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PubMed2001

High selectivity from configurational match/mismatch in carbon-hydrogen insertion reactions of steroidal diazoacetates catalyzed by chiral dirhodium(II) carboxamidates.

Diazo decomposition of steroidal diazoacetates, where the point of attachment is the 3-position of the steroid A-ring, catalyzed by chiral dirhodium(II) carboxamidates results in products from carbon-hydrogen insertion in high yield and selectivities. Use of S-configured catalysts shows a distinctive preference for insertion into the 3-position to form beta-lactone products. The R-configured catalysts direct insertion preferentially to the equatorial C-H bond at the 2-position. Substituents or functional groups at the 5/6-position prevent C-H insertion from taking place at the 4-position. Even in the best case with the 5/6-positions fully saturated, however, insertion into the 3-position remains competitive with insertion into the 4-position. Corresponding 3-substituted phenyldiazoacetates give only beta-lactone products, and selectivity here is highest with chiral dirhodium(II) prolinate catalysts. A model is presented to explain these results. Overall, this methodology is versatile for functionalization of the steroid A-ring at positions 2 and 3.

Azo Compounds↗
PubMed2004

Anomalous intramolecular C-H insertion reactions of rhodium carbenoids: factors influencing the reaction course and mechanistic implications.

The intramolecular insertion of rhodium carbenoids into the alpha-C-H bonds of allylic ethers to give 3(2H)-furanones has been explored. Cyclopropanation is favored irrespective of the complex used for carbenoid generation or the substitution pattern of the allylic ether, unless a substituent is placed on the tether connecting the ether to the alpha-diazo ketone. Unusual acetal products resulting from an anomalous C-H insertion process are obtained in addition to the expected 3(2H)-furanones formed by conventional carbenoid C-H insertion. These acetals are the favored C-H insertion products in certain circumstances and particularly in cases where carbenoid generation is effected using an electron-deficient rhodium complex. Experiments with simple deuterium labeled substrates reveal that anomalous C-H insertion products arise by a mechanism that is distinct from that leading to the formation of conventional C-H insertion products. The formation of acetal products and the outcome of reactions performed using deuterium-labeled substrates suggest that a mechanism involving hydride migration to the rhodium center of the carbenoid is operative.

Journal Article↗
PubMed2006

Dynamics of the interaction of vapor-deposited copper with alkanethiolate monolayers: bond insertion, complexation, and penetration pathways.

We have investigated the interaction of vapor-deposited copper with -CH3, -OH, -OCH3, -COOH, and -CO2CH3 terminated alkanethiolate self-assembled monolayers (SAMs) adsorbed on polycrystalline Au using time-of-flight secondary ion mass spectrometry and density functional theory calculations. For -OH, -COOH, and -CO2CH3 terminated SAMs measurements indicate that for all copper coverages there is a competition between Cu atom bond insertion into C-O bonds, stabilization at the SAM/vacuum interface, and penetration to the Au/S interface. In contrast, on a -OCH3 terminated SAM Cu only weakly interacts with the methoxy group and penetrates to the Au substrate, while for a -CH3 terminated SAM deposited copper only penetrates to the Au/S interface. The insertion of copper into C-O terminal group bonds is an activated process. We estimate that the barriers for Cu insertion are 55 +/- 5 kJ mol(-1) for the ester, 50 +/- 5 kJ mol(-1) for the acid, and 55 +/- 5 kJ mol(-1) for the hydroxyl terminated SAMs. The activation barrier for the copper insertion is much higher for the -OCH3 SAM. Copper atoms with energies lower than the activation barrier partition between complexation (weak interaction) with the terminal groups and penetration through the monolayer to the Au/S interface. Weakly stabilized copper atoms at the SAM/vacuum interface slowly penetrate through the monolayer. In contrast to the case of Al deposition, C-O bond insertion is favored over C=O, C-H, and C-C bond insertion.

Journal Article↗
PubMed2002

Oral clonidine premedication reduces the EC50 of propofol concentration for laryngeal mask airway insertion in male patients.

