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PubMed1983

Insertional mutagenesis of the cauliflower mosaic virus genome.

A series of small insertions has been introduced into the various translational reading frames of the DNA of a "severe" strain of cauliflower mosaic virus (CaMV). A selectable gene (the kanamycin phosphotransferase gene of Tn903), flanked by a series of symmetrically arranged cloning sites taken from M13mp7, was used to prepare the site-specific mutants. In-phase insertions of 12 or 30 bp, which introduced unique SalI sites into reading regions I, III, IV, V and into the amino-proximal portion of region VI, destroyed infectivity. Insertions in the amino-distal portion of region VI, in the large intergenic region, and in region II retained infectivity. The amino-distal insertions in region VI reduced the severity of symptoms in plants. The insertion in region II destroyed aphid transmissibility. Longer DNA segments when inserted into region II or into the amino-distal portion of region VI destroyed infectivity, but similar insertions in the intergenic region were without effect on virus infection or development.

Animals↗
PubMed1988

Insertional restoration of beta-galactosidase alpha-complementation (white-to-blue colony screening) facilitates assembly of synthetic genes.

The assembly of synthetic genes from oligodeoxynucleotides can be an inefficient process. Upon ligation of a synthetic assembly into a plasmid vector and transformation of an Escherichia coli host, it is often found that only a minor fraction of the putative recombinant plasmids contains synthetic sequences. Moreover, the synthetic sequences cloned are often altered versions of those originally designed. We have designed a biological test to detect those plasmids that contain synthetic sequences of the proper length, termini and reading frame. The test is the reversal of the beta-galactosidase alpha-complementation (blue-to-white) test used to detect the insertion of DNA segments into the polylinker sequences of the phage M13 mp, plasmids pUC, and related vectors. We begin with a modified vector defective in alpha-complementation and use insertion of the synthetic DNA segment to restore alpha-complementation. The alpha-complementation activity of the original vector (e.g., pUC18) was first abolished by a frameshift or DNA insertion within the polylinker sequence of the lacZ' gene segment. The alpha-complementation was then restored by insertion of the synthetic DNA sequence between the cohesive ends generated by digestion of two polylinker restriction sites. Formation of blue colonies requires the insertion of a DNA segment of appropriate length and termini to reconstruct the lacZ' open reading frame and thus is much more selective than the usual insertional inactivation strategy. We show that this 'insertional restoration' screening method markedly enhances the proper assembly of synthetic genes and describe manipulations to readily and reliably frameshift various polylinker sequences.

Amino Acid Sequence↗
PubMed1995

Vagal response to feeding tube insertion in preterm infants: has the key been found?

We prospectively studied the vagal response to feeding tube insertion in eight healthy preterm infants, on three occasions in each infant during the first three weeks of life. Heart rate, oxygen saturation, respiration and cerebral blood flow velocities were assessed before, during and immediately after insertion of an orogastric feeding tube. The whole procedure was recorded on video. The duration and quality of tube insertion and the behaviour of the infant were evaluated from the recordings. Twenty-one measurements in eight infants were evaluated. The heart-rate decrease observed immediately after tube insertion correlated significantly with the duration of tube insertion, the quicker the manipulation, the greater the heart-rate decrease (P = 0.000). The maximal decrease of oxygen saturation after tube insertion correlated with the degree of heart rate deceleration (P = 0.009). Significant alterations of the flow velocities were observed only when the heart rate fell below 80 beats/min. We speculate that such episodes of bradycardia can be avoided by carefully inserting the feeding tube over a period of at least 15 s.

Blood Flow Velocity↗
PubMed2004

Emergency medicine-trained physicians are proficient in the insertion of transvenous pacemakers.

