Crystallization of macromolecular heparin

X-ray fibre-diffraction photographs were obtained from oriented films of the sodium salt of macromolecular heparin (molecular weight approx. 10(6)) prepared from rat skin. Two distinct molecular chain conformations corresponding to two different crystal lattices were observed as a function of relative humidity. The first conformation, obtained at 78% relative humidity, has a layer-line spacing of 1.73nm, which can be interpreted as an approximate twofold helix. On increasing the relative humidity to 84% a second phase with a layer-line repeat of 1.65nm is obtained with the reflexions indexing on a triclinic unit cell similar to that obtained previously (Nieduszynski & Atkins, 1973) for pig mucosal heparin.     

Structure of the filamentous bacteriophage,Pf3, by X-ray fiber diffraction

Fiber diffraction data have been obtained for the filamentous bacteriophage Pf3. The virus crystallizes on a hexagonal net with lattice constants a = b = 5.61 nm and c = 7.50 nm at 0% relative humidity and a = 6 = 6.20 nm and c = 7.99 nm at 98% relative humidity. The X-ray diffraction pattern resembles those of two other bacteriophages, Pf 1 and Xf, and therefore belongs to class II. The data are consistent with a 27₅ helix symmetry with an axial rise of 0.277 (dry) to 0.296 (wet) nm.     

Transfer Efficiency of Bacteria and Viruses from Porous and Nonporous Fomites to Fingers under Different Relative Humidity Conditions

Fomites can serve as routes of transmission for both enteric and respiratory pathogens. The present study examined the effect of low and high relative humidity on fomite-to-finger transfer efficiency of five model organisms from several common inanimate surfaces (fomites). Nine fomites representing porous and nonporous surfaces of different compositions were studied. Escherichia coli, Staphylococcus aureus, Bacillus thuringiensis, MS2 coliphage, and poliovirus 1 were placed on fomites in 10-μl drops and allowed to dry for 30 min under low (15% to 32%) or high (40% to 65%) relative humidity. Fomite-to-finger transfers were performed using 1.0 kg/cm² of pressure for 10 s. Transfer efficiencies were greater under high relative humidity for both porous and nonporous surfaces. Most organisms on average had greater transfer efficiencies under high relative humidity than under low relative humidity. Nonporous surfaces had a greater transfer efficiency (up to 57%) than porous surfaces (<6.8%) under low relative humidity, as well as under high relative humidity (nonporous, up to 79.5%; porous, <13.4%). Transfer efficiency also varied with fomite material and organism type. The data generated can be used in quantitative microbial risk assessment models to assess the risk of infection from fomite-transmitted human pathogens and the relative levels of exposure to different types of fomites and microorganisms.     

Binding of heparin to antithrombin III: The use of dansyl and rhodamine labels

Low molecular weight heparin of low-anticoagulant activity and high molecular weight heparin of correspondingly high activity were prepared by chromatography on protamine-Sepharose; preparations subjected to limited N-desulfation (5–10% free amino groups) by solvolysis were labeled with 5-dimethylaminonaphthalene-1-sulfonyl chloride (dansyl chloride) or rhodamine B isothiocyanate (RITC). The fluorescent heparins retained approximately 50% of the original anticoagulant activities. Dansyl-heparin on binding to antithrombin III (ATIII) exhibited a 2.5-fold enhancement of dansyl fluorescence intensity. This effect could be prevented by excess unlabeled heparin. A 7900 molecular weight dansyl-heparin preparation bound to ATIII with a stoichiometry of close to 2:1 and with an apparent association constant for binding (K_a) of 4.9 × 10⁵, m^?1, whereas a 21,600 molecular weight fraction bound at 0.7:1 with the protein and with an apparent K_a = 7.9 × 10⁵, m^?1. When ATIII reacted with a mixture of low molecular weight dansyl-heparin and low molecular weight RITC-heparin, there was enhancement of RITC fluorescence emission when excited at the dansyl excitation maximum; this effect was not observed when either of the labeled heparin species was prepared from high molecular weight material. The results are consistent with the proposal that a single molecule of high molecular weight, high-activity heparin occupies two sites when it binds to ATIII, whereas low molecular weight, low-activity heparin binds to the two sites separately.     

