Synthesis, characterization and self-assembly of biosurfactants based on hydrophobically modified inulins.

Biosurfactants have been synthesized using a low energy, environmentally friendly process by the derivatization of inulin with octenyl (OSA) and dodecenyl (DDSA) succinic anhydrides in aqueous solution. The inulin and its derivatives have been characterized using gel permeation chromatography/multi angle light scattering (GPC/MALLS), high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), Fourier transform infrared spectroscopy (FTIR), and NMR, and the reaction efficiency was found to be between 59 and 95%. The efficiency was generally higher for OSA derivatives compared to DDSA derivatives. The hydrophobic derivatives were found to aggregate in solution and the critical aggregation concentration (CAC) was determined using dye solubilization, surface tension, dynamic light scattering (DLS), and conductivity. There was reasonable agreement in the CAC values obtained by the different techniques except for conductivity. It was found that the CAC decreased with increasing alkenyl chain length and degree of modification, and the values were significantly lower for the DDSA derivatives compared to the OSA derivatives. GPC elution profiles for the DDSA-inulin using 12 mol % reagent confirmed the presence of aggregates with a molecular mass of ∼2.5 × 10(6) g/mol and a radius of gyration of ∼25 nm corresponding to ∼550 inulin molecules. DLS study was undertaken to determine the hydrodynamic radius, and values obtained for the DDSA (12%) derivative were 30 nm in both water and 0.1 M sodium nitrate, while for the OSA (12%) derivative values of 13 and 7 nm were obtained. The derivatives have potential application in the stabilization of particulate dispersions and emulsions and also in the encapsulation and delivery of drugs.

Extended magnetic reconnection at the Earth's magnetopause from detection of bi-directional jets

Magnetic reconnection is a process that converts magnetic energy into bi-directional plasma jets; it is believed to be the dominant process by which solar-wind energy enters the Earth's magnetosphere. This energy is subsequently dissipated by magnetic storms and aurorae. Previous single-spacecraft observations revealed only single jets at the magnetopause--while the existence of a counter-streaming jet was implicitly assumed, no experimental confirmation was available. Here we report in situ two-spacecraft observations of bi-directional jets at the magnetopause, finding evidence for a stable and extended reconnection line; the latter implies substantial entry of the solar wind into the magnetosphere. We conclude that reconnection is determined by large-scale interactions between the solar wind and the magnetosphere, rather than by local conditions at the magnetopause.

Clinical significance of intramedullary Gd-DTPA enhancement in cervical myelopathy.

STUDY DESIGN: Prospective multicenter study. OBJECTIVE: To clarify the significance of intramedullary Gd-DTPA enhancement in cervical myelopathy, the prevalence, morphologic features, clinical relevance and postoperative change were investigated. SETTING: Four hospitals in Japan. METHODS: A total of 683 patients with cervical myelopathy who underwent decompressive surgery were consecutively examined. T1, 2 and Gd-DTPA-enhanced MRI were taken before surgery. Fifty consecutive cases without intramedullary enhancement were allocated in the non-enhancement group. The following variables were investigated: prevalence of the enhancement, the morphologic feature, the relationship between the enhancement and T2 high-intensity areas, the change of the Japanese Orthopedic Association (JOA) score for cervical myelopathy and the change of the enhancement after surgery. RESULTS: Intramedullary enhancement was observed in 50 cases (7.3%). The enhancements were observed between the most severely compressed disc and the cranial half of the lower vertebral body. On axial images, they were observed at the posterior or posterolateral periphery of the spinal cord. Enhancement areas were observed within T2 high-intensity areas and smaller than them. The preoperative JOA score was 9.8+/-2.8 points in the enhancement group and 9.8+/-3.3 points in the non-enhancement group (NS). The postoperative JOA score was 12.7+/-2.9 points in the enhancement group and 14.2+/-2.4 in the non-enhancement group (P=0.006). Intramedullary enhancement disappeared in 60% of the patients 1 year after surgery. CONCLUSION: Intramedullary enhancement indicated not the severity of preoperative symptoms, but a sign of a worse prognosis.

A region of the muscarinic-gated atrial K+ channel critical for activation by G protein beta gamma subunits.