BACKGROUND: Oral clonidine, an alpha2-adrenergic receptor agonist, reduces the dose of propofol required for laryngeal mask airway (LMA) insertion. Target-controlled infusion (TCI) is becoming increasingly popular for propofol infusion. There is no information, however, on the propofol blood concentrations required for LMA insertion and the effect of oral clonidine premedication on these values. METHODS: Propofol at target effect-site concentrations from 4.0 to 12.0 microg/ml were randomly administered using TCI in three groups of healthy male patients (n=35 each) who were undergoing elective orthopedic surgery: control, 2.5 microg/kg clonidine, and 5.0 microg/kg clonidine groups. Nothing was administered to the control group. Clonidine(2.5 microg/kg or 5.0 microg/kg) was administered orally 90 min before arrival at the operating room in the clonidine groups. After equilibration between the blood- and effect-site for 15 min, insertion of the LMA was attempted. The EC50 for LMA insertion (measured propofol serum concentration in equilibrium with the effect-site at which 50% of patients do not respond to the insertion of the LMA) was determined by logistical regression. RESULTS: EC50+/-standard error values in the control, 2.5 microg/kg clonidine, and 5.0 microg/kg clonidine groups were 8.72+/-0.55, 7.76+/-0.60, and 5.84+/-0.58 microg/ml, respectively. The EC50 in the 5.0 microg/kg clonidine group was significantly lower than that in the control group (P < 0.01). CONCLUSIONS: The propofol concentration required for LMA insertion in healthy male patients is reduced by premedication with 5.0 microg/kg oral clonidine.

Adrenergic alpha-Agonists↗
PubMed1999

Root debridement of molars with furcation involvement using diamond-coated sonic scaler inserts during flap surgery -- a pilot study.

In our laboratory, we modified sonic scaler inserts to increase effectivity of root surface instrumentation of molars with furcation involvement during flap surgery. The working end of conventional sonic scaler inserts was thickened ellipsoidally and diamond coated. Inserts with variously angled shafts were designed in order to facilitate insertion into the furcation. A clinical, split-mouth study examined the debridement of molars with furcation involvement during flap surgery using either hand instruments or diamond-coated sonic scaler inserts with 15 patients. Average treatment time was cut by ca. 50%, using diamond-coated inserts as compared to hand instruments for root surface debridement during surgery. 2 years after surgery, the clinical parameters had not been influenced by type of instrument used.

Adult↗
PubMed1993

How amino-acid insertions are allowed in an alpha-helix of T4 lysozyme.

Studies of extant protein sequences indicate that amino-acid insertions and deletions are preferentially located in loop regions, which has traditionally been explained as the result of selection removing deleterious mutations within secondary structural elements from the population. But there is no a priori reason to discount the possibility that insertions within secondary structure could either be tolerated until compensatory mutations arise, or have effects that are propagated away from secondary structure into loops. Earlier studies have indicated that insertions are generally tolerated, although much less well within secondary structure elements than in loop regions. Here we show that amino-acid insertions in an alpha-helix of T4 lysozyme can be accepted in two different ways. In some cases the inserted amino acids are accommodated within the helix, leading to the translocation of wild-type residues from the helix to the preceding loop. In other cases the insertion causes a 'looping-out' in the first or last turn of the helix. The individual structural responses seem to be dominated by the maintenance of the interface between the helix and the rest of the protein.

Amino Acid Sequence↗
PubMed2006

Insertion reactions of alkynes and organic isocyanides into the palladium-carbon bond of dimetallic Fe-Pd alkoxysilyl complexes.

