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PubMed2000

Long nucleotide insertions between the HN and L protein coding regions of human parainfluenza virus type 3 yield viruses with temperature-sensitive and attenuation phenotypes.

Recombinant parainfluenza virus 3 (rPIV3) is being developed as a vector to express foreign genes as a bivalent or multivalent live attenuated virus vaccine. In the present study, we examined the effect of inserted foreign sequence on virus replication in vitro and in vivo, focusing on the parameter of insert length. In one type of construct, foreign sequence of increasing length was flanked by PIV3 transcription signals and inserted as an additional gene unit (GU insert) between the HN and L genes, so that one additional mRNA would be made. In a second type of construct, foreign sequence was inserted into the downstream NCR (NCR insert) of the HN gene, so that the number of encoded mRNAs remained unchanged. In each case, the foreign sequence was designed to lack any significant open reading frame, which permitted an evaluation of the effect of insert length on replication independent of an effect of an expressed protein. The GU or NCR insert sizes ranged from 168 nucleotides (nt) to 3918 nt. rPIV3s containing GU insertions of up to 3918 nt in length, the largest size tested, were viable and replicated efficiently at permissive temperatures in vitro, but a reduction in plaque size was seen at 39 degrees C and 40 degrees C. The rPIV3 with a 3918-nt GU insertion was restricted in replication in the upper (fivefold) and lower (25-fold) respiratory tracts of hamsters. Although a 1908-nt GU insertion did not significantly modify replication of wild-type PIV3 in vitro or in vivo, its introduction significantly augmented the level of temperature sensitivity (ts) and attenuation (att) specified by three mutations in the L protein of a cold-passaged attenuated PIV3 vaccine virus. rPIV3s bearing a 3126- or 3894-nt NCR insertion exhibited in vitro and in vivo phenotypes like those of the rPIV3s bearing similar-sized GU insertions. These findings indicate that rPIV3s whose genome length has been increased by more than 3000 nt by either a GU or an NCR insertion exhibit an unexpected host-range phenotype, that is, efficient replication in vitro but restricted replication in hamsters, especially in the lower respiratory tract. Furthermore, these effects were greatly enhanced when the rPIV3 backbone contained other ts or att mutations. The implications of these findings for the use of single-stranded, negative-sense RNA viruses as vectors for vaccines are discussed.

Animals↗
PubMed1999

Disruption of coding regions by IS6110 insertion in Mycobacterium tuberculosis.

SETTING: The insertion sequence IS6110 is widely used as a DNA fingerprinting probe for the classification of Mycobacterium tuberculosis strains. This study has focused on the characterization of regions disrupted by insertion of the IS6110 element. OBJECTIVE: To characterize IS6110 insertion loci in clinical isolates of M. tuberculosis, in terms of their genomic location and genetic identity, to ascertain whether IS6110 transposition could be a mechanism driving phenotypic change. DESIGN: Thirty-three IS6110 insertion loci were cloned from 8 clinical isolates of M. tuberculosis. Clones representing DR locus insertions were identified by hybridization (n = 4), and all other clones were characterized by DNA sequencing (n = 29). The sequence data was analyzed in conjunction with that of 43 other insertion loci identified in published literature and DNA sequence databases. RESULTS: The 76 sequences analyzed represented 66 unique insertion loci (including 9 unique insertions into the ipl locus). When mapped to the H37Rv genome, the majority of unique insertion loci demonstrated disruption of coding regions by IS6110 (n = 42; including the ipl insertions), while the remainder either occurred within intergenic regions (n = 17), or could not be mapped to the H37Rv genome sequence (n = 7). Mapping of the insertion loci reveals distribution throughout the chromosome, with isolated preferential insertion loci. CONCLUSIONS: This study has demonstrated the occurrence of 66 unique IS6110 insertion loci dispersed throughout the M. tuberculosis genome, with an unexpectedly high incidence of IS6110 insertions occurring within coding regions. However, the IS6110-mediated coding region disruptions identified here may only have limited impact on phenotype, as most of the coding regions disrupted are members of multiple gene families. Disruption of individual members of a family of genes may have no effect on phenotype or could have a minor or major impact, depending on the specificity and activity of the encoded protein.

