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PubMed2004

Ligating DNA with DNA.

Cloning DNA typically involves the joining of target DNAs with vector constructs by enzymatic ligation. A commonly used enzyme for this reaction is bacteriophage T4 DNA ligase, which requires ATP as the energy source to catalyze the otherwise unfavorable formation of a phosphodiester bond. Using in vitro selection, we have isolated a DNA sequence that catalyzes the ligation of DNA in the absence of protein enzymes. We have used the action of two catalytic DNAs, an ATP-dependent self-adenylating deoxyribozyme (AppDNA) and a self-ligating deoxyribozyme, to create a ligation system that covalently joins oligonucleotides via the formation of a 3',5'-phosphodiester linkage. The two-step process is conducted in separate reaction vessels wherein the products of deoxyribozyme adenylation are purified before their use as substrates for deoxyribozyme ligation. The final ligation step of the deoxyribozyme-catalyzed sequence of reactions mimics the final step of the T4 DNA ligase reaction. The initial rate constant (k(obs)) of the optimized deoxyribozyme ligase was found to be 1 x 10(-)(4) min(-)(1). Under these conditions, the ligase deoxyribozyme promotes DNA ligation at least 10(5)-fold faster than that generated by a simple DNA template. The self-ligating deoxyribozyme has also been reconfigured to generate a trans-acting construct that joins separate DNA oligonucleotides of defined sequence. However, the sequence requirements of the AppDNA and that of the 3' terminus of the deoxyribozyme ligase limit the range of sequences that can be ligated.

Base Sequence↗
PubMedDate not supplied

Fertility of mares after unilateral laparoscopic tubal ligation.

OBJECTIVE: To develop a technique for laparoscopic tubal (oviductal) ligation and to evaluate pregnancy rates for mares that ovulated ipsilateral or contralateral to the ligated oviduct. STUDY DESIGN: Randomized prospective clinical trial comparing pregnancy rates after unilateral laparoscopic tubal ligation. ANIMALS: Twelve mares of light horse breeds. METHODS: One oviduct in each of 6 mares was surgically ligated with a laparoscopic technique; 6 other mares served as nonligated controls. Mares with unilateral tubal ligations (UTL) were inseminated with 500 million progressively motile sperm during 1 cycle when the dominant follicle was ipsilateral to the ligation site and 1 cycle when the dominant follicle was contralateral to the ligation site. Control mares were bred during 2 cycles regardless of the side of the dominant follicle. Pregnancy examinations were performed on days 12, 14, and 16 after ovulation by transrectal ultrasonography. RESULTS: None of the mares became pregnant when ovulations occurred from the ovary adjacent to the ligated oviduct. All 6 mares became pregnant on the first cycle when an ovulation occurred from the opposite ovary. Control mares became pregnant on 10 of 12 cycles (83.3 %). CONCLUSIONS: UTL was completely effective in preventing pregnancy when ovulation occurred ipsilateral to the ligation site. The surgical procedure did not interfere with the establishment of pregnancy when ovulation occurred from the contralateral ovary. CLINICAL RELEVANCE: UTL may be a clinically useful procedure for preparing a recipient mare for gamete intrafallopian transfer. The recipient mare could be allowed to ovulate and UTL would prevent fertilization of her oocyte but would not interfere with normal corpus luteum formation. The donor oocyte could be placed into the oviduct contralateral to the UTL site.

Animals↗
PubMed2006

Structural role of (bacterio)chlorophyll ligated in the energetically unfavorable beta-position.

Chlorophyll is attached to apoprotein in diastereotopically distinct ways, by beta- and alpha-ligation. Both the beta- and alpha-ligated chlorophylls of photosystem I are shown to have ample contacts to apoprotein within their proteinaceous binding sites, in particular, at C-13 of the isocyclic ring. The H-bonding patterns for the C-13(1) oxo groups, however, are clearly distinct for the beta-ligated and alpha-ligated chlorophylls. The beta-ligated chlorophylls frequently employ their C-13(1) oxo in H-bonds to neighboring helices and subunits. In contrast, the C-13(1) oxo of alpha-ligated chlorophylls are significantly less involved in H-bonding interactions, particularly to neighboring helices. Remarkably, in the peripheral antenna, light harvesting complex (LH2) from Rhodobacter sphaeroides, a single mutation in the alpha-subunit, introduced to eliminate H-bonding to the beta-bacteriochlorophyll-B850, which is ligated in the "beta-position," results in significant thermal destabilization of the LH2 in the membrane. In addition, in comparison with wild type LH2, the expression level of the LH2 lacking this H-bond is significantly reduced. These findings show that H-bonding to the C-13(1) keto group ofbeta-ligated (bacterio)-chlorophyll is a key structural motif and significantly contributes to the stability of bacteriochlorophyll proteins in the native membrane. Our analysis of photosystem I and II suggests that this hitherto unrecognized motif involving H-bonding to beta-ligated chlorophylls may be equally critical for the stable assembly of the inner core antenna of these multicomponent chlorophyll proteins.

