heparitin-sulfate and Body-Weight

ORG 10172 is a heparinoid with mean molecular weight 6500 daltons. Intravenous bolus injections of ORG 10172 were compared with placebo and heparin injections in 91 separate studies in 83 healthy male subjects. 6400 units ORG 10172 produced a mean maximum change of 14.7 s in kaolin cephalin time (c.f. greater than 120 s for 5000 units heparin). Changes in prothrombin time were minimal (1.6 s for 6400 units ORG 10172 and 4.5 s after 5000 units heparin). A dose-related increase in bleeding time occurred after ORG 10172 and at high doses (greater than 3200 units) some secondary bleeding, which was never serious, occurred at between 1 and 4 h after incision. A dose-dependent reduction in ex vivo platelet adhesiveness was found at 10 min after ORG 10172 injection. ORG 10172 promoted a much smaller release of lipoprotein lipase as compared with heparin. The effect of ORG 10172 on plasma factor Xa activity (one measure of its action) was described by a biexponential decay with a mean distribution half-life of 2.34 (s.e. mean 0.16) h and mean disposition half-life of 17.6 (s.e. mean 1.1) h. It thus has a much longer duration of effect than heparin. There was a linear relationship of plasma anti-Xa response to increasing dose although there was some variability only partly explained by differences in body weight or surface area. ORG 10172 administration by bolus intravenous injection was well tolerated and there was no evidence of adverse effects on clinical chemistry or haematology tests.

Topics: Adolescent; Adult; Bleeding Time; Blood Coagulation Tests; Body Weight; Chondroitin Sulfates; Dermatan Sulfate; Factor X; Factor Xa; Fibrinolytic Agents; Glycosaminoglycans; Half-Life; Heparitin Sulfate; Humans; Kinetics; Lipoprotein Lipase; Male; Partial Thromboplastin Time; Platelet Adhesiveness; Prothrombin Time

Hospitalized cancer patients are at increased risk of thrombosis and prophylaxis with heparin is recommended. Heparanase is a protein capable of degrading heparan sulfate (HS) chains. The first objective of the study was to examine the effects of weight on anti-Xa levels in cancer patients treated with a fixed dose of enoxaparin as thromboprophylaxis. The second aim was to assess a potential correlation between plasma pre-treatment coagulation parameters and anti-Xa levels in an assumption that heparanase degradation activity towards heparins and HS chains could affect anti-Xa levels. Two blood samples (prior to and 3 h after drug injection) of 76 cancer patients with an indication for prophylaxis with enoxaparin (40 mg) were evaluated for coagulation markers. Sub-prophylactic levels of anti-Xa (< 0.2 U/ml) were found in 16/76 (21%) patients; in 13/76 (13%) patients the values were supra-prophylactic (> 0.5 U/ml). In the subgroup of patients weighing > 80 kg, 7/14 (50%) individuals had a sub-prophylactic level. Overall, anti-Xa levels appeared to correlate with patient's weight (r = - 0.48, p < 0.0001), pre-treatment partial thromboplastin time (PTT), D-dimer, HS, heparanase levels and procoagulant activity. We concluded that plasma anti-Xa levels correlated with patient's weight. A substantial portion of cancer patients receiving enoxaparin prophylaxis was undertreated. For patients > 80 kg, a weight-adjusted prophylactic dose of enoxaparin could be considered. Elevated enoxaparin anti-Xa levels correlated with pre-treatment parameters of coagulation system activation. High pre-treatment HS and elevated plasma anti-Xa levels may potentially serve as biomarkers for the identification of patients at increased thrombosis risk.

