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1.
Adv Med Sci ; 58(2): 185-95, 2013.
Article in English | MEDLINE | ID: mdl-24327530

ABSTRACT

In recent years, a constant growth of knowledge and clinical applications of stem cells have been observed. Mesenchymal stromal cells, also described as mesenchymal stem cells (MSCs) represent a particular cell type for research and therapy because of their ability to differentiate into mesodermal lineage cells. The most investigated source of MSCs is bone marrow (BM). Yet, collection of BM is an invasive procedure associated with significant discomfort to the patient. The procedure results in a relatively low number of these cells, which can decrease with donor's age. Therefore, it seems to be very important to find other sources of mesenchymal stem cells nowadays. A human placenta, which is routinely discarded postpartum, in spite of its natural aging process, is still a rich source of stem cells capable to proliferate and in vitro differentiate in many directions. Besides homing and differentiation in the area of injury, MSCs there elicit strong paracrine effects stimulating the processes of repair. In this review, we focus on the biology, characteristics and potential clinical applications of cells derived from human fetal membranes: amnion and chorion.


Subject(s)
Amnion/cytology , Chorion/cytology , Extraembryonic Membranes/cytology , Mesenchymal Stem Cells/cytology , Placenta/cytology , Cell Differentiation , Cell- and Tissue-Based Therapy , Female , Humans , Mesoderm/cytology , Pregnancy
2.
Bone ; 51(3): 546-52, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22627031

ABSTRACT

Peri-implant tissue reactions in failed total ankle replacement (TAR) are characterized by early developing peri-implant osteolysis. The hypothesis of the study was that this reaction is mediated by receptor activator of nuclear factor kappa B ligand (RANKL). Samples of peri-prosthetic tissues from failed TAR implants were stained for macrophages, RANKL, its receptor RANK and osteoprotegerin (OPG), and compared to control samples. The failed TAR implants were surrounded by implant capsule, synovial lining-like interface membrane or necrotic tissues. Infiltrating scavenger receptor I positive CD163(+) macrophages were frequent, in particular around necrotic soft tissues or bone sequestrate, and possibly in part formed due to ischemia and mechanical factors. In contrast, implant-derived wear debris was scanty. Still many RANK(+) macrophages were often seen in close contact with RANKL(+) mesenchymal cells, whereas OPG was mostly located at a distance in vascular endothelial cells. Foreign body giant cells were frequent. RANKL seems to stimulate locally accumulated CD163(+) RANK-expressing cells to fusion, which leads to the local formation of multinuclear foreign body giant cells (and probably of osteoclasts). Therefore, peri-implant osteolysis in early TAR implant failure seems to be caused by the RANKL-driven chronic foreign body inflammation directed against, not implant-derived particles, but against necrotic autologous tissues.


Subject(s)
Arthroplasty, Replacement, Ankle , Osteolysis/metabolism , Prostheses and Implants , RANK Ligand/metabolism , Aged , Ankle/pathology , Antigens, CD/metabolism , Antigens, Differentiation, Myelomonocytic/metabolism , Humans , Immunohistochemistry , Middle Aged , Osteolysis/pathology , Prosthesis Failure , Receptor Activator of Nuclear Factor-kappa B/metabolism , Receptors, Cell Surface/metabolism
3.
Arthritis Rheum ; 64(8): 2663-8, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22488408

ABSTRACT

OBJECTIVE: The conventional H(1) and H(2) histamine receptors have >10,000-fold lower avidity for histamine than H(4) histamine receptor, which has been implicated in autoimmune diseases. This study was undertaken to compare H(4) histamine receptor levels in the salivary glands (SGs) of healthy controls with those in the SGs of patients with primary Sjögren's syndrome (SS). METHODS: H(4) histamine receptor messenger RNA (mRNA) was analyzed using real-time quantitative polymerase chain reaction, and the receptor protein was examined using immunostaining. Effects of the H(4) histamine receptor agonist ST-1006 on cytokine synthesis by human SG (HSG) cells were analyzed using xMAP technology and enzyme-linked immunosorbent assay. RESULTS: Healthy SGs contained H(4) histamine receptor mRNA. The receptor protein was localized to the acinar and ductal epithelial cells. H(4) histamine receptor agonist stimulated HSG cells to produce the cytokines interleukin-8 and vascular endothelial growth factor. SS patients had low H(4) histamine receptor levels. CONCLUSION: H(1) and H(2) histamine receptor antagonists are not effective in the treatment of autoimmune diseases. However, such antagonists do not affect the newly discovered H(4) histamine receptor. Dendritic cells and lymphocytes are nonprofessional histamine-producing cells, which produce histamine at 100-1,000-fold lower rates than mast cells do. Saliva contains only 0.31-12.4 ng/ml histamine, which is too low to stimulate H(1) or H(2) histamine receptor, but stimulates H(4) histamine receptor half maximally. Our findings show that H(4) histamine receptor is strongly expressed in tubuloacinar SG cells, which emphasizes the role of these cells in the pathogenesis of SS.


