Fixative-Stain Sequences for Selective Demonstration of Juxtaglomerular Cells

The effects of 31 fixatives, containing alcohol, acids, formalin and metallic salts, and representing many of the standard fixatives, were observed for selectivity and intensity of staining of juxtaglomerular granules in mouse kidney. Four staining methods: 1:400,000 aqueous methyl violet 2B; Bowie's ethyl violet-Biebrich scarlet; 1:200,000 aldehyde fuchsin; and periodic acid-Schiff were used. Fixatives containing HgCl₂, trichloroacetic acid or formalin were found to be the most satisfactory for subsequent staining of the granules.     

Long-Term Bioavailability of Single Doses of Intramuscular Vitamin D2

Objective: We sought to determine the long-term bioavailability of single doses of intramuscular (IM) vita-min D₂ (D₂) in healthy adults.Methods: Forty healthy volunteers with hypovitaminosis D received a single dose of 200,000, 400,000, or 600,000 IU intramuscular D₂ or no treatment. Levels of 25-hydroxyvitamin D₂ (25[OH]D₂) and 25-hydroxyvitamin D₃ (25[OH]D₃) in serum were measured by liquid chromatography-tandem mass spectrometry. Vitamin D binding protein (DBP) and intact parathyroid hormone (iPTH), bone turnover markers (BTMs), and serum and urinary calcium were also measured.Results: After a single dose of D₂ injection, the level of 25(OH)D₂ increased slowly and reached a plateau at 8 weeks. The plateau remained stable for 12 weeks. The mean increase in 25(OH)D₂ was 6.8, 9.6, or 15.6 ng/mL after injection of 200,000 IU, 400,000 IU, or 600,000 IU D₂. Although endogenous 25(OH)D₃ levels were reduced by IM D₂, the total 25(OH)D levels increased by 5.0, 7.0, or 10.3 ng/mL in average after injection of 200,000 IU, 400,000 IU, or 600,000 IU D₂. The iPTH levels were also decreased by IM D₂. However, levels of serum calcium, BTMs, and DBP and urinary calcium were not altered by IM D₂.Conclusion: A single dose of 200,000 IU, 400,000 IU, or 600,000 IU IM D₂ raises total 25-hydroxyvitamin D levels by 5.0, 7.0, or 10.3 ng/mL on average for at least 12 weeks and reduces iPTH and endogenous 25(OH)D₃ levels without affecting levels of serum calcium, BTMs, DBP, and urinary calcium.     

Structure of proteoheparan sulfates from fibroblasts. Confluent and proliferating fibroblasts produce at least three types of proteoheparan sulfates with functionally different core proteins

[3H]Leucine- and [35S]sulfate-labeled proteoheparan sulfates were isolated from postconfluent or proliferating cultures of human skin fibroblasts. Cell layers were solubilized by Triton X-100, and transferrin-binding macromolecules were isolated by affinity chromatography. Proteoglycans with no affinity for transferrin were purified by using ion-exchange and gel permeation chromatography. Postconfluent cells synthesize a proteoheparan sulfate of Mr 350,000 (as determined by gel permeation chromatography) which has affinity for transferrin as well as for octyl-Sepharose. Its core protein (Mr 180,000) consists of two disulfide-bonded polypeptides of Mr 90,000. This species was not detected in cultures of proliferating cells. Proliferating and confluent cells also synthesize other forms of proteoheparan sulfates (Mr 200,000-400,000) which have no affinity for transferrin. However, most of them have affinity for octyl-Sepharose. The core protein of proteoheparan sulfates made by proliferating cells has Mr 50,000. A smaller form (Mr 250,000) of this proteoglycan was solubilized by Triton X-100, whereas a larger form (Mr 400,000) remained associated with the pericellular matrix. A third type of proteoheparan sulfate (Mr 200,000) without affinity for transferrin nor octyl-Sepharose was associated with postconfluent cell layers but not with proliferating ones. Its core protein has Mr 35,000. Heparan sulfate oligosaccharides (Mr 6,000 or higher) were found in proliferating cells but not in postconfluent ones.     

Randomised,double blind,placebo controlled clinical trial of efficacy of vitamin A treatment in non-measles childhood pneumonia.

