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Biomedical subjects

J Hradecký

Publications and source records attributed to J Hradecký.

At least 19 recordsLinked to original sources

Immune characterization of the Lewis rats inoculated with K2 sarcoma cell line and newly derived R5-28 malignant cells.

Sarcoma is a relatively rare malignant disease with high mortality, bad prognosis and response to conventional therapy. Two possible models of this disease were tested: the K2 rat sarcoma cell line, which was described previously, and the new rat R5-28 cell line derived from a spontaneously growing rat neoplasm with sarcoma morphology. While all rats inoculated with K2 cells developed tumours at 22th-25th day after inoculation (D = 22-25), only 60%-75% of R5-28-inoculated rats were affected by tumours. The frequency and progress of the disease depended on the number of inoculated cells. No metastases were detected in both cases. All affected animals showed large splenomegaly. A possible response of some immune system components to tumours was tested. No tumour-infiltrating lymphocytes were revealed in the tumour tissue. Anti-tumour antibodies were not found in tumour-bearing animal sera. Appropriate changes in peripheral blood lymphocyte subsets were explored. While the relative numbers of both NK cells and Tc were impaired, no changes were noted in numbers of CD4+CD8- T helper cells. Leukocytosis with highly increased numbers of CD11b+ myeloid cells displaying variable expression of CD4 was detected in terminal stages of the disease.

Animals↗

[Melanoblastoma in laboratory minipigs: a model for studying human malignant melanoma].

Human malignant melanoma is a progressively growing tumour mostly with a fatal prognosis during short time. Its frequency has doubled in some states of the USA during the last few years. Surgical treatment, which is still generally used, has very low efficiency due to the numerous metastases in different organs. An animal model is a necessary prerequisite to develop a new therapy methods. We have recorded primary cutaneous malignant melanomas as well as extensive metastases in the visceral organs in darkly pigmented laboratory minipigs. The primary skin tumours were mostly multiple and they showed a tuberous nodular form. The regular occurrence of tumours in a progeny of three specific mating types suggests the existence of genetic predisposition to malignant transformation. We discuss the use of this animal model for further studies and treatment of human malignant melanoma by our ischemization technique developed previously.

Animals↗

Two new common alleles of erythrocyte adenosine deaminase (ADA) in pigs.

New adenosine deaminase variants ADA C and ADA D were found by means of agarose gel electrophoresis in pig erythrocytes. Family data supported the hypothesis that these are controlled by codominant alleles ADAC and ADAD. The ADAC allele was present in Large White (q = 0.076), Landrace (q = 0.037) and their crosses with other breeds. The ADAD allele was present in Duroc (q = 0.067) and its crosses. Allele frequencies for six pig breeds are given.

Adenosine Deaminase↗

New blood group factors Es, Et and a new allele Ebdgjmt (E16) in the pig E blood group system.

By immunizing a miniature sow a monovalent reagent (later designated anti-Es) was prepared, detecting an alternative antigen to the Ej. Blood group factors Ej and Es thus form the fifth genetically closed E subsystem. Analyses of selected raw sera containing anti-Ej led to the determination of a further Ej subgroup (designated Et) which is antithetical, mutually excluding with the blood factor Er. New blood factor Et is inherited by alleles Edeghjmnt (= E9) and (= E16). The investigation in pig breeds kept in CSFR indicated that allele Edeghjmnt occurs in Black and White Prestice breed (qE9 = 0.076 +/- 0.010) while allele Edeghjmnr (= E14) only in miniature pigs (qE9 = 0.147 +/- 0.011) and in wild pigs. In an élite herd of Swedish Landrace kept in CSFR a new complex allele Ebdgjmt (= E16) was found. Its frequency in the population studied was 0.058 +/- 0.022.

Alleles↗

Polymorphism of pig serum alpha-protease inhibitor-3 (PI3) and assignment of the locus to the Pi1, Po1A, Po1B, Pi2, Igh linkage group.