DNA Fingerprinting↗
PubMed2004

Cardiovascular changes following insertion of oropharyngeal and nasopharyngeal airways.

BACKGROUND: The cardiovascular responses following the insertion of oropharyngeal airways in anaesthetized patients have been found to be of little consequence. However, those following the insertion of nasopharyngeal airways have not been studied. The aim of this investigation was to compare the cardiovascular responses to the insertion of oropharyngeal and nasopharyngeal airways in anaesthetized patients. METHODS: Twenty-four ASA I or II patients aged 16-65 yr, requiring nasotracheal intubation as part of their anaesthetic management, received a standardized general anaesthetic and were randomly allocated to receive either a nasopharyngeal or an oropharyngeal airway. RESULTS: The two groups were similar with respect to age, weight and gender. There was a significant decrease in systolic pressure following the induction of anaesthesia in both groups. Following nasopharyngeal airway insertion, there was a significant rise in systolic pressure above pre-insertion levels (P< or =0.001), though not above pre-induction levels (P=0.808). There was no significant change in the oropharyngeal airway insertion group (P=0.619). At 1 min post-insertion, the mean (sd) systolic pressure in the nasopharyngeal insertion group, 122 (21.6) mm Hg, was significantly greater than that in the oropharyngeal insertion group, 103 (15.3) mm Hg (P=0.017). Diastolic pressure followed a similar pattern. In both groups, heart rate fell after induction and fell further after insertion, and at 4 min post-insertion was significantly lower than pre-induction and pre-insertion levels. There was no significant difference in heart rates between the two groups (P=0.372). CONCLUSIONS: The pressor response following the insertion of nasopharyngeal airways in anaesthetized patients is significantly greater than that following the insertion of oropharyngeal airways. However, the mean rise in arterial pressure does not exceed pre-induction level, and thus the response is less severe than that likely to be associated with orotracheal or nasotracheal intubation.

Adolescent↗
PubMed1991

Electrocardiographic body surface potential mapping in the Wolff-Parkinson-White syndrome. Noninvasive determination of the ventricular insertion sites of accessory atrioventricular connections.

BACKGROUND: A reliable, noninvasive procedure to determine the location of accessory atrioventricular connections in patients with Wolff-Parkinson-White syndrome would add an important diagnostic tool to the clinical armamentarium. METHODS AND RESULTS: Body surface potential mapping (BSPM) using 180 electrodes in various-sized vests and displayed as a calibrated color map was used to determine the ventricular insertion site of the accessory atrioventricular (AV) connections in 34 patients with Wolff-Parkinson-White syndrome. Attempts were made to determine the 17 ventricular insertion sites described by Guiraudon et al. All 34 patients had an electrophysiologic study (EPS) at cardiac catheterization, and 18 had surgery so the ventricular insertion sites could be accurately located using EPS at surgery. A number of physiologic observations were also made with BSPM. CONCLUSIONS: The following conclusions were drawn: 1) BSPM using QRS analysis accurately predicts the ventricular insertion site of accessory AV connections in the presence of a delta wave in the electrocardiogram; 2) the ventricular insertion sites of accessory AV connections determined by BSPM and by EPS at surgery were identical or within one mapping site (1.5 cm or less) in all but four of 18 cases; three of the four exceptions had more than one accessory AV connection, and the other had a very broad ventricular insertion; 3) BSPM and EPS locations of the accessory AV connections correlated very well in the 34 cases despite the fact that BSPM determines the ventricular insertion site and EPS determines the atrial insertion site of the accessory AV connection; 4) as suggested by the three cases of multiple accessory AV connections, EPS and BSPM may be complementary since BSPM identified one pathway and EPS identified the other (in the case with a broad ventricular insertion, BSPM and EPS demonstrated different proportions of that insertion); 5) BSPM using ST-T analysis is very much less accurate in predicting the ventricular insertion site of accessory AV connections unless there is marked preexcitation; 6) standard electrocardiography using the Gallagher grid methodology (but with no attempt at stimulating maximal preexcitation) was not as accurate as QRS analysis of BSPM in predicting the ventricular insertion site of the accessory AV connection; however, exact comparison is hampered by the different number and size of the Gallagher and Guiraudon insertion sites; 7) BSPM using QRS analysis appears to be very accurate in predicting right ventricular versus left ventricular posteroseptal accessory AV connections; 8) typical epicardial right ventricular breakthrough, indicative of conduction via the specialized AV conduction system, occurs in all patients with left ventricular free wall accessory AV connections; 9) epicardial right ventricular breakthrough was not observed in cases with right ventricular free wall or anteroseptal accessory AV connections; 10) epicardial right ventricular breakthrough can occur in the presence of posteroseptal accessory AV connections, whether right or left ventricular; and 11) the delay in epicardial right ventricular breakthrough in cases with left ventricular insertion may provide a marker to estimate the degree of ventricular preexcitation.