OBJECTIVE: To evaluate the impact on clinical recovery and severity of the addition of large doses of vitamin A to the standard treatment for childhood pneumonia. DESIGN: A randomised, double blind, placebo controlled trial. SETTING: Study children were recruited at a public hospital in Recife, north east Brazil, an area of marginal vitamin A deficiency. SUBJECTS: 472 children aged 6 to 59 months with clinical diagnosis of pneumonia. INTERVENTIONS: 200,000 IU (infants) or 400,000 IU (1-4 year olds) of vitamin A in oil or similar capsules of placebo divided into two daily oral doses, in addition to the standard treatment. MAIN OUTCOME MEASURES: Duration of the episode and incidence of adverse outcomes. RESULTS: The groups were similar with respect to overall duration of pneumonia and incidence of adverse outcomes. Children who received vitamin A, however, were less likely to have fever by day 3 (P = 0.008) and were 29% less likely to fail to respond to the first line antibiotic (P = 0.054). CONCLUSION: There was little evidence for an effect of vitamin A treatment on the immediate outcome of the pneumonia episode.     

Serotonin or its agonist 5-methoxytryptamine can stimulate hamster sperm acrosome reactions in a more direct manner than catecholamines

Serotonin (50 microM) or its agonist 5-methoxytryptamine (5 microM) stimulated the acrosome reactions of golden hamster sperm within 15 min after addition to sperm capacitated in vitro for 4.5 h. The stimulation was inhibited by the serotonin receptor antagonists quipazine or cyproheptadine. Epinephrine (70 microM), norepinephrine (50 microM), and dopamine (25 microM) were unable to stimulate acrosome reactions even at 30 min under the same conditions, even though previous studies had demonstrated stimulation by these catecholamines at the same concentrations when present from the start of the capacitation time course. Epinephrine (5 microM) also was unable to stimulate at 30 min. These results demonstrate that serotonin and its agonist have a more direct effect on the hamster sperm acrosome reaction than other biogenic amines and that the effect is receptor-mediated.     

Epidural Analgesia in Dogs with Special Reference to the Intraarterial Blood Pressure

In order to investigate whether epidural analgesia in dogs employing mepivacaine chloride gives rise to complications, mainly changes in blood pressure, such analgesia was carried out in 35 dogs brought to the clinic for orthopedic, abdominal or perineal operations. Intra-arterial blood pressure measurements were made in 16 dogs. Three different solutions of mepivacaine chloride were used: 1 % solution with adrenaline 1:200,000; 2% solution with adrenaline 1:100,000 and 2% solution with adrenaline 1:200,000. The various solutions showed no significant difference in analgesic effect. Nor was any blood pressure change found during the epidural blockade. As a solution with a higher concentration causes a higher blood level of the analgesic and also greater toxic effects it is recommended that first preference be given to use of a 1 % solution with adrenaline 1:200,000; second choice is a 2 % solution with adrenaline 1:200,000 and third choice the 2 % solution with adrenaline 1:100,000.     

颈交感神经阻滞对严重烧伤大鼠的救治作用及其机制研究

目的:观察颈交感神经阻滞(CSB)对严重烧伤大鼠的救治作用,并对其可能机制进行初步探讨。方法:将大鼠随机分为正常对照组、烧伤组和CSB组,烧伤组与CSB组均制作20%体表面积Ⅲ0烧伤模型,CSB组于致伤后进行颈交感神经阻滞,观察动脉血压和心率变化;测定大鼠血中皮质酮、肾上腺素浓度;观察伤后21天动物伤死情况。结果:1.颈交感神经对严重烧伤大鼠救治效果显著,烧伤组和CSB组动物21天死亡率分别为73.33%和53.33%;2.创伤后大鼠血浆内肾上腺素浓度在伤后24小时有明显上升,然后迅速下降,但是CSB组肾上腺素浓度上升幅度远远低于烧伤组;3.与正常组相比,烧伤后血清GC的水平升高非常显著,CSB治疗组虽较正常组也升高,但显著低于烧伤组。结论:颈交感神经阻滞对严重创伤动物具有明显保护效应,其保护作用机制可能与调节创伤后神经-内分泌-免疫系统功能紊乱有关。     

