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Haemodynamic studies in dogs with chronic bile-duct ligation.

1. Dogs with bile-duct ligation retain salt and water and form ascites. The present study was under-taken to examine the role of haemodynamic factors in the aetiology of this sodium retention. 2. Haemodynamic studies were performed in five dogs before and 5 weeks after bile-duct ligation. 3. After the operation there was an insignificant fall in mean arterial pressure, a significant rise in mean cardiac index and significant fall in mean total peripheral resistance. 4. It is concluded that heart failure is not a factor in renal sodium retention of the dog with bile-duct ligation, since the central venous pressure was not elevated. 5. The haemodynamic pattern and the tendency to salt retention in the dog with chronic bile-duct ligation closely resemble findings reported in patients with cirrhosis of the liver, and it is suggested that oedema formation in patients with cirrhosis of the liver and dogs with chronic bile-duct ligation shares a common aetiology.

Animals↗

Blunted pressor response to angiotensin and sympathomimetic amines in bile-duct ligated dogs.

1. The pressor and heart rate response to angiotensin II, noradrenaline, isoprenaline, tyramine and beta-phenylethylamine were examined before and after bile-duct ligation in conscious trained dogs. 2. The pressor response to angiotensin II was markedly suppressed after bile-duct ligation, although responses to noradrenaline and the indirectly acting sympathomimetic amines were only slightly reduced. 3. Bradycardia, in response to the pressor drugs, and tachycardia, in response to isoprenaline, were unchanged after bile-duct ligation. 4. Refractoriness to the pressor action of angiotensin after chronic bile-duct ligation in the present study is similar to that reported previously by others in patients with liver cirrhosis. 5. No evidence was found in the bile-duct ligated dog to support a role of false neurotransmitters as mediators of the circulatory disturbances associated with liver injury.

Angiotensin II↗

Guidelines for the ligation of the sigmoid or transverse sinus during large petroclival meningioma surgery.

The authors present guidelines for the ligation of the transverse or sigmoid sinus during the surgical removal of petroclival meningiomas. The medical records and venograms of 14 patients with a petroclival meningioma requiring transverse or sigmoid sinus ligation treated in the Department of Neurosurgery, Seoul National University Hospital between 1986 and 1999 were reviewed. All patients successfully received a sinus trial clamping during the operation. The drainage pattern of the confluens of Herophili was classified into four types: Type A, confluens and equal on both transverse sinuses; Type B, confluens and nondominant transverse sinus on the tumor side; Type C, confluens and dominant transverse sinus on the tumor side; and Type D, unilateral transverse sinus only. Of the 14 cases, four were Type A, five were Type B, and two were Type C. There was no brain swelling after intraoperative test clamping of the sinus for more than 30 minutes. None of the cases developed postoperative complications related to the sinus ligation. Patients with Type A, B, or C drainage patterns were ideal candidates for sinus ligation, especially transverse sinus ligation, if the test clamping proved to be safe. The sinus was cut proximal to the superior petrosal sinus, distal to the vein of Labbé.

Journal Article↗

Comparison of resistance to sliding between different self-ligating brackets with second-order angulation in the dry and saliva states.

Resistance to sliding was investigated for 3 self-ligating brackets having passive slides and 3 self-ligating brackets having active clips. Four of these products are currently marketed, and 2 are of historic interest. For all cases, an 0.018 x 0.025-in stainless steel archwire was drawn through each bracket at a rate of 10 mm/min over a distance of 2.5 mm. For each bracket, the resistances to sliding were measured at 14 second-order angulations, which ranged from -9 degrees to +9 degrees. Both the dry and the wet (human saliva) states were evaluated at 34 degrees C. From dimensional measurements, the critical contact angles for binding were determined for all products and ranged from 3 degrees to 5 degrees. Below each characteristic critical angle, brackets with passive slides exhibited negligible friction; brackets with active clips exhibited frictional forces as great as 50 cN (50 g). Above each critical angle, all brackets had elastic binding forces that increased at similar rates as angulation increased and were independent of bracket design. Generally speaking, at second-order angulations that exceeded the critical angle, brackets with active clips that had a low critical angle had more resistance to sliding than did brackets with active clips that had a higher critical angle. Brackets with passive slides that had a high critical angle exhibited the lowest resistance to sliding, but could do so at a cost of some loss of control. Nonetheless, self-ligating brackets represent a compromise between friction and control; ie, self-ligating brackets produce frictional forces that are more reproducible than do conventionally ligated stainless steel brackets but without the potential control problems associated with Begg-style brackets.

