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Response of human REV1 to different DNA damage: preferential dCMP insertion opposite the lesion.

REV1 functions in the DNA polymerase zeta mutagenesis pathway. To help understand the role of REV1 in lesion bypass, we have examined activities of purified human REV1 opposite various template bases and several different DNA lesions. Lacking a 3'-->5' proofreading exonuclease activity, purified human REV1 exhibited a DNA polymerase activity on a repeating template G sequence, but catalyzed nucleotide insertion with 6-fold lower efficiency opposite a template A and 19-27-fold lower efficiency opposite a template T or C. Furthermore, dCMP insertion was greatly preferred regardless of the specific template base. Human REV1 inserted a dCMP efficiently opposite a template 8-oxoguanine, (+)-trans-anti-benzo[a]pyrene-N2-dG, (-)-trans-anti-benzo[a]pyrene-N2-dG and 1,N6-ethenoadenine adducts, very inefficiently opposite an acetylaminofluorene-adducted guanine, but was unresponsive to a template TT dimer or TT (6-4) photoproduct. Surprisingly, the REV1 specificity of nucleotide insertion was very similar in response to different DNA lesions with greatly preferred C insertion and least frequent A insertion. By combining the dCMP insertion activity of human REV1 with the extension synthesis activity of human polymerase kappa, bypass of the trans-anti-benzo[a]pyrene-N2-dG adducts and the 1,N6-ethenoadenine lesion was achieved by the two-polymerase two-step mechanism. These results suggest that human REV1 is a specialized DNA polymerase that may contribute to dCMP insertion opposite many types of DNA damage during lesion bypass.

Binding Sites↗

Both amino- and carboxy-terminal portions are required for insertion of yeast porin into the outer mitochondrial membrane.

Yeast porin, the major outer mitochondrial membrane protein, is synthesized without a cleavable extension peptide and post-translationally inserted into the membrane. When inserted into the membrane, it acquires resistance to externally added trypsin. To locate the sequences responsible for membrane insertion and topogenesis in the primary structure of yeast porin, we constructed several deletion and chimeric mutants of the porin cDNA. These cDNAs were expressed in vitro and the products were assayed for capacity to be correctly inserted into isolated mitochondria. It was thus found that deletion of the segment spanning residues 37-98 did not appreciably impair the insertion competence and the inserted protein became resistant to trypsin. On the other hand, the porin mutant lacking the segment consisting of residues 17-98 did not acquire the trypsin resistance, though it could bind to mitochondria specifically. Deletion of the carboxy-terminal 62 amino acid residues also abolished the capacity to be correctly inserted into mitochondria. We conclude that information required for membrane insertion and intramembranous topogenesis of the porin molecule is stored not only in the amino-terminal region but also in the carboxy-terminal portion.

Alkalies↗

Comparison of sevoflurane with propofol for laryngeal mask airway insertion in adults.

UNLABELLED: We performed a prospective, randomized, controlled trial to compare the quality and ease of laryngeal mask airway (LMA) insertion after either rapid inhaled sevoflurane or i.v. propofol induction of anesthesia. Seventy-six unpremedicated ASA physical status I or II patients were anesthetized with either a single vital capacity breath of sevoflurane 8% or i.v. propofol 3 mg/kg, which produced equally rapid loss of consciousness (40.5 +/- 13.9 vs 37.7 +/- 9.9 s; P > 0.05). The LMA was inserted more rapidly in patients in the propofol group (74 +/- 29 vs 127 +/- 35 s; P < 0.01) and required fewer attempts (1.2 vs 1.6; P < 0.05) than the sevoflurane group. There was a greater incidence of initially impossible mouth opening in the sevoflurane group (45% vs 21%; P < 0.05). Once mouth opening was possible, the degree of attenuation of laryngeal reflexes was similar. The overall incidence of complications related to LMA insertion, especially apnea (32% vs 0%; P < 0.01), was more frequent in the propofol group (82% vs 26%; P < 0.01). There were four failures of LMA insertion in the propofol group and none in the sevoflurane group. Both groups had stable hemodynamic profiles and good patient satisfaction. We conclude that sevoflurane vital capacity breath induction compares favorably with i.v. propofol induction for LMA insertion in adults. However, prolonged jaw tightness after the sevoflurane induction of anesthesia may delay LMA insertion. IMPLICATIONS: In this randomized, controlled trial, we compared the ease of insertion of the laryngeal mask airway in adults after induction of anesthesia with either a sevoflurane vital capacity breath technique or propofol i.v.. We conclude that sevoflurane compares favorably with propofol, although prolonged jaw tightness may delay laryngeal mask airway insertion.