Amino Acid Motifs↗
PubMed1997

Ligation of major histocompatability complex (MHC) class I molecules on human T cells induces cell death through PI-3 kinase-induced c-Jun NH2-terminal kinase activity: a novel apoptotic pathway distinct from Fas-induced apoptosis.

Ligation of major histocompatability complex class I (MHC-I) molecules expressed on T cells leads to both growth arrest and apoptosis. The aim of the current study was to investigate the intracellular signal pathways that mediate these effects. MHC-I ligation of human Jurkat T cells induced a morphologically distinct form of apoptosis within 6 h. A specific caspase inhibitor, which inhibited Fas-induced apoptosis, did not affect apoptosis induced by MHC-I ligation. Furthermore, MHC-I-induced apoptosis did not involve cleavage and activation of the poly(ADP- ribose) polymerase (PARP) endonuclease or degradation of genomic DNA into the typical fragmentation ladder, both prominent events of Fas-induced apoptosis. These results suggest that MHC-I ligation of Jurkat T cells induce apoptosis through a signal pathway distinct from the Fas molecule. In our search for other signal pathways leading to apoptosis, we found that the regulatory 85-kD subunit of the phosphoinositide-3 kinase (PI-3) kinase was tyrosine phosphorylated after ligation of MHC-I and the PI-3 kinase inhibitor wortmannin selectively blocked MHC-I-, but not Fas-induced, apoptosis. As the c-Jun NH2-terminal kinase (JNK) can be activated by PI-3 kinase activity, and has been shown to be involved in apoptosis of lymphocytes, we examined JNK activation after MHC-I ligation. Strong JNK activity was observed after MHC-I ligation and the activity was completely blocked by wortmannin. Inhibition of JNK activity, by transfecting cells with a dominant-negative JNKK- MKK4 construct, led to a strong reduction of apoptosis after MHC-I ligation. These results suggest a critical engagement of PI-3 kinase-induced JNK activity in apoptosis induced by MHC-I ligation.

Androstadienes↗
PubMed1997

Selective suppression of interleukin-12 induction after macrophage receptor ligation.

Interleukin (IL)-12 is a monocyte- and macrophage-derived cytokine that plays a crucial role in both the innate and the acquired immune response. In this study, we examined the effects that ligating specific macrophage receptors had on the induction of IL-12 by lipopolysaccharide (LPS). We report that ligation of the macrophage Fcgamma, complement, or scavenger receptors inhibited the induction of IL-12 by LPS. Both mRNA synthesis and protein secretion were diminished to near-undetectable levels following receptor ligation. Suppression was specific to IL-12 since IL-10 and tumor necrosis factor-alpha (TNF-alpha) production were not inhibited by ligating macrophage receptors. The results of several different experimental approaches suggest that IL-12 downregulation was due to extracellular calcium influxes that resulted from receptor ligation. First, preventing extracellular calcium influxes, by performing the assays in EGTA, abrogated FcgammaR-mediated IL-12(p40) mRNA suppression. Second, exposure of macrophages to the calcium ionophores, ionomycin or A23187, mimicked receptor ligation and inhibited IL-12(p40) mRNA induction by LPS. Finally, bone marrow-derived macrophages from FcR gamma chain-deficient mice, which fail to flux calcium after receptor ligation, failed to inhibit IL-12(p40) mRNA induction. These results indicate that the calcium influxes that occur as a result of receptor ligation are responsible for inhibiting the induction of IL-12 by LPS. Hence, the ligation of phagocytic receptors on macrophages can lead to a dramatic decrease in IL-12 induction. This downregulation may be a way of limiting proinflammatory responses of macrophages to extracellular pathogens, or suppressing the development of cell-mediated immunity to intracellular pathogens.