Insertion of MeO(2)C-C[triple bond]C-CO(2)Me (DMAD) into the Pd-C bond of the heterodimetallic complex [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d(dmba-C)] (2) (dppm = Ph(2)PCH(2)PPh(2), dmba-C = metallated dimethylbenzylamine) and [(OC)(3){(MeO)(3)Si}F[upper bond 1 start]e(mu-dppm)P[upper bond 1 end]d(8-mq-C,N)] (3) (8-mq-C,N = cyclometallated 8-methylquinoline) yielded the sigma-alkenyl complexes [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(CO(2)Me)=C(CO(2)Me)(o-C(6)H(4)CH(2)NMe(2))}] (7) and [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(CO(2)Me)[double bond, length as m-dash]C(CO(2)Me)(CH(2)C(9)H(6)N)}] (8), respectively. The latter afforded the adduct [(OC)(3){(MeO)(3)Si}F[upper bond 1 start]e(mu-dppm)P[upper bond 1 end]d{C(CO(2)Me)=C(CO(2)Me)(CH(2)C(9)H(6)N)}(CNBu(t))] (9) upon reaction with 1 equiv. of Bu(t)NC. The heterodinuclear sigma-butadienyl complexes [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(Ph=C(Ph)C(CO(2)Me)=(CO(2)Me)(o-C(6)H(4)CH(2)NMe(2))}] (11) and [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(Ph)=C(CO(2)Et)C(Ph)=C(CO(2)Et)(CH(2)C(9)H(6)N)}] (13) have been obtained by reaction of the metallate K[Fe{Si(OMe)(3)}(CO)(3)(dppm-P)] (dppm = Ph(2)PCH(2)PPh(2)) with [P[upper bond 1 start]dCl{C(Ph)=C(Ph)C(CO(2)Me)=C(CO(2)Me)(o-C(6)H(4)CH(2)N[upper bond 1 end]Me(2))}] or [P[upper bond 1 start]dCl{C(Ph)=C(CO(2)Et)C(Ph)=(CO(2)Et)}(CH(2)C(9)H(6)N[upper bond 1 end])], respectively. Monoinsertion of various organic isocyanides RNC into the Pd-C bond of 2 and 3 afforded the corresponding heterometallic iminoacyl complexes. In the case of complexes [(OC)(3){(MeO)(3)Si}F[upper bond 1 start]e(mu-dppm)P[upper bond 1 end][upper bond 1 start]d{C=(NR)(CH(2)C(9)H(6)N[upper bond 1 end])}] (15a R = Ph, 15b R = xylyl), a static six-membered C,N chelate is formed at the Pd centre, in contrast to the situation in [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(=NR)(o-C(6)H(4)CH(2)NMe(2))}] (14a R = o-anisyl, 14b R = 2,6-xylyl) where formation of a mu-eta(2)-Si-O bridge is preferred over NMe(2) coordination. The outcome of the reaction of the dimetallic alkyl complex [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]dMe] with RNC depends both on the stoichiometry and the electronic donor properties of the isocyanide employed for the migratory insertion process. In the case of o-anisylisocyanide, the iminoacyl complex [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{C(=N-o-anisyl)Me}] (16) results from the reaction in a 1 : 1 ratio. Addition of three equiv. of o-anisylisocyanide affords the tris(insertion) product [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{[C(=N-o-anisyl)](3)Me}] (18). After addition of a fourth equivalent of o-anisylNC, exclusive formation of the isocyanide adduct [(OC)(3){(MeO)(3)Si}F[upper bond 1 start]e(mu-dppm)P[upper bond 1 end]d{[C(=N-o-anisyl)](3)Me}(CN-o-anisyl)] (19) was spectroscopically evidenced. In the complex [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]d{[C(=N-o-C(6)H(4)COCH(2))](2)Me}] (20), the sigma-bound diazabutadienyl unit is part of a 12-membered organic macrocyle which results from bis(insertion) of 1,2-bis(2-isocyanophenoxy)ethane into the Pd-Me bond of the precursor complex [(OC)(3)F[upper bond 1 start]e{mu-Si(OMe)(2)([lower bond 1 start]OMe)}(mu-dppm)P[lower bond 1 end][upper bond 1 end]dMe]. In contrast, addition of two equivalents of tert-butylisocyanide to a solution of the latter afforded [(OC)(3){(MeO)(3)Si}F[upper bond 1 start]Fe(mu-dppm)P[upper bond 1 end]d{C(=NBu(t))Me}(CNBu(t))] (21) in which both a terminal and an inserted isocyanide ligand are coordinated to the Pd centre. In all cases, there was no evidence for competing CO substitution at the Fe(CO)(3) fragment by RNC. The molecular structures of the insertion products 8 x CH(2)Cl(2) and 16 x CH(2)Cl(2) have been determined by X-ray diffraction.