Subject(s)
Receptors, G-Protein-Coupled/metabolism , Receptors, Histamine/metabolism , Salivary Glands/metabolism , Sialadenitis/etiology , Sialadenitis/metabolism , Sjogren's Syndrome/complications , Adult , Case-Control Studies , Cells, Cultured , Female , Humans , Interleukin-8/metabolism , Male , Middle Aged , Piperazines/pharmacology , Pyrimidines/pharmacology , RNA, Messenger/metabolism , Receptors, Histamine H4 , Salivary Glands/cytology , Sialadenitis/pathology , Vascular Endothelial Growth Factor A/metabolism
4.
Oral Dis ; 16(7): 577-85, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20412448

ABSTRACT

Usually no distinction is made between female and male salivary glands although cyclic changes of and / or differences in serum and salivary sex steroid concentrations characterize women and men. Moreover, sexual dimorphism is well recognized in salivary glands of rodents.Salivary glands contain estrogen and androgen receptors and are, according to modern high throughput technologies,subjected to gender differences not explainable by gene dose effects by the X chromosome alone. Because sex steroids are lipophilic, it is often thought that approximately 10% of them passively diffuse from plasma to saliva. Indeed, saliva can find use as sample material in sports medicine, pediatrics, veterinary medicine and behavioral sciences. Last but not least, humans and other primates are unique in that they have a reticular zone in their adrenal cortex, which produces dehydroepiandrosterone and androstendione pro-hormones. These are processed in peripheral tissues, not only in female breast and uterus and male prostate, but also in salivary glands by an intracrine enzymatic machinery to active 17b-estradiol,dihydrotestosterone and others, to satisfy and buffer against a constantly changing needs caused by circadian,menstrual, pregnancy and chronobiological hormonal changes in the systemic circulation. Female dominance of Sjögren's syndrome and certain forms of salivary gland cancer probably reflect these gender-based differences.


Subject(s)
Gonadal Steroid Hormones/metabolism , Salivary Glands/metabolism , Aging/metabolism , Androstenedione/metabolism , Dehydroepiandrosterone/metabolism , Female , Humans , Male , Pregnancy , Receptors, Steroid/metabolism , Saliva/metabolism , Salivary Gland Diseases/metabolism , Sex Characteristics , Zona Reticularis/metabolism
5.
Vet Pathol ; 46(6): 1292-300, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19605905

ABSTRACT

The aim of this study was to check the balance between tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and interleukin-10 (IL-10) in well-developed end-stage disk disease in the disk itself as well as in paradiskal spine. In 6 domestic pigs the cranial bony end plate of the L4 vertebra was perforated to the nucleus pulposus. At 3 months the degenerated experimental and contiguous control disks, together with the adjoining bony and cartilaginous vertebral end plates, bone marrow, and spinal ligaments, were excised and used for immunohistochemical analysis. In general, there were more TNF-alpha and in particular IL-10 positive cells in the degenerated disks than in the control disks, whereas the number of IL-6 labeled cells did not differ among sites or between control and experimental intervertebral disks. These results suggest that TNF-alpha and IL-10 are involved in the late reparatory phases of the experimental disk lesion. Use of an experimental model showed that strictly disk-directed manipulation and degeneration are also reflected in the contiguous vertebrae, including adjoining cartilage, bone, marrow, and ligaments.


Subject(s)
Interleukin-10/metabolism , Interleukin-6/metabolism , Intervertebral Disc Degeneration/veterinary , Swine Diseases/metabolism , Tumor Necrosis Factor-alpha/metabolism , Animals , Immunohistochemistry , Interleukin-10/genetics , Interleukin-6/genetics , Intervertebral Disc/metabolism , Intervertebral Disc/pathology , Intervertebral Disc Degeneration/metabolism , Intervertebral Disc Degeneration/pathology , Swine , Swine Diseases/pathology
6.
Clin Exp Rheumatol ; 27(2): 299-306, 2009.
Article in English | MEDLINE | ID: mdl-19473572

