Type II alveolar epithelial cells and free alveolar cells after intratumor TNF-alpha administration.

The experiment used Morris hepatoma 5123 series growing in muscles of the Buffalo rats. A suspension of 3 x 10(6) neoplastic cells was injected into the right hind leg of the animals. After fourteen days, TNF-alpha was administered into the tumour in a dose of 1.5 x 10(4) U/24 hours in 0.5 ml PBS solution. The group I animals were injected for 4 days and group II for 8 days. Control groups consisted of rats with injected Morris hepatoma which were given PBS solution instead of TNF-alpha (group III A and B) and animals without the hepatoma, given 4 or 8 TNF-alpha, respectively (groups IV A and B). In the present study, we have explored the effect of intratumor TNF-alpha administration on the composition of cells isolated from the lungs through multiple bronchoalveolar lavages (BAL). Ultrastructural evaluation of the pulmonary tissue was done using a transmission electron microscope (TEM), with special attention paid to type II alveolar epithelial cells and free alveolar cells. Examinations in TEM in groups I, II and IV (A and B) found, in the lumen of alveoli, an increase in the number of alveolar macrophages (AM) with morphological features of intensified activity and AM with numerous secondary lysosomes containing material of phospholipid structure. Also, numerous type II alveolar epithelial cells with emptied lamellar bodies were observed. The above mentioned changes were especially marked after eightfold TNF-alpha administration. In groups I, II and IV (A and B), compared with group III, a significant increase was found in the total number of cells isolated by BAL as well as in the number of cells with positive reaction in staining according to Beckstead's method. It may indicate that the changes in the parameters mentioned above are related to TNF-alpha action. The results obtained indicate the possibility of systemic effect of TNF-alpha after its administration into the experimental Morris hepatoma.

Regression of Morris hepatoma in response to intralesional treatment with tumor necrosis factor muteins.

We examined the antitumor effects of human recombinant tumor necrosis factor alpha (rhTNF-alpha) and its muteins with the N-terminal amino acid sequence altered by point mutations against transplantable Morris hepatoma 5123 in rats. In vivo studies showed antiproliferative activity of the drugs in the dose range tested. For in vivo studies rhTNF-alpha and muteins were administered intratumorly (i.t.). The preparations were given at a dose of 10 microg/rat, once daily for eight days. Although the therapy was significantly effective in inhibiting tumor growth, complete growth inhibition could not be achieved. Nevertheless, there was a significant increase in survival time of tumor-bearing rats.

The antitumor effect of intraperitoneal treatment with rhTNF-alpha muteins on Ehrlich ascites tumor growth.

The intraperitoneal (i.p.) treatment with recombinant human tumor necrosis factor-alpha (rhTNF-alpha) is one of the possible therapies for tumors that are confined to the abdominal cavity. Clinical trials aiming at the exploitation of the antitumor effects of rhTNF-alpha have been largely disappointing. In this model the activity of some rhTNF-alpha derivatives was studied. Ehrlich's ascites tumor (EAT) bearing Swiss albino male mice were treated i.p. three times a week with 10 micrograms/mice of rhTNF-alpha, mutein V or mutein VI for two weeks, starting on the 4th day after tumor inoculation. Control mice received PBS. The effect of the rhTNF-alpha derivatives on the course of EAT was evaluated basing on: total ascites volume (TAV); packed cell volume (PCV); total packed cell volume (TPCV); inhibitory growth rate (IGR); cellular population of EAT fluid; morphological EAT cell changes and mean survival time (MST). In the study mutein VI had only a slight effect on MST but significant on TAV- and TPCV-IGR (p < 0.001). In mice treated with rhTNF-alpha and mutein V the enhancement of MST (p < 0.01) was accompanied by TAV- and TPCV-IGR (p < 0.001). The number of EAT cells in ascites decreased after rhTNF-alpha and mutein V administration (p < 0.001). We conclude that treatment with high-dose of this modified molecule lacking the possibility of binding with p75R and not producing so intensified side effects is likely to find wider application in therapy and prevent the ascites growth just as rhTNF-alpha dosage.

Effect of the cytokine rhTNF-alpha on the population of mast cells in the growth of MethA fibrosarcoma--a TEM study.