Structure of polar pili from Pseudomonas aeruginosa strains K and O

The polar pili of Pseudomonas aeruginosa strains K and O are hollow cylinders with 52 Å outer diameter and 12 Å inner diameter. There is a girdle of low electron density (interpreted as due to a local concentration of hydrophobic amino acid side-chains) centred at 31 Å diameter. Similar X-ray diffraction patterns are obtained from oriented fibres of the two types of pili, to a resolution of 7 Å in the equatorial direction and 4 Å in the meridional direction. The two types of pilin protein subunits have a similar molecular weight, and their sequences contain a number of homologous regions. They form a helical array with 4.06 to 4.08 units per turn of a basic helix that has a pitch of 40.8 Å for strain K pili and 41.3 Å for strain O pili at 75% relative humidity. A method is described for distinguishing between very similar diffraction patterns.There is strong intensity at 10 Å near the equator and at 5 Å near the meridian on the diffraction patterns. This intensity distribution is characteristic of α-helical rods running roughly in the direction of the fibre axis. The orientation of these rods was established by the fit between the transform of an α-helical polyalanine model and the strong near-equatorial layer-line.     

Characterization of the A in equilibrium B transition of DNA in fibers and gels by laser Raman spectroscopy.

Both Raman spectra and X-ray diffraction patterns have been obtained from oriented fibers of sodium deoxyribonucleic acid (Na-DNA) as a function of salt content and relative humidity. We have confirmed the previously reported X-ray results that, for oriented fibers, the A-form always exists between 75 and 92% relative humidity and that the conformation will change to the B-form at 92% relative humidity only if an excess (3–5%) of added salt is present. Oriented fibers containing low amounts of added salt remain in the A-type conformation at 92% relative humidity and higher. An exact correlation has been found between the familiar A- and B-type X-ray diffraction patterns of DNA fibers and the Raman spectra previously reported without X-ray verification from this laboratory for the A- and B-forms. In particular, a band at 807 cm^?1 was always present when a fiber showed the A-type diffraction pattern, and this band shifts to 790 cm^?1 in the B-form. Using the Raman spectrum to determine the specific conformation of DNA in samples less amenable to X-ray analysis, we have studied the A ? Btransformation in unoriented fibrous masses of DNA and in concentrated, oriented gels. We find that in unoriented fibrous masses, the A ? B transition always occurs at 92% relative humidity even at very low salt concentration (0–4%). However, in oriented DNA gels at low salt, the A-form can persist as a metastable state to concentration as low as 20% DNA. The origin of the bands at 807 and 790 cm^?1 and the possible biological implications of these findings are discussed.     

Salinity and the Hydraulic Conductance of Roots

The effect of salinity on hydraulic conductance of intact roots of tomato (Lycopersicon esculentum Mill.) and sunflower (Helianthus annuus L.) was determined in split-root experiments using salinized nutrient solutions. Experiments were conducted in controlled climate chambers under two or three relative humidity levels and four solution osmotic potential levels. The relationship between water flux through roots (J_v) and total water potential difference between the leaves and the root medium (Δψ) was linear, usually with a small intercept. Thus, the root hydraulic conductance (L) was not affected by salinity within the range of fluxes obtained in these experiments, with L= 0.036 mm h^?1 bar^?1 for tomato and L= 0.0167 mm h^?1 bar^?1 for sunflower. Our results agreed with theoretical analysis of coupled water and ion uptake. From Cl^? and Na⁺ uptake data, the reflection coefficient (o) for tomato roots was calculated as 0.956, which was compatible with the near-zero intercept. A large intercept for sunflower could not be readily explained. Relative humidity strongly affected root growth, with more rapid growth under low humidity conditions. Transpiration of sunflower plants was reduced by 20% when the relative humidity was increased from 34% to 84%, whereas transpiration in tomato was reduced 50%.     

Hyaluronic acid: molecular conformations and interactions in two sodium salts.