Vascular endothelial growth factor expression in pig latissimus dorsi myocutaneous flaps after ischemia reperfusion injury

Exogenous administration of vascular endothelial growth factor (VEGF) improves long-term viability of myocutaneous flaps. However, endogenous expression of this substance in flaps following ischemia-reperfusion injury has not been reported previously. Endogenous production of VEGF was measured in myocutaneous pig latissimus dorsi flaps after ischemia-reperfusion injury. Latissimus dorsi myocutaneous flaps (15 x 10 cm) were simultaneously elevated bilaterally in six Yorkshire-type male pigs (25 kg). Before elevation, three flap zones (5 x 10 cm) were marked according to their distance from the vascular pedicle. After isolation of the vascular pedicle, ischemia-reperfusion injury was induced in one flap by occlusion of the thoracodorsal artery and vein for 4 hours, followed by 2 hours of reperfusion. The contralateral flap served as a control. Perfusion in each zone was monitored by laser Doppler flowmetry at baseline, during ischemia, and during reperfusion. At the end of the protocol, skin and muscle biopsies of each flap zone and adjacent tissues were obtained for later determination of VEGF protein levels. VEGF concentrations were quantified using the Quantikine human VEGF immunoassay. Skin perfusion was similar among all flap zones before surgery. Flow fell in all flaps immediately after flap elevation. After 4 hours of ischemia, blood flow in the ischemic flaps was significantly decreased (p < 0.05) compared with nonischemic control flaps. After 2 hours of reperfusion, flow in ischemic flap skin recovered to levels similar to those in control flaps. VEGF protein concentrations in muscle tissue exceeded concentrations in skin and decreased from zones 2 to 3 in control and ischemic flaps. No significant differences in VEGF concentrations between ischemic and control muscle zones were observed. However, the concentration of VEGF in all muscle zones was significantly higher (p < 0.05) than muscle adjacent to the flap. Concentrations in skin zones 1 and 2 were significantly higher (p < 0.05) in ischemic flaps than in control flaps, but levels in zone 3 (most ischemic flaps) showed no significant difference.     

Differentiation between carrier-bound forms of non-suppressible insulin-like activity (NSILA) in serum

• 1.1. Gel-permeation chromatography of serum on Sephacryl S-300 at pH 7.4 has shown that NSILA was detected over a range of MW 50,000–400,000 with a peak at about MW 200,000.
• 2.2. When fractions from the above chromatography were rechromatographed on Sephadex G-75 at pH 2.4 major amounts of acid-stable NSILA were found in a fraction of MW 200,000–600,000 (77% of the fraction NSILA or 28% of total serum NSILA).
• 3.3. Further evidence was obtained for the presence of an active acid-dissociable complex in serum. This was present in both the MW 100,000–200,000 and 35,000–100,000 fractions and corresponded to 37% of total serum NSILA.
• 4.4. Con-A Sepharose affinity chromatography of the serum fractions from Sephacryl S-300 chromatography, followed by Sephadex G-75 chromatography under acid conditions, showed that the acid-stable complex was consistently found in weakly bound materials. The active acid-dissociable complex was found in the bound fractions, especially in the Sephacryl S-300 pool of MW 35,000–100,000.
• 5.5. Low MW NSILA (<15,000) was also released on acid treatment from an otherwise inactive high MW complex(es) of MW 35,000–600,000. This complex was not bound by Con-A Sepharose.

     

Effect of hyperbaric oxygen and medicinal leeching on survival of axial skin flaps subjected to total venous occlusion