The aim of the present study was the ultrastructural characteristics of mast cell (MC) involved in host antitumor responses induced by local (i.t.) administration of recombinant human tumor necrosis factor alpha (rhTNF-alpha) in the primary focus of methA fibrosarcoma. MC were involved in tumor interstitium remodeling. Numerous mitochondria, well-developed RER and Golgi apparatus, clusters of polyribosomes, considerable polymorphism of granules and differentiated lamellar structures which frequently presented myelinic forms were observed after rhTNF-alpha application. In the study numerous fibres of the fibrous tissue, richly vascularized, occurred in the peripheral and intermediate tumor zones. Cluster of MC and tumor cells were seen on the border of the necrotic foci. However, proteolytic enzymes released by MC cause interstitial lysis, ensuring the place for tumor growth, and are involved in angiogenesis. Thus, it is not clear whether MC contribute to the inhibition of tumor growth or have an adjunctive role in tumor progression.

Malignant neoplasms of the pancreas. A study based on autopsy data from 1953 to 1982 in Bialystok, Poland. I. Frequency, age and sex distribution.

Among 18 751 autopsies in persons over 20 years of age 195 primary malignant neoplasms of pancreas (MNP) were stated (117 in males, 78 in females; the ratio being 1.5 : 1). In the period of the investigation (1953--1982) MNP comprised 1.0% of all autopsies, 4.1% of all malignant neoplasms (MN), and 11.0% of MN of alimentary system (MNAS). The frequency of MNP in all autopsies was from 0.5% during the years 1953--1957 to 1.2% in 1978--1982, in all MN from 2.7% to 4.5%, and in all MNAS from 5.7% to 12.7%, respectively. MNP were observed most frequently at the age of 61--70. The mean age was 63.6 and was higher in females than in males (66.3 and 61.8, respectively), and increased in the period under study from 54.6 to 65.6 (in males from 50.8 to 64.8, in females from 59.3 to 66.8). The significant increase in the incidence of MNP in the population of patients who died in all hospitals in Bialystok (the north-eastern Poland) between 1953--1982 and were autopsied in our Department correlates with data indicating the rise in the incidence of MNP in many countries of the world.

Malignant neoplasms of the pancreas. A study based on autopsy data from 1953 to 1982 in Bialystok, Poland. II. A survey of 195 cases.

Among 195 malignant neoplasms of the pancreas (MNP) diagnosed at autopsy in 1953--1982 the exocrine carcinomas comprised 85.6% and inmular carcinomas 13.8% of cases. MNP were localized most frequently in the head of the pancreas (54.4%), then in the whole pancreas in 14.9%, in the head and the body or in the body and the tail in 20.5%, in the body or in the tail in 10.3% of MNP. There were no infiltrations and no metastases in other organs only in 16 patients (8.2% of MNP). The biliary tracts and duodenum were the most frequent sites of secondary infiltrations. Metastases were most frequent in the liver, then in the liver hilus and mesentery lymph nodes. The concordance of clinical and post mortem diagnosis of MNP was stated in 40.5% cases.

Evaluation of biometric parameters of Morris hepatoma after application of human recombinant tumor necrosis factor.

The effect of human recombinant tumor necrosis factor alpha (h rec TNF-alpha) on the growth of Morris hepatoma 5123 implanted in the skeletal muscles of the thigh of Buffalo rats was investigated. The cytokine was repeatedly given in an intratumor administration (i.t.) in dose of 1.5 x 10(4) U once a day in regimens of four or eight days. Comparative groups consisted of animals which were given saline i.t. Control groups included healthy rats subjected to local cytokine effect. The experiments revealed an inhibitory effects of the preparation on the growth of tumors. Biometric parameters of the tumors induced indicated that the inhibition of Morris hepatoma was most effective after the eighth dose of h rec TNF-alpha. The administration of fourfold dose resulted in an initial loss of body mass increase. However, when injected eight times, the factor produced a relative tolerance reflected in minor reduction of actual body mass. The estimation of survival time in rats injected i.t. with h rec TNF-alpha, compared to those given saline, revealed statistically significant differences at the eighth repeated dose.

Dynamics and phases of changes in the primary focus of Morris hepatoma under local application of cytokine hrecTNF-alpha.

Biological activity of cytokine hreeTNF-alpha was tested on a transplantable Morris hepatoma 5123 in Buffalo rats. Local effects of hrecTNF-alpha activity were evaluated basing on morphological examinations. The cytokine effects on tumor biometric parameters and body mass of experimental animals were analyzed. It has been found that hrecTNF-alpha considerably decreases body mass and reduces studied parameters of the tumor, the changes being statistically significant. Three consecutive phases of the neoplasm regression have been distinguished. Phase I revealed prevailing hemorrhagic necrosis of the central and intermediate zones, with numerous thrombi in the vascular lumen and fibrinoid necrosis of vascular walls. Phase II presented a number of cellular infiltrations, clearance of necrotic foci and initial angiogenesis and fibroplasia. Phase III showed intensified angiogenesis and proliferation of the connective tissue in the peripheral and intermediate zones.