A detailed structure for the tetragonal form (a = b = 0.989 nm, c, fibre axis, = 3.394 nm) of sodium hyaluronate has been obtained by analysing X-ray fibre diffraction data using new molecular modelling techniques. Two polysaccharide chains pass through each unit cell, one at the corner and one at the centre. The chains are anti-parallel to one another. Each chain is a left-handed, 4-fold helix of disaccharide units. There are intramolecular hydrogen bonds stabilising each glycosidic linkage. Octahedrally co-ordinated sodium ions link, by O … Na⁺ … O bridges, neighbouring polysaccharide chains that are further linked by hydrogen bonds. No double-helix model (as originally proposed for this structure) has been found to be free of unacceptable non-bonded contacts or to fit the diffraction intensities as closely.The tetragonal form, which is stable at zero relative humidity, contains no detectable water molecules. At higher relative humidities a related orthorhombic form is observed in which only the a dimension of the lattice is different (a = 1.153 nm, b = 0.989 nm, c = 3.386 nm). In this form the hyaluronate helix is 2-fold with tetrasaccharide units conformationally similar to the 4-fold helix of the tetragonal form. The Na⁺ … O binding and hydrogen bonds lost on expansion of the tetragonal lattice are all replaced in the orthorhombic structure by bridges through water molecules, four of which associated with each tetrasaccharide.     

Conformation of heparin studied with macromolecular hydrodynamic methods and X-ray scattering

The hydrodynamic characteristics of heparin fractions in a 0.2 M NaCl solution have been determined. Experimental values varied over the following ranges: the sedimentation coefficient (at 20.0 °C), 1.3<s₀×10¹³<3.2 s; the Gralen coefficient (sedimentation concentration-dependence parameter), 10<k_s<70 cm³ g^–1; the translational diffusion coefficient, 3.9<D₀×10⁷<15.4 cm² s^–1; the intrinsic viscosity, 7.9<[]<40 cm³ g^–1. Combination of s₀ with D₀ using the Svedberg equation yielded molecular weights in the range 3.9<M×10^–3<37 g mol^–1. The value of the mass per unit length of the heparin molecule, M_L, was determined using the theory of hydrodynamic properties of a weakly bending rod, giving M_L=570±50 g nm^–1 mol^–1. The equilibrium rigidity, Kuhn segment length (A=9±2 nm) and hydrodynamic diameter (d=0.9±0.1 nm) of heparin were evaluated on the basis of the worm-like coil theory without the excluded volume effect, using the combination of hydrodynamic data obtained from fractions of different sizes. Small-angle X-ray scattering for three heparin fractions allowed an estimate for the cross-sectional radius of gyration as 0.43 nm; from the evolution with the macromolecule contour length of the radius of gyration, a value for the Kuhn segment length of 9±1 nm was obtained. A good correlation is thus observed for the conformational parameters of heparin from hydrodynamic and X-ray scattering data. These values describe heparin as a semi-rigid polymer, with an equilibrium rigidity that is essentially determined by a structural component, the electrostatic contribution being negligible in 0.2 M NaCl.Presented at the conference for Advances in Analytical Ultracentrifugation and Hydrodynamics, 8–11 June 2002, Grenoble, France     

Interactions between climatic and production factors on returns of female pigs to service during summer in Japanese commercial breeding herds

The objectives of this study were to determine the associations between climatic factors and production factors for returns to service of female pigs during summer and to quantify the associations between these factors and occurrences of returns to service. The factors that were assessed were as follows: maximum temperature (HT), relative humidity, age of gilts at first mating, parity, weaning-to-first-mating interval (WMI), and lactation length. The study analyzed records of 18,307 gilts in 99 herds and 78,135 parity records of 56,322 sows in 103 herds; all the females were first-serviced between June and September, 2007 to 2009. Average daily HT and relative humidity for 15 days post-service of a female were obtained from 21 local weather stations and coordinated with the performance data of the respective local herds. The returns to service were categorized into three groups: regular returns (RRs: 18–24 days), irregular returns (IRs: 25–38 days), and late returns to service (LRs: 39 days or later). Two-level mixed-effects models were applied to the data by using a herd at level 2 and an individual record at level 1. In mated gilts, the occurrences (%) of RRs, IRs, and LRs were 4.8%, 1.8%, and 5.3%, respectively. In mated sows, the respective occurrences were 3.3%, 1.8%, and 4.2%. Mean values (ranges) of HT and relative humidity were 28.4 °C (13.6 °C–39.8 °C) and 73.4% (35.0%–98.0%), respectively. In gilts, as HT increased from 25 °C to 30 °C, the occurrence of RR increased from 3.7% to 4.4% (P < 0.05). However, there was no association between the occurrence of RR and either relative humidity (P = 0.17) or age at first mating (P = 0.23). In addition, there were no associations between the occurrences of either gilt IR or LR and HT (P ≥ 0.05), relative humidity (P ≥ 0.46), or age at first mating (P ≥ 0.32). In sows, greater occurrences of RRs, IRs, and LRs were associated with higher HT, lower parity, and a WMI of 7 days or longer (P < 0.05), but they were not associated with relative humidity (P ≥ 0.62) or lactation length (P ≥ 0.13). The occurrence of RRs in sows of all WMI groups increased 1.22 (1.04⁵) times for each 5 °C increase in HT. For sows with WMI 0 to 6 days, each 5 °C rise in HT increased the occurrence of IRs and LRs by 1.36 (1.06⁵) and 1.27 (1.05⁵) times, respectively. However, there was no association between increased HT and occurrences of IRs or LRs for sows with WMI 7 days or longer (P ≥ 0.38). Therefore, in order to prevent returns to service, it is recommended that producers apply cooling management for females during the post-service periods in summer.     