ABSTRACT

OBJECTIVE: This study was designed to clarify the role of the receptor activator of nuclear factor kappa B ligand (RANKL) in the process of discus degeneration and spondylarthrosis. It was hypothesized that experimental discus lesion would initiate not only local bone remodelling but also increased osteoclast formation on a location remote to the injury site due to altered spinal biomechanics. It was speculated that these changes in vertebral bone remodelling could be reflected in an increased RANKL expression. METHODS: The presence of RANKL in the spine was studied in an experimental perforating lesion of the cranial endplate of L4 and the adjoining disc in six domestic pigs and in one human herniated disc. After three months, the experimental and contiguous control vertebrae, complete with intervertebral discs, were subjected for immunohistochemistry. RESULTS: This is the first study to show that RANKL was locally seen (produced) in osteoblasts, fibroblasts replacing annulocytes and mesenchymal bone marrow cells and, in part, apparently bound to the surface of osteoclasts and macrophage-like prefusion macrophages. Such RANKL induction was also seen at sites remote from the experimental lesion driving the whole process. More RANKL-positive cells were found in close proximity to the endplate than in the central parts of the vertebrae. Osteocytes in bone matrix and most bone marrow cells in the marrow microenvironment showed no RANKL staining. Human annulus fibrosus also contained RANKL, RANK and OPG. CONCLUSIONS: We have demonstrated that RANKL is produced locally, also in soluble form, at the site of injury and also in intact vertebrae and bony structures likely due to altered biomechanics. It seems to be engaged in bone healing and remodelling, essentially proving our working hypothesis. These secondary bone changes could represent part of the degenerative spine disease (spondylarthrosis). RANKL inhibitors, like recombinant human osteoprotegerin (OPG), could be interesting drugs to test, not only in osteoporosis, but also in spondylarthrosis.


Subject(s)
Intervertebral Disc Displacement/metabolism , Intervertebral Disc/metabolism , Lumbar Vertebrae/metabolism , RANK Ligand/metabolism , Spondylarthropathies/metabolism , Animals , Bone Remodeling/physiology , Disease Models, Animal , Fibroblasts/metabolism , Humans , Intervertebral Disc/injuries , Intervertebral Disc/pathology , Lumbar Vertebrae/injuries , Lumbar Vertebrae/pathology , Mesenchymal Stem Cells/metabolism , Osteoblasts/metabolism , Sus scrofa
7.
Clin Exp Rheumatol ; 26(6): 1018-26, 2008.
Article in English | MEDLINE | ID: mdl-19210865

ABSTRACT

OBJECTIVE: To analyze immunohistochemically the localization of the VEGF receptors in experimental intervertebral disc degeneration tissues in a pig model. MATERIALS AND METHODS: In six domestic pigs, the cranial bony endplate of the L4 vertebra were perforated into the nucleus pulposus. Three months postoperatively, the animals were sacrificed and the experimental and control vertebrae, complete with intervertebral discs, were excised and subjected for immunohistochemical staining of vascular endothelial growth factor receptors (VEGFR) along with VEGF - A, -C, -D and blood and lymphatic vessel markers vWF and LYVE-1. RESULTS: The results of immunohistochemical analysis of experimental samples showed VEGFR-1 (Flt-1) expression in intervertebral disc and all paradiscal tissues studied. In control samples expression of VEGFR-1 was lower and absent in the intervertebral discs. Comparatively less of VEGFR-2 (KDR/Flk-1) and VEGFR-3 (Flt-4) than VEGFR-1was found in degenerated intervertebral discs and paradiscal tissues. In contiguous control intervertebral discs and control paradiscal tissues VEGFR-2 and-3 receptors were expressed to a lower extent than in experimental tissues or were even totally absent. Also growth factors VEGF-A, -C, -D, as well as von Willebrand factor and to a much lower extent LYVE-1 were differently expressed in experimental and control intervertebral discs. CONCLUSION: In experimental intervertebral disc degeneration, VEGF receptors were expressed in the damaged disc and paradiscal tissues. In the same tissues, VEGF-A, -C, and -D, signs of blood and lymphatic vessel in-growth and reactive/adaptive vertebral bone remodelling were found.


Subject(s)
Bone Remodeling/physiology , Intervertebral Disc Displacement/metabolism , Intervertebral Disc Displacement/physiopathology , Neovascularization, Physiologic/physiology , Receptors, Vascular Endothelial Growth Factor/metabolism , Animals , Antibodies , Blood Vessels/metabolism , Immunohistochemistry , Intervertebral Disc/blood supply , Intervertebral Disc/pathology , Intervertebral Disc Displacement/pathology , Lumbar Vertebrae/blood supply , Lumbar Vertebrae/pathology , Lymphatic Vessels/metabolism , Receptors, Vascular Endothelial Growth Factor/immunology , Sus scrofa , Vascular Endothelial Growth Factor A/metabolism , Vascular Endothelial Growth Factor C/metabolism , Vascular Endothelial Growth Factor D/metabolism , Vascular Endothelial Growth Factor Receptor-1/immunology , Vascular Endothelial Growth Factor Receptor-1/metabolism , Vascular Endothelial Growth Factor Receptor-2/immunology , Vascular Endothelial Growth Factor Receptor-2/metabolism , Vascular Endothelial Growth Factor Receptor-3/immunology , Vascular Endothelial Growth Factor Receptor-3/metabolism , von Willebrand Factor/immunology , von Willebrand Factor/metabolism
8.
Ann Rheum Dis ; 64(8): 1229-32, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16014685