Polymorphism of an alpha-protease inhibitor, PI3, in pig serum samples was detected using 2D agarose gel (pH 5.4)--polyacrylamide gel (pH 9.0) electrophoresis. Evidence was obtained that the five variants observed (A, B1, B2, C and D) are under genetic control by codominant alleles (Pi3A, Pi3B1, Pi3B2, Pi3C and Pi3D) at one autosomal locus. Variants A, B1, B2 and C inhibited chymotrypsin; there was no appreciable inhibition of trypsin and papain. Variant D did not inhibit chymotrypsin, and therefore its classification as a PI3 variant was put in question. PI3 typing was not possible in about 50% of the studied pigs since in those cases the PI3 variants were either too weak or absent. On the basis of backcross matings and haplotyping in complete families for protease inhibitor loci Pi1, Po1A, Pi2 and Pi3 it was proved that the Pi3 locus belongs to the protease inhibitor gene cluster, and the position of the locus in the linkage group was proposed as being Pi1-Po1A-(Po1B)-Pi3-Pi2-(Igh1, Igh2, Igh3, Igh4).

Alleles↗

Experimental devascularization (devitalization) of the rectum and sigmoideum.

In 6 laboratory pigs, devascularization (devitalization) of the rectum and the adjacent part of the sigmoideum was carried out by the vasa rectalia cranialia et media ligatures with simultaneous aboral closure of the rectum at the base of the lesser pelvis and oral closure at the borderline between the rectum and the sigmoideum. The devascularized rectum was not removed but left in situ. Terminal colostomy was established. Laparotomic revision was carried out after 12 weeks, colostomy was removed and continuity of the intestine was reestablished by insertion anastomosis, by suture of the end of the colon to the anal mucosa and skin. In further two pigs, devascularization of the rectosigmoideum was carried out with colostomy and reconstruction was carried out after 12 weeks by procto-sigmoideo-anastomosis. In another 2 animals, devascularization was carried out on the lower part of the sigmoideum and continuity was immediately renewed by end-to-side sigmoideo-sigmo-ideoanastomosis. All revisions indicated that the devascularized necrotic segment shrank to form a minute fibrous tissue residue, anastomosis was patent and continence was retained for colo-proctoanastomosis. The described techniques are technically simple and could be employed as palliative surgical methods for inoperable tumours in the region of the rectum and lower part of the sigmoideum.

Anal Canal↗

[Use of the tourniquet in gastrointestinal surgery].

Experimentally induced total or partial intestinal obstructions (ileus) in pigs were surgically treated by means of a tourniquet technique. A lace used in obstetrics for ligation of umbilicus served as the tourniquet. The lace was protracted around an intestine, both ends were threaded through 7-10 cm long rubber tube and fixed by Pean's forceps. During application of the tourniquet we performed suction of the ileus content, devitalization or resection of the affected intestinal segments and intestinal anastomoses. Since the tourniquet provides a flexible compression of intestine with minimum mechanical damage, manipulation without danger of instrument release, and it has negligible cost, this tool gives supposition for the possible use in human surgical praxis.

Animals↗

Changes of clinical biochemical values after bilateral nephrectomy and allotransplantation of the kidney in pigs.

In a series of 10 healthy pigs, the normal ranges of the following biochemical parameters in blood serum were assessed: creatinine, urea, mineralogram (Na, K, Cl), uric acid, bilirubin, total protein, cholesterol, blood lipid level, thymol turbidity reaction and activities of alkaline phosphatase, alpha-amylase and transaminases. The changes of these values were followed up in two pigs after bilateral nephrectomy and in 18 pigs subjected to orthotopic allotransplantation of the kidneys. The examination confirmed that creatinine and urea are the most important values with a prognostic impact. However, other biochemical values signalize also renal dysfunction. In the mineralogram, values of Na and K ions are important indicators. Deterioration of renal functions was associated as a rule with a decline of alpha-amylase and alkaline phosphatase. The other examinations did not display any marked changes during renal damage.

Alkaline Phosphatase↗

Polymorphism of the SLB serologically defined (non-MHC) alloantigenic system in pigs.

Six new antigenic specificities (designated SLB-2 to SLB-7) detectable with polyclonal reagents in the complement-dependent lymphocytotoxic test were determined in the SLB alloantigenic system of pigs. The SLB specificities (including the previously described SLB-1) are controlled by 13 alleles. An analysis of a further 121 double backcross matings (with 817 offspring) confirmed that the products of SLB alleles segregated independently of the SLA haplotypes (swine MHC). In addition, in 112 families of the same mating type (with 746 progeny) the map distance of the SLB locus and the highly polymorphic L blood group locus was found to be 7.82 centimorgans in the heterogametic sex and 12.57 centimorgans in the homogametic sex.