Adult↗
PubMed2006

Clinical manifestations and risk factors of children receiving triple ventilating tube insertions for treatment of recurrent otitis media with effusion.

OBJECTIVE: Our goal was to determine risk factors for children receiving additional ventilating tube insertions after initial tube insertion and to determine the changes of clinical manifestations in children who have undergone 3 ventilating tube insertion procedures. STUDY DESIGN: We retrospectively analyzed medical and operation records of 423 young patients who had ventilating tube insertion because of chronic otitis media with effusion from January 1993 to December 1998. The single-operation group included patients who had 1 ventilating tube insertion only, and the triple-operation group included patients who received ventilating tube insertion 3 times because of recurring chronic otitis media with effusion. RESULTS: At the first operation, there were significant differences between the single- and triple-operation groups in mean age, the proportion who received a concurrent adenoidectomy, the mean indwelling period of the first ventilating tube, the proportion who developed postoperative otorrhea within 1 month, and the proportion who had early extrusion of the ventilating tube within 3 months of surgery. In the triple-operation group, the accumulated number of adenoidectomies, the indwelling period of the ventilating tube, and the time interval before subsequent ventilating tube insertion after ventilating tube extrusion significantly increased as ventilating tube insertion procedures were performed repeatedly. Although there was no difference when compared with the single-operation group, the proportion of glue-like effusion significantly decreased as ventilating tube insertion procedures were performed repeatedly. There were no significant differences between the single- and triple-operation groups in male/female ratio, site of ventilating tube insertion, and the proportion of patients with glue-like effusion at the first ventilating tube insertion. CONCLUSIONS: The probability of receiving additional ventilating tube insertion because of recurrent otitis media with effusion significantly increased in younger patients at the time of first ventilating tube insertion. The concurrent adenoidectomy, duration of the ventilating tube, postoperative otorrhea within 1 month, and early extrusion of the ventilating tube also influenced the probability of additional ventilating tube insertion.

Child, Preschool↗
PubMed2003

Intrauterine device insertion-related complications: can they be predicted?

OBJECTIVE: To determine risk factors affecting the incidence of intrauterine device (IUD) insertion-related complications and failures and, in particular, whether postcoital IUD insertions have a higher incidence of complications than routine IUD insertions. DESIGN: Retrospective case notes analysis. PARTICIPANTS: The study examined 545 case notes of patients having IUD insertions at East Cheshire NHS Trust family planning clinics between 1 October 1997 and 31 December 2000. MAIN OUTCOME MEASURES: The incidence of complications at insertion, or up to 12 weeks after insertion, was determined and included failed insertion, cervical problems, syncope, bradycardia, convulsions, early perforation and early expulsion. Fourteen potential risk factors were examined to determine the effect on incidence of complications. RESULTS: Failed insertions were statistically more likely in women who had never previously had a vaginal delivery and also when a less experienced doctor performed the insertion. Nulliparous women were at statistically increased risk of cervical problems and bradycardia. Cervical problems at insertion also increased significantly with age. Patients who were amenorrhoeic at insertion were more likely to suffer an early IUD expulsion. CONCLUSIONS: Of the potential risk factors, nulliparity was the most important. IUD insertion failures and complications were no more common in postcoital than routine IUD insertions. In general, complications were unpredictable, indicating the need for constant vigilance and the inserting doctor being trained and prepared to deal with any complication arising.