Adult↗

Anatomical landmarks for femoral nerve block: a comparison of four needle insertion sites.

UNLABELLED: The site for needle insertion in femoral nerve block varies significantly among various descriptions of the technique. To determine the site with the highest likelihood of needle-femoral nerve contact, femoral nerve block was simulated in a human cadaver model (17 femoral triangles from 9 adult cadavers). Four 20-gauge 50-mm-long styletted catheters were inserted at four frequently suggested insertion sites for femoral nerve block. At the levels of inguinal ligament and the inguinal crease, the catheters were inserted adjacent to the lateral border of the femoral artery and 2 cm lateral to the femoral artery. During anatomical dissection, we studied the number of catheter-nerve contacts for each of the four insertion sites, and relationships between the femoral nerve and other anatomical structures of relevance to femoral nerve block. Insertion of the needle at the level of the inguinal crease, next to the lateral border of the femoral artery resulted in the highest frequency of needle-femoral nerve contacts (71%). Of note, the femoral nerve was significantly wider (14.0 vs 9.8 mm) and closer to the fascia lata (6.8 vs 26.4 mm) at the inguinal crease than at the inguinal ligament level. We conclude that needle insertion at the inguinal crease level immediately adjacent to the femoral artery produced the highest rate of needle-femoral nerve contacts. The main factors influencing this result include the greater width of the femoral nerve and the more predictable femoral artery-femoral nerve relationship at the inguinal crease level, compared with the inguinal ligament level. IMPLICATIONS: Insertion of a needle at the inguinal crease level and immediately adjacent to the lateral border of the femoral artery results in a high rate of needle-femoral nerve contact.

Adult↗

Gum elastic bougie-guided insertion of the ProSeal laryngeal mask airway is superior to the digital and introducer tool techniques.

BACKGROUND: The authors compare three techniques for insertion of the ProSeal laryngeal mask airway. METHODS: Two hundred forty healthy patients aged 18-80 yr were randomly allocated for ProSeal laryngeal mask airway insertion using the digital, introducer tool (IT), or gum elastic bougie (GEB)-guided techniques. The digital and IT techniques were performed according to the manufacturer's instructions. The GEB-guided technique involved priming the drain tube with the GEB, placing the GEB in the esophagus under direct vision, and inserting the ProSeal laryngeal mask airway using the digital technique with the GEB as a guide. Failed insertion was defined by any of the following criteria: (1) failed pharyngeal placement; (2) malposition (air leaks, negative tap test results, or failed gastric tube insertion if pharyngeal placement was successful); and (3) ineffective ventilation (maximum expired tidal volume < 8 ml/kg or end-tidal carbon dioxide > 45 mmHg if correctly positioned). Any visible or occult blood was noted. Sore throat, dysphonia, and dysphagia were assessed 18-24 h postoperatively. RESULTS: Insertion was more frequently successful with the GEB-guided technique at the first attempt (GEB, 100%; digital, 88%; IT, 84%; both P < 0.001), but success after three attempts was similar (GEB, 100%; digital, 99%; IT, 98%). The time taken to successful placement was similar among groups at the first attempt but was shorter for the GEB-technique after three attempts (GEB, 25 +/- 14 s; digital, 33 +/- 19 s; IT, 37 +/- 25 s; both: P < 0.003). There were no differences in the frequency of visible blood, but occult blood occurred less frequently with the GEB-guided technique (GEB, 12%; digital, 29%; IT, 31%; both: P < 0.02) but was similar among techniques if insertion was successful at the first attempt. There were no differences in postoperative airway morbidity. CONCLUSION The GEB-guided insertion technique is more frequently successful than the digital or IT techniques. The authors suggest that the GEB-guided technique may be a useful backup technique for when the digital and IT techniques fail.

