair of changes induced in the rabbit tracheal epithelium by saline lavage of the airways. The tracheal epithelium was examined 2, 24, 48 and 72 hours after treatment. Saline lavage stimulated the goblet cells to instantaneous discharge of their secretion. 2 hours after treatment 98 +/- 3% of the goblet cells were completely exhausted and had degenerated. Repair of the changes began 24 hours after lavage and was associated with massive differentiation of new goblet cells resulting in hyperplasia of the mucus-secreting elements with formation of endoepithelial mucous glands. The most pronounced injury to the ciliated cells was apparent 2 hours after lavage, then the degree of alteration of these cells gradually decreased. Saline lavage markedly impaired the ciliary border. The mean number of kinocilia per micron2 fell to 1.5 +/- 0.3. In subsequent phases the number of kinocilia rose gradually to 7.5 +/- 0.5/micron2. This value was still significantly lower (P less than 0.005) compared with controls. The first signs of impairment of the self-cleaning ability of the epithelium were recorded 2 hours after lavage. The most pronounced disturbances of the mucus flow were observed after 24 hours. At the end of the experimental period small clumps of condensed mucus and rather numerous bacteria were still present in the area of the ciliary border.

[Reaction of the respiratory tract epithelium to bronchoalveolar lavage]. / Reakce epitelu dýchacích cest na bronchoalveolární laváz.

The authors investigated experimentally the reaction of the airway epithelium to the bronchoalveolar lavage. The epithelium of the rabbit trachea was examined immediately after rinsing with saline and 2, 24, 48 and 72 hours after rinsing. Lavage causes excessive stimulation of goblet cells. Immediately after lavage we find in the epithelium 99% goblet cells which after releasing the secretion degenerate and are gradually expelled from the epithelium. Regeneration of goblet cells begins 24 hours after lavage and is associated with massive differentiation of new goblet cells which leads to hyperplasia of mucus secreting elements and the formation of endoepithelial mucous glands. Signs of pathological alteration of ciliated cells are most marked 2 hours after impairs subsequently the degree of alteration of these cells declines gradually. Bronchoalveolar lavage reduces markedly the ciliary border above the epithelium, the number of kinocilia declines on average to 1.5/microns 2. In the subsequent stages the number of kinocilia increases again to 7.5/microns 2. This value is still significantly lower (p less than 0.005) than in controls. In the impaired ciliary border as a morphological sign of the impaired self-cleaning ability of the airway epithelium, numerous bacteria and condensed mucus are founded. Signs of local disorders of mucus flow are apparent still three days after bronchoalveolar lavage.