Adolescent↗
PubMed2005

Site-specific reflex response of ubiquitin to loop insertions.

Predicting the structural effects of insertions in proteins by homology modeling remains a challenge. To investigate the molecular basis for conformational adaptations to insertions, ten mutants of ubiquitin were generated by introducing five different inserts, varying from five to 11 residues in size, at two different sites. Most insertion sequences were derived from homologous positions in structurally homologous ubiquitin-like proteins; to test sequence specificity, insertions were made into both homologous and non-homologous sites in ubiquitin. Structural inferences from NMR data suggest that each insertion site shows a reflex response to insertions: the sequence of the insertion has much less impact on structural adaptations than does the site of the insertion. Further, each site responds to insertions in a unique but consistent manner. For a given insertion site, different inserted sequences give rise to different stabilities, but the relationship between stability and sequence is not yet clear. However, the change in stability is similar for all insertions in a given site.

Amino Acid Sequence↗
PubMed2002

A collection of sequenced and mapped Ds transposon insertion sites in Arabidopsis thaliana.

Insertional mutagenesis is a powerful tool for generating knockout mutations that facilitate associating biological functions with as yet uncharacterized open reading frames (ORFs) identified by genomic sequencing or represented in EST databases. We have generated a collection of Dissociation (Ds) transposon lines with insertions on all 5 Arabidopsis chromosomes. Here we report the insertion sites in 260 independent single-transposon lines, derived from four different Ds donor sites. We amplified and determined the genomic sequence flanking each transposon, then mapped its insertion site by identity of the flanking sequences to the corresponding sequence in the Arabidopsis genome database. This constitutes the largest collection of sequence-mapped Ds insertion sites unbiased by selection against the donor site. Insertion site clusters have been identified around three of the four donor sites on chromosomes 1 and 5, as well as near the nucleolus organizers on chromosomes 2 and 4. The distribution of insertions between ORFs and intergenic sequences is roughly proportional to the ratio of genic to intergenic sequence. Within ORFs, insertions cluster near the translational start codon, although we have not detected insertion site selectivity at the nucleotide sequence level. A searchable database of insertion site sequences for the 260 transposon insertion sites is available at http://sgio2.biotec.psu.edu/sr. This and other collections of Arabidopsis lines with sequence-identified transposon insertion sites are a valuable genetic resource for functional genomics studies because the transposon location is precisely known, the transposon can be remobilized to generate revertants, and the Ds insertion can be used to initiate further local mutagenesis.

Arabidopsis↗
PubMed1996

Site-selected insertional mutagenesis of tomato with maize Ac and Ds elements.

Site-selected insertion (SSI) is a PCR-based technique which uses primers located within the transposon and a target gene for detection of transposon insertions into cloned genes. We screened tomato plants bearing single or multiple copies of maize Ac or Ds transposable elements for somatic insertions at one close-range target and two long-range targets. Eight close-range Ds insertions near the right border of the T-DNA were recovered. Sequence analysis showed a precise junction between the transposon and the target for all insertions. Two insertions in separate plants occurred at the same site, but others appeared dispersed in the region of the right T-DNA border with no target specificity. However, insertions showed a preference for one orientation of the transposon. Use of plants with multiple Ac (HiAc) or Ds (HiDs) elements allowed detection of somatic insertions at two single-copy genes, PG (polygalacturonase) and DFR (dihydroflavonol 4-reductase). Certain HiDs plants showed much higher rates of insertion into PG than others. Insertions in PG and DFR were found throughout the gene regions monitored and, with the exception of one insertion in PG, the junctions between transposon and target were exact. SSI analysis of progeny from the HiDs parents revealed that in some cases the tendency to incur high levels of somatic insertions in PG was inherited. Inheritance of this character is an indication that SSI could be used to direct a search for germinal PG insertions in tomato.

Alcohol Oxidoreductases↗
PubMed2006

The insertional footprint of the rotator cuff: an anatomic study.