This study evaluates the effect of hyperbaric oxygen and medicinal leeching on axial skin flaps subjected to total venous occlusion. Axial epigastric skin flaps (3 x 6 cm) were elevated on their vascular pedicles in 40 male Wistar rats. Total venous occlusion was achieved by division of all veins draining the skin flap. Arterial inflow was left intact. Animals were randomly assigned to one of five groups: sham (n = 8); control, total venous occlusion only (n = 8); occlusion with hyperbaric oxygen (n = 8); occlusion with leeching (n = 8); occlusion with leeching and hyperbaric oxygen (n = 8). The hyperbaric oxygen protocol consisted of 90-minute treatments, twice daily, with 100% O2 at 2.5 atmospheres absolute for 4 days. The leeching protocol consisted of placing medicinal leeches on the congested flaps for 15 minutes, once daily, for 4 days. Laser Doppler measurements of flap perfusion were recorded preoperatively, postoperatively, and on postoperative days 1 and 3. The percentage of flap necrosis was evaluated on postoperative day 3. Mean percentage necrosis and mean laser Doppler readings were compared between both groups. The flaps in the sham group demonstrated 99 percent survival, whereas the flaps in the occlusion-only group demonstrated 100 percent necrosis. The flaps in the occlusion with oxygen, the occlusion with leeching, and the occlusion with oxygen and leeching groups demonstrated 1, 25, and 67 percent survival, respectively. Sham laser Doppler readings remained within normal limits. Laser Doppler readings in the occlusion-only and the occlusion with oxygen groups decreased to negligible levels on postoperative day 1, and on postoperative day 3 no perfusion was demonstrated. In both the occlusion with leeching and the occlusion with leeching and oxygen groups, there was also a significant decrease in laser Doppler measurements after surgery, but perfusion remained stable throughout the remainder of the study. This study demonstrates that hyperbaric oxygen alone is not an effective treatment for skin flaps compromised by total venous occlusion. The combination of leeching and hyperbaric oxygen treatment of total venous occlusion results in a significant increase in flap survival above that found with leeching alone. It appears that hyperbaric oxygen is effective because of the venous outflow provided by leeching as demonstrated by laser Doppler flow readings.     

A novel group of sperm activating peptides from the sea urchin Glyptocidaris crenularis

1. Six new sperm activating peptides were purified from the egg jelly of the sea urchin Glyptocidaris crenularis and their amino acid sequences were determined as follows: Ser-Ala-Lys-Leu-Cys-Pro-Gly-Gly-Asn-Cys-Val, Lys-Leu-Cys-Pro-Gly-Gly-Asn-Cys-Val, Leu-Cys-Pro-Gly-Gly-Asn-Cys-Val, Ser-Phe-Lys-Leu-Cys-Pro-Gly-Gly-Gln-Cys-Val, Lys-Leu-Cys-Pro-Gly-Gly-Gln-Cys-Val and Leu-Cys-Pro-Gly-Gly-Gln-Cys-Val. 2. The peptides were specific for G. crenularis spermatozoa and caused significant increases of sperm respiration rates and sperm cyclic nucleotide concentrations at concentrations as low as 10(-10) M. 3. The addition of the peptides to intact spermatozoa resulted in the change of the apparent mol. wt of a sperm protein from 195,000 to 200,000.     

Beta-2-Adrenergic stimulation of ornithine decarboxylase activity in porcine granulosa cells in vitro

Epinephrine, norepinephrine, and isoproterenol produced dose-dependent stimulation of ornithine decarboxylase (EC 4.1.1.17) activity in isolated porcine granulosa cells maintained under defined conditions in vitro. beta- but not alpha-receptor-blocking agents prevented enzyme stimulation by catecholamines. Application of preferential beta-1 and beta-2-receptor antagonists and agonists localized the epinephrine effect to beta-2-adrenergic mediation. Epinephrine action was enhanced by the phosphodiesterase inhibitor, 1-methyl-3-isobutyl-xanthine, but not by saturating concentrations of the cyclic AMP analogue, 8-bromocyclic AMP, of follicle-stimulating hormone, or of prostaglandin E2. However, stimulation by epinephrine was additive to that of luteinizing hormone. Follicular fluid obtained from immature Graafian follicles contained concentrations of norepinephrine and epinephrine active in vitro. Thus, catecholamines may participate in the regulation of ornithine decarboxylase activity in the ovary. Catecholamine effects may be mediated by beta-2-receptors linked to the adenylate cyclase system.     