The effect of local hrec TNF-alpha administration upon the spontaneous lung metastases in rats with Morris 5123 hepatoma.

The experiment used Morris hepatoma 5123 series growing in muscles of the right hind limb of Buffalo rats. The group I animals were given intratumor 4 doses of TNF-alpha and group II-8 doses of TNF-alpha (10 micrograms/day). Control groups (III and IV) consisted of rats with injected Morris hepatoma, which were given PBS solution instead of TNF-alpha. A decrease in the volume of neoplastic metastases was observed in groups I and II, compared with groups III and IV. At the same time an increase was found in the volume of metastatic tumors in group II (8 x TNF-alpha), compared with group I (4 x TNF-alpha). Histological and ultrastructural analysis of the pulmonary tissue revealed intensified fibrotic reactions and inflammatory infiltrations around the metastatic tumors. The change were much more enhanced in group II, which might affect the results of neoplastic metastatic volume measurements. We concluded that multiple human recombinant TNF-alpha, hrec TNF-alpha, local injections inhibited dissemination of tumor cells and prolonged the survival time of rats up to the 76th day of the follow-up.

The effect of repeated BCG vaccine administration associated with surgery upon the growth of lung metastases in rats with transplantable Morris 5123 hepatoma.

Eighty four male Buffalo rats bearing transplantable Morris 5123 hepatoma were used for the experiment. Half of the animals were given into the tumor BCG vaccine (final dose 2.2 X 10(6) living mycobacteria), the other 42 rats after BCG underwent surgical removal of the hind leg together with the tumor. Though the immunotherapy reduced the size of lung metastases of the tumor it did not, however diminish the survival rate of the animals. Multiple BCG doses associated with the surgical procedure prevented dissemination of the tumor cells and prolonged the survival of rats up to the 100th day of the follow-up.

AgNORs in duct epithelial lesions in chronic pancreatitis and in pancreas cancer cells.

BACKGROUND/AIMS: Argyrophilic nucleolar organizer regions (AgNORs) reflect the proliferative activity of cells. Since the majority of pancreatic cancers are ductal carcinomas, the aim of the study was to determine the AgNORs expression of potential pre-neoplastic ductal epithelial lesions in advanced chronic pancreatitis compared with pancreatic cancer cells. METHODOLOGY: Histological preparations obtained from 24 patients with chronic pancreatitis and 16 patients with pancreatic cancer were used to estimate the number of AgNORs per nucleus. Four types of AgNORs were distinguished and histograms with cell percentage of each type were performed for all forms of epithelial anomalies. RESULTS: In simple hyperplasia, squamous and mucous metaplasia the number of AgNORs ranged from 1.92 to 2.23; type I was predominant. In papillary hyperplasia, dysplasia and in situ carcinoma the number ranged from 2.98 to 3.34, with a predominance of type II-IV. In invasive carcinoma the number was 4.29 and 74% of cells were of type II-IV. CONCLUSIONS: Both counts of AgNORs and the percentage of type II-IV cells showed a gradual increase from simple hyperplasia through papillary hyperplasia and dysplasia to invasive carcinoma which in this respect differs significantly from all forms of the epithelial anomalies examined.

Epithelial anomalies in chronic pancreatitis as a risk factor of pancreatic cancer.

BACKGROUND/AIMS: The relationship between chronic pancreatitis and the development of pancreatic cancer is still a matter of dispute. Our aim was to determine the frequency of hyperplastic, metaplastic and dysplastic epithelial anomalies in the course of chronic pancreatitis and the potential steps in their development to malignancy. METHODOLOGY: The study was based on biopsy material of 70 patients with clinically diagnosed advanced chronic pancreatitis, who underwent partial or total pancreatectomy, as well as other operations. The patients were assigned to 2 groups: Group I (n = 41) with calcifying chronic pancreatitis; Group II (n = 29) with other forms of the disease. Histological sections were stained with hematoxylin-eosin, Mallory-azan, Gomori's silver method, and glycosaminoglycans (PAS and Alcian blue staining). Special interest was focused on the type and incidence of epithelial ductal and acinar cell anomalies, and on the degree of parenchymal scarring. RESULTS: Hyperplasia of the ductal epithelium was present in 31.4%, focal squamous metaplasia in 21.4%, mucous metaplasia in 11.1%, cellular dysplasia in 8.6%, dysplastic acinar cell nodules in 21.4%, and "tubular complexes" in 30.0% of all cases. The differences in the frequency of these changes, except for ductal epithelial hyperplasia, were not statistically significant in two comparable groups. Advanced pancreatic fibrosis was associated with epithelial anomalies in 65.7% of all cases. CONCLUSIONS: From the morphological point of view, the adequate prerequisites for the consideration of advanced forms of chronic pancreatitis, independent of type, as a risk factor of pancreatic cancer exist, necessitating the surgical removal of pathological lesions.