Animals↗
PubMed2002

CD40 ligation-induced cytokine production in human skin explants is partly mediated via IL-1.

CD40 ligation by CD40 ligand(+) CD4(+) T cells has been claimed to be involved in inflammatory responses in human skin. However, these data are derived from in vitro cell culture systems and immunohistochemistry, and the mechanisms involved have not been fully elucidated. We previously observed that cells in intact normal human skin secrete high levels of IL-6 and IL-8 upon stimulation with IL-1 beta. In vitro studies have shown that CD40 ligation on human keratinocytes results in the production of IL-6 and IL-8 as well. We used a novel tissue culture system with intact normal human skin, and show that antibody ligation of CD40 results in the induction of several pro- and anti-inflammatory cytokines. IL-6, IL-8, tumor necrosis factor (TNF)-alpha, IL-12 and IL-1 beta were induced upon CD40 ligation and IFN-gamma stimulation, while IL-10 could be induced by CD40 ligation alone and was reduced again by the addition of IFN-gamma. Since CD40 ligation on monocytes and dendritic cells in vitro results in the secretion of IL-1, which is pre-stored in high concentrations in normal human keratinocytes, we subsequently investigated whether CD40 induced IL-6 and IL-8 production in skin is mediated via IL-1. Indeed IL-1 receptor antagonist inhibited the CD40 ligation-induced IL-6 and IL-8 production, while TNF-alpha and IL-10 production were not affected. These data show that CD40 ligation-induced secretion of IL-6 and IL-8, but not TNF-alpha and IL-10, is partially mediated via IL-1 and that IL-1 plays a prominent role in the inflammatory response initiated by CD40 ligation in intact human skin.

CD40 Antigens↗
PubMed1998

Chemical ligation of unprotected peptides directly from a solid support.

In this article we describe a new, convenient procedure to carry out intramolecular (cyclization) and intermolecular native chemical ligations of unprotected peptides directly from a solid support. Our solid-phase ligation approach eliminates the need to manipulate peptide (alpha)thioacid and peptide (alpha)thioester intermediates in aqueous solution before the ligation step, thereby leading to a reduction in handling losses and significantly increasing the overall efficiency of the chemical ligation strategy. A key step in our ligation scheme is the ability to generate fully unprotected peptides tethered to a solid support through an (alpha)thioester linkage. This can be achieved efficiently using optimized Boc-solid-phase peptide synthesis on a 3-mercaptopropionamide-polyethylene glycol-poly-(N,N-dimethylacrylamide) copolymer support (HS-PEGA). Once the synthesis is complete, the fully protected peptide (alpha)thioester resin is treated with HF to give the corresponding fully unprotected peptide (alpha)thioester resin. Using this procedure several polypeptides ranging from 15 to 47 residues were synthesized successfully. These peptide-resins were then used to perform both intramolecular (head-to-tail cyclizations) and intermolecular solid-phase ligations. The intramolecular solid-phase ligations proceeded much faster than their intermolecular counterparts, but in both cases the reactions were observed to be remarkably clean. The presence of aromatic thiol cofactors significantly accelerated the relatively slow intermolecular ligations. This novel methodology was then extended to provide a general method for performing sequential intermolecular ligations, allowing easy access to much larger polypeptide and protein systems.

Amino Acid Sequence↗
PubMed1983

Portal vein ligation selectively lowers hepatic cytochrome P450 levels in rats.

