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PubMed2005

No correlation between trapeziometacarpal arthritis and abductor pollicis longus insertion.

The primary hypothesis of this study is that supernumerary slips of the abductor pollicis longus (particularly those slips inserting distal to the trapeziometacarpal joint) increase the risk of trapeziometacarpal osteoarthritis on account of increased transarticular forces. Other hypotheses surmised that age and female gender were directly correlated with severity of arthritis. We did cadaveric dissection of the distal first dorsal compartment in 61 specimens, noting the cadaveric age, gender, and number and insertion sites of abductor pollicis longus tendon slips. Each variable was statistically correlated with visual grade of trapeziometacarpal arthritis. The median number of abductor pollicis longus tendon slips in these cadaveric hands was three (range, 1-4). Seventy-nine percent of the hands had a digastric-type insertion into the abductor pollicis brevis. Ninety percent had an insertion into the trapezium. All hands possessed an insertion into the base of the first metacarpal. Age and female gender were directly correlated with severity of arthritis. No other correlations existed. We conclude that trapeziometacarpal joint arthritis progresses with age and occurs independently of any aspect of abductor pollicis longus insertion. Based on our results, we do not recommend surgical release of these supernumerary abductor pollicis longus tendon slips for the treatment of trapeziometacarpal osteoarthritis.

Adult↗
PubMed1985

Retroviral insertional mutagenesis in murine mammary cancer.

We are attempting to identify cellular oncogenes activated in mammary tumours by using the mouse mammary tumour virus (MMTV) as an insertional mutagen. MMTV, a retrovirus lacking a host cell-derived viral oncogene, induces adenocarcinomas of the mammary gland after a long latency period. The tumours are clonal outgrowths of cells carrying one or more integrated MMTV proviral copies. We have cloned an integrated MMTV provirus with its adjacent chromosomal DNA and we have established that the insertion site was part of a domain of the mouse genome in which MMTV proviruses are inserted in many different tumours. A gene within this domain, called int-1 is transcriptionally activated as a consequence of proviral integration. We have proposed that int-1 is a cellular oncogene for mammary tumours. Proviral activation of int-1 occurs in cis, over distances of up to 10 kilobases and is presumably caused by the transcriptional enhancer present on the MMTV long terminal repeat. The putative int-1 mammary oncogene has been subjected to a detailed structural analysis by S1 mapping and DNA sequencing. It encodes a protein that is highly conserved between mouse and man. The protein encoding domain of the gene is distributed over four exons which are demarcated by the insertion sites of MMTV proviruses found in mammary tumours. Some insertions, however, are found in the transcriptional unit of int-1, but these insertions do not disrupt the protein encoding domain of the gene.

Adenocarcinoma↗
PubMed1991

Chromosomal insertion of TOL transposons in Pseudomonas aeruginosa PAO.

Insertions of the TOL plasmid transposons Tn4651 and Tn4653 into the Pseudomonas aeruginosa PAO chromosome were isolated by a temperature selection technique. The locations and orientations of 16 insertions were determined by pulsed field gel electrophoresis and Southern hybridization with genomic and TOL DNA probes. All insertions occurred within a 334 kb region of the chromosome (representing less than 6% of the genome) with nine of the inserts clustered within a 10 kb area. Each transposon was able to insert in either orientation. An internal duplication of the 39 kb excisable region of pWW0 was seen in two independent insertions.

Chromosome Mapping↗
PubMed1996

IS1236, a newly discovered member of the IS3 family, exhibits varied patterns of insertion into the Acinetobacter calcoaceticus chromosome.