Topics: Anticoagulants; Biomarkers, Pharmacological; Blood Coagulation; Body Weight; Drug Dosage Calculations; Enoxaparin; Factor Xa; Female; Fibrin Fibrinogen Degradation Products; Heparitin Sulfate; Humans; Male; Middle Aged; Neoplasms; Thrombosis

Mucopolysaccharidosis type VII (MPS VII, Sly syndrome) is a very rare lysosomal storage disease caused by a deficiency of the enzyme β-glucuronidase (GUS), which is required for the degradation of three glycosaminoglycans (GAGs): dermatan sulfate, heparan sulfate, and chondroitin sulfate. Progressive accumulation of these GAGs in lysosomes leads to increasing dysfunction in numerous tissues and organs. Enzyme replacement therapy (ERT) has been used successfully for other MPS disorders, but there is no approved treatment for MPS VII. Here we describe the first human treatment with recombinant human GUS (rhGUS), an investigational therapy for MPS VII, in a 12-year old boy with advanced stage MPS VII. Despite a tracheostomy, nocturnal continuous positive airway pressure, and oxygen therapy, significant pulmonary restriction and obstruction led to oxygen dependence and end-tidal carbon dioxide (ETCO2) levels in the 60-80mmHg range, eventually approaching respiratory failure (ETCO2 of 100mmHg) and the need for full-time ventilation. Since no additional medical measures could improve his function, we implemented experimental ERT by infusing rhGUS at 2mg/kg over 4h every 2 weeks for 24 weeks. Safety was evaluated by standard assessments and observance for any infusion associated reactions (IARs). Urinary GAG (uGAG) levels, pulmonary function, oxygen dependence, CO2 levels, cardiac valve function, liver and spleen size, and growth velocity were assessed to evaluate response to therapy. rhGUS infusions were well tolerated. No serious adverse events (SAEs) or IARs were observed. After initiation of rhGUS infusions, the patient's uGAG excretion decreased by more than 50%. Liver and spleen size were reduced within 2 weeks of the first infusion and reached normal size by 24 weeks. Pulmonary function appeared to improve during the course of treatment based on reduced changes in ETCO2 after off-ventilator challenges and a reduced oxygen requirement. The patient regained the ability to eat orally, gained weight, and his energy and activity levels increased. Over 24 weeks, treatment with every-other-week infusions of rhGUS was well tolerated with no SAEs, IARs, or hypersensitivity reactions and was associated with measurable improvement in objective clinical measures and quality of life.

Topics: Administration, Intravenous; Body Weight; Child; Dermatan Sulfate; Enzyme Replacement Therapy; Glucuronidase; Glycosaminoglycans; Heparitin Sulfate; Hepatomegaly; Humans; Male; Mucopolysaccharidosis VII; Quality of Life; Splenomegaly; Therapies, Investigational

Heparan sulfate mimetic polymers promotes tissue repair when injected locally in doses of 1-2mg/kg by various routes. These biopolymers, have been extensively studied for their diverse biological activities. However, there is no detailed report investigating the toxicity of OTR4120. In this study, the acute and subchronic (30 days) toxicity of varying levels of OTR4120 was investigated in mice after intraperitoneal administration. The results showed that no significant toxicological changes were observed when 50mg/kg body weight per day OTR4120 was administered to mice. But when the dose was increased to 60 and 70 mg/kg body weight per day, the clotting time was significantly prolonged. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities were reduced female and male at dose 70 mg/kg body weight per day. These blood biochemistry data suggest that OTR4120 have a hepatoprotective effect. Based on these results, it can be concluded that the no adverse effect level of OTR4120 is 50 mg/kg body weight per day.

Topics: Animals; Biomimetics; Blood Cell Count; Blood Coagulation; Body Weight; Dose-Response Relationship, Drug; Enzymes; Female; Glycosaminoglycans; Heparitin Sulfate; Injections, Intraperitoneal; Male; Mice; Organ Size