Dental Stress Analysis↗

Selective inhibition of protein kinase C isozymes by Fas ligation.

Activation of protein kinase C (PKC) can protect cells from apoptosis induced by various agents, including Fas ligation. To elucidate a possible interaction between Fas-mediated apoptotic signals and activation-related protective signals, we investigated the impact of Fas ligation on PKC activity. We demonstrate that engagement of Fas on human lymphoid Jurkat cells triggered apoptosis, and Fas ligation resulted in partial blockade of cellular PKC activity. The phorbol 12-myristate 13-acetate-mediated translocation of PKCtheta from the cytoplasm to the membrane was inhibited by treatment with anti-Fas antibody, whereas the translocation of PKCalpha or epsilon was not affected. In vitro kinase assay of PKCalpha or epsilon phosphotransferase activity demonstrated that Fas ligation inhibited the ability of PKCalpha to phosphorylate histone H1 as substrate but did not inhibit epsilon isozyme activity. This inhibition of PKCalpha activity mediated by Fas ligation was reversed by okadaic acid, a phosphatase inhibitor, suggesting the involvement of a member of the protein phosphatase 2A subfamily in this component of Fas signaling. Identical patterns of PKC isozyme inhibition were obtained using mouse thymoma cells overexpressing the fas gene (LF(+)). These results suggest that the selective inhibition of a potentially protective, PKC-mediated pathway by Fas activation may, to some extent, contribute to Fas-induced apoptotic signaling.

Animals↗

E-cadherin homophilic ligation directly signals through Rac and phosphatidylinositol 3-kinase to regulate adhesive contacts.

Classical cadherins mediate cell recognition and cohesion in many tissues of the body. It is increasingly apparent that dynamic cadherin contacts play key roles during morphogenesis and that a range of cell signals are activated as cells form contacts with one another. It has been difficult, however, to determine whether these signals represent direct downstream consequences of cadherin ligation or are juxtacrine signals that are activated when cadherin adhesion brings cell surfaces together but are not direct downstream targets of cadherin signaling. In this study, we used a functional cadherin ligand (hE/Fc) to directly test whether E-cadherin ligation regulates phosphatidylinositol 3-kinase (PI 3-kinase) and Rac signaling. We report that homophilic cadherin ligation recruits Rac to nascent adhesive contacts and specifically stimulates Rac signaling. Adhesion to hE/Fc also recruits PI 3-kinase to the cadherin complex, leading to the production of phosphatidylinositol 3,4,5-trisphosphate in nascent cadherin contacts. Rac activation involved an early phase, which was PI 3-kinase-independent, and a later amplification phase, which was inhibited by wortmannin. PI 3-kinase and Rac activity were necessary for productive adhesive contacts to form following initial homophilic ligation. We conclude that E-cadherin is a cellular receptor that is activated upon homophilic ligation to signal through PI 3-kinase and Rac. We propose that a key function of these cadherin-activated signals is to control adhesive contacts, probably via regulation of the actin cytoskeleton, which ultimately serves to mediate adhesive cell-cell recognition.

Androstadienes↗

Investigation of the structural requirements in the lipopolysaccharide core acceptor for ligation of O antigens in the genus Salmonella: WaaL "ligase" is not the sole determinant of acceptor specificity.

The ligation of O antigen polysaccharide to lipid A-core oligosaccharide is a late step in the formation of the complex glycolipid known as lipopolysaccharide. Although the process has been localized to the periplasmic face of the inner membrane, details of the ligation mechanism have not been resolved. To date, there is only one gene product (WaaL, often referred to as "ligase") known to be required. There exists a requirement for a specific lipid A-core oligosaccharide acceptor structure for ligation activity, and it has been proposed that the WaaL protein imparts this acceptor specificity. Here the structural requirements in the core oligosaccharide acceptor for O antigen ligation are investigated in prototype serovars of Salmonella enterica. Complementation experiments in mutants with defined core oligosaccharide structure indicate that the specificity of the ligation reaction for a particular core oligosaccharide structure is not dependent on the WaaL protein alone. The data provide the first indication of a more complicated recognition process involving additional cellular components.