ABSTRACT

OBJECTIVES: To analyse some scavenging related molecules in Wegener's granulomatosis (WG) macrophages. METHODS: Immunohistochemical staining of lung, nasopharynx, and skin for macrophage markers related to scavenging (macrophage scavenger receptor MARCO, collagenase-1 and gelatinase-B), formation of multinuclear foreign body giant cells (ADAM 9/meltrin-gamma and ADAM 12/meltrin-alpha), and cell debris derived from neutrophils, endothelial cells and mast cells (specific granule protein 28 (SGP28), von Willebrand factor (vWF) and mast cell tryptase, respectively). TechMate staining robot and biotin-streptavidin protocol were used. RESULTS: Some macrophages were activated and expressed collagenase-1 and gelatinase-B. Approximately 5% of macrophages expressed scavenger receptor, whereas 20-30% were meltrin positive. Interstitial and granuloma associated macrophages and giant cells contained partly undigested, immunoreactive SGP28-, vWF- and tryptase-positive cell rests and collagenous matrix. Lymphocytic follicles with germinal centres were found in the same areas. CONCLUSION: In WG tissue lesions macrophage and giant cells seem to be overwhelmed by the bulk to be scavenged. Despite cellular activation and continuing maturation to professional scavenger receptor (MARCO) and meltrin positive multinuclear giant cells combined with an organisation into granulomas, macrophages still contain partially undigested cell and tissue rests. This necrotic and damaged self may be the driving force for the formation of giant cell ("foreign body") granulomas. This, together with the local formation of secondary lymphatic follicles (with germinal centres), indicates active local antigen processing and presentation.


Subject(s)
Granulomatosis with Polyangiitis/metabolism , Macrophages/metabolism , Collagenases/metabolism , Giant Cells, Foreign-Body/metabolism , Giant Cells, Foreign-Body/pathology , Granulomatosis with Polyangiitis/pathology , Humans , Macrophage Activation , Macrophages/pathology , Macrophages, Alveolar/metabolism , Macrophages, Alveolar/pathology , Matrix Metalloproteinase 9/metabolism , Nasopharynx/metabolism , Nasopharynx/pathology , Phagocytosis , Receptors, Immunologic/metabolism , Receptors, Scavenger , Skin/metabolism , Skin/pathology
10.
Clin Rheumatol ; 22(3): 196-202, 2003 Sep.
Article in English | MEDLINE | ID: mdl-14505210

ABSTRACT

The aim of this study was to analyse microvascular damage and compensatory angiogenesis in skin from patients with systemic sclerosis (SSc) compared with systemic lupus erythematosus (SLE), Raynaud's phenomenon (RP) and healthy controls. Immunohistochemistry was used for skin biopsies (9 SSc, 10 SLE, 9 RP and 12 healthy controls) using von Willebrand factor and beta3 integrin subunit specific antibodies, TechMate immunostaining robot and biotin-streptavidin protocol. In the early stages of SSc, vWF was found in the perivascular space and interstitial matrix in papillary but not in the reticular dermis, in particular around small oedematous blood vessels infiltrated by mononuclear cells. The extravascular release of vWF in SSc specimens was associated with weak or even a total lack of immunoreactivity within the associated endothelial cells. Late stages of SSc were characterised by loss of the dermal papillae, subepidermal fibrosis, hypovascularity and strong endothelial vWF expression without extravascular leakage. In all SSc patients studied only a few vascular profiles were weakly immunostained for beta3 integrin subunit. This work demonstrates that vWF is not only released into the systemic circulation, but is also leaked to the perivascular space/matrix. This local release and deposition of vWF is probably a sensitive and early marker of microvascular involvement in SSc pathogenesis. Local vWF release may play a role in platelet adhesion, aggregation, thrombogenesis and dermal connective tissue remodelling. In spite of some attempts towards compensatory angiogenesis in SSc, as evidenced by beta3 integrin subunit expression, it was evident that the angiogenic response was not able to prevent the development of hypovascularity during the advanced stages of the disease.