Journal Article↗
PubMed2004

Parasite-specific inserts in the bifunctional S-adenosylmethionine decarboxylase/ornithine decarboxylase of Plasmodium falciparum modulate catalytic activities and domain interactions.

Polyamine biosynthesis of the malaria parasite, Plasmodium falciparum, is regulated by a single, hinge-linked bifunctional PfAdoMetDC/ODC [ P. falciparum AdoMetDC (S-adenosylmethionine decarboxylase)/ODC (ornithine decarboxylase)] with a molecular mass of 330 kDa. The bifunctional nature of AdoMetDC/ODC is unique to Plasmodia and is shared by at least three species. The PfAdoMetDC/ODC contains four parasite-specific regions ranging in size from 39 to 274 residues. The significance of the parasite-specific inserts for activity and protein-protein interactions of the bifunctional protein was investigated by a single- and multiple-deletion strategy. Deletion of these inserts in the bifunctional protein diminished the corresponding enzyme activity and in some instances also decreased the activity of the neighbouring, non-mutated domain. Intermolecular interactions between AdoMetDC and ODC appear to be vital for optimal ODC activity. Similar results have been reported for the bifunctional P. falciparum dihydrofolate reductase-thymidylate synthase [Yuvaniyama, Chitnumsub, Kamchonwongpaisan, Vanichtanankul, Sirawaraporn, Taylor, Walkinshaw and Yuthavong (2003) Nat. Struct. Biol. 10, 357-365]. Co-incubation of the monofunctional, heterotetrameric approximately 150 kDa AdoMetDC domain with the monofunctional, homodimeric ODC domain (approximately 180 kDa) produced an active hybrid complex of 330 kDa. The hinge region is required for bifunctional complex formation and only indirectly for enzyme activities. Deletion of the smallest, most structured and conserved insert in the ODC domain had the biggest impact on the activities of both decarboxylases, homodimeric ODC arrangement and hybrid complex formation. The remaining large inserts are predicted to be non-globular regions located on the surface of these proteins. The large insert in AdoMetDC in contrast is not implicated in hybrid complex formation even though distinct interactions between this insert and the two domains are inferred from the effect of its removal on both catalytic activities. Interference with essential protein-protein interactions mediated by parasite-specific regions therefore appears to be a viable strategy to aid the design of selective inhibitors of polyamine metabolism of P. falciparum.

Adenosylmethionine Decarboxylase↗
PubMed1998

Requirements for target-controlled infusion of propofol to insert the laryngeal mask airway.

The success rate for insertion of the laryngeal mask airway was assessed in 60 patients using a target-controlled infusion of propofol. Eleven of 15 patients who received a target propofol concentration of 5 micrograms.ml-1 had the laryngeal mask inserted successfully within 3 min of induction of anaesthesia. Fourteen of 15 patients who received 6 micrograms.ml-1 had successful insertion within 3 min and all 15 patients who received 7 micrograms.ml-1 had successful insertion within 3 min. Patients who received 8 micrograms.ml-1 had successful insertion within 90s of induction of anaesthesia. The increased success rate of LMA insertion with the higher target propofol concentrations was achieved without major haemodynamic effects in this group of fit patients. However, adverse cardiovascular effects may occur with these doses in elderly or unfit patients. It is important in individual patients to titrate the target propofol concentration to achieve the required clinical endpoint.

Adult↗
PubMed1998

Insertion of the Combitube airway with the cervical spine immobilised in a rigid cervical collar.

The Combitube is a twin lumen device designed to establish the airway after blind insertion. Under general anaesthesia a rigid cervical collar was used to immobilise the neck in 15 ASA 1 and 2 patients. Insertion of the Combitude airway was then attempted. In 10/15 (66%) patients, blind insertion was not possible. In 5/15 (33%) successful blind insertions the Combitube entered the oesophagus on each occasion. In 8/10 of the failures, re-insertion of the Combitude was attempted with the aid of a Macintosh laryngoscope. In 6/8 cases (75%) satisfactory placement was then possible with the Combitube again entering the oesophagus on each occasion. Ventilation was satisfactory in all patients when insertion was successful. Blood staining of the Combitube was present in 7/15 (47%) patients. The Combitude cannot be recommended for use in patients whose necks are immobilised in rigid cervical collars.