Analysis of spontaneous mutations in Acinetobacter calcoaceticus revealed a 1237 bp insertion sequence named IS1236 and possessing a nucleotide sequence resembling those of members of the lS3 family. The chromosome of A. calcoaceticus strain ADP1 contains seven copies of IS1236 which appears to insert preferentially into pobR, the transcriptional activator of the structural gene for p-hydroxybenzoate hydroxylase. IS1236 creates tandem 3 bp DNA duplications flanking the sites of its insertion in pobR. Different duplication patterns are found following insertion of IS1236 into pcaH, a structural gene for protocatechuate 3,4-dioxygenase. Therefore the insertion of properties of IS1236 appear to be influenced by its DNA target. Amino acid sequences associated with the apparent transposase function have been conserved in ORFB of IS1236 whereas the presumed DNA-binding helix-turn-helix region of IS1236 ORFA exhibits substantial amino acid sequence divergence from its IS3 counterparts. IS1236 ORFA and ORFB coding sequences overlap considerably, and sequence evidence indicates mechanisms for ORFB expression in IS1236 may resemble those employed by other members of the IS3 family. Portions of the IS1236 terminal repeats exhibit substantial sequence divergence from other members of the IS3 family, but evolution appears to have conserved a mechanism preventing expression of the insertion sequence genes as a consequence of transcriptional readthrough.

Acinetobacter calcoaceticus↗
PubMed1998

Vector-hexamer PCR isolation of all insert ends from a YAC contig of the mouse Igh locus.

We have developed a simple PCR strategy, termed vector-hexamer PCR, that is unique in its ability to easily recover every insert end from large insert clones in YAC and BAC vectors. We used this method to amplify and isolate all insert ends from a YAC contig covering the mouse Igh locus. Seventy-seven ends were amplified and sequenced from 36 YAC clones from four libraries in the pYAC4 vector. Unexpectedly, 40% of the insert ends of these YACs were LINE1 repeats. Nonrepetitive ends were suitable for use as probes on Southern blots of digested YACs to identify overlaps and construct a contig. The same strategy was used successfully to amplify insert ends from YACs in the pRML vector from the Whitehead Institute/MIT-820 mouse YAC library and from BACs in pBeloBAC11. The simplicity of this technique and its ability to isolate every end from large insert clones are of great utility in genomic investigation. [The nucleotide sequence data reported in this paper are accessible in GenBank under accession nos. B07512-B07598.]

Animals↗
PubMed1991

Embryonic Lethals and T-DNA Insertional Mutagenesis in Arabidopsis.

T-DNA insertional mutagenesis represents a promising approach to the molecular isolation of genes with essential functions during plant embryo development. We describe in this report the isolation and characterization of 18 mutants of Arabidopsis thaliana defective in embryo development following seed transformation with Agrobacterium tumefaciens. Random T-DNA insertion was expected to result in a high frequency of recessive embryonic lethals because many target genes are required for embryogenesis. The cointegrate Ti plasmid used in these experiments contained the nopaline synthase and neomycin phosphotransferase gene markers. Nopaline assays and resistance to kanamycin were used to estimate the number of functional inserts present in segregating families. Nine families appeared to contain a T-DNA insert either within or adjacent to the mutant gene. Eight families were clearly not tagged with a functional insert and appeared instead to contain mutations induced during the transformation process. DNA gel blot hybridization with internal and right border probes revealed a variety of rearrangements associated with T-DNA insertion. A general strategy is presented to simplify the identification of tagged embryonic mutants and facilitate the molecular isolation of genes required for plant embryogenesis.

Journal Article↗
PubMedDate not supplied

Modification of the peritoneoscopic technique of peritoneal dialysis catheter insertion: experience of an interventional nephrology program.