Subject(s)
Endothelium, Vascular/pathology , Lupus Erythematosus, Systemic/pathology , Neovascularization, Physiologic/physiology , Raynaud Disease/pathology , Scleroderma, Systemic/pathology , Biopsy, Needle , Case-Control Studies , Female , Humans , Immunohistochemistry , Lupus Erythematosus, Systemic/physiopathology , Male , Prognosis , Raynaud Disease/physiopathology , Reference Values , Risk Assessment , Scleroderma, Systemic/physiopathology , Severity of Illness Index , von Willebrand Factor/analysis
11.
Clin Exp Rheumatol ; 21(1): 41-8, 2003.
Article in English | MEDLINE | ID: mdl-12673888

ABSTRACT

OBJECTIVE: To analyse the expression of factors potentially involved in skeletal muscle degeneration and regeneration in dermatomyositis (DM), systemic sclerosis (SSc), polymyositis (PM), systemic lupus erythematosus (SLE) and non-inflammatory myopathies. METHODS: Immunohistochemical staining of skeletal muscle biopsies (10 DM, 10 SSc, 10 PM, 10 SLE, 10 non-inflammatory myopathies) for tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), activated caspase-1, pan-macrophage marker CD68, inducible nitric oxide synthase (NOS2) and nerve growth factor receptor (NGFR). TechMate staining robot and biotin-streptavidin protocol were used. RESULTS: Expression of TNF-alpha, IL-1 beta, caspase-1 and NOS2 was found in the cytoplasm and sarcolemma of dystrophic skeletal muscle fibres. TNF-alpha and IL-1 beta immunoreactive profiles were faint and few and close to satellite nuclei-containing regenerating muscle fibres both in inflammatory and non-inflammatory myopathies. NGFR expression was found in comparable areas. In non-inflammatory inherited myopathies more nuclei were caspase-1 immunoreactive whereas caspase-1 expression was rarely seen in inflammatory myopathies, implying regeneration of the affected muscle fibres. CONCLUSION: Prominent expression of the proinflammatory factors TNF-alpha, IL-1 beta and NOS2 and caspase-1 is associated with muscle fibre damage, albeit when expressed to a low degree these factors may, like NGFR, contribute to muscle regeneration and healing.


Subject(s)
Caspase 1/metabolism , Dermatomyositis/metabolism , Interleukin-1/metabolism , Polymyositis/metabolism , Receptor, Nerve Growth Factor/metabolism , Tumor Necrosis Factor-alpha/metabolism , Adolescent , Adult , Aged , Biomarkers/analysis , Dermatomyositis/etiology , Dermatomyositis/pathology , Female , Humans , Immunohistochemistry , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/metabolism , Lupus Erythematosus, Systemic/pathology , Male , Middle Aged , Muscle, Skeletal/metabolism , Muscle, Skeletal/pathology , Polymyositis/etiology , Polymyositis/pathology , Regeneration/physiology , Scleroderma, Systemic/complications , Scleroderma, Systemic/metabolism , Scleroderma, Systemic/pathology
12.
Clin Exp Rheumatol ; 20(5): 641-6, 2002.
Article in English | MEDLINE | ID: mdl-12412194

ABSTRACT

OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) and its vascular and lymphatic receptors in skin in systemic sclerosis (SSc) compared to systemic lupus erythematosus (SLE), Raynaud's phenomenon (RP) and normal healthy control skin. METHODS: Staining was performed using rabbit anti-human antibodies in DAKO TechMate Horizon staining robot programmed for the biotin-streptavidin protocol. RESULTS: VEGF was sporadically and weakly expressed in normal skin, but in spite of vascular damage in diseased skin, VEGF expression was only slightly upregulated. In contrast, its vascular receptors VEGFR-1 (Flt-1) and VEGFR-2 (Flk-1), were clearly upregulated. Finally, the lymphatic VEGFR-3 (Flt-4) receptor was also upregulated in diseased skin and ectopically expressed also in blood vessels. Negative staining and positive sample controls confirmed the specificity of the staining. CONCLUSION: The imbalanced expression of VEGF and its vascular receptors suggest that the compensatory efforts to angiogenesis fail in SSc, in part due to insufficient local production of VEGF, which was low compared to VEGFR expression. This is compatible with the recent observations on the lack of alpha V beta 3+ newly formed blood vessels in SSc skin. Since microvascular angiogenic stimuli normally induce first VEGF and then VEGFR, these findings also suggest that the angiogenic cascade is turned on, but there is a defect in the finalization of its effects. Normalization of angiogenic cascade in SSc could provide a future therapeutic target.