Adolescent↗

Assessment of backside wear from the analysis of 55 retrieved tibial inserts.

Fifty-five retrieved tibial inserts with four different locking mechanisms were evaluated for evidence for polyethylene wear between the inferior surface of the tibial insert and metal tray. This type of wear will be referred to as backside wear. Backside wear was assessed by evaluating manufacturer's stamped markings on the inferior polyethylene surface. Because these markings are embossed into the polyethylene surface, they were used as indicators of backside wear. Decreases in the depths of markings indicated that backside wear was clearly evident, regardless of design, in 24 (44%) of the inserts. In eight of these 24 inserts, the manufacturer's stamped markings were removed completely. The amount of polyethylene wear was as a high as 591 mg from the inferior surface. This corresponds to a polyethylene wear rate from the backside of the tibial insert of greater than 100 mg per year, which is two to four times higher than wear rates associated with total hip replacements. The current work provides direct evidence of backside wear in all four tibial insert designs. Backside wear of tibial inserts can be a significant contributor to polyethylene wear in total knee arthroplasty. Close attention should be given to the fixation of tibial inserts to metal trays by manufacturers and surgeons.

Arthroplasty, Replacement, Knee↗

The causes of insert backside wear in total knee arthroplasty.

Wear of the insert backside occurs ostensibly because of micromotion at the undersurface articulation that occurs with loading. When a cyclic axial load was applied to contemporary knee implants, all inserts tested moved 2 to 25 microm in the shear plane relative to the metal backing suggesting that undersurface motion may be inevitable. Variables that increase the forces between the insert and metal backing can worsen relative micromotion and backside wear. Forces at the undersurface articulation, created during physiologic loading, are influenced by insert type, articular design, and surgical technique. Increasing articular insert constraint can cause forces at the main articulation to be resisted and transferred to this and the other interfaces. Designs with a cam post mechanism that force rollback at a certain flexion angle create a significant force in this shear plane. Inserts with highly conforming articular geometries can have a similar affect if used to inhibit anteroposterior or mediolateral motion of the femur on the tibial insert. Component alignment and position, and ligament balance also may influence backside wear as suggested by the great variability of wear patterns seen on like insert retrievals and by kinematic differences observed in fluoroscopic studies of the same implant design. Only by understanding these potential causes of backside motion and subsequent wear, can backside wear be mitigated.

Arthroplasty, Replacement, Knee↗

The effect of insertion rates on fill length and adaptation of a thermoplasticized gutta-percha technique.

The purpose of this in vivo study was to determine the effect of the insertion rate of a thermoplasticized gutta-percha obturation technique (Thermafil Plus) on the quality of the root canal obturation. A calibrated practitioner obturated the tooth in each of three groups with insertion rates of 18, 6, and 3 mm/s, respectively. The quality of the obturation was assessed by measuring the length of fill and the replication of induced canal irregularities, consisting of dimples and grooves. All measurements were made on a single tooth, with a working length of 18 mm from the canal orifice, embedded in a split mold, with 20 measurements for each insertion rate. Measurements were made under a low-power microscope, with the images recorded on videotape. The length of fill decreased with decreasing the insertion rate, from a mean overextension of +0.88 mm (SD 1.12 mm) for the 18 mm/s rate to a mean underfill of -0.13 mm (SD 1.03 mm) for the 3 mm/s rate. A trend of decreasing fill with decreasing insertion rate was observed, with the difference between the insertion rates of 18 mm/s and 3 mm/s being statistically significant at p < 0.05 using a single comparison test. Replication of dimples decreased with decreasing the insertion rate from a mean reproduction of 99% for the 18 mm/s rate to a mean reproduction of 90% for the 3 mm/s rate. The replication of grooves also decreased with decreasing the insertion rate from a mean reproduction of 100% for the 18 mm/s rate to a mean reproduction of 88% for the 12 mm/s rate. Statistical analysis of the data for both the dimples and the grooves using a paired comparison post hoc Scheffe test found these differences are significant at p < 0.05.