In rats, surgical creation of a portacaval shunt leads to hepatic atrophy and lowered levels of cytochrome P450, the key component of liver enzymes involved with drug metabolism. These effects are largely attributable to diversion of portal blood away from the liver and not to decreased hepatic blood flow. The present study has established a simpler model of portal blood diversion in order to examine the role of portal blood constituents in the regulation of hepatic cytochrome P450. Portal vein ligation was performed on male Wistar rats in which portasystemic anastomoses had been produced by subcutaneous transposition of the spleen. Portal vein ligation resulted in portal hypertension, as evidenced by splenomegaly, and in hepatic atrophy. In liver of rats with portal vein ligation, microsomal cytochrome P450 levels were significantly less than in sham-operated control rats, but cytochrome b5, NADPH-cytochrome c reductase, and glucose-6-phosphatase were unaltered. The activities of four mixed function oxidases also were reduced significantly in the liver of rats with portal vein ligation, the changes being greatest for ethylmorphine N-demethylase, a prototype substrate for the phenobarbital-inducible isoenzyme of cytochrome P450. In contrast, the activity of microsomal heme oxygenase, the rate-limiting step in catabolism of heme to bilirubin, was enhanced after portal vein ligation. Experiments in pair-fed rats showed that the changes observed in liver from rats with portal vein ligation could not be attributed to caloric deprivation. Administration of phenobarbital increased liver mass, cytochrome P450 levels, and mixed function oxidase activities both in rats with portal vein ligation and in controls, indicating that the liver of the ligated rats retained considerable protein synthetic capacity. It appears that hepatic atrophy and lowering of cytochrome P450 levels that follow portal vein ligation are consequences of altered exposure of the liver to factors normally present in portal blood, and that the same alterations may also enhance heme oxygenase activity.

Animals↗
PubMed1993

Oxygen equilibrium studies of cross-linked iron-cobalt hybrid hemoglobins. Models for partially ligated intermediates of cobalt hemoglobin.

To probe the molecular mechanism of allosteric function of cobaltous protoporphyrin-substituted hemoglobin (CoHb), a series of alpha, alpha-cross-linked symmetric and asymmetric Fe-Co hybrid hemoglobins, which contain (1Co porphyrin/3Fe porphyrins), (2Co porphyrins/2Fe porphyrins), and (3Co porphyrins/1Fe porphyrin) per tetramer, have been prepared. Because only Fe porphyrin-containing subunits react with CO, these Fe-Co hybrids are converted to mono-, di-, and tri-CO-ligated states in the presence of CO, respectively, and are proposed to stand as models for mono-, di-, and tri-ligated intermediates of CoHb, respectively. The oxygen binding properties of these Fe-Co hybrids were investigated by measuring oxygen binding isotherms in the presence of CO as a function of pH in the presence and absence of IHP. The ligation of CO to a beta subunit causes larger changes in the oxygen affinity and the Bohr effect than that to an alpha subunit, indicating that the ligation to a beta subunit induces larger affinity-related structural changes in cross-linked CoHb. Di- and tri-CO-ligated intermediates exhibited substantially increased oxygen affinity, reduced Bohr effect, and reduced IHP effect, indicating that they are in high affinity states. Calculation of the Adair equilibrium constants for the first and last oxygenation steps for each of these intermediates permitted the determination of the level of free energy of cooperation. The intermediately ligated species of cross-linked CoHb are distributed in multiple levels of free energy of cooperation within the free energy difference of 1.14 kcal mol-1 between deoxy and fully ligated states of cross-linked CoHb at pH 7.4. The ligation process in CoHb is determined by the number and distribution of the bound ligands, and ligation takes place through steps that require minimal free energy changes.

Cobalt↗
PubMed1991

Effects of maternal bilateral ureteral ligation on the differentiation of glomerular anionic sites in fetal rat kidney.

Effects of maternal bilateral ureteral ligation on the differentiation of glomerular anionic sites in fetal kidney were studied with the colloidal iron method. The ligation was performed on days 17, 19, and 21 of pregnancy, and the animal were euthanized 1 day after each ligation. On fetal day 20, in the fetuses from ligated mothers, colloidal iron was concentrated in laminae rarae interna and externa of the glomerular basement membrane. In the fetuses from sham-ligated mothers, however, a different pattern of iron distribution was noted. In most areas of the glomerular basement membrane, colloidal iron was randomly distributed as in both fetuses of fetal day 18. In the remaining areas, the same pattern of colloidal iron distribution as was noted in the fetuses from ligated mothers of fetal day 20 was observed. On fetal day 22, colloidal iron was densely accumulated in laminae rarae interna and externa of the glomerular basement membrane in both groups. However, the iron-free zone was slightly thicker in the fetuses from the ligated mothers. These findings suggest that maternal bilateral ureteral ligation accelerates the differentiation of glomerular anionic sites in the kidney of fetal rats.