Complementary DNAs encoding two types of inwardly rectifying K+ channels, GIRK1 and IRK1, have been cloned from rat atrium and mouse macrophage, respectively. GIRK1 expressed in Xenopus oocytes was activated by acetylcholine when m2 muscarinic acetylcholine receptor was coexpressed. The acetylcholine-induced activation of GIRK1 was enhanced by coexpression with the G protein beta 1 gamma 2 subunit but not the beta 1 gamma 1 or alpha subunits. Deletion of the C-terminus of GIRK1 impaired the channel activation associated with the beta 1 gamma 2 subunit. Moreover, replacement of the C-terminus of IRK1 with that of GIRK1 produced a chimera channel that was activated by the beta 1 gamma 2 subunit, whereas intact IRK1 was not activated by the beta 1 gamma 2 subunit. These findings define the C-terminus of GIRK1 as a regulatory region for the G protein beta gamma subunit.

The interfacial, emulsification and encapsulation properties of hydrophobically modified inulin.

Octenyl- and dodecenyl succinic anhydride derivatives (OSA- and DDSA-) of inulin have been synthesised and their solution and interfacial properties have been determined and compared to a commercially available alkylated inulin, Inutec SP1. All samples formed micellar aggregates in solution above a critical concentration (critical aggregation concentration) and were able to 'dissolve' a hydrophobic dye. They were also able to form stable oil-in-water (O/W) emulsions as assessed by measurements of their droplet size as a function of time. DDSA-inulin with a high degree of substitution was found to be effective at encapsulating beta carotene using the solvent evaporation method which yielded a solid which dissolved readily in simulated gastric fluid. The results confirm the potential application of these materials in a number of areas including, drug delivery, pharmaceuticals, neutraceuticals, cosmetics and personal care.

One-stage lateral rhachotomy and posterior spinal fusion with compression hooks for Pott's paralysis in the elderly.

PURPOSE: To evaluate the safety and effectiveness of one-stage lateral rhachotomy and posterior fusion with compression hooks, for the treatment of Pott's paralysis in the elderly. METHODS: 11 elderly patients underwent lateral rhachotomy (costotransversectomy and pediculectomy) to debride the tuberculosis focus extending into the epidural space and to decompress the spinal cord. After debridement, the interbody cavity was packed with autologous iliac bone chips. For stabilisation, posterior fusion was performed using a compression lamina hook system. Patients were followed up for at least 2 years for complications. Neurological status was assessed using the Frankel score. The kyphotic deformity was measured on lateral radiographs taken before surgery and at follow-up. RESULTS: During separation of the adhesion around the abscess, a dural tear occurred in one patient and a pleural tear in another. Both tears were successfully repaired. One patient had mild pneumonia after surgery. The Frankel scores of the 11 patients improved from C or D before surgery to D or E after surgery. No relapse of spinal tuberculosis was encountered. The mean deformity angle was 25.5 degrees before surgery and 23.2 degrees at the final follow-up. Spinal fusion was achieved in all patients. CONCLUSION: Without the need of thoracotomy, one-stage lateral rhachotomy with posterior spinal fusion using compression hooks was an effective option for treating Pott's paralysis in the elderly.

The role of sex hormones in the kinetics of chondrocytes in the growth plate. A study in the rabbit.

Sex hormones play important roles in the regulation of the proliferation, maturation and death of chondrocytes in the epiphyseal growth plate. We have investigated the effects of male castration on the cell kinetics of chondrocytes as defined by the numbers of proliferating and dying cells. The growth plates of normal rabbits and animals castrated at eight weeks of age were obtained at 10, 15, 20 and 25 weeks of age. Our study suggested that castration led to an increase in apoptosis and a decrease in the proliferation of chondrocytes in the growth plate. In addition, the number of chondrocytes in the castrated rabbits was less than that of normal animals of the same age.

Ossification of the Achilles tendon--a case report.

Ossification of the Achilles tendon is a very unusual condition. We report a case with bilateral Achilles tendon ossification followed up for twelve years, treated twice surgically on the left side and conservatively on the right side. At the age of 51, the patient's plain radiogram showed bilateral ossifications of the Achilles tendon (right 55 mm, left 15 mm in length). The left side small mass was removed. The right side mass was decided to be followed up because of its large size. At the age of 61, ossification recurred on the left side. Conservative treatment with etidronate disodium for two years failed to prevent ossification from progressing. At the age of 63, the mass on the right and left sides increased to 70 mm and 45 mm in length, respectively. The mass of the left side was excised again, and dull pain disappeared postoperatively on the left side.