Gutta-Percha↗

Effect of cutting flute design on cortical bone screw insertion torque and pullout strength.

OBJECTIVE: To determine the effect of the number and length of cutting flutes on the insertion torque and pullout strength for self-tapping 4.5-millimeter cortical bone screws. DESIGN: Screws were self-tapped in the diaphysis of human cadaver femurs. Each of the six screw types studied had different designs with varying cutting flute lengths and numbers. Bone mineral density, insertion torque, and pullout strength were measured. SETTING: The study was conducted at an experimental biomechanics laboratory associated with a university medical center. OUTCOME MEASUREMENTS: Insertion torque and pullout strength were normalized by the local bone mineral density. RESULTS: The mean normalized insertion torque of the design with four full-length cutting flutes was less than the design with three full-length flutes and the two designs with one-third length flutes (p < 0.05). The mean normalized pullout strength of the screw with four full-length flutes was significantly greater than that of all screws with fewer than three flutes (p < 0.05). CONCLUSIONS: Priorities for a cutting flute design should ideally include ease of screw insertion, minimal soft tissue irritation, and maximal screw holding power. Screws with more than two flutes were easier to insert and did not cause cortical damage during insertion. The screw with four full-length flutes showed a trend toward being the easiest to insert and having the greatest holding strength.

Biomechanical Phenomena↗

Bedside insertion of inferior vena cava filters in the intensive care unit.

BACKGROUND: Injured patients are at significant risk for venous thromboembolic complications. Multiple studies have reported a benefit of prophylactic inferior vena cava filter (IVCF) insertion in selected high-risk trauma patients. Often, these high-risk patients reside in the intensive care unit (ICU) and require mechanical ventilation, intracranial pressure monitoring, multiple intravenous infusions, and other invasive monitoring modalities. This puts these patients at risk for transport from the ICU. METHODS: We prospectively studied a series of consecutive patients undergoing bedside preinsertion contrast cavagram and IVCF insertion in the ICU. RESULTS: Thirty-two patients received IVCF. There were no failures to insert IVCF. One insertion-site hematoma occurred; however, there were no documented insertion-site deep venous thromboses. One patient death was unrelated to the IVCF, and one potential contrast-related acute renal failure occurred in an unstable patient who underwent IVCF insertion for a pulmonary embolus. CONCLUSION: Bedside IVCF insertion with a preinsertion cavagram is a percutaneous procedure that can be safely performed in the ICU. Bedside insertion of IVCF avoids the potential complications of transporting critically ill patients and may reduce costs.

Adolescent↗

Office-based insertion of pressure equalization tubes: the role of laser-assisted tympanic membrane fenestration.