Factors Affecting Dalapon Absorption and Translocation in Johnsongrass

The factors affecting the absorption and translocation of ¹⁴C-dalapon (2.2-dichloropropionic acid) in johnsongrass were studied. Johnsongrass [Sorghum halepense (L.) Pers.] was first pot-grown in a greenhouse and then treated and placed in controlled-environment chambers. Absorption of ¹⁴C-dalapon into johnsongrass leaves and subsequent translocation occurred continuously within the plant during a 48-h period after treatment. Gas chromatographic analysis of johnsongrass extracts showed that the dalapon molecule was translocated intact. Absorption and translocation of ¹⁴C-dalapon increased as the droplet volume of the diluent was increased from 0.2 to 5.0 μl per treated spot. At 21 and 32°C, translocation of ¹⁴C-dalapon from a 2-cm treated leaf section into the plant was greater at 100% than at 35% relative humidity. At 38°C, translocation was greater at 35% than at 100% relative humidity. The addition of 0.5% surfactant to the dalapon solution increased translocation under all environmental conditions studied. The addition of 0.1 M KH₂PO₄ to dalapon-surfactant solutions increased ⁴-dalapon translocation under high temperature (38°C), especially at 35% relative humidity.     

The Solution Structure of Heparan Sulfate Differs from That of Heparin: IMPLICATIONS FOR FUNCTION*

The highly sulfated polysaccharides heparin and heparan sulfate (HS) play key roles in the regulation of physiological and pathophysiological processes. Despite its importance, no molecular structures of free HS have been reported up to now. By combining analytical ultracentrifugation, small angle x-ray scattering, and constrained scattering modeling recently used for heparin, we have analyzed the solution structures for eight purified HS fragments dp6–dp24 corresponding to the predominantly unsulfated GlcA-GlcNAc domains of heparan sulfate. Unlike heparin, the sedimentation coefficient s_20,w of HS dp6–dp24 showed a small rotor speed dependence, where similar s_20,w values of 0.82–1.26 S (absorbance optics) and 1.05–1.34 S (interference optics) were determined. The corresponding x-ray scattering measurements of HS dp6–dp24 gave radii of gyration R_G values from 1.03 to 2.82 nm, cross-sectional radii of gyration R_XS values from 0.31 to 0.65 nm, and maximum lengths L from 3.0 to 10.0 nm. These data showed that HS has a longer and more bent structure than heparin. Constrained scattering modeling starting from 5,000 to 12,000 conformationally randomized HS structures gave best fit dp6–dp24 molecular structures that were longer and more bent than their equivalents in heparin. Alternative fits were obtained for HS dp18 and dp24, indicating their higher bending and flexibility. We conclude that HS displays bent conformations that are significantly distinct from that for heparin. The difference is attributed to the different predominant monosaccharide sequence and reduced sulfation of HS, indicating that HS may interact differently with proteins compared with heparin.     