Cyclic AMP-dependent phosphorylation and regulation of the cardiac dihydropyridine-sensitive Ca channel.

A polyclonal antibody, CR2, prepared using the C-terminal peptide of the alpha 1 subunit of the rabbit cardiac DHP-sensitive Ca channel, specifically immunoprecipitated the [3H]PN200-110-labeled Ca channel solubilized from cardiac microsomes. The antibody recognized 250 and 200-kDa cardiac microsomal proteins as determined by immunoblotting, and cAMP-dependent protein kinase phosphorylated the 250-kDa, but not the 200-kDa protein in vitro. CHO cells, transfected with the cardiac alpha 1 subunit cDNA carried by an expression vector, synthesized a 250-kDa protein which was recognized by CR2. Adding db-cAMP or forskolin to the transformed CHO cells induced phosphorylation of the 250-kDa protein and stimulated the DHP-sensitive Ba current under patch-clamp conditions. These results suggested that the cardiac DHP-sensitive Ca channel was regulated by cAMP-dependent phosphorylation of the alpha 1 subunit.

Ectopic bone formation by electroporatic transfer of bone morphogenetic protein-4 gene.

Orthopedic surgeons have long awaited the clinical application of bone morphogenetic proteins (BMPs) for bone regeneration. However, such possible applications involving proteins or genes transferred with virus vectors have encountered many problems, including high cost, immunological reactions, viral infection, etc. We adopted a new gene transfer system of in vivo electroporation with a plasmid expression vector. A solution of plasmid DNA containing mouse BMP-4 (pMiw-BMP4) was injected into the gastrocnemius of BALB/cA mice, and electric pulses were applied through paired-needle electrodes inserted percutaneously. As a control plasmid, LacZ-containing plasmid (pMiwZ) was transferred by electroporation. A control group in which pMiw-BMP4 was injected and not electroporated was also introduced. In these groups, the gastrocnemius was harvested at 7, 14, 21, and 28 days after electroporation (n = 6 in each). As nonplasmid controls, electroporation with saline injection (n = 6), electroporation without injection (n = 6), and saline injection only (n = 3) were prepared. In these groups, the mice were killed 7 days after experimentation. Ectopic calcification or ossification was examined by histology as well as soft X-ray. In all electroporated groups (pMiwZ, pMiw-BMP4, saline injection, and without injection), dystrophic calcification of muscle bundles and infiltration of mesenchymal cells were observed histologically. Ectopic bone formation was observed only in the pMiw-BMP4 electroporation group. At 7 days after pMiw-BMP4 electroporation, extracellular eosinophilic matrix in a collection of mesenchymal cells was observed. Between 14 and 28 days after electroporation, ectopic bone was observed in 44% of mice, and bone marrow-like cells observed in 22%. The newly formed bone was woven. Injection of pMiw-BMP4 or saline induced neither calcification nor ossification. Our findings indicate that BMP-4 transferred by electroporation can induce in vivo and in situ ectopic bone formation in skeletal muscle.

Subtypes of purinoceptors in rat and dog urinary bladder smooth muscles.

1. Both adenosine and adenosine 5'-triphosphate (ATP) (10 microM and 100 microM) relaxed 10 microM acetylcholine (ACh)-induced contraction of rat bladder strips, which was completely antagonized by 100 microM 8-(p-sulphophenyl) theophylline. In dog bladder neither adenosine nor ATP inhibited ACh-induced contraction. 2. P2x-purinoceptor agonists contracted both rat and dog bladder strips with the potency order of alpha,beta-MeATP > ATP > ADP. 3. Alpha,beta-MeADP (100 microM) induced a contraction of the rat bladder strip even after desensitization of P2x-purinoceptors but failed to contract the dog bladder strip. 4. 2-MeSATP (1 microM to 300 microM) concentration-dependently induced contraction of rat bladder strips; this contraction was significantly inhibited after desensitization of P2x-purinoceptors. Cibacron blue 3GA (100 microM) antagonized the drug at concentrations lower than 30 microM, whereas it augmented the response to the drug at concentrations above 30 microM. 5. ADP beta S (1 microM to 1 mM) concentration-dependently induced contraction of rat bladder strips after desensitization of P2x-purinoceptors; a contraction which was significantly antagonized by cibacron blue 3GA (100 microM). 6. It is concluded that three subtypes of purinoceptors, P1 (mediating relaxation), and P2x and another type of P2 (mediating contraction), exist in rat urinary bladder smooth muscle, whereas a single subtype of the receptor, P2x-purinoceptor (mediating contraction) occurs in dog urinary bladder smooth muscle.