PURPOSE: The purpose of this study was to define the entire rotator cuff footprint and relate it to known, easily identifiable landmarks as a guide for both open and arthroscopic rotator cuff repair. Anatomic: Gross and microscopic. METHODS: The myotendinous units of the rotator cuff and their insertions onto the humerus were dissected in 20 fresh-frozen cadavers. The separate tendon insertions were identified, and their length and width measured. The character and exact anatomy of the tendons were also noted. The entire insertion was measured and referenced to the articular surface, biceps groove, and bare area of the humerus. In a separate part of the study, 6 cadavers were decalcified and thin-sliced through the supraspinatus tendon insertion. This insertion was evaluated via scanning electron microscopy (SEM). RESULTS: Our findings demonstrated a consistent pattern at the insertion of the rotator cuff. The horseshoe-shaped insertion tapers away from the articular surface in a superior-to-inferior direction. Interdigitation of the muscle units may be noted, particularly between the supraspinatus and the infraspinatus. Average maximum insertional lengths and widths were as follows: subscapularis (SC): 40 x 20 mm; infraspinatus (IS): 29 x 19 mm; supraspinatus (SS): 23 x 16 mm; and teres minor (TM): 29 x 21 mm. The SC inserted on the lesser tuberosity adjacent to the biceps groove at the edge of the articular surface. It tapered away 18 mm at its inferior border. The SS inserted at the articular surface along its entire insertion from the bicipital groove to the top of the bare area. The IS wrapped the posterior border of the SS superiorly at the articular surface and tapered away inferiorly, framing the bare area. SEM microscopy showed the SS to be adherent to the edge of the articular surface medially. As it filled the sulcus, its lateral edge extended over the edge of the greater tuberosity. CONCLUSIONS: A consistent pattern was noted at the insertional anatomy of the rotator cuff. This anatomy was related to known, easily identifiable landmarks and may serve as a guide for evaluation of size, location, and propagation patterns of rotator cuff tears, as well as for their repair. CLINICAL RELEVANCE: Knowledge of the insertional anatomy of the rotator cuff can facilitate grading and repair of rotator cuff tears.

Cadaver↗
PubMed2003

One-side-coated insert as a unique ophthalmic drug delivery system.

We newly prepared a unique one-side-coated insert that releases drug from only uncoated side. The purpose of this study is to determine whether ocular and systemic absorption of ophthalmic drug could be altered by an inserting direction of the insert in rabbit eyes. One-side-coated insert was prepared by attaching a polypropylene tape on the one side of the polymer disc of poly(2-hydroxypropyl methacrylate) (HPM) containing tilisolol as a model ophthalmic drug. The insert was applied in the lower conjunctival cul-de-sac of albino rabbits with the uncoated side facing bulbar conjunctiva/sclera (SC insert) or palpebral conjunctiva (CJ insert). At the adequate intervals, the tear fluid, plasma, aqueous humor, conjunctiva, and sclera were collected and the drug concentrations were determined by an HPLC. A release of tilisolol from the one-side-coated insert was twice slower than from the uncoated insert. Ocular application of the one-side-coated insert produced the constant concentrations of tilisolol in the tear fluid over 180 min. SC insert showed higher drug concentrations in the aqueous humor and sclera, and lower drug concentrations in the plasma and conjunctiva than CJ insert.The one-side-coated insert can alter the ocular and systemic absorption of drug by an inserting direction.

Adrenergic beta-Antagonists↗
PubMed2003

Variation of biomechanical, structural, and compositional properties along the tendon to bone insertion site.

The tendon to bone insertion site is a complex transitional region that links two very different materials. The insertion site must transfer a complex loading environment effectively to prevent injury and provide proper joint function. In order to accomplish this load transfer effectively, the properties of the insertion site were hypothesized to vary along its length. The quasilinear viscoelastic (QLV) Model was used to determine biomechanical properties, polarized light analysis was used to quantitate collagen orientation (structure), and in situ hybridization was used to determine the expression of extracellular matrix genes (composition). All assays were performed at two insertion site locations: the tendon end of the insertion and the bony end of the insertion. Biomechanically, the apparent properties of peak strain, the coefficients (A and B) that describe the elastic component of the QLV model, and one of the coefficients (tau(1)) of the viscous component of the model were significantly higher, while another of the coefficients (C) of the viscous component was significantly lower at the tendon insertion compared to the bony insertion. The collagen was significantly more oriented at the tendon insertion compared to the bony insertion. Finally, collagen types II, IX, and X, and aggrecan were localized only to the bony insertion, while decorin and biglycan were localized only to the tendon insertion. Thus, the tendon to bony insertion site varies dramatically along its length in terms of its viscoelastic properties, collagen structure, and extracellular matrix composition.