Mucopolysaccharidosis type IIIA is a neurodegenerative lysosomal storage disorder characterized by progressive loss of learned skills, sleep disturbance and behavioural problems. Absent or greatly reduced activity of sulphamidase, a lysosomal protein, results in intracellular accumulation of heparan sulphate. Subsequent neuroinflammation and neurodegeneration typify this and many other lysosomal storage disorders. We propose that intra-cerebrospinal fluid protein delivery represents a potential therapeutic avenue for treatment of this and other neurodegenerative conditions; however, technical restraints restrict examination of its use prior to adulthood in mice. We have used a naturally-occurring Mucopolysaccharidosis type IIIA mouse model to determine the effectiveness of combining intravenous protein replacement (1 mg/kg) from birth to 6 weeks of age with intra-cerebrospinal fluid sulphamidase delivery (100 microg, fortnightly from 6 weeks) on behaviour, the level of heparan sulphate-oligosaccharide storage and other neuropathology. Mice receiving combination treatment exhibited similar clinical improvement and reduction in heparan sulphate storage to those only receiving intra-cerebrospinal fluid enzyme. Reductions in micro- and astrogliosis and delayed development of ubiquitin-positive lesions were seen in both groups. A third group of intravenous-only treated mice did not exhibit clinical or neuropathological improvements. Intra-cerebrospinal fluid injection of sulphamidase effectively, but dose-dependently, treats neurological pathology in Mucopolysaccharidosis type IIIA, even when treatment begins in mice with established disease.

Topics: Analysis of Variance; Animals; Antibodies; Body Weight; Brain; Chromatography, High Pressure Liquid; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Administration Routes; Exploratory Behavior; Heparitin Sulfate; Hydrolases; Lysosomal Storage Diseases; Male; Maze Learning; Mice; Mice, Inbred C57BL; Mice, Knockout; Mucopolysaccharidosis III; Necrosis; Proteins; Tandem Mass Spectrometry; Time Factors

We have generated homozygous transgenic mice (hpa-tg) overexpressing human heparanase (endo-beta-D-glucuronidase) in all tissues and characterized the involvement of the enzyme in tissue morphogenesis, vascularization, and energy metabolism. Biochemical analysis of heparan sulfate (HS) isolated from newborn mice and adult tissues revealed a profound decrease in the size of HS chains derived from hpa-tg vs. control mice. Despite this, the mice appeared normal, were fertile, and exhibited a normal life span. A significant increase in the number of implanted embryos was noted in the hpa-tg vs. control mice. Overexpression of heparanase resulted in increased levels of urinary protein and creatinine, suggesting an effect on kidney function, reflected also by electron microscopy examination of the kidney tissue. The hpa-tg mice exhibited a reduced food consumption and body weight compared with control mice. The effect of heparanase on tissue remodeling and morphogenesis was best demonstrated by the phenotype of the hpa-tg mammary glands, showing excess branching and widening of ducts associated with enhanced neovascularization and disruption of the epithelial basement membrane. The hpa-tg mice exhibited an accelerated rate of hair growth, correlated with high expression of heparanase in hair follicle keratinocytes and increased vascularization. Altogether, characterization of the hpa-tg mice emphasizes the involvement of heparanase and HS in processes such as embryonic implantation, food consumption, tissue remodeling, and vascularization.

Topics: Animals; Body Weight; Cattle; Cornea; Epithelial Cells; Feeding Behavior; Female; Glucuronidase; Hair; Hair Follicle; Heparitin Sulfate; Humans; Keratinocytes; Mammary Glands, Animal; Mice; Mice, Transgenic; Morphogenesis; Neovascularization, Physiologic; Phenotype; Transgenes

Heparan sulfate proteoglycans (HSPG) are a group of extracellular matrix molecules that link skeletal muscle cells to their extrinsic environment. To investigate if HSPG expression is affected by muscle growth and gender, a turkey line (F) selected for increased 16-wk BW and its unselected random-bred control line, RBC2, were used in the present study. Heparan sulfate (HS) and HSPG levels were measured in embryonic and posthatch pectoralis major muscle. HS levels plateaued at embryonic day (ED) 16 in both lines. A significant decrease of HS occurred at ED 18 in F males and females, and at ED 20 and 22 in the RBC2 males and females, respectively. Embryonic HSPG levels peaked at ED 18, and were significantly higher from ED 14 through 18 in F males and females compared with those of the RBC2 line. Male pectoralis major muscle had more HSPG at early embryonic stages than female muscle in both lines. During 1 to 16 wk posthatch, F male and female pectoralis major muscle contained more HSPG than the RBC2 samples, and HSPG levels in F males were higher than those of the females. Myogenic satellite cells derived from F and RBC2 male and female pectoralis major muscle were cultured to measure HSPG expression during proliferation and differentiation. No significant difference in HSPG level was found between the RBC2 and F line cells. However, in both lines, male-derived satellite cells had more HSPG than the female cells during proliferation and differentiation. These data show that HS and HSPG expression are affected by muscle growth properties and sex.