Species differences in epinephrine concentration and norepinephrine N-methyltransferase activity in hypothalamus and brain stem

1. The concentration of epinephrine, norepinephrine and dopamine, and the activity of norepinephrine N-methyltransferase, the epinephrine-forming enzyme, were determined in hypothalamus and brain stem in several species. 2. Epinephrine concentration in hypothalamus, a nerve terminal region, varied in the order frog greater than turtle greater than chicken greater than cat greater than dog greater than pigeon greater than rat greater than ferret greater than hamster greater than mouse, with concentrations being undetectable in rabbits, horses and guinea pigs. 3. Epinephrine concentration was lower than norepinephrine concentration in all species except the frog. 4. NMT activity was detected in all species except guinea pigs. 5. Epinephrine concentration was lower in brain stem, a cell body region, than in hypothalamus in all species. Only in the frog brain stem was there more epinephrine than norepinephrine. 6. No epinephrine or NMT activity was detected in either brain region in guinea-pigs.     

High molecular weight soluble neutral maltase-glucoamylases in the intestine of the suckling rat

Previous work from our laboratory has shown that the intestine of the suckling rat, unlike adult rat intestine, contains abundant quantities of at least two soluble neutral maltase-glucoamylases. These enzymes are related antigenically to membrane-bound maltase-glucoamylase, which predominates in adult intestine, but are either more easily solubilized or occupy a different cellular locus. To study the soluble enzymes further, we attempted their isolation from the intestine of 11-day-old suckling rats. Initial attempts were complicated by proteolytic degradation, despite the addition of phenylmethylsulfonyl fluoride, N-ethylmaleimide, leupeptin, pepstatin, and EDTA to buffers used for homogenization and column chromatography. Addition of aprotinin, amastatin, bestatin, and phosphoramidone resulted, however, in the isolation of two stable, high molecular weight maltases (HM1 and HM2). Both enzymes eluted before a papain-solubilized membrane-derived maltase-glucoamylase on Sepharose 4B and were separable by DE-52 and Sepharose 6B - Tris affinity columns. They were further purified on a lentil lectin - Sepharose 4B column. Substrate specificities were almost the same and characteristic of maltase-glucoamylases. Hydrophobic binding properties and pH optima of HM1 and HM2 were also similar. HM1 was resolved by sodium dodecyl sulfate - polyacrylamide gel electrophoresis into approximately equal portions of an endo-beta-N-acetylglucosaminidase H sensitive enzyme of molecular weight (MW) 200,000 and an endo-beta-N-acetylglucosaminidase H resistant but endo-beta-acetylglucosaminidase F sensitive enzyme of MW 400,000. In contrast, most of HM2 consisted of a doublet of MW 200,000 - 210,000 that was endo-beta-N-acetylglucosaminidase H sensitive. The intestine of the suckling rat, therefore, contains two soluble maltase-glucoamylase fractions, with a major portion of high mannose rather than complex oligosaccharides.(ABSTRACT TRUNCATED AT 250 WORDS)     

A map of photolytic and tryptic cleavage sites on the beta heavy chain of dynein ATPase from sea urchin sperm flagella

NH2-terminal analysis of the alpha and beta heavy chain polypeptides (Mr greater than 400,000) from the outer arm dynein of sea urchin sperm flagella, compared with that of the 230,000- and 200,000-Mr peptides formed upon photocleavage of dynein by irradiation at 365 nm in the presence of vanadate and ATP, shows that the NH2 termini of the intact chains are acetylated and that the 230,000- and 200,000 Mr peptides constitute the amino- and carboxy-terminal portions of the heavy chains, respectively. Tryptic digestion of the beta heavy chain is known to separate it into two particles, termed fragments A and B, that sediment at 12S and 6S (Ow, R. A., W.-J. Y. Tang, G. Mocz, and I. R. Gibbons, 1987. J. Biol. Chem. 262:3409-3414). Immunoblots against monoclonal antibodies specific for epitopes on the beta heavy chain, used in conjunction with photoaffinity labeling, show that the ATPase-containing fragment A is derived from the amino-terminal region of the beta chain, with the two photolytic sites thought to be associated with the purine-binding and the gamma-phosphate-binding areas of the ATP-binding site spanning an approximately 100,000 Mr region near the middle of the intact beta chain. Fragment B is derived from the complementary carboxy-terminal region of the beta chain.     