Combination of site-directed mutagenesis and calcium ion addition for enhanced production of thermostable MBP-fused heparinase I in recombinant Escherichia coli

Heparinase I (HepI), which specifically cleaves heparin and heparan sulfate, is one of the most extensively studied glycosaminoglycan lyases. Low productivity of HepI has largely hindered its industrial and pharmaceutical applications. Loss of bacterial HepI enzyme activity through poor thermostability during its expression and purification process in production can be an important issue. In this study, using a thermostabilization strategy combining site-directed mutagenesis and calcium ion addition during its production markedly improved the yield of maltose-binding protein-fused HepI (MBP–HepI) from recombinant Escherichia coli. Substitution of Cys297 to serine in MBP–HepI offered a 30.6 % increase in the recovered total enzyme activity due to a mutation-induced thermostabilizing effect. Furthermore, upon addition of Ca²⁺ as a stabilizer at optimized concentrations throughout its expression, extraction, and purification process, purified mutant MBP–HepI showed a specific activity of 56.3 IU/mg, 206 % higher than that of the wild type obtained without Ca²⁺ addition, along with a 177 % increase in the recovered total enzyme activity. The enzyme obtained through this novel approach also exhibited significantly enhanced thermostability, as indicated by both experimental data and the kinetic modeling. High-yield production of thermostable MBP–HepI using the present system will facilitate its applications in laboratory-scale heparin analysis as well as industrial-scale production of low molecular weight heparin as an improved anticoagulant substitute.     

A Template-Free,Ultra-Adsorbing,High Surface Area Carbonate Nanostructure

We report the template-free, low-temperature synthesis of a stable, amorphous, and anhydrous magnesium carbonate nanostructure with pore sizes below 6 nm and a specific surface area of ∼ 800 m² g⁻¹, substantially surpassing the surface area of all previously described alkali earth metal carbonates. The moisture sorption of the novel nanostructure is featured by a unique set of properties including an adsorption capacity ∼50% larger than that of the hygroscopic zeolite-Y at low relative humidities and with the ability to retain more than 75% of the adsorbed water when the humidity is decreased from 95% to 5% at room temperature. These properties can be regenerated by heat treatment at temperatures below 100°C.The structure is foreseen to become useful in applications such as humidity control, as industrial adsorbents and filters, in drug delivery and catalysis.     

Changes in the pitch of the myosin helix preceding cross-brdige attachment in oscillatory contractions of frog sartorius muscle

When glycerinated fibres in presence of 5·10^?7M Ca²⁺ were oscillated at a frequency of 10 Hz, the X-ray intensities at 429 Å and at the small radial position of the 191 Å layer-line change in an inverse pattern. This rearrangement of the pitch of the myosin helix precedes by at least 10 msec the intensity increase on the 59 Å X-ray layer-line, which provides a measure for cross-bridge attachment to the actin. The perfect match of the new pitch of the myosin helix with the pitch of the actin helix may ensure optimal cross-bridge attachment.     

Effects of light, CO2 and humidity on carnation growth, hyperhydration and cuticular wax development in a mist reactor

Summary Plant survival ex vitro requires functioning stomata, adequate cuticular wax composition and deposition, and normal morphological development. Light intensity, CO₂ and relative humidity were altered inside an acoustic window mist reactor to study their effects on carnation (Dianthus caryophyllus) growth, stomata development, hyperhydration and epicuticular wax content. Increasing the light intensity from 65 to 145 μmol m⁻² s⁻¹ and enrichment of the gas phase with CO₂ (1350 ppm) reduced the number of hyperhydrated plants from 75 to 25% and increased the percentage dry weight of normal (healthy) plants from 17 to 25%. Lowering the relative humidity (≈70% RH) surrounding the plants during the mist-off phase for the last 2 wk of culture reduced the number of hyperhydrated plants from 70 to 9% and also increased the percentage of dry weight of normal plants from 16 to 25%. The stomata on plants grown in conditions of high light or low humidity had smaller apertures and appeared sunken when compared to stomata from plants grown in low light and high relative humidity. The epicuticular wax profiles of plants from the greenhouse or Magenta boxes showed a distinct shift in wax compounds with developmental age, plant type (hyperhydrated or normal), and type of box that was used (vented or not). In addition, very different wax profiles were observed from plants grown in reactors with altered CO₂ and light intensities.     

Effects of Relative Humidity on Carbon Isotope Fractionation in Plants

Four C₃ plants and a C₄ plant were grown from seeds at four levels (30, 45, 60, and 75 %) of relative humidity. All plants were subjected to a 16 h day, at 500 μE/m².s^?1 photon flux density. Mature leaves were analyzed for their carbon isotopic composition. Isotope fractionation decreased by up to 3 ‰ with decreasing relative humidity in all C₃ plants, while the opposite trend was observed in the C₄ plant. The observed shifts in both C₃ and C₄ plants are attributed to decreased stomatal conductance at low relative humidity, resulting in a smaller P_i.     