Bowel perforation is a well-recognized complication of peritoneal dialysis catheter insertion and is associated with increased morbidity and cost of medical care. In this article we describe our 2-year experience (August 2001-October 2003) with a modified peritoneoscopic technique of peritoneal dialysis catheter insertion to minimize the incidence of bowel perforation. Seventy patients underwent 82 consecutive peritoneal dialysis catheter insertions using the innovative technique. The modified technique is very similar to the traditional peritoneoscopic procedure except for the following differences. To gain access to the peritoneal cavity, a Veress insufflation needle (Ethicon Endo-Surgery Inc., Cincinnati, OH) is utilized instead of the trocar. In contrast to the sharp tip of the trocar, the Veress needle has a blunt, self-retracting end. In addition, the Veress needle is only 14 gauge as opposed to the 2.2 mm diameter of the trocar. Upon introduction of the Veress needle into the abdominal cavity, two "pops" are discerned similar to the trocar. After introduction, 400-500 cc of air are infused and the needle is removed. The infusion of air creates a space between the peritoneal surface of the anterior abdominal wall and the bowel loops. At this point, the cannula with trocar is inserted into the space created. The rest of the steps of the procedure are the same as the traditional peritoneoscopic technique. Utilizing the innovative technique, all 82 catheter insertions were performed successfully without a single bowel perforation. No other complications except for catheter migration (n = 2) were noted. The extra cost of the needle (35 USD) should be viewed in the context of the costs associated with management of a bowel perforation. Large-scale studies are needed to confirm the superiority of this innovative technique over the traditional peritoneoscopic insertion found in our case series. In the interim, however, the increased morbidity and cost associated with bowel perforation calls for logical measures to be taken to avoid this dreaded complication.

Adult↗
PubMed2001

Identification of shiga toxin-producing Escherichia coli possessing insertionally inactivated Shiga toxin gene.

We have investigated the Shiga toxin genes of Shiga toxin-producing Escherichia coli (STEC) strains, using polymerase chain reaction (PCR) amplifying the full lengths of these genes. As a result, we found the Shiga toxin 2 gene which was insertionally inactivated by an insertion sequence (IS). This IS element was identical to IS1203v which has been also found in inactivated Shiga toxin 2 genes, and was inserted at the same site as in the previous paper. On the other hand, both Shiga toxin 2 genes were different (98.3% identity). These suggested that IS1203v independently inserted into each Shiga toxin 2 genes, and STEC strains possessing the insertionally inactivated Shiga toxin genes are most likely to have a wide distribution. Amplification of the full length of the Shiga toxin gene is one of the effective methods to detect the gene no matter where the IS element is included, i.e., the insertion can be reflected in the size of amplicon.

Animals↗
PubMed1996

Jaw thrusting as a clinical test to assess the adequate depth of anaesthesia for insertion of the laryngeal mask.

We have studied the efficacy of the loss of response to jaw thrust as a clinical test to assess adequate depth of anaesthesia for insertion of the laryngeal mask in 60 patients. After induction of anaesthesia with propofol (infused using a syringe driver), the patients were randomly allocated to one of two groups. In one group, insertion of the laryngeal mask was attempted immediately after the loss of verbal contact and in the other group, after the loss of motor response to a jaw thrust. Conditions for insertion of the laryngeal mask were assessed. The mean dose of propofol required to obtain loss of verbal contact was 1.94 mg.kg-1 (SD 0.39, 95% confidence intervals (CI) 1.79-2.08 mg.kg-1) and that for the loss of response to jaw thrust was 2.55 mg.kg-1 (SD 0.46, 95% CI 2.38-2.72 mg.kg-1). When depth of anaesthesia was assessed using jaw thrusting, it was always possible to insert the mask and the conditions were optimal in 87% (95% CI 72-95%) of patients. Neither coughing nor gagging occurred. In contrast, conditions were almost always less than optimal when insertion was attempted after the loss of verbal contact. Conditions were significantly better when jaw thrust was used as a clinical test compared with loss of verbal contact (p < < 0.001). No marked haemodynamic depression occurred in any patient. Thus, jaw thrust is a reliable clinical test to assess the adequate depth of anaesthesia for uncomplicated insertion of the laryngeal mask after induction of anaesthesia with propofol.

Adult↗
PubMed2004

Ease of insertion of the laryngeal tube during manual-in-line neck stabilisation.