Adult↗
PubMed2000

Randomised double-blind comparison of fentanyl, mivacurium or placebo to facilitate laryngeal mask airway insertion.

In a double-blind randomised study, we compared conditions during insertion of the laryngeal mask airway in 150 patients who received either fentanyl 1 microg.kg-1, mivacurium 0.04 mg.kg-1 or normal saline, before induction of anaesthesia with propofol 2 mg.kg-1. Insertion conditions, including mouth opening, swallowing, gagging or coughing, head or limb movement and ease of insertion, were each graded using a three-point scale. The median (interquartile range) summed insertion scores were more favourable with the use of fentanyl [8.0 (7.0-9.0)] and mivacurium [7.5 (6.8-8.3)] than with normal saline [9.0 (7.8-10.3); p < 0.01]. Fentanyl and mivacurium decreased swallowing and head or limb movement, and mivacurium improved mouth opening. Insertion conditions were similar between fentanyl and mivacurium, while both prolonged apnoea. Fentanyl and mivacurium are equally effective in facilitating insertion of the laryngeal mask airway following anaesthetic induction with propofol.

Adolescent↗
PubMed1995

Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR.

Thermal asymmetric interlaced (TAIL-) PCR is an efficient technique for amplifying insert ends from yeast artificial chromosome (YAC) and P1 clones. Highly specific amplification is achieved without resort to complex manipulations before or after PCR. The adaptation of this method for recovery and mapping of genomic sequences flanking T-DNA insertions in Arabidopsis thaliana is described. Insertion-specific products were amplified from 183 of 190 tested T-DNA insertion lines. Reconstruction experiments indicate that the technique can recover single-copy sequences from genomes as complex as common wheat (1.5 x 10(10) bp). RFLPs were screened using 122 unique flanking sequence probes, and the insertion sites of 26 T-DNA transgenic lines were determined on an RFLP map. These lines, whose mapped T-DNA insertions confer hygromycin resistance, can be used for fine-scale mapping of linked phenotypic loci.

Arabidopsis↗
PubMed1996

A transposon insertion in the Arabidopsis SSR16 gene causes an embryo-defective lethal mutation.

The SSR16 gene of Arabidopsis has been identified as a gene encoding a ribosomal protein S16 homolog through analysis of a transposon insertion mutation. The insertion mutation is lethal, arresting embryonic development at approximately the transition from the globular to the heart stage of embryonic development. Co-segregation of the mutant phenotype with the transposon-borne drug-resistance marker and loss of the inserted transposon concomitant with phenotypic reversion provided evidence that the transposon had caused the mutation. Sequences flanking the insertion site were amplified from DNA of viable heterozygotes by thermal asymmetric interlaced (TAIL) PCR. The amplified fragment flanking the 3' end of the inserted element was sequenced and found to be identical to an Arabidopsis expressed sequence tag (EST). The EST, in turn, contained a coding sequence homologous to the ribosomal protein S16 (RPS16) of bacteria such as Escherichia coli, Bacillus subtilis and Salmonella typhimurium, as well as Neurospora crassa mitochondria and higher plant plastids. Thus the gene identified by the embryo-defective lethal insertion mutation encodes an RPS16 homolog and has been designated the SSR16 gene.

Amino Acid Sequence↗
PubMed2000

Insertion of light-harvesting chlorophyll a/b protein into the thylakoid topographical studies.

The major light-harvesting chlorophyll a/b-binding protein (Lhcb1,2) of photosystem II is inserted into the thylakoid via the signal recognition particle dependent pathway. However, the mechanism by which the protein enters the membrane is at this time unknown. In order to define some topographical restrictions for this process, we constructed several recombinant derivatives of Lhcb1 carrying hexahistidine tags at either protein terminus or in the stromal loop domain. Additionally, green fluorescent protein (GFP) was fused to either terminus. None of the modifications significantly impair the pigment-binding properties of the protein in the in vitro reconstitution of LHCII. With the exception of the C-terminal GFP fusion, all mutants stably insert into isolated thylakoids in the absence of Ni2+ ions. The addition of low concentrations of Ni2+ ions abolishes the thylakoid insertion of C-terminally His-tagged mutants whereas the other His-tagged proteins fail to insert only at higher Ni2+ concentrations. The C-terminus of Lhcb1 must cross the membrane during protein insertion whereas the other sites of Lhcb1 modification are positioned on the stromal side of LHCII. We conclude that a Ni2+-complexed His tag and fusion to GFP inhibit translocation of the protein C-terminus across the thylakoid. Our observations indicate that the N-terminal and stromal domain of Lhcb1 need not traverse the thylakoid during protein insertion and are consistent with a loop mechanism in which only the C-terminus and the lumenal loop of Lhcb1 are translocated across the thylakoid.