Structural changes in the thin filament during activation studied by X-ray diffraction of highly stretched skeletal muscle

The actin layer-lines were recorded from a frog semitendinosus muscle stretched to a sarcomere length greater than 4.4 microM. On activation of the muscle, the equator, the second layer-line at 1/18 nm-1 and the 5.9 nm layer-line increased in integrated intensity. On the other hand, the integrated intensity of the first layer-line at 1/36 nm-1 decreased markedly on activation. This decrease was not fully attributable to shifts of tropomyosin strands and therefore suggested a structural change in the actin subunit. The decrease may account for the apparent lack of an intensity increase of this layer-line on activation at normal muscle lengths where attachment of myosin heads to actin increases the intensities of other layer-lines.     

Application of Maltodextrin as Chiral Selector in Capillary Electrophoresis for Quantification of Amlodipine Enantiomers in Commercial Tablets

Maltodextrin was investigated as a chiral selector in capillary electrophoresis (CE) analysis of amlodipine (AM) enantiomers. For development of a stereoselective CE method, various effective parameters on the enantioseparation were optimized. The best results were achieved on an uncoated fused silica capillary at 20 °C using phosphate buffer (100 mM, pH 4) containing 10% w/v maltodextrin (dextrose equivalent value 4–7). The UV detector was set at 214 nm and a constant voltage of 20 kV was applied. The range of quantitation was 2.5–250 µg/mL (R² > 0.999) for both enantiomers. Intra‐ (n = 5) and interday (n = 3) relative standard deviation (RSD) values were less than 7%. The limits of quantitation and detection were 1.7 µg/mL and 0.52 µg/mL, respectively. Recoveries of R(+) and S(?) enantiomers from tablet matrix were 97.2% and 97.8%, respectively. The method was applied for the quantification of AM enantiomers in commercial tablets. Also, the enantioseparation capability of heparin was evaluated and the results showed that heparin did not have any chiral selector activity in this study. Chirality 26:394–399, 2014. © 2014 Wiley Periodicals, Inc.     

Isolation and characterization of a heparin with high anticoagulant activity from the clam Tapes phylippinarum: evidence for the presence of a high content of antithrombin III binding site

Heparin with high anticoagulant activity (activated partial thromboplastin time of 347 +/- 56.4 and anti-Xa activity of 317 +/- 48.3) was isolated from the marine clam species Tapes phylippinarum in an amount of approximately 2.1 mg/g dry animals. Agarose-gel electrophoresis showed a high content of the slow-moving heparin component (22 +/- 6.8%) and 78 +/- 5.4% of the fast-moving species. An average molecular mass of 13,600 was calculated by PAGE analysis, whereas a number average molecular weight Mn value of 10,700, a weight average molecular weight Mw of 14,900, and a dispersity index Mn/Mw of 1.386 were obtained by high-performance size-exclusion chromatography. Structural analysis of clam heparin, performed by depolymerizing heparin samples with heparinase (EC 4.2.2.7) and then separating the resulting unsaturated oligosaccharides by strong anion exchange-HPLC revealed the presence of large amounts (more than 130% than standard pharmaceutical heparin obtained from bovine intestine) of the oligosaccharide sequence bearing part of the ATIII-binding region, DeltaUA2S (1-->4)-alpha-D-GlcN2S6S (1-->4)-alpha-L-IdoA (1-->4)-alpha-D-GlcNAc6S (1-->4)-beta-D-GlcA (1-->4)-alpha-D-GlcN2S3S6S in the T. phylippinarum heparin, in comparison with bovine mucosal heparin and a sample of porcine mucosal heparin previously published. Furthermore, as expected from the oligosaccharide compositional analysis, due to the presence of a great mol % (80.6%) of the trisulfated disaccharide DeltaUA2S(1-->4)-alpha-D-GlcN2S6S, mollusc heparin is a more sulfated polysaccharide than bovine mucosal heparin (73.5%) and a sample of porcine mucosal (72.8%) heparin previously reported. To our knowledge, this is the first article describing a clam heparin having the ATIII binding site mainly identical to that of human and porcine intestinal mucosal heparins and bovine intestinal mucosal heparin but different from that found in beef lung heparin.