Aggrecans↗
PubMed1995

Neuronal cell expression of inserted isoforms of vertebrate nonmuscle myosin heavy chain II-B.

Previous work has demonstrated that unique isoforms of nonmuscle myosin heavy chain II-B (MHC-B) are expressed in chicken and human neuronal cells (Takahashi, M., Kawamoto, S., and Adelstein, R. S. (1992) J. Biol. Chem. 267, 17864-17871). These isoforms, which appear to be generated by alternative splicing of pre-mRNA, differ from the MHC-B isoform present in a large number of nonmuscle cells in that they contain inserted cassettes of amino acids near the ATP binding region and/or near the actin binding region. The insert near the ATP binding region begins after amino acid 211 and consists of either 10 or 16 amino acids. The insert near the actin binding region begins after amino acid 621 and consists of 21 amino acids. Using a variety of techniques, we have studied the distribution and expression of the inserted MHC-B isoforms. In the developing chicken brain, mRNA encoding the 10-amino acid insert gradually increases after embryonic day 4, peaks in the 10-14-day embryo, and then declines. In contrast, the mRNA encoding the 21-amino acid insert appears just before birth and is abundantly expressed in the adult chicken cerebellum. There is a marked species difference between the distribution of the inserted isoforms in adult tissues. The mRNA encoding MHC-B containing the 10-amino acid insert near the ATP binding region is expressed at low levels in the adult chicken brain, but makes up most of the MHC-B mRNA expressed in the human cerebrum and approximately 90% of MHC-B in the human retina. It is also expressed in neuronal cell lines. The mRNA encoding MHC-B containing the 21-amino acid insert is abundantly expressed in the chicken cerebellum and human cerebrum, but is absent from the retina and cell lines. Employing human retinoblastoma (Y-79) and neuroblastoma (SK-N-SH) cell lines, an increase in expression of mRNA encoding the 10-amino acid inserted isoform was seen following treatment by a number of agonists or by serum deprivation. In each case, expression of the inserted MHC-B isoform correlated with cell differentiation (neuronal phenotype) and inhibition of cell division. Using a rat pheochromocytoma cell line (PC12), we found that prior to treatment with nerve growth factor (NGF), there was no evidence for either inserted isoform, although noninserted MHC-B was present. NGF treatment resulted in the appearance of mRNA encoding MHC-B containing the 10-amino acid insert, concomitant with neurite outgrowth.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1987

The influenza hemagglutinin insertion signal is not cleaved and does not halt translocation when presented to the endoplasmic reticulum membrane as part of a translocating polypeptide.

The co-translational insertion of polypeptides into endoplasmic reticulum membranes may be initiated by cleavable amino-terminal insertion signals, as well as by permanent insertion signals located at the amino-terminus or in the interior of a polypeptide. To determine whether the location of an insertion signal within a polypeptide affects its function, possibly by affecting its capacity to achieve a loop disposition during its insertion into the membrane, we have investigated the functional properties of relocated insertion signals within chimeric polypeptides. An artificial gene encoding a polypeptide (THA-HA), consisting of the luminal domain of the influenza hemagglutinin preceded by its amino-terminal signal sequence and linked at its carboxy-terminus to an intact prehemagglutinin polypeptide, was constructed and expressed in in vitro translation systems containing microsomal membranes. As expected, the amino-terminal signal initiated co-translational insertion of the hybrid polypeptide into the membranes. The second, identical, interiorized signal, however, was not recognized by the signal peptidase and was translocated across the membrane. The failure of the interiorized signal to be cleaved may be attributed to the fact that it enters the membrane as part of a translocating polypeptide and therefore cannot achieve the loop configuration that is thought to be adopted by signals that initiate insertion. The finding that the interiorized signal did not halt translocation of downstream sequences, even though it contains a hydrophobic region and must enter the membrane in the same configuration as natural stop-transfer signals, indicates that the HA insertion signal lacks essential elements of halt transfer signals that makes the latter effective membrane-anchoring domains. When the amino-terminal insertion signal of the THA-HA chimera was deleted, the interior signal was incapable of mediating insertion, probably because of steric hindrance by the folded preceding portions of the chimera. Several chimeras were constructed in which the interiorized signal was preceded by polypeptide segments of various lengths. A signal preceded by a segment of 111 amino acids was also incapable of initiating insertion, but insertion took place normally when the segment preceding the signal was only 11-amino acids long.(ABSTRACT TRUNCATED AT 400 WORDS)