Topics: Age Factors; Animals; Body Weight; Breeding; Cell Differentiation; Cell Division; Cells, Cultured; Female; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Male; Muscle Development; Pectoralis Muscles; Satellite Cells, Skeletal Muscle; Selection, Genetic; Sex Factors; Turkeys

Topics: Anticoagulants; Body Weight; Chondroitin Sulfates; Contraindications; Dermatan Sulfate; Drug Monitoring; Heparin; Heparitin Sulfate; Humans; Kidney Failure, Chronic; Renal Dialysis; Thrombocytopenia

Glypicans are a family of heparan sulfate proteoglycans that are linked to the cell surface through a glycosyl-phosphatidylinositol anchor. One member of this family, glypican-3 (Gpc3), is mutated in patients with the Simpson-Golabi-Behmel syndrome (SGBS). These patients display pre- and postnatal overgrowth, and a varying range of dysmorphisms. The clinical features of SGBS are very similar to the more extensively studied Beckwith-Wiedemann syndrome (BWS). Since BWS has been associated with biallelic expression of insulin-like growth factor II (IGF-II), it has been proposed that GPC3 is a negative regulator of IGF-II. However, there is still no biochemical evidence indicating that GPC3 plays such a role.Here, we report that GPC3-deficient mice exhibit several of the clinical features observed in SGBS patients, including developmental overgrowth, perinatal death, cystic and dyplastic kidneys, and abnormal lung development. A proportion of the mutant mice also display mandibular hypoplasia and an imperforate vagina. In the particular case of the kidney, we demonstrate that there is an early and persistent developmental abnormality of the ureteric bud/collecting system due to increased proliferation of cells in this tissue element. The degree of developmental overgrowth of the GPC3-deficient mice is similar to that of mice deficient in IGF receptor type 2 (IGF2R), a well characterized negative regulator of IGF-II. Unlike the IGF2R-deficient mice, however, the levels of IGF-II in GPC3 knockouts are similar to those of the normal littermates.

Topics: Abnormalities, Multiple; Animals; Beckwith-Wiedemann Syndrome; Body Weight; Cell Division; Female; Genotype; Glypicans; Growth Disorders; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Insulin-Like Growth Factor II; Kidney Tubules, Collecting; Male; Mandible; Mice; Mice, Inbred C57BL; Mice, Knockout; Organ Size; Phenotype; Proteoglycans; Syndrome

The effect of short term (8 weeks) sodium (Na+) depletion and its repletion on glomerular synthesis of heparan sulfate and urinary excretions of albumin, total protein, heparan sulfate, Na+ and potassium (K+) was studied in spontaneous hypertensive rats (SHR) and their control normotensive Wistar-Kyoto rats (WKY). Na+ depletion in SHRs significantly increased the synthesis of glomerular heparan sulfate and decreased urinary excretions of albumin, Na+ and heparan sulfate when compared with the Na+ repleted group. In WKY rats, Na+ depletion did not cause any of the above changes. These data suggest that Na+ depletion prevents the urinary loss of protein through preservation of glomerular heparan sulfate only in SHRs.

Topics: Albuminuria; Animals; Blood Pressure; Body Weight; Drinking; Glycosaminoglycans; Heparitin Sulfate; Hypertension; Kidney Glomerulus; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Sodium