Subject(s)
Endothelial Growth Factors/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Lupus Erythematosus, Systemic/metabolism , Lymphokines/metabolism , Raynaud Disease/metabolism , Receptors, Vascular Endothelial Growth Factor/metabolism , Scleroderma, Systemic/metabolism , Skin/blood supply , Adult , Animals , Female , Humans , Immunohistochemistry , Male , Middle Aged , Neovascularization, Pathologic , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors
13.
J Vasc Access ; 1(3): 81-3, 2000.
Article in English | MEDLINE | ID: mdl-17638231

ABSTRACT

PURPOSE: The problem of long form fistula for hemodialysis is well known among vascular surgeons because of frequent thromboses in anastomotic place. One of the pro-thrombotic factors is suture in the vessel's lumen. Since the 'sleeve' method prevents this, the patency rate should improve. METHODS: One hundred and eighty-two (182) patients were operated using the "sleeve" method. The operation was done under local anaesthesia and the first step was the dissection of the cephalic vein and the radial artery. Using Prolene 7-0 the artery and the vein were connected as follows: the first suture was placed intramuraly about 5 mm from the artery edge to the vein edge. The second suture (placed in the same way) was on the opposite side. The end of the radial artery was placed into the lumen of the cephalic vein and the sutures were tied. Two additional sutures were placed between them. Finally, four sutures were added to keep the artery in the vein. RESULTS: In our hands the patency rate of 'sleeve' arterio-venous fistulas was high: 95% after one month, 86% after one year, 64% after 5 years and 57% after 10 and 11 years. Every 'sleeve' fistula provided adequate blood flow, which increased during the follow-up period. CONCLUSIONS: The authors consider the "sleeve" fistula method as the primary choice because of its simplicity and excellent long-term patency.

14.
HIV Med ; 1(4): 200-4, 2000 Oct.
Article in English | MEDLINE | ID: mdl-11737349

ABSTRACT

OBJECTIVES: The HLA-Cw3 molecule has been reported to present peptides derived from HIV-1 p24gag protein to a cytotoxic T lymphocyte clone. We have shown previously that the synthetic octapeptide 145-152 derived from the p24gag sequence upregulated cell surface HLA-C expression on HLA-Cw*0303+ cells. Here, we examined the question of whether the nonapeptide 144-152 also exerts a similar effect. METHODS: The HLA-Cw*0303+ B-LCL PAJ and control HLA-Cw3-negative cells B-LCL HAJ and T-LCL 500/C9 were used. HLA expression on peptide-pulsed and non-pulsed cells was evaluated using specific antibodies and flow cytofluorimetry. Binding of dansylated peptides onto different cell lines was measured spectrofluorimetrically. RESULTS: The HIV-1 p24gag octapeptide upregulated cell surface HLA-C on PAJ (Cw*0303+) cells, whereas the nonapeptide did not. HLA-A2 expression was not affected by these peptides. Specificity of the effect of octapeptide was confirmed by the lack of HLA-C upregulation on HLA-Cw3- cells and by lower binding of dansylated peptide to the HLA-Cw3- cells HAJ and 500/C9. CONCLUSIONS: The above results indicate that HLA-Cw*0303 preferentially binds the octapeptide rather than the nonapeptide derived from HIV-1 p24gag protein.


Subject(s)
Gene Expression Regulation, Viral , HIV Core Protein p24/metabolism , HIV-1/metabolism , HLA-C Antigens/biosynthesis , Oligopeptides/metabolism , Antibodies, Monoclonal/genetics , Cell Line , Flow Cytometry , HIV Core Protein p24/genetics , HIV Core Protein p24/immunology , HIV-1/genetics , HIV-1/immunology , HLA-C Antigens/genetics , Humans , Oligopeptides/genetics , Oligopeptides/immunology , Up-Regulation
15.
Acta Microbiol Pol ; 49(3-4): 243-51, 2000.
Article in English | MEDLINE | ID: mdl-11293657

ABSTRACT

The susceptibility (MIC) of 44 strains of anaerobic bacteria isolated from the oral cavity and 3 standard strains to statherin and its C-terminal fragments with sequences QYQQYTF, YQQYTF, QQYTF, QYTF and YTF was determined by means of plate dilution technique in Brucella agar with 5% content of defibrinated sheep's blood, menadione and hemin. The culture was anaerobic. As shown, at concentrations from 12.5 to 100 microg/ml statherin and its C-terminal fragments inhibited the growth of anaerobic bacteria isolated from the oral cavity. Peptostreptococcus strains were the most susceptible to statherin and YTF (MIC < or = 12.5 mg/ml), whereas the most susceptible to the peptides investigated were Fusobacterium necrogenes and Fusobacterium necrophorum strains: QYQQYTF, YQQYTF, QQYTF, QYTF (MIC < or = 12.5 microg/ml). Prevotella oralis, Bacteroides forsythus and Bacteroides ureolyticus strains exhibited the lowest susceptibility (MIC > 100 microg/ml). When analysing the bacteriostatic activity of statherin it should be pointed out that the concentrations of this peptide used in microbiological investigations are within the range of physiological concentrations determined for whole saliva when at rest and stimulated in healthy donors of 19-25 years of age. Since the anaerobes investigated may be involved in the diseases of periodontum, the results presented seem to have also a practical aspect, i.e. a possibility to apply the C-terminal fragments of statherin as a novel therapeutic agent, affecting favourably the oral cavity.