STUDY OBJECTIVE: The insertion and management of a temporary transvenous pacemaker can be a lifesaving procedure in the emergency department setting. We compare the success and complication rates associated with temporary transvenous pacemaker insertion between physicians trained in either emergency medicine or cardiology. METHODS: This was a retrospective medical record review of all patients with a billing code for temporary transvenous pacemaker insertion at our institution between July 1999 and December 2002. Patients were excluded if the temporary transvenous pacemaker was not supervised by an attending physician or was placed under fluoroscopy or if the indication for pacing was asystole. Cases were reviewed by 2 physicians certified in emergency medicine and categorized by the specialty training of the attending physician providing direct supervision. RESULTS: During the review period, 10 emergency medicine faculty and 8 cardiologists directly supervised 141 of the 158 temporary transvenous pacemaker insertions in 154 patients. Twenty-four were placed for asystole, 4 were placed under fluoroscopy, and 13 were placed without direct attending supervision, leaving a total of 117 cases (30 emergency medicine, 87 cardiology) for review. The procedure was successful 97% (95% confidence interval [CI] 90% to 100%) of the time for emergency medicine faculty and 95% (95% CI 91% to 100%) of the time for cardiology faculty. Complications were seen in 23% (95% CI 7% to 39%) of the temporary transvenous pacemakers inserted by emergency medicine attending physicians and 20% (95% CI 11% to 28%) of the temporary transvenous pacemakers inserted by cardiologists. There was no statistical difference in errors between the specialties, and no complication resulted in death or prolonged disability. CONCLUSION: Physicians trained in emergency medicine perform temporary transvenous pacemaker insertions in the acute care setting with a proficiency similar to that of their counterparts in cardiology.

Adult↗
PubMed2005

Reconciling arthroscopic and anatomic morphology of the humeral insertion of the inferior glenohumeral ligament.

PURPOSE: To clarify the morphology of the humeral insertion of the inferior glenohumeral ligament (IGHL). TYPE OF STUDY: Cadaveric and arthroscopic anatomic analysis. METHODS: The morphology of the humeral insertion was studied in 200 nonembalmed cadavers through open dissection (100 specimens), by arthroscopy (50 specimens), or both (50 specimens). In addition, the morphology was studied in 100 living subjects with stable shoulders undergoing shoulder arthroscopy. RESULTS: On the humeral side, the insertion of the inferior capsular fibers is usually in the form of a V, the point of which is covered by the tendon of the latissimus dorsi. When viewed intra-articularly, the inferior insertion usually gives a collar-like impression because the capsular recess is filled with frenula capsulae. CONCLUSIONS: Our description corresponds with that found in the classic literature. Our results are, however, in contrast with those of others who have observed about 50% of V-shaped insertions. This difference may be explained by the method of observation and by the small numbers of specimens studied. The form of the humeral insertion of the IGHL is linked to the formation of a supporting hammock that can accommodate the humeral head during movement as described by several authors. CLINICAL RELEVANCE: We believe that the difference between arthroscopic and anatomic observation of the humeral insertion may have 2 major clinical implications. An observed tear of the frenula capsulae may not necessarily represent a humeral avulsion of the glenohumeral ligaments (HAGL). In the case of a HAGL, the capsule may have to be reattached in its V-form to adequately retension the inferior capsule.

Aged↗
PubMed2006

The subscapularis footprint: an anatomic study of the subscapularis tendon insertion.

PURPOSE: Arthroscopic repair of the subscapularis tendon has become more prevalent in recent years. Tears of the subscapularis insertion can be measured arthroscopically when the size of the average subscapularis tendon insertion is known. This anatomic study was performed to measure the dimensions and describe the anatomy of the subscapularis footprint. METHODS: A total of 6 male and 6 female shoulders were dissected down to the insertion of the subscapularis tendon. The insertion was demarcated, the tendon was detached, and the dimensions of the insertion site were measured. RESULTS: The footprint is the insertion of the subscapularis tendon onto the lesser tubercle. The shape of the footprint was characterized as resembling a human ear. The insertion is broad proximally and tapered distally and has a straight medial border that is almost parallel to the longitudinal axis of the humerus. The total average cephalocaudal height of the footprint was 25.8 mm (+/-3.2 mm). The total average width was 18.1 mm (+/-1.6 mm). Average male cephalocaudal height was 26.7 mm (range, 22 to 32 mm), and width was 18.3 mm (range, 16 to 21 mm). Average female cephalocaudal height was 24.8 mm (range, 22 to 29 mm), and width was 17.8 mm (range, 15 to 19 mm). Although the male footprint was slightly larger than the female footprint, differences were not statistically significant (P = .18 and .31 for height and width, respectively). CONCLUSIONS: An anatomic study was performed to determine the size of the footprint of the subscapularis tendon. We found that the average cephalocaudal height of the footprint was 25.8 mm, and the average width was 18.1 mm. CLINICAL RELEVANCE: Subscapularis tears are now more frequently addressed arthroscopically. This anatomic study was performed to characterize the anatomy of the subscapularis footprint so that the surgeon can achieve a more anatomic repair.