OBJECTIVE: To describe the role of the hand-held otoscope combined with a flashscanner CO2 laser, OtoLAM (ESC/Sharplan, Yokneam, Israel), for pressure equalization tube (PET) insertion in an office setting. STUDY DESIGN: Prospective, multisite, clinical cohort trial (Institutional Review Board approved; informed consent) in the setting of pediatric otolaryngology outpatient departments at four tertiary care children's hospitals. METHODS: Selected for the study were 54 patients (96 ears), ages 6 months to 23 years, who met standard indications for PET insertion using cold-knife myringotomy and tube insertion under general anesthesia. PETs were indicated for recurrent otitis media, chronic otitis media with effusion, and eustachian tube dysfunction-all unresponsive to medical therapy. Topical anesthesia was achieved with iontophoresis (n = 1) or topical anesthesia: 8% tetracaine on an Otowick (Xomed Surgical Products, Jacksonville, FL, catalogue No. 400141) against the tympanic membrane for 45 to 180 minutes (n = 53). Laser-assisted tympanic membrane fenestration was performed with the OtoLAM set at single pulse, 2.0- to 2.6-mm spot size, and between 3 and 18 W. Insertion of grommets was accomplished using the otomicroscope and an "alligator" microforceps. Restraints with papoose were used in 79% of children with a mean age of 34.4 months (SD = 60.9 mo). Clinical, parent/patient, and physician satisfaction and comparative cost impact outcomes are described. RESULTS: All ears but three (3%) underwent successful placement of a PET. Pain was described as "absent" in 39%, "present but tolerable" in 30%, and "severe" in 30% of children at the time of procedure; 5 minutes after the procedure pain was described as "absent" in 75%, "present but tolerable" in 22%, and "severe" in 3%. Tube plugging (3 of 74 available ears; 4%) or persistent otorrhea (1 of 74 ears; 1.4%) occurred infrequently at the 1-month follow-up. Before PET insertion, hearing loss was noted in 66% of cases (mild, 38%; moderate, 22%; and severe, 6%). Mild hearing loss was noted in only 8% and moderate hearing loss in 2% of 47 (50%) of the ears at the 3-month follow-up. Ninety-two percent of parents were highly satisfied with the procedure in preference to PETs in the operating room under general anesthesia, and 97% preferred OtoLAM with PET insertion, rather than further courses of antibiotics; only one parent would rather have had the PET insertion under general anesthesia. Cost savings to health care organizations, particularly payers, and to parents are substantial (32%-48%) and warrant attention. Cost to the physician is manageable only if an appropriate approach to the third party payers results in a substantial increase in reimbursements. CONCLUSIONS: The data indicate excellent clinical effectiveness, reduced risk, and high parent and physician satisfaction. Strong incentives for physicians to use this technique are in all stakeholders' best interests. These incentives need to evolve as soon as possible for the more widespread acceptance of OtoLAM with PET insertion in an office setting for appropriately selected patients.

Adolescent↗

Appropriateness of endotracheal tube size and insertion depth in children undergoing air medical transport.

