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Biomedical subjects

D Kwiatkowska

Publications and source records attributed to D Kwiatkowska.

At least 19 recordsLinked to original sources

The effect of endotoxin administration on cytokine production in obstructive jaundiced rats.

The aim of this study was to evaluate the effect of lipopolysaccharide (LPS) administration, which mimics a surgical intervention, on the immune status of obstructive jaundiced (OJ) and sham-operated control rats. Rats were given 20 microgram LPS intraperitoneally on day 13 following bile duct ligation or sham surgery. We determined serum levels of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) on day 14 after surgery, and spontaneous as well as LPS-induced production of these cytokines in splenocyte and peritoneal exudate cell (PEC) cultures. We found that IL-6, but not TNF-alpha, serum concentrations were significantly elevated (4-fold) in OJ rats treated with LPS compared with LPS-untreated OJ rats. In sham-operated rats the differences between the respective groups were not significant. The production of TNF-alpha by splenocyte and PEC cultures was depressed in OJ rats treated with LPS; in particular, a very deep decline was observed in the case of spontaneous TNF-alpha production in PEC cultures. In contrast, TNF-alpha production in LPS-untreated and LPS-treated sham-operated rats did not differ. In the case of IL-6 production by splenocytes and PEC cultures, we observed a significant suppression of this cellular function in both OJ and sham-operated rats treated with LPS when compared with the respective controls. In conclusion, the results indicate that the already depressed cytokine production in OJ rats leads to even deeper hyporeactivity following LPS challenge. Lack of TNF-alpha suppression upon LPS treatment in sham-operated rats suggests that surgery-elicited hyporeactivity is mediated by a different mechanism than that leading to immune hyporesponsiveness in OJ. Our findings may explain the relatively high mortality rates observed of OJ patients subjected to surgery.

Animals↗

The impairment of wound healing process is correlated with abnormalities of TNF-alpha production by peritoneal exudate cells in obstructive jaundiced rats.

The wound healing process and production of tumour necrosis factor alpha (TNF-alpha) by peritoneal cells of 7-day and 14-day obstructive jaundice (OJ) and sham-operated rats were investigated. In the study the skin wound breaking strength was measured. In addition such histological and biochemical parameters as fibroblast and endothelial cell proliferation, inflammatory cell infiltration and hydroxyproline content were evaluated in polyurethane sponge discs implanted subcutaneously into rats. TNF-alpha production by peritoneal exudate cells (PEC), both spontaneous and lipopolysaccharide (LPS)-induced was determined by a bioassay. In OJ rats the process of both early as well as late phase of healing was impaired. The breaking strength of skin wound was decreased, the fibroblast and endothelial cell proliferation and collagen deposition, as well as hydroxyproline content were diminished. In 7 day OJ the numbers of inflammatory cells in the implants were lowered with a subsequent slight increase on day 14 of OJ. The spontaneous and LPS induced TNF-alpha production by PEC were significantly higher in 7 day OJ as compared with sham-operated controls. On day 14 of OJ the LPS-induced TNF-alpha level was, in contrast, much lower and did not differ much from the spontaneous TNF-alpha production. We conclude that the impairment of wound healing in OJ results from disturbances in functioning of the immune system caused by systemic endotoxaemia.

Animals↗

[Adhesive glycoproteins of the extracellular matrix].

Glycoproteins of the extracellular matrix and basement membrane play crucial role in cell recognition, adhesion, migration and proliferation. Morphogenesis and tissue and organ development in embryo, as well as healing processes in adults are dependent on the interactions of extracellular proteins with cell surface receptors. In this paper the structure and function of major adhesive proteins: fibronectin, tenascin, laminin and nidogen are reviewed. Recent data on the role of small leucine-rich and modular proteoglycans, such as decorin, biglycan, fibromodulin, versican, perlecan, neurocan, agrecan, agrin and other are given. Modifications of the expression and localization of extracellular glycoproteins in pathology are also presented.

Adult↗

Platinum complexes and pyruvate kinase activity.

The interaction of platinum complexes with bovine heart pyruvate kinase (PK) was studied by absorption, CD, fluorescence spectroscopy and enzymic activity test. Our results showed that activity of PK was reduced by cis-DDP and potassium tetrachloroplatinate in a time-and concentration dependent manner. Cis-DDP was less effective than K2PtCl4 in reducing PK activity. The native enzyme showed well defined negative Cotton effect at 222 and 208 nm indicating the presence of alpha-helical and beta structure. Platinum binding lowered the Cotton effect in this region by about 10-20% and 30-50% for the system with cis-DDP and K2PtCl4, respectively. Fluorescence study showed that platinum binding quenched tryptophan fluorescence suggesting that binding occurs at the tryptophan residue or its proximity. PK modifications induced by platinum binding would result in a greater resistance to denaturing agents.

Animals↗

[Metabolism of nicotine--mechanism and clinical effects of toxicity].

Nicotine is a major constituent of tobacco and exerts a number of physiological effects. Metabolism of nicotine in animal and human tissues is reviewed. Clinical considerations regarding the problems of toxic effects of nicotine in human and molecular basis of carcinogen mechanism of nicotine derivatives are discussed.