Aged↗
PubMed2006

Insertion selectivity of antimicrobial peptide protegrin-1 into lipid monolayers: effect of head group electrostatics and tail group packing.

The ability to selectively target the harmful microbial membrane over that of the host cell is one of the most important characteristics of the antimicrobial peptides (AMPs). This selectivity strongly depends on the chemical and structural properties of the lipids that make up the cell membrane. A systematic study of the initial membrane selectivity of protegrin-1 (PG-1), a beta-sheet AMP, was performed using Langmuir monolayers. Constant pressure insertion assay was used to quantify the amount of PG-1 insertion and fluorescence microscopy was employed to observe the effect of PG-1 on lipid ordering. Charge and packing properties of the monolayer were altered by using lipids with different head groups, substituting saturated with unsaturated lipid tail group(s) and incorporating spacer molecules. PG-1 inserted most readily into anionic films composed of phosphatidylglycerol (PG) and lipid A, consistent with its high selectivity for microbial membranes. It also discriminated between zwitteranionic phospholipids, inserting more readily into phosphatidylcholine (PC) monolayers than those composed of phosphatidylethanolamine, potentially explaining why PG-1 is hemolytic for PC-rich human erythrocytes and not for the PE-rich erythrocytes of ruminants. Increased packing density of the monolayer by increased surface pressure, increased tail group saturation or incorporation of dihydrocholesterol diminishes the insertion of PG-1. Fluorescence microscopy shows that lipid packing is disordered upon PG-1 insertion. However, the presence of PG-1 can still affect lipid morphology even with no observed PG-1 insertion. These results show the important role that lipid composition of the cell membrane plays in the activity of AMPs.

Anti-Infective Agents↗
PubMed2006

Comparison of efficacy and complications of IUD insertion in immediate postplacental/early postpartum period with interval period: 1 year follow-up.

OBJECTIVE: This study aimed to compare immediate postplacental (IPP) and early postpartum (EP) intrauterine device (IUD) insertions with interval (INT) IUD insertions with respect to efficacy and complications. METHODS: The study group consisted of 268 women in whom the following TCu 380A IUD insertions were performed: 84 IPP (less than 10 min), 46 EP (10 min to 72 h) and 138 INT (more than 6 weeks). The women were followed up 8 weeks, 6 months and 12 months after insertion. Complications and pregnancies encountered at the end of 1 year following IPP, EP and INT insertions were compared. The chi-square test and Fisher's Exact Test were used for the evaluation of the data. RESULTS: Complications developed in 40.4% of the women in the IPP group, in 74.4% of the women in the EP group and in 19.2% of the women in the INT group (p<.001). Although no statistically significant difference was found between the groups for uterine perforation and infection (p>.001), there was a statistically significant difference between the groups in the incidence of complete and partial expulsion according to the time of IUD insertion. The overall cumulative pregnancy rate and frequency of pregnancy were found to be higher (p>.05 for both), which are both insignificant for the EP group (2 of 43 women), as compared with the INT (4 of 130 women) and IPP groups (2 of 84 women), and pregnancy rates at 1 year for all groups was 3.1% (8 of 257 women). CONCLUSION: IPP and EP insertion of the TCu 380A IUD is an effective and convenient procedure, and expulsion rates in these groups are higher than in the INT group. Further studies are necessary to determine the cause of the higher expulsion rates and to find ways to reduce such rates.

Adolescent↗
PubMed2005

Dexamethasone treatment reduces astroglia responses to inserted neuroprosthetic devices in rat neocortex.