Animals↗
PubMed1999

Tracheal ligation increases mitogen-activated protein kinase activity and attenuates surfactant protein B mRNA in fetal sheep lungs.

BACKGROUND: Tracheal ligation has been shown to accelerate fetal pulmonary growth in normal and hypoplastic lungs. Our aim was to study the effects of tracheal ligation on established molecular markers of growth and differentiation [mitogen-activated protein (MAP) kinase] and maturity [surfactant protein B (SPB) and fatty acid synthase (FAS)]. MATERIALS AND METHODS: Tracheal ligation was performed on four 100-day-gestation fetal sheep, with four age-matched fetuses undergoing maternal laparotomy and hysterotomy as control. Lungs from surviving fetuses (n = 2 in each group) were harvested after 4 days and frozen in liquid nitrogen. Protein lysates were prepared, and MAP kinase enzymatic assays [extracellular signal regulated protein kinase (ERK)-1 and -2] and Western blots were performed. Total RNA was isolated, and a fetal sheep lung cDNA library was created. The sheep SPB and FAS genes were cloned and sequenced. Northern blots were performed with the new clones, normalizing to beta-actin. RESULTS: Tracheal ligation lungs contained a larger volume of fluid (40 ml) compared with age-matched controls (8 ml). MAP kinase enzymatic ERK-1 activity was increased and SPB mRNA expression was reduced in fetal lungs after tracheal ligation. Neither ERK-2 enzymatic activities and FAS mRNA nor ERK protein levels were affected by tracheal ligation, by Western blot analysis. CONCLUSION: Tracheal ligation-induced fetal lung growth may be mediated in part via the MAP kinase pathway. Expression of SPB mRNA is attenuated by tracheal ligation, whereas FAS, one of the key enzymes that synthesizes the lipid portion of surfactant, is not affected.

Animals↗
PubMed1999

Triple rubber band ligation for hemorrhoids: prospective, randomized trial of use of local anesthetic injection.

PURPOSE: Rubber band ligation is a common office procedure for hemorrhoids. Triple rubber band ligation in a single session has been shown to be a safe and economical way of treating hemorrhoids. However, postligation discomfort after triple rubber band ligation is not uncommon. The aim of this study was to evaluate the effectiveness of local anesthetic injection to the banded hemorrhoidal tissue in reducing postligation discomfort. METHODS: Patients attending an outpatient clinic for symptomatic hemorrhoids suitable for triple rubber band ligation were randomly assigned to two groups. In the treatment group rubber band ligation was performed at three columns of hemorrhoids, and 1 to 2 ml of 2 percent lignocaine was injected into the banded hemorrhoidal tissue. In the control group triple rubber band ligation was performed in a similar manner, but local anesthetic was not given. Patients were followed up by telephone at the second week and in the clinic after six weeks. RESULTS: From April to August 1996, 101 patients entered the trial and were treated with triple rubber band ligation. Sixty-two patients were randomly assigned to the local anesthetic injection group and 39 to the control group. Overall good to excellent results occurred in 89 percent of patients, and there was no difference between the two groups. Postligation pain occurred in 26 and 20 percent of patients in the treatment and control groups, respectively (P > 0.05). Postligation tenesmus occurred in 32 and 41 percent of patients in the treatment and control groups, respectively (P > 0.05). No patients suffered from septic complications or bleeding that required transfusion. CONCLUSION: Triple rubber band ligation in a single session is a safe, economical, and effective way of treating symptomatic hemorrhoids. Postligation pain and tenesmus occurred in 24 and 37 percent, respectively. Discomfort was usually tolerable. Local anesthetic injection to the banded hemorrhoidal tissue did not help to reduce postligation discomfort.

Anesthetics, Local↗
PubMed2001

Endoscopic ligation for patients with active bleeding Mallory-Weiss tears.