Contribution of P2-purinoceptors to neurogenic contraction of rat urinary bladder smooth muscle.

1. The contribution of P2-purinoceptors to neurogenic contraction was investigated in rat urinary bladder smooth muscle by measurement of isotonic tension. 2. Contraction of rat urinary bladder smooth muscle induced by electrical stimulation was decreased to 84.19 +/- 3.90% of the control (n = 16) in the presence of atropine (1 microM), which was further decreased to 38.80 +/- 2.75% of the control (n = 49) in the presence of both atropine and 10 microM alpha, beta-methylene adenosine 5'-triphosphate (alpha, beta-Me ATP). 3. The contractile response induced by electrical stimulation in the presence of atropine and alpha, beta-Me ATP was decreased to 27.81 +/- 4.07% (n = 23) and 26.63 +/- 5.01% (n = 15) of the control, by the addition of 100 microM cibacron blue 3GA and 100 microM suramin, respectively. The application of 100 microM adenosine 5'-o-2-thiodiphosphate (ADP beta S) in the presence of atropine and alpha, beta-Me ATP decreased the contractile response induced by electrical stimulations to 17.15 +/- 3.71% (n = 15) of the control. 4. Pretreatment of muscle strips with 100 microM ADP beta S significantly reduced the response to either 200 microM alpha, beta-methylene adenosine 5'-diphosphate or 200 microM ADP beta S. 5. Uridine 5'-triphosphate (100 microM to 1 mM) concentration-dependently contracted muscle strips, and this contraction was significantly antagonized by desensitization of P2-receptors with alpha, beta-Me ATP (10 microM), and completely antagonized by pretreatment of muscle strips with both alpha, beta-Me ATP and ADP beta S (100 microM). 6. Di(adenosine-5') tetraphosphate (30 and 100 microM) contracted muscle strips, whereas it failed to contract after desensitization of P2-receptors.7. It is suggested that about 20% of the neurogenic contraction of rat urinary bladder smooth muscle is mediated via ADP beta S-sensitive purinoceptors.

The signalling pathway which causes contraction via P2-purinoceptors in rat urinary bladder smooth muscle.

1. The signalling pathway which causes contractions to adenosine 5'-O-2-thiodiphosphate (ADPbetaS) and alpha,beta-methylene adenosine 5'-diphosphate (alpha,beta-Me ADP) was investigated in rat urinary bladder smooth muscle by measuring isotonic tension. 2. The responses to 10 microM alpha,beta-methylene adenosine 5'-triphosphate (alpha,beta-Me ATP) in 0 and 3.6 mM Ca2+ were 5.9+/-1.3 (n=10) and 122.2+/-6.4 (n=8) % respectively of those obtained in 1.8 mM Ca2+, whereas those to 100 microM ADPbetaS were 34.6+/-3.3 (n=8) and 96.8+/-7.2 (n = 8) %, in 0 and 3.6 mM Ca2+, respectively. In both experimental conditions, the responses to the two agonists expressed as % of the control responses were significantly different (P<0.01). 3. Indomethacin at high concentrations (>1 microM) decreased the responses to alpha,beta-Me ATP (10 microM), ADPbetaS (100 microM) and alpha,beta-Me ADP (100 microM). However, no significant difference was obtained between the responses to all the agonists at 30 microM indomethacin. 4. 2-Nitro-4-carboxphenyl n,n-diphenylcarbamate (NCDC) at concentrations between 1 microM and 100 microM concentration-dependently decreased the responses to ADPbetaS (100 microM) and alpha,beta-Me ADP (100 microM) and almost completely inhibited them at 100 microM. Although the responses to alpha,beta-Me ATP (10 microM) were also inhibited by the drug, at 50 and 100 microM NCDC the responses to alpha,beta-Me ATP were significantly larger than those to ADPbetaS and alpha,beta-Me ADP (P<0.01). 5.NCDC 100 microM significantly inhibited the KCl-induced contraction to 65.9+/-4.9% (n=6) of the control (P<0.01). 6. It is suggested that the contraction via ADPbetaS-sensitive receptors in the rat urinary bladder smooth muscle mainly depends on Ca2+ ions liberated from intracellular Ca2+ stores, though the contribution of Ca2+ ions from the extracellular space cannot be neglected. The release of Ca2+ ions from stores is mainly mediated by the production of inositol trisphosphate (IP3) via the activation of phospholipase C.