Alleles↗

The devitalization of the stomach by a devascularization technique.

Different parts of the stomach in healthy animals (pigs) were devitalized (devascularized) by eliminating the blood supply and left in situ. In model experiments, parts of the stomach were chosen for the devitalization with respect to most frequent location of carcinomas. The operation technique consisted of a complete ligature of perigastric vessels, bling enclosure of the cranial part of the duodenum with two ligatures with a possible application of a seromuscular suture and transverse cutting of the oral part of the stomach and its enclosing by a suture with a partial invagination of the wall. The passage was renewed by a gastroenteroanastomosis, some of the used modifications of the operation technique according to Billroth II (Schlater-Pólya). A method was mostly proposed with an interposition of the jejunal loop, which bridged the site of the devitalized part of the stomach. In 10 to 20 weeks the stomach tissue was stepwise degraded and it was replaced by fibrous tissue. The operation technique is simple; it could be applicable as a palliative or possibly therapeutic intervention in the case of inoperable tumours in human practice.

Animals↗

A technique of the segmental devitalization of kidneys in experiment.

With the help of partially overlapping parenchymatous sutures, different renal segments were devitalized. After 8 weeks, a fibrous conversion occurred in devitalized part of the kidney. For the orientation during establishing the sutures, a topographic schematic diagram was used, which is simple and quite satisfactory for the use in practice. The study was performed on minipigs (21 x unilaterally, 3 x bilaterally) without any exitus of the experimental animals. A long series of experiments was performed in 37 rats on one kidney and in 40 rats bilaterally. Only in 3 rats the cause of the exitus was obviously in a connection with eliminating more than 70 to 75% of the renal parenchyma from the function. The devitalization technique presents good results in both series of the experiments. Possibilities of its use in the clinical practice in cases, where the resection therapy is being used currently, are discussed.

Animals↗

Porcine fetal antibodies to maternal granulocytes and lymphocytes.

Immunological tests were used to determine the presence of antibodies to maternal lymphocytes and granulocytes in the blood serum of porcine fetuses and 4-week-old young. Enzyme-linked immunosorbent assay showed that the presence of these antibodies in fetal blood was an exception. No cross-reactions of these sera with porcine reproductive tract antigens were demonstrated.

Animals↗

Morphology of regressive changes in the kidney following experimental ischaemia.

Total or partial renal ischaemia was studied in rats and pigs in a series of experiments, after total devascularization (ligature of renal vessels and ureter); ligature of isolated renal artery; experimental embolization of renal artery by non-resorbable material; devascularization of a specific renal segment by deep, strangulating sutures. Morphological studies showed that the resulting necrotization is represented most frequently by coagulation necrosis, less frequently by colliquating necrosis. Occlusion of the renal artery by embolization or ligature leads only to partial necrotization or atrophy. Total devascularization of the renal hilus or total devascularization of a specific renal segment by strangulating sutures leads to total destruction of the devascularized (devitalized) tissues. Within 8-12 weeks, the whole organ changes into a small residue of connective tissue. The possibility of the use of the devascularization technique in the treatment of tumours is discussed.

Animals↗

Localization of the Po2 locus in the S, Phi, Hal, H, Po2, Pgd linkage group in pigs.

The localization of the Po2 locus controlling a polymorphic serum postalbumin was studied in 41 families of the Czech Landrace breed. The haplotypes involving six closely linked loci (S, Phi, Hal, H, Po2, Pgd) were determined for each family member. The crossovers observed between the H, Po2 and Pgd loci indicated that Po2 is located between H and Pgd. The Po2 locus appears to be closer to H [theta = 0.54% (0.06%-1.92%)] than to Pgd [theta = 4.02% (1.67%-7.96%)]. A strong Ha-Po2S association (r = 0.96, P less than 0.001) and H-PO2 linkage disequilibrium (D = 0.2218, P less than 0.01, D/DMax = 0.98) were found.

Alpha-Globulins↗