BACKGROUND: Only a few patients with active nonesophageal variceal upper gastrointestinal bleeding have been treated with endoscopic ligation. To further address this issue, four patients with active bleeding Mallory-Weiss tears who underwent endoscopic band ligation are presented. PATIENTS AND METHODS: Endoscopic ligation was performed in four patients with a median age of 52 years (range, 40-93 years) after a diagnosis of active bleeding Mallory-Weiss tears (MWTs). A 45-year-old man with massive persistent upper gastrointestinal bleeding as a cause of a MWT underwent therapeutical endoscopic band ligation after an unsuccessful endoscopic injection trial. On the contrary, injection therapy should have been performed on a 93-year-old woman with multiple myeloma because of an actively bleeding MWT caused by the fibrotic tissue after an unsuccessful endoscopic ligation trial, although her other actively bleeding MWT lesion had been ligated successfully. RESULTS: After endoscopic ligation, all patients achieved complete hemostasis, and rebleeding did not occur. They were discharged without complications after a control endoscopy. CONCLUSIONS: Endoscopic ligation can be performed easily and without any complications such as perforation or delayed hemorrhage in patients with actively bleeding nonfibrotic MWTs.

Adult↗
PubMed2001

A novel ligation forceps can be used as a ligature carrier and knot pusher during laparoscopic surgery.

BACKGROUND: To extend the usefulness of laparoscopic operations, a secure and easy method for the ligation of large vessels is needed. Herein we describe a novel ligation forceps that can be used as a ligature carrier and knot pusher. METHODS: A 2-0 suture thread with a knot already tied near one end is hooked in the upper jaw of a novel ligation forceps. After the lower jaw is passed under the vessel or cystic duct, the forceps is closed. When one end of the thread is withdrawn, the knot is trapped in the indentation built into the lower jaw; the ligature is then passed under the pedicle. An extracorporeal ligation can then be performed continuously by the same forceps. RESULTS: The origins of large vessels were ligated safely and easily with this device during 65 laparoscopic procedures (four total colectomies, 12 colectomies, and 49 gastrectomies). Following temporary hemostasis of accidental bleeding with clamping forceps, ligation hemostasis can also be performed using this instrument. CONCLUSION: This novel ligation forceps permits the secure ligation of vessels or a cystic duct without the need for another device. The proposed method is both easy and inexpensive.

Gastrectomy↗
PubMed2003

Randomized trial of rubber band ligation vs. stapled hemorrhoidectomy for prolapsed piles.

PURPOSE: The introduction of stapled hemorrhoidectomy may replace local techniques such as rubber band ligation as a first-line treatment for Grade III and small Grade IV piles. We conducted a randomized trial to determine the role of rubber band ligation in the era of stapled hemorrhoidectomy. METHODS: Fifty-five patients with Grade III or small Grade IV hemorrhoids were randomly allocated to either rubber band ligation or stapled hemorrhoidectomy. Patient demographics and procedure-related details were recorded. Follow-up was at two weeks and two and six months to assess complications, symptom relief, incontinence scores, quality of life, and patient satisfaction. RESULTS: Twenty-five patients were randomly assigned to rubber band ligation and 30 to stapled hemorrhoidectomy. The groups were equally matched for age, gender, grade of piles, continence scores, and quality of life. Stapled hemorrhoidectomy was associated with increased pain and analgesia usage at both 2-week and 2-month follow-up (P < 0.001). Rubber band ligation and stapled hemorrhoidectomy were equally effective in controlling symptomatic prolapse, but rubber band ligation was associated with an increased incidence of recurrent bleeding (P = 0.002). There were 6 procedure-related complications in the stapled hemorrhoidectomy group compared with none in the rubber band ligation group (P = 0.027). There was no difference between the two groups in terms of continence scores, patient satisfaction, or quality of life. CONCLUSION: Stapled hemorrhoidectomy is associated with more pain and minor morbidity than rubber band ligation in the treatment of Grade III and small Grade IV piles. However, for those patients who do not want the risk of further intervention procedures, stapled hemorrhoidectomy offers the better chance of a symptomatic cure.

Adult↗
PubMed1994

The cerebrovascular response to prolonged hypoxia with carotid artery and jugular vein ligation in the newborn lamb.