OBJECTIVES: Guidelines for pediatric endotracheal tube (ETT) size and insertion depth are important in the helicopter EMS (HEMS) setting, where intubated patients are frequently transported by a non-physician flight crew providing protocol-based care in an environment noted for limitations in clinical airway assessment. The objectives of this study were to characterize, in a HEMS pediatric population, the frequency of compliance with guideline-recommended ETT size and insertion depth, and to test for association between guideline noncompliance and subsequent receiving hospital adjustment of ETT size or insertion depth. DESIGN: This retrospective review analyzed 216 consecutive pediatric (age <14) scene and interfacility HEMS transports, of patients intubated before or during HEMS transport, by an urban two-helicopter HEMS service providing protocol-based care with a nurse/paramedic crew configuration. Patients were transported to one of three receiving academic pediatric referral centers. Pediatric Advanced Life Support (PALS) criteria for ETT size and insertion depth were used to assess guideline-appropriateness of pediatric ETTs. Receiving hospital records were reviewed to determine if post-transport ETT size or lipline adjustment were associated with guideline-appropriateness of size and lipline during HEMS transport. Univariate (chi-square and Fisher's exact) and multivariate (logistic regression) statistics were used to assess and control for the following covariates: intubator group (physician, flight crew, ground EMS), transport year, sex, age, transport type (scene versus interfacility), and receiving hospital. For all analyses, statistical significance was set at the 0.05 level. RESULTS: The initial ETT size was within 0.5 mm of guideline-recommended sizes in 178 (83.6%) of the 213 patients for whom this data were available. Inappropriate sized ETTs were nearly always (32 of 35, 91.4%) too small. Compared to initial ETTs placed by ground EMS personnel, initial ETTs placed by flight crew or physicians were more likely to be appropriate as defined by guidelines (P = .008 and .032, respectively). Receiving hospitals changed the ETT size in 18 (8.3% of 216) cases. Receiving hospital ETT size change was more likely with later transport year (P = .018) and less likely in patients over 2 years of age (P = .03); there was no significant association between receiving hospital ETT size change and intubator group (P > .22) or guideline-appropriateness of ETT size (P = 0.94). The initial ETT insertion depth was within 1 cm of the guideline-recommended lipline in 86 (43.2%) of the 199 patients for whom this data were available. Inappropriate liplines were almost always (109 of 113, 96.5%) too deep. Compared to initial ETT liplines determined by ground EMS personnel, initial liplines determined by flight crew (P = .007), but not physician (P = .47) were more likely to be appropriate as defined by guidelines. Receiving hospitals changed the ETT insertion depth in 72 (33.3% of 216) cases. Receiving hospital lipline change was more likely (P = .03) in patients older than 2 years of age, but was not associated with intubator group (P = .75) or lipline guideline-appropriateness (P = .35). CONCLUSIONS: As judged by frequently used guidelines, pediatric ETTs are often too small and commonly inserted too deep. However, this retrospective study, limited by lack of clinical correlation for ETT size and insertion depth, failed to find an association between lack of ETT size or lipline guideline compliance and subsequent ETT adjustment at receiving pediatric centers. This study's findings, which should be confirmed with prospective investigation, cast doubt upon the utility of pediatric ETT size/lipline guidelines as strict clinical or quality assurance tools for use in pediatric airway management.

Age Factors↗

Pedicle screw pullout strength. Correlation with insertional torque.

This study was designed to correlate several parameters regarding pedicle screw bone/metal interface strength. The insertional torque measured during tapping and placement of pedicle screws was correlated with the bone mineral density of the vertebral body, the dimensions of the pedicle, the method of preparation of the pedicle, and the amount of load and number of cycles to failure of the bone/metal interface. Thirty human cadaveric lumbar vertebrae were instrumented with 6.5-mm pedicle screws. The maximum torque achieved during insertion was digitally recorded. A cyclic pedicle screw pullout test was performed. A linear correlation existed between both the insertional torque when tapping or when inserting a screw and the number of cycles to ultimate pedicle screw pullout. An inverse linear relationship was found between the pedicle width and cycles to failure. There was no linear correlation found when comparing the number of cycles to failure to bone mineral density. These findings suggest that insertional torque is a good predictor of bone-metal interface failure. Bone mineral density of the vertebral body was less effective as a predictor of failure. Smaller pedicle width correlated with increased insertional torque and cycles to failure. This may explain why patients with osteoporosis on radiography may still obtain stable fixation with pedicle screws. Other factors, such as pedicle dimensions and shape, affect screw purchase as much as vertebral body bone density. Insertional torque less than 4.0 inch-pounds led to early pedicle screw pullout. This study forms the basis for the authors' clinical use of an instrumented torque screwdriver to measure insertional torque in the operating room.

Aged↗

Insertional torque and pull-out strengths of conical and cylindrical pedicle screws in cadaveric bone.