Biological Transport↗
PubMed2002

Localization of the ventricular insertion site of concealed left-sided accessory pathways using ventricular pace mapping.

The aim of the present study was to localize the ventricular insertion site of concealed accessory pathway (APs) by using the ventricular pace mapping and examined if the analysis of the timing of retrograde atrial electrogram recorded at the ventricular side of the mitral annulus is useful in identifying the ventricular insertion site of the AP. In 39 patients with concealed left-sided APs, ventricular pacing was delivered along the mitral annulus at a cycle length of 500 ms while measuring the conduction interval from the pacing stimulus to the earliest retrograde atrial electrogram recorded in the coronary sinus (St-A). The ventricular insertion site of the AP was localized by identifying the shortest St-A. Also the interval between the onsets of QRS and atrial electrograms (QRS-A) and presence of continuous electrical activity (CEA) between the ventricular and atrial electrograms were evaluated at each mapping site during atrioventricular reciprocating tachycardia. Initial radiofrequency energy application to the site with the shortest St-A (46 +/- 15 ms) eliminated the AP conduction in all patients, suggesting the accurate localization of the ventricular insertion site by ventricular pace mapping. The QRS-A and the percentage of the presence of CEA at the shortest St-A site were 79 +/- 19 ms and 64%, respectively. However, the earliest retrograde atrial activation site did not coincide with the shortest St-A site in 19 of 39 patients, suggesting an oblique course of AP. Thus, in these 19 patients, there was a significant difference in St-A (47 +/- 16 vs 59 +/- 15 ms, P < 0.0001), QRS-A (83 +/- 13 vs 72 +/- 12 ms, P < 0.0001) and the presence of CEA (32 vs 74%, P < 0.01) between the shortest St-A site and the earliest retrograde atrial activation site, respectively. These indicate that the earliest retrograde atrial activation is not necessarily indicative of the ventricular insertion site of AP. However, ventricular pace mapping was considered to be useful for identifying and ablating the ventricular insertion site of AP, irrespective of the course of AP.

Adolescent↗
PubMed2003

Effects of simultaneous insertion of 66 plunge needle electrodes on myocardial activation, function, and structure.

Transmural recordings using plunge needle electrodes are useful in mapping ventricular tachyarrhythmia, but they interfere with activation sequences or damage the myocardium. This study evaluated the effects of insertion of 66 transmural needles on myocardial activation, structure, and function. Epicardial maps were performed at thoracotomy using a 40-electrode plaque in five mongrel dogs. Sixty-six transmural plunge needles were introduced into the anterior aspect of the septum and left ventricle. Transmural maps of unipolar electrograms were recorded every 15 minutes via 124 electrodes over a 2-hour period. Epicardial maps were repeated after the needles were removed. All recordings were performed during sinus rhythm and ventricular pacing at 300- and 200-ms cycle lengths. Gated heart pool studies were performed preoperatively and 2 weeks after thoracotomy. Programmed ventricular stimulation was performed 2 weeks after thoracotomy. In total, 15,996 electrograms were analyzed. Maximum negative dV/dt of each electrogram and the activation time at each electrode did not change significantly over the 2 hours of needle insertion. After removal of the needles, epicardial maps were unchanged compared to before needle insertion. Mean left ventricular ejection fraction 2 weeks after needle insertion was 59% versus 58% before needle insertion (P=0.9). No dogs had inducible ventricular tachycardia. Histology showed contraction bands of 0.8-mm diameter adjacent to the needle tracks but no scarring. Insertion of 66 closely spaced plunge needles did not distort epicardial or transmural maps. Multiple needles did not result in myocardial scarring, left ventricular dysfunction, or predispose to ventricular tachycardia.