Biomechanical and viscoelastic properties of skin,SMAS, and composite flaps as they pertain to rhytidectomy

Previous studies have focused on biomechanical and viscoelastic properties of the superficial musculoaponeurotic system (SMAS) flap and the skin flap lifted in traditional rhytidectomy procedures. The authors compared these two layers with the composite rhytidectomy flap to explain their clinical observations that the composite dissection allows greater tension and lateral pull to be placed on the facial and cervical flaps, with less long-term stress-relaxation and tissue creep. Eight fresh cadavers were dissected by elevating flaps on one side of the face and neck as skin and SMAS flaps and on the other side as a standard composite rhytidectomy flap. The tissue samples were tested for breaking strength, tissue tearing force, stress-relaxation, and tissue creep. For breaking strength, uniform samples were pulled at a rate of 1 inch per minute, and the stress required to rupture the tissues was measured. Tissue tearing force was measured by attaching a 3-0 suture to the tissues and pulling at the same rate as that used for breaking strength. The force required to tear the suture out of the tissues was then measured. Stress-relaxation was assessed by tensing the uniformly sized strips of tissue to 80 percent of their breaking strength, and the amount of tissue relaxation was measured at 1-minute intervals for a total of 5 minutes. This measurement is expressed as the percentage of tissue relaxation per minute. Tissue creep was assessed by using a 3-0 suture and calibrated pressure gauge attached to the facial flaps. The constant tension applied to the flaps was 80 percent of the tissue tearing force. The distance crept was measured in millimeters after 2 and 3 minutes of constant tension. Breaking strength measurements demonstrated significantly greater breaking strength of skin and composite flaps as compared with SMAS flaps (p < 0.05). No significant difference was noted between skin and composite flaps. However, tissue tearing force demonstrated that the composite flaps were able to withstand a significantly greater force as compared with both skin and SMAS flaps (p < 0.05). Stress-relaxation analysis revealed the skin flaps to have the highest degree of stress-relaxation over each of five 1-minute intervals. In contrast, the SMAS and composite flaps demonstrated a significantly lower degree of stress-relaxation over the five 1-minute intervals (p < 0.05). There was no difference noted between the SMAS flaps and composite flaps with regard to stress-relaxation. Tissue creep correlated with the stress-relaxation data. The skin flaps demonstrated the greatest degree of tissue creep, which was significantly greater than that noted for the SMAS flaps or composite flaps (p < 0.05). Comparison of facial flaps with cervical flaps revealed that cervical skin, SMAS, and composite flaps tolerated significantly greater tissue tearing forces and demonstrated significantly greater tissue creep as compared with facial skin, SMAS, and composite flaps (p < 0.05). These biomechanical studies on facial and cervical rhytidectomy flaps indicate that the skin and composite flaps are substantially stronger than the SMAS flap, allowing significantly greater tension to be applied for repositioning of the flap and surrounding subcutaneous tissues. The authors confirmed that the SMAS layer exhibits significantly less stress-relaxation and creep as compared with the skin flap, a property that has led aesthetic surgeons to incorporate the SMAS into the face lift procedure. On the basis of the authors' findings in this study, it seems that that composite flap, although composed of both the skin and SMAS, acquires the viscoelastic properties of the SMAS layer, demonstrating significantly less stress-relaxation and tissue creep as compared with the skin flap. This finding may play a role in maintaining long-term results after rhytidectomy. In addition, it is noteworthy that the cervical flaps, despite their increased strength, demonstrate significantly greater tissue creep as compared with facial flaps, suggesting earlier relaxation of the neck as compared with the face after rhytidectomy.     

Alkaline phosphatase isozymes of Xenopus laevis embryos and tissues.