STUDY DESIGN: Insertion torque and pull-out strengths of conical and cylindrical pedicle screws were compared in human cadaveric vertebral bodies. OBJECTIVES: To compare the performance of the conical design with the cylindrical design, and to determine whether insertional torque correlates with pull-out strength. SUMMARY OF BACKGROUND DATA: A tapered pedicle screw design may lessen the likelihood of implant failure. Its effect on thread purchase is not known. Previous studies of cylindrical designs on the relation between insertion torque and pull-out strength have been conducted in bovine and synthetic bone. METHODS: Seventy-eight pedicles were assigned randomly to one of the following pedicle screw: Texas Scottish Rite Hospital (Sofamor-Danek, Memphis, TN), Steffee VSP (Acromed, Cleveland, OH), Diapason (Dimso, Paris, France), AO Schanz (Synthes, Paoli, PA), or Synthes USS (Synthes, Paoli, PA). Pedicle screws were inserted with a torque screwdriver. Each screw was extracted axially from the pedicle at a rate of 1.0 mm/sec until failure using an MTS machine (Bionix 858, Minneapolis, MN). Force data were recorded. RESULTS: The conical design had the highest insertion torque. There were no significant differences in pull-out between any of the screw types. Correlation between insertional torque and pull-out strength was statistically significant only with the Texas Scottish Rite Hospital and Steffee VSP in L4 and AO Schanz in L5. CONCLUSIONS: A conical screw profile increases insertion torque, although insertional torque is not a reliable predictor of pull-out strength in cadaveric bone. Screw profile (with similar dimensions) has little effect on straight axial pull-out strengths in cadaveric bone.

Aged↗

A comparison of various angles of halo pin insertion in an immature skull model.

STUDY DESIGN: A basic science biomechanical study involving an animal model. OBJECTIVES: To evaluate the effect of varying angles of halo pin insertion on the force generated at the pin-bone interface, and thereby the stability of the halo pin-bone interaction during insertion. BACKGROUND DATA: Because of variations in the shape and size of the pediatric skull, halo pins often are inserted at various angles rather than perpendicular to the skull. Concern exists that the high complication rate associated with pediatric halo use may result in part from less than ideal structural properties at the halo pin-bone interface. METHODS: The authors used a fetal calf skull model to simulate the thickness and structural properties of the pediatric skull. Halo pins were inserted at angles of 0 degree (perpendicular), 10 degrees, 15 degrees, and 30 degrees into skull segments via a halo ring. Load generated at the pin-bone interface was measured using a modified mechanical testing device. Twenty trials were conducted per angle, with the endpoint being specimen failure, pin penetration, or maximum load. RESULTS: Mean maximum loads per unit thickness were 82.15 +/- 7.54 N/mm at 0 degree, 68.80 +/- 4.79 N/mm at 10 degrees, 51.49 +/- 5.08 N/mm at 15 degrees, and 42.38 +/- 3.51 N/mm at 30 degrees, There was a significant difference between perpendicular insertion (0 degree) and 15 degrees angles of insertion. There was also a significant difference between the 10 degrees and 30 degrees angles of insertion. CONCLUSIONS: Perpendicular halo pin insertion in an immature skull model was shown to result in increased load at the pin-bone interface. This improved structural behavior may help to reduce the incidence of complications of halo application in children.

Animals↗

Measurement of air emboli during central venous access: do "protective" sheaths or insertion techniques matter?