The laryngeal tube has a potential role in airway management during anaesthesia or cardiopulmonary resuscitation. In patients with unstable necks, the head and neck may need to be stabilised manually (manual in-line stabilisation), but it is not known whether this procedure affects the ease of insertion of the laryngeal tube. We studied, in a cross-over study, 21 adult patients to compare the success rate of ventilation through the laryngeal tube between the Magill position (a pillow under the occiput and the head extended) or the manual in-line position of the head and neck (without a pillow under the occiput). After induction of anaesthesia and neuromuscular blockade, the laryngeal tube was inserted in turn in the two positions. The ease of insertion was scored with four categories (easy, moderately difficult, difficult and impossible), and adequacy of ventilation through the device was assessed. Ventilation was adequate in all 21 patients in the Magill position, but only in two of 21 patients during manual in-line positioning (p < 0.01; 95%CI for difference: 68-94%). In the Magill position, insertion of the laryngeal tube was easy in 16 patients and moderately difficult in the remaining five patients; in the manual in-line stabilisation position, insertion was moderately difficult in two patients and impossible in the remaining 19 patients. Stabilisation of the patient's head and neck by the manual in-line method made insertion of the laryngeal tube either difficult or impossible.

Adolescent↗
PubMed2006

Evaluation of four airway training manikins as simulators for inserting the LMA Classic*.

Airway manikins have traditionally been used for teaching mask ventilation and tracheal intubation. There is an increasing need to use manikins for training in procedures such as insertion of the laryngeal mask airway. We have assessed four new airway training manikins (latest versions of the Airway Trainer (Laerdal, Norway), Airway Management Trainer (Ambu, UK), 'Bill 1' (VBM, Germany) and Airsim (Trucorp, Ireland)) as simulators for insertion of the LMA Classic laryngeal mask airway. Twenty volunteer anaesthetists inserted a size-4 laryngeal mask airway five times into each of the four manikins, in random order. Each insertion was assessed using objective and subjective tests. Subjective assessment varied widely but overall assessment indicated that the Airway Management Trainer was the poorest simulator for insertion of the laryngeal mask airway. The 'Bill 1' and Airsim manikins performed best as simulators for insertion of the laryngeal mask airway, although realistic ventilation with 'Bill 1' was not possible.

Anesthesiology↗
PubMed2007

Insertion polymorphism of transposable elements and population structure of Anopheles gambiae M and S molecular forms in Cameroon.

The insertion polymorphism of five transposable element (TE) families was studied by Southern blots in several populations of the M and S molecular forms of the mosquito Anopheles gambiae sensu stricto from southern Cameroon. We showed that the mean TE insertion site number and the within-population insertion site polymorphism globally differed between the M and S molecular forms. The comparison of the TE insertion profiles of the populations revealed a significant differentiation between these two molecular forms (0.163 < Phi(ST) < 0.371). We cloned several insertions of a non-LTR retrotransposon (Aara8) that were fixed in one form and absent in the other one. The only insertion that could be clearly located on a chromosome arm mapped to cytological division 6 of chromosome X, confirming the importance of this region in the ongoing speciation between the M and S molecular forms.

Animals↗
PubMed1994

IS231A insertion specificity: consensus sequence and DNA bending at the target site.

In its natural host, Bacillus thuringiensis, the insertion sequence IS231A is preferentially inserted into the terminal inverted repeats of the transposon Tn4430. Using a novel transposition assay, we demonstrate that the Tn4430 ends behave as insertion hot spots for IS231A in Escherichia coli. Sequence analysis reveals that IS231A insertion sites match the 5'-GGG(N)5CCC-3' consensus. However, this consensus is not the only determinant of IS231A insertion specificity. Although both Tn4430 ends have identical sequences, one is strongly preferred to the other and the orientation of insertion into this end is not random. We demonstrate that this preference is determined by the flanking regions of the site. These regions display a conserved periodic organization of their sequence which, by conferring anisotropic flexibility, would induce the DNA to bend in a roughly 'S'-shaped structure centered on the target consensus. DNA conformation analysis by polyacrylamide gel electrophoresis indeed shows that the preferred target site of IS231A is flanked by DNA segments curved in opposite directions. We present a model in which DNA bendability and curvature would contribute to the positioning of IS231A transposase on the target DNA.