Alkaline phosphatase was obtained by treating embryos of Xenopus laevis with n-butanol at different developmental stages from gastrula to tadpole; the enzyme was also obtained from adult kidney, liver, and intestinal mucosa. Purification was carried out by gel filtration and polyacrylamide gel electrophoresis. The enzyme activity is chromatographically spearated into two peaks, with molecular weights of approximately 200,000 and 400,000. Alternatively, two groups may be characterized on the basis of their electrophoretic mobilities, which correspond to the different molecular weight classes. Effects of pH, temperature, inhibitors, and substrate concentration were studied. The kinetic and physical properties of the two alkaline phosphatase isozymes are similar, and are comparable to the properties reported for this enzyme from other vertebrates. Alkaline phosphatase activity increased sharply at the gastrula stage and reached a plateau at the late tailbud stage. During this period there was an 18-fold increase in activity.     

L-精氨酸对任意型皮瓣成活的影响

目的　探讨L 精氨酸对任意型皮瓣的成活的影响。方法　以Wistar大鼠为实验对象 ,在其背部设计 7cm×2cm任意型皮瓣 ,于术后给予腹腔注射L 精氨酸 30 0mg/kg ,对照组给予 0 9%生理盐水。术后 7d ,通过图像分析技术观察皮瓣成活率 ;通过生物化学技术、组织学和免疫组织化学技术对不同时间皮瓣组织中一氧化氮含量、组织形态学变化、白细胞计数以及ICAM 1的表达进行观测。结果　外源性L 精氨酸可提高皮瓣组织一氧化氮含量 ,L 精氨酸组ICAM 1表达呈弱阳性、术后 12h皮瓣组织真皮中性粒细胞计数减少 ,L 精氨酸组皮瓣成活率明显高于对照组 (P <0 0 1)。结论　外源性L 精氨酸可提高皮瓣组织NO含量 ,ICAM 1表达下调 ,中性粒细胞浸润减少 ,提高任意皮瓣成活率。     

The aggregation state of bovine heart cytochrome c oxidase and its kinetics in monomeric and dimeric form

The monomeric and dimeric forms of bovine cytochrome c oxidase (EC 1.9.3.1) were obtained from gel filtration chromatography on Ultrogel AcA 34 and analyzed. Both species contained all 12-13 subunits described for this enzyme. In the dimer 320 molecules [3H]dodecyl-beta-D-maltoside were bound per heme aa3 and in the monomer 360 molecules per heme aa3. The monomers contained 10 mol of tightly bound phospholipid/mol heme aa3 and the dimers 14. Sedimentation coefficients of 15.5-18 S for the dimer and 9.6 S for the monomer were calculated from sucrose density centrifugation analysis and analytical centrifugation. By the laser beam light-scattering technique a Stokes radius of 70 A for the dimeric detergent-lipid-protein complex was measured. From those parameters and the densitometric determined partial specific volumes of the detergent and the enzyme, the molecular weights of 400,000 for the protein moiety of the dimer and 170,000-200,000 for the monomer were calculated. Under very low ionic strength conditions the monomer/dimer equilibrium was found to be dependent on the protein concentration. At low enzyme concentrations (10(-9) M) monomers were predominant, whereas at concentrations above 5 X 10(-6) M the amounts of dimers and higher aggregates were more represented. The cytochrome c oxidase activity, measured spectrophotometrically and analyzed by Eadie-Hofstee plot, was biphasic as a function of cytochrome c concentration for the dimeric enzyme. Pure monomers gave monophasic kinetics. The data, fitting with a homotropic negative cooperative mechanism for the dimer of cytochrome c oxidase, are discussed and compared with other described mechanisms.     

Effects of epinephrine on the net hepatic uptake of lactate, pyruvate, and glycerol in sheep

Epinephrine caused hyperglycemia in part by increasing gluconeogenesis. However, the mechanism of its gluconeogenic effects has not been studied in ruminants. This study was undertaken to examine the effect of epinephrine on the net hepatic uptake of selected glucose precursors in sheep. The major abdominal blood vessels of the sheep were catheterized in normal and alloxan diabetic sheep. Glucose production, metabolic clearance of glucose, and the hepatic removal of certain glucose precursors were determined before, during, and after epinephrine infusion. Epinephrine increased the hepatic glucose output, the concentrations of lactate and glycerol in plasma, and the net hepatic uptake and fractional hepatic extraction of lactate and glycerol. These effects were independent of changes in the concentrations of insulin and glucagon in plasma. These results show that epinephrine directly stimulates hepatic gluconeogenesis in sheep.