PURPOSE: Clinically significant air emboli during placement of central venous catheters are rare yet potentially fatal events. An in vitro model was developed to measure the volume of air emboli during catheter placement under a variety of experimental conditions. MATERIALS AND METHODS: The volume of air emboli during catheter insertion with use of a standard sheath was measured using "open," "finger-pinch," and "hemostat" techniques. Corresponding experiments were performed with newly designed protective sheaths. Additional experiments evaluated the air emboli related to specific steps of catheter insertion, a sheathless insertion technique, and two commonly used dialysis catheters. RESULTS: Under physiologic conditions, the volumes of air emboli were 9.1 mL +/- 3.6, 8.6 mL +/- 3.9, and 10.0 mL +/- 4.1 for dialysis catheter insertions with open, finger-pinch, and hemostat techniques, respectively. In the open and closed positions, the sliding-valve protective sheath yielded 5.8 mL +/- 2.3 and 4.4 mL +/- 2.3 of air emboli, respectively, and the slide-clamp protective sheath yielded 5.6 mL +/- 2.0 and 5.4 mL +/- 2.1 of air emboli, respectively. The standard sheath demonstrated air emboli volumes of 14.4 mL +/- 12.8, 17.3 mL +/- 3.9, and 32.3 mL +/- 10.9 during cumulative steps of catheter insertion. The sliding-valve and slide-clamp protective sheaths yielded air emboli measuring 4.4 mL +/- 2.0, 10.9 mL +/- 5.2, and 8.6 mL +/- 1.5, and 4.4 mL +/- 1.8, 10.9 mL +/- 1.4, and 9.4 mL +/- 4.0, respectively. The sheathless insertion technique resulted in air emboli measuring 12.2 mL +/- 5.4. Split-tip and step-tip catheters resulted in air emboli volumes of 16.1 mL +/- 4.5 and 15.3 mL +/- 7.6, respectively, in the open position and 11.3 mL +/- 3.1 and 12.9 mL +/- 5.0, respectively, in the closed position. CONCLUSIONS: The newly designed protective sheaths result in smaller volumes of air emboli compared with standard sheaths in most situations evaluated. There was no significant difference in the volume of air emboli with use of protective clinical maneuvers. In some cases, the volume of the air emboli continued to increase during catheter insertion and sheath removal. There was no statistically significant difference between the use of protective sheaths and the use of the sheathless insertion technique.

Catheterization, Central Venous↗

Distribution and genomic location of active insertion sequences in the Burkholderia cepacia complex.

This study aimed firstly to establish the distribution and copy number within the Burkholderia cepacia complex of three insertion sequences (IS402, IS407 and IS1416) that possess the ability to activate transcription and hence influence gene expression. A second aim was to map the genomic insertion sites of one of the active insertion sequences (IS407) to establish putative links between insertion site and downstream gene activation. The resulting data revealed that all three insertion sequences were present in one-third of the 66 isolates tested. The three insertion sequences were prevalent across the nine B. cepacia complex species, although IS402 was absent from the 16 Burkholderia anthina strains tested and IS407 was absent from all 10 Burkholderia pyrrocinia strains. IS407 copies from six strains (two Burkholderia cenocepacia strains and one strain each of Burkholderia multivorans, Burkholderia stabilis, Burkholderia vietnamiensis and B. anthina) were mapped to the genome using hemi-nested inverse PCR. Insertions were found upstream of genes with wide-ranging functions. This study suggests that the abundance and distribution of these active insertion sequences is likely to affect genomic plasticity, and potentially gene transcription and pathogenicity.

Burkholderia Infections↗

Variants of Peach latent mosaic viroid inducing peach calico: uneven distribution in infected plants and requirements of the insertion containing the pathogenicity determinant.

Previous characterization of Peach latent mosaic viroid (PLMVd) variants from a single peach calico (PC) isolate showed that PC symptoms are induced by variants with a 12-13 nt insertion at a specific position and folding into a hairpin with a U-rich loop. Here, this study was extended to two other PC isolates. PLMVd variants with insertions similar to those reported previously (type 1), predominated in one isolate (PC-P2). The second (PC-P1), in addition to these variants, contained others with insertions in the same position and of the same size, but with the hairpin capped by a GA-rich loop (type 2). When symptomatic and non-symptomatic tissues from both isolates were used to inoculate GF-305 peach seedlings, they reproduced the phenotype of the inoculum source, indicating that variants differing in pathogenicity are unevenly distributed within single plants. Moreover, characterization of the progeny from inoculations with the PC-P1 source showed that variants with insertions of type 1 and 2 were predominant in the symptomatic and non-symptomatic seedlings, respectively, confirming the association between PC and variants with type 1 but not type 2 insertions. Inoculations with dimeric in vitro transcripts from PLMVd variants with type 1, type 2 and with a chimeric insertion showed that the variant with type 2 insertion was latent and established that the U-rich capping loop has a major role in PC, although the adjacent stem may also have some influence. Insertions can be acquired and lost during infection, suggesting that latent variants can evolve into pathogenic variants and vice versa.

Genetic Variation↗