Amino Acid Motifs↗

Dynamic ligation properties of the Escherichia coli heme chaperone CcmE to non-covalently bound heme.

The cytochrome c maturation protein CcmE is an essential membrane-anchored heme chaperone involved in the post-translational covalent attachment of heme to c-type cytochromes in Gram-negative bacteria such as Escherichia coli. Previous in vitro studies have shown that CcmE can bind heme both covalently (via a histidine residue) and non-covalently. In this work we present results on the latter form of heme binding to a soluble form of CcmE. Examination of a number of site-directed mutants of E. coli CcmE by resonance Raman spectroscopy has identified ligands of the heme iron and provided insight into the initial steps of heme binding by CcmE before it binds the heme covalently. The heme binding histidine (His-130) appears to ligate the heme iron in the ferric oxidation state, but two other residues ligate the iron in the ferrous form, thereby freeing His-130 to undergo covalent attachment to a heme vinyl group. It appears that the heme ligation in the non-covalent form is different from that in the holo-form, suggesting that a change in ligation could act as a trigger for the formation of the covalent bond and showing the dynamic and oxidation state-sensitive ligation properties of CcmE.

Amino Acid Sequence↗

Ischemia, whether from ligation or torsion, causes ultrastructural changes on the contralateral testis.

OBJECTIVE: Unilateral testicular torsion results with a decrease in contralateral testicular blood flow caused by a reflexive sympathetic response. The aim of this study was to investigate whether twisting of the spermatic cord, or testicular ischemia without twisting, activates this reflex mechanism and causes ultrastructural changes in the contralateral side. MATERIALS AND METHODS: Thirty adult male albino Wistar rats were randomly allocated into three groups of sham, torsion, and ligation. Right testes were twisted 720 degrees counterclockwise in the torsion group. Right spermatic cords were ligated permanently with a silk suture including the vas deferens in the ligation group. After 24 h of testicular ischemia, contralateral left testes were removed for electron microscopic evaluation. RESULTS: Contralateral testes showed similar ultrastructural changes in the torsion and ligation groups. The fibrous tunica propria enveloping the seminiferous tubule was thickened due to increased collagen fibers. The basal lamina was continuous but thickened and showed several foldings. The gap between basal lamina and the germ cells was increased because of collagen fibers. Leydig cells showed mitochondrial degeneration with the loss of its cristae. Leydig cells lost their contact with its neighborhood cells in some areas, and these gaps were filled with collagen fibers. Germ cells showed dilated cisternae of smooth endoplasmic reticulum, cytoplasmic electron-dense bodies and clear regions. CONCLUSIONS: Similar electron microscopic findings observed in the torsion and ligation groups indicate that testicular ischemia rather than twisting of the spermatic cord is responsible for the ultrastructural changes in the contralateral side.

Animals↗

Effects of aortic ligation on the renin angiotensin system in hydronephrotic mice.

A model of aortic ligation in mice with a hydronephrotic kidney (absence of macula densa) was used to determine the effects of aortic ligation on the renal renin-angiotensin system (RAS). Blood pressure increased from 83 +/- 2 to 133 +/- 8 mmHg within 7 days after aortic ligation (p < 0.01). Aortic ligation increased plasma renin (p < 0.01); renin (p < 0.05) and renin mRNA levels (p < 0.001) rose in the ischaemic kidney. In mice with a left hydronephrotic kidney without ischaemia, blood pressure did not change significantly. Plasma renin levels from the left renal vein were lower than from the contralateral vein, but renin (p < 0.01) and renin mRNA levels (p < 0.05) in the hydronephrotic kidney were higher than in the contralateral kidney. In mice with hydronephrosis that had an aortic ligature, blood pressure increased from 81 +/- 2 to 135 +/- 6 mmHg (p < 0.01). Plasma renin increased; renin and renin mRNA levels increased significantly in the ischaemic hydronephrotic kidney (p < 0.01), but not in the contralateral kidney. Thus, the presence of the macula densa is critical for renin release but not for renin gene expression. Aortic ligation results in a significant rise in blood pressure and the activity of the RAS. The mechanisms may involve a baroreceptor and/or an unknown factor.