Animals↗

Structure and expression of the cDNA for the C isozyme of phosphofructo-1-kinase from rabbit brain.

Rabbit brain contains three phosphofructo-1-kinase (PFK) isozymic subunits designated A, B, and C. The primary structures of the first of these two isozyme types have been determined previously. The isozyme C of rabbit brain was isolated by immunoaffinity chromatography and subjected to proteolytic and chemical digestion. A large number of peptides were sequenced, the total number of amino acids identified being equal to about 80% of the total structure. The sequence of the cDNA derived from brain mRNA for C isozyme was determined from polymerase chain reaction fragments synthesized using oligonucleotides designed on the basis of the peptide sequences. The deduced size of the C isozyme was 86,371 Da, slightly larger than PFKs described previously. The amino acid sequence identity with the rabbit A isozyme was 68.9% and a range of identity to other sequenced mammalian PFKs was 67-69%. Using these data plus previously published data on chemical modification, assignments of the 6 organic ligand binding sites of PFK were inferred. The full-length cDNA was cloned into and expressed in Escherichia coli. Phosphofructokinase C was purified to homogeneity from the bacterial extracts.

Amino Acid Sequence↗

[Proteins modified in the nonenzymatically glycosylation reaction (AGE-proteins)--new markers for diabetes?].

Paper present a recent review on the formation and clinical significance of advanced glycosylation end products, produced in nonenzymatically glycosylation, called Maillard reaction. The special attention was paid to AGEs role in diabetic and aging processes. Instant of occurring of AGEs in circulation or increase of AGE receptor concentration are many years faster than clinical pathology of vessels, nervous or kidneys connect with diabetes or aging. May be in the future it will be possible to decrease the consequence of Maillard reaction by using pharmacology drugs.

Aging↗

[Surgical treatment of ventricular tachycardia in patients with post-infarction aneurysms].

4 patients (P) with recurrent, sustained ventricular tachycardia (VT) resistant to medical treatment, underwent surgery for cure of this arrhythmia. Each P had episodes of VT lasting 30 or more seconds, 3 of them had episodes of ventricular fibrillation. In all cases rhythm disturbances were secondary to post myocardial infarction aneurysm. Coronary angiography showed in all P total occlusion of LAD, in 2 cases significant lesion in RCA were found. 1 P had lung cancer. All P underwent aneurysmectomy and an excision of the altered endocardium by Harken's method. The endocardial excision was performed without endocardial mapping. 2 P had concomitant CABG to RCA. In the P with lung cancer lobectomy was performed. There were 2 ++non-arrhythmic death. The P with lung cancer died because of sepsis due to lung abscess. One P died because of heart failure (preoperative EF 10%), 6 months after the surgery. The 2 survivors remained free of VT during a follow-up period 8 months. In conclusion, endocardial excision by Harken's method is efficient in treating recurrent sustained VT, resistant to medical treatment, in patients with post myocardial infarction aneurysm. The surgical procedure can be performed without intraoperative endocardial mapping.

Endocardium↗

[Analysis of causes for hospitalization in the pediatric department of a regional hospital in Trzcianka in the years 1981-1985].

The authors analysed 2,575 case histories of children hospitalised in 1981-1985 at pediatric department of regional hospital in Trzcianka (Piła region) taking to account: age, sex, permanent residence, duration of stay at a hospital, disease causing hospitalisation. Within the period studied both decrease of number of neonates (1-6 months old) and young children of (1-3 years old) admitted to the hospital and a tendency toward shortening time of hospitalisation were found. Among causes of hospitalisation the first were respiratory tract infections (88.3% of all admissions in 1981 but 69.8% in 1985), and acute diarrheas (22.1% of all admissions in 1981, 17.1% in 1985).

Child↗

The sites of phosphorylation of rabbit brain phosphofructo-1-kinase by cyclic AMP-dependent protein kinase.

The three isozymic subunits of phosphofructo-1-kinase present in rabbit brain and designated A, B and C were phosphorylated in vitro by cyclic AMP-dependent protein kinase with 32P-labeled ATP. Limited digestion of the labeled enzymes with trypsin or with Staphylococcus aureus V8 proteinase led to the solubilization of radiolabeled peptides derived from the three isozymic subunits. Limited digestion by V8 proteinase was accompanied by a slight reduction in the apparent sizes of the subunits, indicating that the phosphorylated sites are located near either the amino or carboxyl termini of the protein. V8 proteinase digestion led to no change in the maximal activity of the enzyme but did abolish sensitivity to ATP inhibition. The phosphopeptides of the tryptic and the V8 digests were purified by chromatography and their amino acid sequences were determined and compared to the previously established sequence from rabbit muscle isozyme A. PFK-A E H I S R K R S G E A T V PFK-B H V T R R S L S M A K G F PFK-C V S A S P R G S Y R K F L In each instance, the phosphorylated serine, underlined in the above sequences, was found to be one or two residues toward the C-terminus of one or more basic residues. No other similarities in structure were noted.