Bacillus thuringiensis↗
PubMed1987

Insertion mutagenesis on a cell-surface-exposed region of outer membrane protein PhoE of Escherichia coli K-12.

Amino acid residue arginine-158 of the outer membrane protein PhoE of Escherichia coli K-12 has been shown to be cell-surface-exposed [Korteland et al. (1985) Eur. J. Biochem. 152, 691-697]. To study the effects of small insertions in this region of the protein on its biogenesis and characteristics, a unique restriction site was created by site-directed mutagenesis in a plasmid carrying the phoE gene and oligonucleotides of 12-74 bp were inserted. The insertions did not interfere with incorporation into the outer membrane since (a) several monoclonal antibodies, directed against the cell-surface-exposed part of PhoE protein, bound to whole cells producing the altered proteins and (b) the proteins formed functional pores for the uptake of beta-lactam antibiotics. The binding of one monoclonal antibody and of the PhoE-specific phages TC45 and TC45hrN3 was disturbed by the insertions, showing that this region of the protein is immunogenic and is involved in the binding of both of these phages. The functioning of the mutant pores was characterized both in vivo by studying the uptake of beta-lactam antibiotics and in vitro after the reconstitution of the proteins in black lipid films. The pore characteristics changed depending on the nature of the inserted amino acids. Addition of a negatively charged amino acid resulted in decreased anion-selectivity, whereas insertion of a positive charge and deletion of a negative charge had only a small influence.

Amino Acid Sequence↗
PubMed1992

An additional exon in the human vinculin gene specifically encodes meta-vinculin-specific difference peptide. Cross-species comparison reveals variable and conserved motifs in the meta-vinculin insert.

We have analyzed the structure, origin and expression of the high-molecular-mass muscle-specific variant of vinculin, called meta-vinculin. The meta-vinculin-specific inserts from the human and avian molecules have been isolated and sequenced and the sequences confirmed via cloning of the corresponding cDNA. Comparison of the human, avian and determined porcine sequences revealed cross-species identity in the C-terminal half of the insert. Human and porcine meta-vinculin were highly similar in the insert region, showing only five amino acid exchanges; avian meta-vinculin showed 22 exchanges in the same region compared to human meta-vinculin and exhibited, in addition, one extra amino acid, making 69 in all. Each insert was flanked by characteristic KWSSK motifs. Evidence for two vinculin mRNA species in human uterus smooth muscle was provided by reverse transcription combined with the polymerase chain reaction, as well as by ribonuclease-mapping analysis of cDNA/mRNA hybrids. One of the mRNA species contained an additional 204-nucleotide insert that precisely encoded the meta-vinculin-specific peptide. Sequence analysis of the appropriate portion of the human vinculin gene showed that the section coding for the meta-vinculin-specific insert is present as a discrete exon. Thus, meta-vinculin and vinculin mRNA are generated by alternative splicing.

Amino Acid Sequence↗
PubMed2006

Reproductive performance of ewes after 5-day treatment with intravaginal inserts containing progesterone in combination with injection of prostaglandin f2alpha.

Three experiments were conducted with a total of 1579 ewes to examine reproductive performance in response to synchronization of oestrus during the breeding season, using controlled internal drug releasing (CIDR-G) inserts in regimens designed to provide high concentrations of circulating progesterone. In experiment 1, treatment with two CIDR-G inserts for 12 days produced conception rate (79%) and prolificacy (1.9) to first service equivalent to breeding at natural oestrus (56% and 2.0, respectively). Pregnancy rates to two service periods were 90 and 79%, respectively. In experiments 2 and 3, progesterone was delivered by a single CIDR-G insert for 5 days in combination with prostaglandin F2alpha (PGF2alpha; 5 mg i.m., twice, 3 h apart) the day before (experiment 2), or at insert removal (experiment 3). The combined treatments improved rates of synchronization of oestrus (p<0.01) by 23 and 20% points, respectively, and pregnancy rates to the first service period by 19 (p<0.05) and 13 (p<0.01) percentage points, respectively, compared to treatment with PGF2alpha alone. It is concluded that the combination of treatment for 5 days with a CIDR-G insert and two injections of 5 mg PGF2alpha, the day before, or the day of insert removal, were effective treatments to obtain high fertility at synchronized oestrus in ewes during the breeding season.