Animals↗

Ligation of CD38 suppresses human B lymphopoiesis.

CD38 is a transmembrane glycoprotein expressed in many cell types, including lymphoid progenitors and activated lymphocytes. High levels of CD38 expression on immature lymphoid cells suggest its role in the regulation of cell growth and differentiation, but there is no evidence demonstrating a functional activity of CD38 on these cells. We used stroma-supported cultures of B cell progenitors and anti-CD38 monoclonal antibodies (T16 and IB4) to study CD38 function. In cultures of normal bone marrow CD19+ cells (n = 5), addition of anti-CD38 markedly reduced the number of cells recovered after 7 d. Cell loss was greatest among CD19+ sIg- B cell progenitors (mean cell recovery +/- SD = 7.2 +/- 11.7% of recovery in control cultures) and extended to CD19+CD34+ B cells (the most immature subset; 7.6 +/- 2.2%). In contrast, CD38 ligation did not substantially affect cell numbers in cultures of normal peripheral blood or tonsillar B cells. In stroma-supported cultures of 22 B-lineage acute lymphoblastic leukemia cases, anti-CD38 suppressed recovery of CD19+ sIg- leukemic cells. CD38 ligation also suppressed the growth of immature lymphoid cell lines cultured on stroma and, in some cases, in the presence of stroma-derived cytokines (interleukin [IL] 7, IL-3, and/or stem cell factor), but did not inhibit growth in stroma- or cytokine-free cultures. DNA content and DNA fragmentation studies showed that CD38 ligation of stroma-supported cells resulted in both inhibition of DNA synthesis and induction of apoptosis. It is known that CD38 catalyzes nicotinamide adenine dinucleotide (NAD+) hydrolysis into cyclic ADP-ribose (cADPR) and ADPR. However, no changes in NAD+ hydrolysis or cADPR and ADPR production after CD38 ligation were found by high-performance liquid chromatography; addition of NAD+, ADPR, or cADPR to cultures of lymphoid progenitors did not offset the inhibitory effects of anti-CD38. Thus, anti-CD38 does not suppress B lymphopoiesis by altering the enzymatic function of the molecule. In conclusion, these data show that CD38 ligation inhibits the growth of immature B lymphoid cells in the bone marrow microenvironment, and suggest that CD38 interaction with a putative ligand represents a novel regulatory mechanism of B lymphopoiesis.

ADP-ribosyl Cyclase↗

Reversal of proinflammatory responses by ligating the macrophage Fcgamma receptor type I.

Macrophages can respond to a variety of infectious and/or inflammatory stimuli by secreting an array of proinflammatory cytokines, the overproduction of which can result in shock or even death. In this report, we demonstrate that ligation of macrophage Fcgamma receptors (FcgammaR) can lead to a reversal of macrophage proinflammatory responses by inducing an upregulation of interleukin (IL)-10, with a reciprocal inhibition of IL-12 production. IL-10 upregulation was specific to FcgammaR ligation, since the ligation of the Mac-1 receptor did not alter IL-10 production. The identification of the specific FcgammaR subtype responsible for IL-10 upregulation was determined in gene knockout mice. Macrophages from mice lacking the FcR gamma chain, which is required for assembly and signaling by FcgammaRI and FcgammaRIII, failed to upregulate IL-10 in response to immune complexes. However, mice lacking either the FcgammaRII or the FcgammaRIII were fully capable of upregulating IL-10 production, implicating FcgammaRI in this process. The biological consequences of FcgammaRI ligation were determined in both in vitro and in vivo models of inflammation and sepsis. In all of the models tested, the ligation of FcgammaR promoted the production of IL-10 and inhibited the secretion of IL-12. This reciprocal alteration in the pattern of macrophage cytokine production illustrates a potentially important role for FcgammaR-mediated clearance in suppressing macrophage proinflammatory responses.

Animals↗

Compliance with Papanicolaou smear screening following tubal ligation in women with cervical cancer.