Inhibitory effect of the human recombinant tumor necrosis factor on the growth of the Morris hepatoma in rats.

The effect of the human recombinant tumor necrosis factor alpha (h rec TNF-alpha) on the transplantable Morris hepatoma 5123 was studied in Buffalo rats. The cytokine was repeatedly administered intratumorly (i.t.) in a dose of 1.5 x 10(4) U once a day in a cycle of four and eight days. The control groups consisted of animals given saline i.t. The experiments revealed an inhibitory effect of the h rec TNF-alpha upon the growth of neoplastic tumors. The biometric parameters of the tumors indicated that the inhibition of the Morris hepatoma was most effective after eight repeated doses of TNF. After injections of TNF-alpha, the tumors presented extensive hemorrhagic necrosis, the regressive alterations being found mainly in the central and intermediate tumor zones. In the early phase of the tumor growth, neoplastic tissue necrosis prevailed, as well as hemorrhages within the necrotic masses, necrosis of the blood vessel walls and thrombi in their lumina. In the later period, numerous fibres of the fibrous tissue, richly vascularized, occurred in the peripheral and intermediate zones. Clusters of eosinophilic granulocytes and macrophages with apoptotic bodies in the cytoplasm were seen on the border of the necrotic foci.

Alveolar epithelial cells in experimental lung emphysema. Ultrastructural analysis of cells in situ in TEM.

A study was made of ultrastructural changes in alveolar epithelial cells after a single intratracheal papain injection. The experiment was carried out on male Wistar rats, of 180-220 g body weight. The animals were sacrificed after 1, 7, 14 and 28 days that followed proteolytic enzyme administration. The study revealed that the development of emphysematous changes in the rat lungs was accompanied by alterations in the epithelium of alveoli. In early stages of emphysema (up to 1 week), destructing changes dominated within the epithelial cells. Similar changes were observed in the endothelium of adjacent blood vessels. In later periods an increased number of type II pneumocytes were seen and focal accumulation of elastic and collagenic fibres was noticed in alveolar septa. Correlation was found between changes in the number of type II cells and their ultrastructure as well as between alveolar epithelium damage and alveolar septal interstitium alterations.

Comparative studies on the ultrastructure of the rat lungs after intratumor treatment of Morris hepatoma with rhTNF-alpha and its muteins.

The aim of the present study was the comparative analysis of morphological changes found in the lungs of Buffalo rats in the course of Morris hepatoma 5123 after i.t. treatment with recombinant human TNF-alpha (rhTNF-alpha) and its muteins. Modification of the native TNF-alpha molecule and synthesis of mutagenized analogues can prevent undesirable symptoms observed in the case of therapeutic administration of rhTNF-alpha. TNF-alpha has been shown to interact with two distinct membrane receptors (TNF-R): p55R and p75R. Mutagenized mutein V binds selectively with p55R. Mutein VI fails to recognize either TNF-R. The cytokines were applied in a dose of 10 micrograms protein in a cycle of 8 days. The control group consisted of tumor-bearing animals which were given PBS. Ultrastructural examinations were based on transmission electron microscope (TEM). Mutein VI-receiving animals showed enhanced changes of cytotoxic nature. Severe damage to endothelial cells (necrosis inclusive) was observed. Blood vascular lumen showed accumulation of neutrophils and monocytes. Features of enhanced activity of endothelial cells were noted. Focally, within pulmonary alveoli conglomerates of fibrin and fragments of damaged cells were found, with erythrocytes, neutrophils and macrophages in their vicinity. The epithelium of pulmonary alveoli showed signs of considerable damage, including necrosis. The lumen of pulmonary capillaries in rhTNF-alpha-treated animals showed a predominance of eosinophils and monocytic cells. Features of endothelial stimulation were observed, although without a tendency to form microthrombi. Much less pronounced changes both in the lung capillary bed and in the alveolar epithelial cells were noted in the mutein V-given animals. Our findings confirm the possibility of peripheral activation of cells involved in the cytokine-induced antitumor response. Mutein V with the smallest effect on the lung tissue rebuilding seems to be a rhTNF-alpha derivative which can delimit the undesirable symptoms in the course of antitumor therapy reduced to i.t. injections.

Ultrastructural analysis of pulmonary capillaries in the course of experimental lung emphysema.