Analysis of Variance↗
PubMed2000

Effects of ageing on the insertion zones of the human vocal fold.

The vocal ligaments insert at the anterior and posterior commissures of the larynx. These structures fulfil biomechanical functions, balancing the different elastic moduli of tendon, cartilage or bone and undergo age-related changes that may be responsible for voice changes with increasing age. The aim of this study was to analyse the insertion structures of the vocal ligaments by means of macroscopic, histological, immunohistochemical and electron-microscopic methods and to draw conclusions from age-related structural changes on a functional basis. Investigations were carried out on the larynges of 22 males and 15 females (aged 1-95 y). In adolescence, the insertion zone of the vocal ligament tendon, a dense network of connective tissue rich in sulphated glycosaminoglycans at the thyroid cartilage, is characterised by a layer between tendon and cartilage comparable to fibrocartilage. The insertion zone lacks a perichondrium. Collagen fibrils of the vocal ligament tendon penetrate directly into the thyroid cartilage. In the insertion area, the chondrocytes are surrounded by collagen fibrils, which show positive reactivity to antibodies against type I and type III collagen. Sulphated glycosaminoglycans are integrated between the collagen fibrils. In the area of the posterior glottis, elastic cartilage rests like a cap on the hyaline base of the arytenoid cartilage. There is no distinctive border between the structures. With increasing age, ossification of the laryngeal skeleton occurs, involving hyaline cartilage at the posterior glottis and hyaline and fibrocartilage at the anterior commissure. At the same time, a loss or sulphated glycosaminoglycans is observed inside the vocal ligament tendon. Advanced ossification of the laryngeal skeleton, particularly in the area of the commissures, an increasing loss of glycosaminoglycans in the vocal ligament tendon and changes in the elastic tissue reduce the elastic modulus between tendon, cartilage and bone, thus 'stiffening' the insertion zones, which could be one factor among others favouring voice changes with advancing age.

Aging↗
PubMed2005

Push-and-pull enteroscopy using the double-balloon technique: method of assessing depth of insertion and training of the enteroscopy technique using the Erlangen Endo-Trainer.

BACKGROUND AND STUDY AIMS: This study was conducted to test a method of measuring the depth of insertion into the small bowel during push-and-pull enteroscopy using the Erlangen Endo-Trainer. Furthermore, the Erlangen Endo-Trainer model for training in the new method of push-and-pull enteroscopy using the double-balloon technique was also evaluated. MATERIALS AND METHODS: Specially prepared packages of porcine upper visceral organs were used, including the esophagus, stomach, duodenum, and small bowel, implanted into the Erlangen Endo-Trainer. In the first step of this study, all of the modifications needed to obtain a model useful for training in the new enteroscopy technique were tested, including different lengths of small bowel. In the next step, the Erlangen Endo-Trainer was used to evaluate a special method of measuring the depth of insertion during push-and-pull enteroscopy by comparing estimated insertion depths of 100 cm and 200 cm, marked on the porcine small bowel, with endoscopic insertion depths determined afterwards by measuring the length of the small bowel from the pylorus to the mark on the porcine small bowel. Additionally, the Endotrainer was used to demonstrate the principle of double-balloon enteroscopy and to provide training in this new enteroscopy technique with a lifelike simulation. RESULTS: The modified Erlangen Endo-Trainer proved its value for demonstrating the principle of push-and-pull enteroscopy and for providing training in this enteroscopic technique as well as for testing the measurement method. The evaluation of the measurement method showed that the estimation of the insertion depths was accurate, with a mean deviation of less than 10 %. A total of 13 workshops (seven national and six international), including a total of 97 participants, were carried out between January and August 2004. Under the supervision of one of the authors, pairs of trainees were able to reach the ileal valve or the end of the small bowel from the oral route. CONCLUSIONS: The modified Erlangen Endo-Trainer is useful for training in the push-and-pull enteroscopy technique. The new specially developed method of measuring the depth of insertion during push-and-pull enteroscopy seems to be valid.

Animals↗