A coarctation hypertensive rat model was used to examine the effects of elevated blood pressure on basement membrane component synthesis by cardiac myocytes and aorta using immunohistochemistry and Northern blot analysis. Carotid arterial pressure increased immediately on coarctation, and left ventricular hypertrophy was maximal within 5 days. In immunohistochemical studies, fibronectin and laminin were increased and the basement membrane chondroitin sulfate proteoglycan decreased in both the subendothelial space and smooth muscle cell basement membranes of the aorta above the clip compared with controls, whereas only fibronectin was elevated in the aorta below the clip. No change in basement membrane staining intensity for the cardiac myocytes was observed. Alterations in steady-state mRNA levels for fibronectin and laminin in the aorta paralleled those observed by immunohistochemical analysis with regard to protein and tissue type affected as well as intensity of the changes. However, changes in mRNA levels (but not protein deposition) for perlecan and type IV collagen were also observed in aortas from hypertensive rats compared with controls. Increases in steady-state mRNA levels for all basement membrane components in the heart and vasculature peaked before maximal cardiac hypertrophy (5 days). These studies indicate that alterations in basement membrane component deposition in the hypertrophied vasculature occur at both transcriptional and translational levels and suggest that the cell attachment glycoproteins fibronectin and laminin may be important factors in the vascular response to elevated transmural pressure.

Topics: Animals; Aorta, Abdominal; Aortic Coarctation; Basement Membrane; Blood Pressure; Body Weight; Carotid Arteries; Collagen; DNA Probes; Fibronectins; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Hypertension; Laminin; Male; Muscle, Smooth, Vascular; Organ Size; Proteoglycans; Rats; Rats, Sprague-Dawley; Reference Values; Time Factors

We studied changes in glycosaminoglycan content and concentration during postresectional compensatory lung growth in adult male rats. After right trilobectomy, left lung dry weight was normal at 4 days, increased 74% between 4 and 7 days, and more slowly over the next week. Total glycosaminoglycan content per milligram dry lung weight increased early and rapidly, reaching 189% of the control value at 4 days postresection. The magnitude and temporal pattern of increase was different for different glycosaminoglycan subtypes. Hyaluronate and chondroitin sulfate content were increased by 198 and 113%, respectively, at 4 days, with no further increases subsequently. Heparan sulfate content increased more slowly and steadily, and dermatan sulfate concentrations did not change. At 4 days, the percent of total glycosaminoglycans that was hyaluronate was almost doubled, whereas the percent that was heparan sulfate was decreased; by day 7 the percent compositions had returned to normal. We conclude that changes in glycosaminoglycans occur early in postresectional lung growth and speculate that they may play a facilitatory role.

Topics: Animals; Body Weight; Chondroitin Sulfates; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Lung; Male; Organ Size; Rats; Rats, Inbred Strains

The effect of rapid eye movement (REM) sleep deprivation on the total content and proportion of different mucopolysaccharides (AMPS) containing uronic acid in rat brain was studied. REM sleep deprivation was induced by the water tank methods. Five experimental groups of animals were used: control, stressed, REM sleep deprived, post-stress sleeping and post-deprivation sleeping rats. No changes of AMPS were observed in any of the experimental groups when the whole brain was analysed. A significant increase of AMPS was found in the cerebral hemispheres of stressed and REM deprived rats. A significant decrease of AMPS was observed in the cerebellum and brain stem. A further increase of AMPS was found in the cerebral hemispheres after the rebound of REM sleep following its deprivation, and after the recovery sleep following the stress. A significant increase of AMPS was found in the brain stem of rats allowed to recuperate after REM deprivation or stress as compared with the stressed and REM deprived animals. Recovery sleep induced a significant increase of AMPS in the cerebellum in previously stressed rats, while previously REM deprived rats exhibited a further decrease of AMPS from control values. The possible functional meaning of these results is discussed in relation to the role of REM sleep in protein synthesis and learning and memory processes. Intriguing, well-controlled positive findings and the fact that no experimental design is known where stress is minimal while REM deprivation is 100 per cent, justify and encourage continued efforts in studying the biochemical state of the brain during sleep and/or its alterations.

Topics: Adrenal Glands; Animals; Body Weight; Brain Chemistry; Brain Stem; Cerebellum; Chondroitin Sulfates; Glycosaminoglycans; Heparitin Sulfate; Hyaluronic Acid; Male; Organ Size; Rats; Sleep Deprivation; Sleep, REM; Stress, Physiological; Telencephalon