Effects of purinoceptor agonists on cytosolic Ca2+ concentration in swine tracheal smooth muscle cells in culture.

1. The effects of various purinoceptor agonists on intracellular Ca2+ concentration ([Ca2+]i) in swine tracheal smooth muscle cells in primary culture were examined to investigate the subtype of purinoceptors in these cells. 2. ATP (1 microM to 1 mM) concentration-dependently increased [Ca2+]i which was measured by monitoring the fluorescence signal of fura2. 3. alpha, beta-Me ATP at concentrations higher than 10 microM increased [ca2+]i in the presence of extracellular Ca2+. Responses to the drug were 12 +/- 5 and 61 +/- 4% of responses to ATP (100 microM) at 100 microM and 1 mM, respectively (n = 7). The response to 100 microM ATP was inhibited by 62% in the presence of 1 mM alpha, beta-Me ATP (n = 8), though the drug at concentrations lower than that did not affect the response to ATP. 4. ATP increased [Ca2+]i in the absence of extracellular Ca2+. The response to ATP in this condition was 40% of that in the presence of extracellular Ca2+ (n = 8). 5. Neither cibacron blue 3GA (10 microM) (n = 8) nor suramin (10 and 100 microM) (n = 10) affected the response to ATP (1 microM to 100 microM). 6. The rank order of potency in the absence of extracellular Ca2+ was UTP > ATP > adenosine 5'-o-(3-thiotriphosphate) > > ADP = alpha, beta-methylene adenosine 5'-triphosphate > 2-(methylthio)-adenosine 5'-(tetrahydrogen triphosphate). 7. UTP (1 microM to 100 microM) concentration-dependently increased inositol 1,4,5-triphosphate (IP3) production. 8. These results suggest that the increase in [Ca2+]i induced by purinoceptor agonists is mediated mainly via a nucleotide receptor in swine tracheal smooth muscle cells in primary culture.

Piroxicam suppositories in the treatment of osteoarthritis of the knee joint.

The subjects were 231 patients aged 16 to 75 years with osteoarthritis of the knee joint. Each patient received 20 mg of piroxicam daily as a suppository administered before sleep; 75% of the patients were treated for 14 days or longer. Overall treatment outcome was excellent in 34% according to physicians' ratings and in 36% according to the patients' self-ratings, good in 39% and 41%, fair in 22% and 17%, and unimproved in 5% and 7%, respectively. Side effects were reported by 3% of the patients. It is concluded that treatment of osteoarthritis with piroxicam suppositories is safe and effective.

Clinical evaluation of indomethacin-containing patches for osteoarthritis and extremity trauma.

Seventy-eight patients, aged 12-87 years, with osteoarthritis or extremity trauma, attached one indomethacin patch (containing 96 mg indomethacin) to the most symptomatic site twice daily. The final overall treatment outcome was excellent in 52% of the osteoarthritic patients and in 82% of the trauma patients, good in 27% and 13%, and fair in 3% and 0%, respectively. Side effects were reported by 4% of the patients. It is concluded that indomethacin-containing patches were effective and well-tolerated for the treatment of osteoarthritis and extremity trauma.

The long-term efficacy and tolerability of the new anti-inflammatory agent zaltoprofen in rheumatoid arthritis.

This paper reports on a non-comparative multicentre study designed to assess the usefulness of the non-steroidal anti-inflammatory agent zaltoprofen in the long-term treatment of rheumatoid arthritis. The efficacy and safety of zaltoprofen in the long-term treatment of rheumatoid arthritis were evaluated on the bases of changes of symptoms and presentation signs in the patients including the following: averaged bilateral grip strength, blood sedimentation rate, the duration of morning stiffness, joint pain for Lansbury scoring, restriction of activities of daily living (ADL) and pain score, and on patients' impressions at each hospital on the first day of attendance and then at 6 and 12 months after the start of administration of zaltoprofen, in a total of 46 patients. Overall improvements were recorded as 'marked' in 9%, 'moderate' or better in 60%, 'slight' or better in 80%, 'no change' in 17% and 'worse' in 3%, after 12 months' treatment. The overall safety was recorded as 'safe' in 86% and 'almost safe' or better in 97% after 12 months' treatment. On the basis of these results it can be concluded that, in the long-term treatment of rheumatoid arthritis, zaltoprofen is a well-tolerated and safe non-steroidal anti-inflammatory agent.