To evaluate patient compliance with Papanicolaou (Pap) smear screening after tubal ligation compared with other methods of birth control in patients who develop cervical cancer, a retrospective review of 262 women with cervical cancer diagnosed at age < or = 70 years was undertaken at the Albert Einstein College of Medicine from January 1987 to December 1995. Demographic data, stage of the disease, histologic type, history of smoking, history of sexually transmitted disease (STD), and birth control use were recorded. The Pap screening history was obtained from all the patients. Women who had a bilateral tubal ligation (BTL) were compared with those who did not have this form of birth control. The date and result of their last Pap test prior to their diagnosis of cervical cancer was noted. Two hundred fourteen women with cervical cancer were evaluable. The clinical stage, mean age, history of smoking, and history of STD were similar for both groups. Gravidity among the BTL group was higher than in the non-BTL group (p < 0.01). Forty-eight (22.4%) women had a previous BTL. Twenty-seven of these 48 patients (56.3%) did not have a Pap smear within 3 years prior to the diagnosis of cervical cancer. Of the 166 patients, 61 (36.7%) did not have a Pap test within 3 years (p < 0.05). Fourteen women (29.2%) in the tubal ligation group never returned for a Pap test following the BTL. An average of 6.2+/-5.9 years elapsed since the last Pap test in the BTL group, with 4.0+/-5.1 years in the nontubal ligation group (p < 0.05). There was a correlation between the number of years since BTL (14.2+/-7.7) to the number of years since the last Pap test (6.2+/-5.9) (p < 0.05). Women who have had a BTL should be considered high risk because of poor screening compliance. A Pap test every 3 years is not adequate in this high-risk population group. We advocate improved counseling regarding the importance of continued annual Pap screening for women who are considering tubal ligation.

Adult↗

Maturation of human myelomonocytic leukemia cells following ligation of the low affinity receptor for IgE (Fc epsilon RII/CD23).

The regulation of the low affinity receptor for IgE (Fc epsilon RII/CD23) expression and its role were investigated in U937 cell line and in leukemic cells from a patient (Amb) with acute myeloblastic leukemia. Both cell populations were CD23- but could acquire CD23 expression following treatment with IL-4. CD23+ cells, however, remained blastic and did not show any significant phenotypical and functional modifications. Following ligation of the CD23 on U937 and Amb cells by anti-CD23 mAb, these leukemic cells differentiated into mature monocyte/macrophage-like cells. CD23 ligation promoted the expression of the monocyte marker, CD14, increased the expression of the common beta chain of the LFA-1 family (CD18), and down-regulated the expression of the promonocytic marker CD33. Morphological and phenotypical changes were associated with functional modifications as CD23 ligation allowed the acquisition of the oxidative metabolism in leukemic cells as revealed by luminol-dependent chemiluminescence. As in mature monocytes, CD23 ligation induced an accumulation of intracellular cAMP in leukemic cells. These data indicate that ligation of CD23 may induce the maturation of myelomonocytic cells into monocytic-like cells.

Cyclic AMP↗

Differential sensitivity of B lymphocyte populations to IgM receptor ligation is determined by local factors.

Ligation of surface IgM on B cells responding to lipopolysaccharide (LPS) suppresses terminal differentiation and high-rate Ig secretion with no effect on proliferation. As shown here, different B cell populations show characteristic mean values of ligand concentration required for 50% inhibition, with Gaussian distributions of sensitivity to IgM receptor ligation that reflect cellular heterogeneity of 'al-or-none' inhibitions in single cells. Differential sensitivity of B cell populations to IgM ligation seems to be locally determined by the cellular environment and unrelated to the 'maturity' of the responding cells. Thus, while long-lived peritoneal B cells are 3- to 5-fold more resistant than splenic B cells, there is no difference in sensitivity between short- and long-lived B cells in the spleen. Furthermore, while B cells in bone marrow and spleen differ in sensitivity by two orders of magnitude, B cells differentiated in vitro from bone marrow pre-B cells are as resistant as splenic B cells. Moreover, bone marrow cell culture supernatants restore a high level of sensitivity in such cell populations. Differential sensitivity to IgM receptor ligation is reproduced by multivalent nominal antigen, in cell populations that show identical dose-response inhibition curves to direct activation of protein kinase C by phorbol esters. We conclude that the level of sensitivity to IgM ligation is independent of the life span or maturity of the B cell, but differentially regulated in vivo by putative tissue factors.

Animals↗

A minor population of splenic dendritic cells expressing CD19 mediates IDO-dependent T cell suppression via type I IFN signaling following B7 ligation.