This study was designed to evaluate the effect of ligation of the carotid artery and/or jugular vein, after exposure to prolonged (4 hours) hypoxia, and the effect of acute normalization of PaO2 after prolonged hypoxia with vessel ligation, on the cerebral circulation. Twelve 1- to 7-day-old lambs were anesthetized with pentobarbital. Catheters were placed in the femoral artery and vein, left ventricle, lingual artery, and sagittal sinus. Cerebral blood flow (CBF) was determined using the radiolabeled microsphere technique. After baseline studies, the animals were made hypoxic with a nitrogen/air mixture, to lower PaO2 to 36 +/- 5 mm Hg for 4 hours, followed by 1 hour of normoxia. After four hours of hypoxia, studies were performed. The animals were divided into two groups to evaluate carotid artery and jugular vein ligation separately. In group I, the carotid artery was ligated first, with studies performed after 5 minutes; this was followed by ligation of the jugular vein, with studies after 5 minutes. In group II, the jugular vein was ligated first, with studies after 5 minutes; this was followed by ligation of the carotid artery, with studies after 5 minutes. With regard to physiological variables, there were no differences between the groups. CBF increased 106% (P < .001 compared with the baseline value) after 4 hours of hypoxia, maintaining cerebral oxygen consumption (CMRO2) and oxygen transport (OT) constant in both groups. Ligation of either the carotid artery or jugular vein after 4 hours of hypoxia, did not alter CBF responses to hypoxia.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2004

Venous ligation-mediated bone adaptation is NOS 3 dependent.

Interstitial fluid flow (IFF) in bone has been hypothesized to mediate bone modeling in the absence of mechanical strain. The mechanism of this effect has not been clearly defined, though previous studies indicate that nitric oxide (NO) may play an important role in mediating IFF. In the current study, mice with a targeted disruption of the NOS 3 gene were used according to a previously established model of altered interstitial fluid flow in bone. Femoral vein ligation was performed in one limb to increase intramedullary pressure and consequently its IFF; a sham operation was performed on the contralateral limb. The mice were then hindlimb suspended to uncouple the effects of altered flow in the limb from mechanical loading. Differences in radiographic bone density and bone strength were compared for the sham and venous-ligated femurs in wild-type (WT) mice and NOS 3 knockout (KO) mice. Suspension-induced bone loss in the femurs, as evidenced by a loss in radiographic bone mineral density (BMD), was seen in both groups. Differences between sham and venous-ligated femurs were significant only for the WT mice, in which there appeared to be a protective effect of venous ligation against bone loss [-6.69% (ligated) vs. -12.36% (sham), P<0.05]. Furthermore, the difference in bone density between sham and venous-ligated femurs was muted by NOS 3 knockout, suggesting that the protective effect of venous ligation against bone loss observed in the WT group was NO dependent. The differences in relative BMD were mirrored in the mechanical testing experiments, where maximum load to fracture was significantly higher in the venous-ligated limbs relative to the sham limbs of the WT mice, but not in the NOS 3 group. Taken together, these data further support the hypothesis that fluid flow can modulate bone modeling and suggest that IFF-mediated bone adaptation is NOS 3 dependent.

Animals↗
PubMed2005

Effects of temporary bilateral ligation of the internal carotid arteries on the low- and high-frequency somatic evoked potentials in the swine.

OBJECTIVE: We studied effects of a temporary bilateral ligation of the internal carotid arteries on the subcortical and cortical structures of the somatosensory system by examining the thalamic input and postsynaptic cortical responses contained in the somatic evoked potentials (SEPs) recorded from the primary somatosensory cortex (SI) of the juvenile piglets in vivo. We predicted that the ligation should differentially affect these structures due to differences in blood supply. METHODS: The SEPs between 1 and 3000 Hz were measured in the SI cortex with a multichannel electrode array before, during and after a 20 min bilateral ligation of the internal carotid arteries in the swine under a barbiturate anesthesia. RESULTS: The ligation differentially affected the thalamic input and the cortical responses contained in the high-frequency signals (HFSs) between 400 and 2000 Hz. The amplitude of the thalamic input did not change, but the amplitudes of the cortical HFS postsynaptic to the thalamic inputs decreased immediately after start of ligation, recovering over the next 30-90 min. The latency showed a small, but significant increase for several minutes after the start of ligation for both the thalamic input and cortical responses. The ligation increased the latency and reduced the amplitude of the peak of the first cortical response in the wideband SEP corresponding to human N20. CONCLUSIONS: The HFS is useful for distinguishing selective effects of the temporary ligation on the subcortical and cortical structures of the somatosensory system. Since the porcine N20 starts after the presynaptic HFS, it was not useful in differentiating thalamic and cortical effects. SIGNIFICANCE: The HFS may open a new window in studying the cortical physiology in humans.

Animals↗