Subject(s)
Bacteria, Anaerobic/drug effects , Gram-Negative Anaerobic Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Mouth/microbiology , Salivary Proteins and Peptides/pharmacology , Amino Acid Sequence , Gram-Negative Anaerobic Bacteria/growth & development , Humans , Microbial Sensitivity Tests , Molecular Sequence Data , Periodontal Diseases/microbiology
16.
Thromb Haemost ; 81(1): 26-31, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9974369

ABSTRACT

The aim of this study was to determine the efficacy and safety of subcutaneous weight-adjusted dose low molecular weight heparin (LMWH) compared with oral anticoagulant (OA) in the prevention of recurrent venous thromboembolism. In a prospective multicenter trial, 202 patients with symptomatic proximal deep vein thrombosis (DVT) were included. As soon as the diagnosis of DVT was confirmed by phlebography, 101 were randomly assigned to receive LMWH (nadroparin) for secondary prophylaxis and 101 to receive OA (acenocoumarol). Patients in both groups were initially treated with nadroparin in a dose of 85 anti-Xa IU/kg s.c. every 12 h. Secondary prophylaxis with either nadroparin, 85 anti-Xa IU/kg s. c. once daily, or acenocoumarol was continued for at least 3 months. Three patients in the LMWH group and 6 in the OA group were excluded from analysis for various reasons. During the one-year combined secondary prophylaxis and surveillance period, 7 of of the 98 evaluable patients (7.1%) in the LMWH group and 9 of the 95 evaluable patients (9.5%) in the OA group had a documented recurrence of venous thromboembolism (Fisher's exact test, p = 0.61). Of these, 2 patients who received LMWH and 7 patients on acenocoumarol had recurrences in the 3-month period of secondary prophylaxis. Four patients (4.1%) in the LMWH group developed bleeding complications during this study period, as compared with 7 (7.4%) in the OA group (Fisher's exact test, p = 0.37). There were two major bleedings, one in the LMWH group and one in the OA group. Eleven patients died, 5 (5.1%) in the LMWH group and 6 (6.3%) in the OA group. It is concluded that nadroparin in a dose of 85 anti-Xa IU/kg s.c. once daily provides an effective and safe alternative to oral anticoagulants in the secondary prophylaxis of DVT.


Subject(s)
Acenocoumarol/administration & dosage , Anticoagulants/administration & dosage , Heparin, Low-Molecular-Weight/administration & dosage , Thrombophlebitis/prevention & control , Administration, Oral , Humans , Injections, Subcutaneous , Nadroparin/administration & dosage , Secondary Prevention , Thrombophlebitis/physiopathology
17.
Immunol Lett ; 64(2-3): 57-62, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9870655

ABSTRACT

Human major histocompatibility complex class I antigens, HLA-C, are expressed on the cell surface at approximately a tenfold lower level than HLA-A and -B. We hypothesized that the expression of HLA-C is limited by the quantity of high affinity peptides which bind to these molecules, thus allowing only a small fraction of HLA-C molecules to be transported and/or to remain stable on the cell surface. If this assumption is correct, then the addition of exogenous peptide should increase cell surface HLA-C expression. To verify the hypothesis, we pulsed lymphoblastoid cell line PAJ (HLA-Cw3+) with synthetic HIV-1 p24gag 145-152 peptide, known to be presented to T-lymphocytes by HLA-Cw3 molecule. PAJ (HLA-Cw3+) cells bound approximately two times more of the peptide than HAJ (HLA-Cw3-), and four times more than 500/C9 (HLA-Cw3-) cells. Accordingly, overnight pulsing of PAJ cells with the p24gag 145-152 peptide caused an increase in class I HLA expression detected on the cell surface by flow cytofluorimetric analysis with anti-HLA-B,C monoclonal antibodies but not by anti-HLA-A antibody. In contrast, HLA-Cw3- cells treated in the same manner did not show any increase of HLA class I expression. Our data suggest that low concentration of high affinity peptides within the cell may be one of the factors limiting cell surface expression of HLA-C molecules.