Clinical use of etodolac for the treatment of lumbar disc herniation.

This paper reports a non-comparative multicentre study designed to assess the usefulness of a non-steroidal anti-inflammatory agent--etodolac--for the treatment of lumbar disc herniation. The efficacy and safety of etodolac were evaluated based on the changes of symptoms and signs of the patients, including the following parameters: low back pain, leg pain and tingling, gait, the straight-leg-raising test, sensory disturbance, motor disturbance, restriction of activities of daily living (ADL), urinary bladder function, and patients' impressions at each hospital on the initial day, and at one week and two weeks after the start of administration of etodolac, in a total of 88 patients. The overall improvement was noted as a 'marked' in 5% and 13%, 'moderate' or better in 31% and 54%, 'slight' or better in 82% and 84%, 'no change' in 12% and 9%, and 'aggravated' in 6% and 7%, at one and two weeks after administration, respectively. The only adverse drug reaction reported was stomach pain in one of the 81 patients. It can be concluded that etodolac is an effective and well-tolerated non-steroidal anti-inflammatory agent in the treatment of lumbar disc herniation.

Mechanisms underlying the slow onset of action of a new dihydropyridine, NZ-105, on a cultured smooth muscle cell line.

The inhibitory effect of a new dihydropyridine derivative, (+/-)-2-[benzyl(phenyl)amino]ethyl-1,4-dihydro-2,6-dimethyl-5- (5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-4-(3-nitrophenyl )-3- pyridinecarboxylate hydrochloride (NZ-105), on whole cell Ca2+ current (ICa) in cultured vascular smooth muscle cells was investigated with the patch clamp technique. NZ-105 blocked ICa in a concentration-dependent manner when the command pulse ranged from +10 mV to -50 mV. The inhibitory effect of NZ-105 appeared at concentrations higher than 10 pmol/l and it blocked ICa completely at a concentration of 1 nmol/l. The concentration which produced the half-maximal inhibitory effect was estimated to be around 20 pmol/l. NZ-105 (500 pmol/l) completely blocked ICa elicited by depolarization to +10 mV at a holding potential of -40 mV, whereas it blocked ICa by only 67% at a holding potential of -90 mV. NZ-105 (100 pmol/l) shifted the steady-state inactivation curve by 40 mV to more negative potentials without affecting its slope factor. The blocking time constant of 500 pmol/l NZ-105 was 57.6 +/- 9.9 s at a holding potential of -70 mV. These results indicate that NZ-105 has characteristics typical of dihydropyridines and binds to Ca2+ channels of vascular smooth muscle cells with a high affinity. They also suggested that the slow onset of its action is due to the slow binding of the drug to Ca2+ channels.

The microsatellite alleles on chromosome 1 associated with essential hypertension and blood pressure levels.

Essential hypertension (EH) is thought to be a polygenic disease. Several candidate genes of this disease have been investigated in studies using polymorphic genetic markers, but some studies have failed to show any association of EH with these genes. In this experiment, we used microsatellite markers on chromosome 1, and performed an association study between EH and control subjects. Genomic DNA was amplified with fluorescently labelled primers from the Applied Biosystems PRISM linkage mapping set HD-5 comprising 63 highly polymorphic microsatellite markers with an average spacing of 4.5 cM. We isolated three loci showing significant differences: D1S507, D1S2713 and D1S2842. The P-values of the allele with the greatest post hoc contributions in D1S507, D1S2713 and D1S2842 were 0.0008, 0.0062 and 0.0084, respectively. All these values were significant after Bonferroni correction. Furthermore, we found that the three microsatellite alleles were associated with the levels of systolic blood pressure. These data suggest that there are at least the three susceptibility loci for EH on chromosome 1, and that a case-control study using microsatellite markers on genomewide basis is a useful method for isolating the susceptibility loci of multifactorial disorders.