Adenosine Triphosphate↗

Identification of the AMP binding sites of rabbit phosphofructo-1-kinase isozymes B and C.

Rabbit liver phosphofructo-1-kinase, designated isozyme B, and rabbit brain phosphofructokinase, which contains all three isozymes as heteropolymers, have been modified by [14C]fluorosulfonylbenzoyladenosine (FSBAdo). Several lines of evidence supported modification at the binding site for AMP. The modification proceeded to the extent of 2 to 4 mol of reagent incorporated per mol of tetramer, and AMP protected against the reaction. The kinetic properties of modified isozymes A and B and of modified brain phosphofructokinase were examined and compared to their unmodified forms. It was observed that modification greatly diminished ATP inhibition of all of the isozymes. Furthermore, equilibrium binding studies of modified phosphofructokinase B showed a greatly diminished capacity and affinity for cyclic AMP. Cyclic AMP had little or no influence on the properties of modified A isozyme or brain phosphofructokinase, but was capable of further deinhibiting modified B isozyme, apparently at sites remaining unmodified by FSBAdo. Phosphofructokinase B, modified by radiolabeled FSBAdo, was digested by trypsin, and the digest separated by high-pressure liquid chromatography. The labeled peptide was isolated and sequenced to provide the sequence: Asn-Tyr-Gly-Thr-Lys-Leu-Gly-Val-Lys, with the lysine in the fifth position being the site of modification. To isolate isozyme C, a monoclonal antibody to this isozyme was produced by injecting purified rabbit brain phosphofructokinase into mice, and subsequently selecting for those clones that recognized brain phosphofructokinase but not purified phosphofructokinases A and B. The selected monoclonal was specific for native rabbit isozyme C and would not recognize mouse or rat brain phosphofructokinases. Linking the antibody to an inert phase provided an efficient means of purifying rabbit isozyme C from rabbit brain. The enzyme so recovered retained little of its original activity, but the method provided a simple technique for the preparation of enzyme for protein chemistry studies. The modified C isozyme was isolated on the immuno-affinity column and digested with trypsin. A tryptic peptide bearing the label was isolated and sequenced to provide the structure: Asn-Phe-Gly-Thr-Lys-Ile-Ser-Ala-Arg, with position 5 being the site of modification. The sequences of isozymes B and C are homologous to the site of modification of the A isozyme by FSBAdo.

Adenosine↗

Inorganic pyrophosphate: fructose-6-phosphate 1-phosphotransferase of the potato tuber is related to the major ATP-dependent phosphofructokinase of E. coli.

A procedure was developed for the purification of inorganic pyrophosphate: fructose-6-phosphate 1-phospho-transferase (PPi-PFK) from potato tubers. The enzyme has the structure alpha 4 beta 4 with a subunit of 68 kDa and a beta subunit of 60 kDa. The structural relationship of this enzyme to other PFKs and to fructose bisphosphatase was examined by immunoprecipitation and immunoblotting. Antibodies to the plant enzyme did not react with E. coli PFK. No cross-reaction was seen among the following enzymes or their antibodies: yeast fructose bisphosphatase; rabbit PFKs A, B, or the enzyme from brain; and the two subunits of the potato PPi-PFK. On the other hand, antibody to E. coli PFK-1 strongly cross-reacts with the 60 kDa polypeptide but not 68 kDa peptide.

Escherichia coli↗

Phosphofructokinase in the rat nervous system: regional differences in activity and characteristics of axonal transport.

The distribution of phosphofructokinase (PFK) in gray and white matter regions of the rat nervous system was evaluated. Determinations of PFK activity revealed that cell body enriched regions (sensorimotor cortex) had a significantly higher level of activity than axonal regions (sciatic nerve, dorsal roots, and optic nerve). The level of PFK activity was also significantly higher in central axons (optic nerve) than in peripheral axons (sciatic nerve). Differences in PFK activity could be largely attributed to differences in tissue content of the enzyme rather than to differences in the types of PFK isozymes present. Cortex contained significantly larger amounts of PFK relative to total protein than did peripheral nerve. However, purification of PFK revealed that all three of the PFK isozymes, C (86 kd), A (84 kd), and B (80 kd), were present in both cortex and sciatic nerve. Both SDS/PAGE and immunoblotting studies using PFK isozyme-specific antibodies demonstrated that the relative proportions of the three PFK isozymes were similar in cell body and axonal regions of the nervous system. The PFK-C and PFK-A isozymes each comprised about half the total and only small amounts of the PFK-B isozyme were present in both regions. However, immunoprecipitation experiments suggested that quantitatively different proportions of the possible PFK hybrids (tetramers) may be distributed between axonal and cell body regions. The transport of PFK was examined in this study and PFK was identified in slow component b (SCb) of axonal transport. SCb moves at a rate of 2-4 mm/day in rat axons and is known to contain several other enzymes of intermediary metabolism as well as actin. The finding that PFK, the rate limiting enzyme in glycolysis, is present in SCb lends support to the hypothesis that glycolytic enzymes are not freely diffusing proteins in axons but, instead, are present as organized assemblies that have long-term, yet flexible, associations with structural elements of the cytoplasm.

Animals↗