Subject(s)
HIV Core Protein p24/metabolism , HLA-C Antigens/biosynthesis , Antibodies, Monoclonal/immunology , Cell Line , Female , Flow Cytometry , Fluorescent Antibody Technique , HLA-A Antigens/metabolism , HLA-B Antigens/metabolism , Humans , Lymphocytes , Peptide Fragments/chemical synthesis , Peptide Fragments/metabolism
18.
J Mal Vasc ; 23(5): 368-70, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9894192

ABSTRACT

OBJECTIVE: About 40% of patients with ruptured abdominal aortic aneurysm (AAA) die before admission to the hospital. The next 40-50% of patients who reach a hospital die in the perioperative period or within 30 days after surgery. Two groups of patients with ruptured AAA can be distinguished: first-with intra-abdominal rupture, second-with retro-peritoneal rupture. The aim of the study was a retrospective analysis of the treatment results in patients with retro-peritoneal rupture of AAA. MATERIAL AND METHODS: 78 patients underwent a surgical procedure between 1.01.1985 and 30.10.1996. 78 patients (68 men and 10 women), mean age 67.6 (53-94) were included in this study. Based on diagnostic and surgical procedures, two periods of treatment can be distinguished. In the first period of the time from 1.01.1985 to 31.12.1992 patients were operated on immediately after admission to the hospital. In the second period from 1.01.1993 to 31.10.1996 patients were admitted to Intensive Care Unit. In this unit patients were intensively treated and prepared for surgical procedure. During this time the computerized tomography scanning (CT) was performed. RESULTS: In the first period the perioperative mortality was 75%. In the second period the perioperative mortality was 41.3%. DISCUSSION: The 1.5-2 hours postponement of operative procedure of the patients with retro-peritoneal rupture of AAA can decrease perioperative mortality. During the time patients were intensively treated and prepared for surgical procedure and CT examination enabled to choose proper surgical technique.


Subject(s)
Aortic Aneurysm, Abdominal/diagnosis , Aortic Aneurysm, Abdominal/surgery , Aortic Rupture/diagnosis , Aortic Rupture/surgery , Aged , Aged, 80 and over , Aortic Aneurysm, Abdominal/mortality , Aortic Rupture/mortality , Diagnosis, Differential , Female , Humans , Male , Middle Aged , Retrospective Studies , Time Factors , Tomography, X-Ray Computed , Treatment Outcome , Vascular Surgical Procedures/mortality
19.
Przegl Lek ; 54(1): 15-7, 1997.
Article in Polish | MEDLINE | ID: mdl-9190627

ABSTRACT

Protein C is the important regulating factor of coagulation and fibrinolytic system. It is known that surgery causes disturbances of haemostasis. The aim of work was to study the influence of different kind of surgery on the protein C activity. The study contained 102 operated patients in whom vascular operation (20), operation of prostate because of carcinoma (20) and hypertrophy (40), cholecystectomy (12) and operation of hernia (10) were performed. Protein C activity was measured using Kabi Diagnostika test applying chromogenic substrate. In the most of patients surgery caused on the 0 and 1st postoperative day the decrease of protein C activity and the return to the preoperative values in the next few days. The exception was the hernia operation which did not diminish protein C activity. It seems that the decrease of protein C after surgery was the effect of consumption of it in the extremely activated coagulation and fibrinolytic system during operation.


Subject(s)
Protein C/metabolism , Surgical Procedures, Operative/adverse effects , Aged , Blood Coagulation/physiology , Cholecystectomy/adverse effects , Fibrinolysis/physiology , Hernia/metabolism , Herniorrhaphy , Humans , Male , Middle Aged , Prostatic Diseases/metabolism , Prostatic Diseases/surgery , Vascular Surgical Procedures/adverse effects
20.
Wiad Lek ; 50 Suppl 1 Pt 1: 378-81, 1997.
Article in Polish | MEDLINE | ID: mdl-9446389

ABSTRACT

Failure rate in standard groin hernia repair varies from 3 to 10%. Polypropylene mesh implantation based on Lichtenstein "tension free" method in 1986 year reduced the failure rate to less than 1%. From Feb. '95 to Dec.'96, 115 patients were operated on with 127 groin hernias repair. The average age of patients was 58 years 52 direct hernias, 74 indirect hernias and 1 pantaloon hernia have been diagnosed in examined material, 101 primary repairs and 26 repairs of recurrent hernia have been performed. The operations were performed in subarachnoid anaesthesia--66 patients, in general anaesthesia--11 patients in local anaesthesia--38 patients. After having opened the inguinal canal estimated the type of its wall defect. In case of direct hernia the sac usually was invaginated by absorbing suture. In case of indirect hernia sac was cut and peritoneal cavity left opened. The patch made of polypropylene monofilament mesh (size 6 x 8 cm) was sewn with "tension free" method under spermatic funiculus. As a complication 6 patients had haematomas in operating wounds. Four of the patients had wound infections. One of these patients was operated again and the patch was removed. The patients had no recurrence of hernia during the previous 10.6 months of observation. We haven't confirmed recurrence in examined material, yet it was too short time to estimate the efficiency of repair. The proposed way of groin hernia repair is easy and simple in every-day surgery practice.


Subject(s)
Hernia, Inguinal/surgery , Prostheses and Implants , Surgical Mesh , Adult , Aged , Aged, 80 and over , Humans , Middle Aged , Polypropylenes , Suture Techniques
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