By ligating CD80/CD86 (B7) molecules, the synthetic immunomodulatory reagent CTLA4-Ig (soluble synthetic CTLA4 fusion protein) induces expression of the enzyme indoleamine 2,3-dioxygenase (IDO) in some dendritic cells (DCs), which acquire potent T cell regulatory functions as a consequence. Here we show that this response occurred exclusively in a population of splenic DCs co-expressing the marker CD19. B7 ligation induced activation of the transcription factor signal transducer and activator of transcription (STAT1) in sorted CD19+, but not CD19(NEG), DCs. STAT1 activation occurred even when DCs lacked receptors for type II IFN (IFNgamma); however, STAT1 activation and IDO up-regulation were not observed when DCs lacked receptors for type I IFN (IFNalphabeta). Thus, IFNalpha, but not IFNgamma, signaling was essential for STAT1 activation and IDO up-regulation in CD19+ DCs following B7 ligation. Consistent with these findings, B7 ligation also induced sorted CD19+, but not CD19(NEG), DCs to express IFNalpha. Moreover, recombinant IFNalpha induced CD19+, but not CD19(NEG), DCs to mediate IDO-dependent T cell suppression, showing that IFNalpha signaling could substitute for upstream signals from B7. These data reveal that a minor population of splenic DCs expressing the CD19 marker is uniquely responsive to B7 ligation, and that IFNalpha-mediated STAT1 activation is an essential intermediary signaling pathway that promotes IDO induction in these DCs. Thus, CD19+ DCs may be a target for regulatory T cells expressing surface CTLA4, and may suppress T cell responses via induction of IDO.

Animals↗

Arsenite retards DNA break rejoining by inhibiting DNA ligation.

Arsenic has been shown to inhibit methyl methane-sulphonate (MMS)-induced DNA repair but the exact mechanism remains controversial. The purpose of this investigation is to examine which step of DNA repair is most sensitive to arsenite (As) and how As inhibits it. The results from single-cell alkaline electrophoresis, showing post-treatment with As increased DNA strand breaks in MMS-treated cells, suggest that that the excision step seems to be less sensitive to As than later steps. To test this hypothesis, hydroxyurea (Hu) plus cytosine-beta-D-arabinofuranoside (AraC) were used to block DNA polymerization, allowing the DNA strand breaks to accumulate. These experiments indicated that As had weak inhibitory effects on DNA strand break accumulation. However, As inhibited the rejoining of those DNA strand breaks which could be rejoined within 4 h after release from blockage by Hu plus AraC. To further elucidate this mechanism, a cell extract was used to compare the relative sensitivity of the various steps in DNA repair to As. The potency of the As inhibitory effect as deduced from concentration-response curves were: ligation of poly(rA).oligo(dT) > ligation of poly(dA).oligo(dT) approximately DNA polymerization > or = DNA repair synthesis > excision. As is known to inhibit the activity of pyruvate dehydrogenase by interacting with vicinal dithiol groups. Dithiothreitol could effectively remove As inhibition of both the ligation of poly(rA).oligo(dT) and the activity of pyruvate dehydrogenase but had no obvious effect on As inhibition of poly(dA).oligo(dT) ligation. Since DNA ligase III contains vicinal dithiol groups, we postulate that As may inhibit DNA break rejoining by interacting with the vicinal dithiols to inactivate DNA ligation in MMS-treated cells.

Animals↗

Kinetic analysis for optimization of DNA ligation reactions.

Kinetic equations describing ligation of DNA to circular recombinant forms were developed and solved for four types of reactions: (a) a homogeneous population of singly restricted DNA fragments, (b) insertion of singly restricted insert into vector, (c) forced directional insertion of doubly restricted insert into vector, and (d) insertion of singly restricted insert into phosphatased vector. The effects of varying vector and insert sizes, starting concentrations, and phosphatase treatment on the yield of circular 1:1 recombinants were analyzed. Selected theoretical predictions were experimentally tested and verified. Our suggestions on optimizing ligation reactions in several cases are at variance with common practice. For example, optimum conditions in case (b) and (d) ligations are best specified as individual insert and vector concentrations rather than as insert/vector molar ratios, except in case (d) ligations involving very small insert size. In case (c) ligations, highest efficiencies are obtained when both vector and insert are at relatively low concentration.

Cloning, Molecular↗