Processes which occur within the capillaries of interalveolar septa play a major role in many lung diseases. In the present study, ultrastructural analysis of pulmonary capillaries was made at 7 days after intratracheal instillation of a proteolytic enzyme - papain, that is when macrophage cumulation within pulmonary tissue was maximal. Experimental animals (male Wistar rats) were injected with BCG vaccine, in doses that would stimulate the macrophage system to produce TNF-alpha. All the experimental groups, compared with the control, showed cumulation and increased adherence of monocytes, granulocytes and blood platelets to endothelium. Changes within endothelial cells manifesting their activation were observed. In the animals receiving papain or BCG, focal destruction of endothelial cells was found. The picture of vascular changes induced with papain and by simultaneous stimulation with BCG vaccine revealed a number of features similar to those found in the adult respiratory distress syndrome. The study proved usefulness of vascular perfusion as a method to analyze the vascular system, particularly in the processes progressing with activation and increased adhesion of inflammatory cells to the endothelium of pulmonary capillaries.

Tumor necrosis factor-alpha induces emphysema-like pulmonary tissue rebuilding. Changes in type II alveolar epithelial cells.

The effect of repeated doses of TNF-alpha on the histological picture of the pulmonary tissue was analyzed in the present study. Special attention was paid to the lung rebuilding processes. TNF-alpha was applied intraperitoneally for two weeks in a dose of 10 micrograms/0.5 ml PBS/24h. Morphological analysis of the pulmonary tissue was performed after 1 and 28 days following the last TNF-alpha dose. The study revealed focal pulmonary tissue rebuilding with emphysema-like changes twenty eight days following termination of TNF-alpha administration. The rebuilding processes included interalveolar septal atrophy, collagen accumulation and damage-repair changes in type II alveolar epithelial cells. It has been demonstrated that apart from the protease-antiprotease hypothesis of the lung emphysema, the inflammatory-repair hypothesis should be considered. Both hypotheses are complementary to each other and interpret the emphysema-like changes as complications of various pathological conditions of the pulmonary tissue.

Alveolar epithelial cells in experimental lung emphysema. Analysis of methods for type II pneumocyte identification among cells isolated from the respiratory tract of rats.

Total and differential counts of cells isolated from the lungs of Wistar rats with experimental emphysema were performed. Emphysema was induced by a single intratracheal dose of papain solution (20 mg/kg b.w.). Rats were sacrificed 1, 7, 14 and 28 days later. Cells were isolated from the lungs through multiple bronchoalveolar lavage. An attempt to identify type II pneumocytes was undertaken. Our observations suggest that none of the methods applied for identification of type II pneumocytes are completely specific. At the same time they stain type II cells and subpopulations of alveolar macrophages heavily loaded with phagocytized surfactant elements. However those methods are simple and enable an approximate evaluation of damage and regeneration processes in the extracellular alveolar lining layer and epithelium.

[Hypertrophy of the lateral head of the gastrocnemius muscle with spontaneous electrical activity]. / Przerost gøowy bocznej miesnia brzuchatego øydki z samoistna czynnoscia elektryczna--niezwykøy przypadek radikulopatii.

A patient with chronic S-1 radiculopathy is discussed, in whom, after previous slight weakening of calf muscles, the lateral head of the gastrocnemius muscle was enlarged. Histopathological examination revealed in this muscle a true hypertrophy, mainly second type of muscle fibres. However electromyographic examination showed a continuous spontaneous electrical activity at rest (pseudomyotonic discharges) which, according to the authors, explain the found signs.

[The effect of pentoxifylline on the degree of lung parenchyma injury after papain]. / Wplyw pentoksyfiliny na stopien uszkodzenia tkanki plucnej po papainie.

In the last years pentoxifylline (PTXF) has been successfully used in reducing acute injury of the lung parenchyma in ARDS. The authors have studied the effect of PTXF on degree of papain induced lung injury. Papain was administered intratracheally in a dose of 2 mg, 4 mg, 6 mg in 1 ml of PBS. The effect of papain on bronchoalveolar lavage characteristics was also evaluated. Wistar rats of both sexes were used in the experiment. The degree of lung destruction, reflected by interstitial hemorrhage was assessed by measuring hemoglobin content in the fluid of the lavaged lungs. The hemoglobin levels were assessed spectrofluorometrically with the use of the 414 nm wave length. A reduction of hemoglobin content was seen after PTXF administration only in animals receiving the lowest dosage of papain. However in all animals a decrease in the BAL neutrophil count was demonstrated. The protective effect of PTXF on pulmonary tissue in papain induced injury models and situations leading to development of ARDS may suggest a similar pathomechanism in both entities.