Cytochrome P-450 Enzyme System↗
PubMed1992

Effects of P element insertions on quantitative traits in Drosophila melanogaster.

P element mutagenesis was used to construct 94 third chromosome lines of Drosophila melanogaster which contained on average 3.1 stable P element inserts, in an inbred host strain background previously free of P elements. The homozygous and heterozygous effects of the inserts on viability and abdominal and sternopleural bristle number were ascertained by comparing the chromosome lines with inserts to insert-free control lines of the inbred host strain. P elements reduced average homozygous viability by 12.2% per insert and average heterozygous viability by 5.5% per insert, and induced recessive lethal mutations at a rate of 3.8% per insert. Mutational variation for the bristle traits averaged over both sexes was 0.03Ve per homozygous P insert and 0.003Ve per heterozygous P insert, where Ve is the environmental variance. Mutational variation was greater for the sexes considered separately because inserts had large pleiotropic effects on sex dimorphism of bristle characters. The distributions of homozygous effects of inserts on the bristle traits were asymmetrical, with the largest effects in the direction of reducing bristle number; and highly leptokurtic, with most of the increase in variance contributed by a few lines with large effects. The inserts had partially recessive effects on the bristle traits. Insert lines with extreme bristle effects had on average greatly reduced viability.

Animals↗
PubMed1995

Biomechanical properties of threaded inserts for lumbar interbody spinal fusion.

STUDY DESIGN: Calf and human cadaveric spines were used to determine motion segment stiffness and laxity after implantation of threaded inserts (the Ray Threaded Fusion Cage, Surgical Dynamics, Inc., Concord, CA), comparing direction of placement, number of implants, shape of the device, and integrity of anterior spine structures. Stiffness and laxity of spines with inserts were compared with those with bone grafts, with and without posterior fixation plates. OBJECTIVES: To determine the mechanical stabilizing properties of a threaded insert used for lumbar and lumbosacral fusion and the factors affecting stability. SUMMARY OF BACKGROUND DATA: Limited biomechanical information has shown that implantation of these devices adds stiffness to the lumbar spine, but little information is available concerning stiffness in loading directions other than flexion and extension, the effect on stiffness of position and number of implants, and the effect of this device on motion segment laxity. METHODS: Mechanical properties were determined by testing lumbar vertebral motion segments in flexion, extension, lateral bending, and torsion combined with axial compressive loading. Stiffness (slope of the load/deflection curve) and neutral zone angle or laxity (angular displacement of the vertebra from no load to 1.0 Nm moment) were determined. Initial tests were performed on calf lumbar vertebrae to determine the effects of placement and number of inserts. Comparisons of bone grafts and inserts with and without supplemental plates were made using human lumbar spines. Cylindrical- and conical-shaped inserts, when placed from anterior, were tested in calf spines. The load-bearing capacity of the insert supported in calf vertebral body bone was determined. RESULTS: There was no significant effect of placement of inserts in different orientations (lateral, posterolateral, or posterior) on stiffness, except in torsion where posterior placement damaged facets or lamina, reducing stiffness. Placement of two inserts from posterior decreased flexion and lateral bending laxity compared with the intact motion segment. Compared with intact, bone grafts produced more stiffness only in lateral bending and had no effect on laxity. Supplemental posterior plates fixed by pedicle screws across the fusion segment increased flexion and lateral bending stiffness and reduced laxity in flexion, extension, and lateral bending. Conical-shaped inserts placed from anterior into cylindrical holes distracted soft tissue structures, decreasing laxity. Cutting the anterior structures increased laxity by relieving some tissue tension caused by distraction. The mean maximum compressive load that could be supported by the insert was 2998 N (standard deviation = 980 N). Structural failure occurred in the supporting bone. CONCLUSIONS: Threaded inserts increase vertebral motion segment stiffness and decrease laxity by distracting intervertebral structures. They are not sensitive to placement, except if vertebral structures are injured during insertion and produce constructs with more consistent mechanical properties than bone grafts.