Administration, Intravaginal↗
PubMed2005

The optimal insertion length of central venous catheter via the femoral route for open-heart surgery in infants and children.

BACKGROUND: The recommended insertion length of central venous (CV) catheter via the internal jugular or subclavian vein has been determined in infants and children. However, the insertion length via the femoral vein has not been well-studied. This study determined the optimal insertion length of CV catheter via the femoral vein. METHODS: Infants and children, who had undergone cardiac catheterization via the right femoral vein, were the subjects of the study. After routine cardiac catheterization, the distance from the femoral puncture site to the third lumbar vertebral body (L3) level, was measured and recorded. The femoral-L3 length was termed as the optimal insertion length. RESULTS: This length was measured in 78 infants and children (age: 1-101 months, weight: 3.1-33.8 kg). The body weight of the patient and the length correlated well: the optimal insertion length (cm) = 0.45 x body weight (kg) + 8.13, coefficient of determination (R2) = 0.84. CONCLUSIONS: It has been recommended to place the tip of the catheter below the level of renal veins to avoid blocking free flow of those veins. Therefore, we chose the mid-point, L3 level as the optimal tip position of the femoral venous catheter. The length derived from the above formula could be used as a guideline for CV catheter insertion via the femoral vein in infants and children.

Anesthesia, General↗
PubMed2006

Laryngeal mask airway insertion requires less propofol than endotracheal intubation in dogs.

OBJECTIVE: To compare the doses of propofol required for insertion of the laryngeal mask airway (LMA) with those for endotracheal intubation in sedated dogs. STUDY DESIGN: Randomized prospective clinical study. Animals Sixty healthy dogs aged 0.33-8.5 (3.0 +/- 2.3, mean +/- SD) years, weighing 2.2-59.0 (23.4 +/- 13.6, mean +/- SD) kg, presented for elective surgery requiring inhalation anaesthesia. METHODS: Animals were randomly assigned to receive either a LMA or an endotracheal tube. Pre-anaesthetic medication was intravenous (IV) glycopyrrolate (0.01 mg kg(-1)) medetomidine (10 microg kg(-1)) and butorphanol (0.2 mg kg(-1)). Repeated IV propofol injections (1 mg kg(-1) in 30 seconds) were given until LMA insertion or endotracheal intubation was achieved, when the presence or absence of laryngospasm, the respiratory rate (fr) and the total dose of propofol used were recorded. RESULTS: The total propofol dose (mean +/- SD) required for LMA insertion (0.53 +/- 0.51 mg kg(-1)) was significantly lower than for endotracheal intubation (1.43 +/- 0.57 mg kg(-1)). The LMA could be inserted without propofol in 47% of dogs; the remainder needed a single 1 mg kg(-1) bolus (n = 30). Endotracheal intubation was possible without propofol in 3.3% of the dogs, 47% needed one bolus and 50% required two injections (n = 30). The f(r) (mean +/- SD) was 18 +/- 6 and 15 +/- 7 minute(-1) after LMA insertion and intubation, respectively. CONCLUSION AND CLINICAL RELEVANCE: Laryngeal mask airway insertion requires less propofol than endotracheal intubation in sedated dogs therefore propofol-induced cardiorespiratory depression is likely to be less severe. The LMA is well tolerated and offers a less invasive means of securing the upper airway.

Anesthesia, General↗