Animals↗
PubMed2005

Ceramic inserts do not generally improve resin composite margins.

summary Ceramic inserts are reported to possibly reduce polymerization shrinkage for posterior resin composite fillings. The aim of the present investigation was to evaluate the effect of different insert systems before and after thermomechanical loading. Sixty sound human third molars received occlusomesial Class II cavities, 40 with proximal margins 2 mm above and 20 with proximal margins 1 mm below the cementum-enamel junction. The specimens were randomly assigned to one of the six experimental groups (n = 10). The enamel-bordered cavities were restored with Syntac classic and Tetric Ceram (ST), Syntac classic, Tetric Ceram and beta-quartz inserts (TB), Syntac classic, Tetric Ceram and Cerana inserts (TC), Syntac classic, Tetric flow and SonicSys approx inserts (TS). The dentin-limited cavities were filled with Syntac classic and Tetic Ceram (DT), Syntac classic, Tetric flow and SonicSys approx inserts (DS). Before and after thermomechanical loading (100 000 x 50 N, 2500 x 5 degrees C/55 degrees C), replicas were made and both interfaces tooth/composite and insert/composite were examined under a scanning electron microscope at 200x. The Cerana and SonicSys insert groups showed significantly less gaps in enamel (P < 0.05). With beta-quartz inserts, no reduction of gaps was found (P > 0.05). Marginal integrity in dentine-bordered specimens could not be improved with SonicSys inserts (P > 0.05). The bonding performance insert/composite was promising for all IPS Empress inserts (Cerana, SonicSys enamel) but worse for beta-quartz inserts. Regarding gap formation between resin composite and tooth, Cerana and SonicSys inserts significantly reduced gaps. The use of SonicSys inserts in deep proximal cavities cannot be recommended.

Ceramics↗
PubMed2006

A laboratory study to determine the effects of universal and rotating ultrasonic inserts on wrist movement and scaling time efficiency of dental hygienists.

PURPOSE: The purpose of this study was to determine if differences existed in range of wrist movements and scaling time efficiency of dental hygienists using a rotating ultrasonic insert when compared with a standard universal insert. METHODS: A convenience sample of 32 consenting experienced dental hygienists who met inclusion criteria was invited to participate. Using a cross-over research design, the 32 participants were randomly assigned to one of the two subgroups. Subgroup A (n = 16) used the rotating universal ultrasonic insert on a typodont, rested for 15 min and the standard universal insert on a different typodont. Subgroup B (n = 16) used the standard universal ultrasonic insert on a typodont, rested for 15 min and the rotating universal ultrasonic insert on a different typodont. Each participant used the rotating and standard universal ultrasonic scaling inserts to remove 2 cm3 artificial calculus from two different typodonts for up to 15 min per insert. Scaling time efficiency was determined using a Modified Volpe-Manhold Calculus Index, measuring the amount of artificial calculus remaining after ultrasonic scaling. While scaling, each participant wore the WristSensor goniometry gloves, which determined changes in wrist movements (flexion and extension and ulnar and radial deviations), measured as a deviation from the neutral position. RESULTS: A paired t-test (P = 0.05) using 30 subjects with useable data, revealed no statistically significant differences between the two different inserts in terms of wrist movements and scaling time efficiency. A multivariate analysis of variance revealed no statistically significant differences in the percentage of time dental hygienists were in high, medium or low-risk posture categories while using the rotating ultrasonic insert compared with the standard universal insert. Based on the results of this laboratory study, dental hygienists using a rotating ultrasonic insert appear to experience no ergonomic advantage in terms of wrist postures or timesavings over a standard insert.

Cross-Over Studies↗