Microfabricated neural prosthetic devices hold great potential for increasing knowledge of brain function and treating patients with lost CNS function. Time-dependent loss of brain-device communication limits long-term use of these devices. Lost CNS function is associated with reactive responses that produce an encapsulating cellular sheath. Since early reactive responses may be associated with injuries produced at the time of device insertion, for example, vascular damage and disruption of the blood-brain barrier, we tested the effectiveness of the synthetic glucocorticoid, dexamethasone, in controlling insertion- and device-associated reactive responses. Dexamethasone (200 microg/kg) was administered as subcutaneous injections for 1 or 6 days beginning on the day of device insertion. Single shank microfabricated silicon devices were inserted into pre-motor cortex of adult rats. Reactive responses were assessed by immunohistochemistry for glial fibrillary acidic protein (astrocytes), CD11b (microglia), and laminin that labeled extracellular protein deposited around the insertion site and in association with vascular elements. Data were collected by confocal microscopy imaging of 100-microm-thick tissue slices. Reactive responses in vehicle control animals were similar to non-injected control animals. Dexamethasone treatment profoundly effected early and sustained reactive responses observed 1 and 6 weeks following device insertion, respectively. Dexamethasone treatment greatly attenuated astroglia responses, while microglia and vascular responses appeared to be increased. The 6-day treatment was more effective than the single injection regime. These results suggest that anti-inflammatory agents can be used to control reactive responses around inserted neural prosthetic devices and may provide a means to insure their long-term function.

Animals↗
PubMed2005

Short peripheral venous catheters: effect of evidence-based guidelines on insertion, maintenance and outcomes in a university hospital.

This study was designed to evaluate the impact of hospital-wide guidelines for short peripheral venous catheters (SPVC) insertion on the frequency of local catheter-related complications. In a 1051-bed Parisian university hospital, two observational, point prevalence surveys were undertaken in 1996 and in 1998, separated by implementation of written guidelines for SPVC insertion. The outcomes of SPVC insertion were defined as the presence or absence of local catheter-related complications (erythema, purulence around the insertion point, tenderness or induration along the cannulated vein). The proportion of polyurethane catheter materials used (56% vs. 81%, P<0.001), correct and sterile fixation (80% vs. 92%, P<0.05), non-movable catheters (92% vs. 98%, P=0.03) and insertion record (58% vs. 76%, P<0.01) increased between 1996 and 1998. The frequency of local catheter-related complications decreased (15% vs. 4%, P<0.01). Age >73 years [odds ratio (OR) 6.0, 95% confidence interval (CI) 1.28-28.05] was positively associated with local catheter-related complications, whereas duration of insertion (until 72 h) (OR 0.29, 95% CI 0.09-0.89) and the implementation of guidelines (OR 0.26, 95% CI 0.09-0.67) were negatively associated with local catheter-related complications. The implementation of guidelines was independently negatively associated with local catheter-related complications (OR 0.31, 95% CI 0.09-0.97). The results suggest that hospital guidelines for SPVC insertion can improve catheter care and significantly reduce local catheter-related complications.

Adolescent↗
PubMed2005

The N-terminal cleavable extension of plant carrier proteins is responsible for efficient insertion into the inner mitochondrial membrane.

A subset of mitochondrial carrier proteins from plants contain a cleavable N-terminal extension. We have used a reconstituted protein import assay system into intermembrane space-depleted mitochondria to study the role of the cleavable extension in the carrier import pathway. Insertion of carrier proteins into the inner membrane can be stimulated by the addition of a soluble intermembrane space fraction isolated from plant mitochondria. Greater stimulation of import of the adenine nucleotide carrier (ANT) and phosphate carrier (Pic), which contain N-terminal cleavable extensions, was observed compared to the import of the oxoglutarate malate carrier (OMT), which does not contain a cleavable extension. Removal of the N-terminal cleavable extension from ANT and Pic resulted in loss of stimulation of insertion into the inner membrane. Conversely, addition of the N-terminal extension from ANT or Pic to OMT resulted in significantly enhanced insertion into the inner membrane. The polytopic inner membrane proteins TIM17 and TIM23 that are imported via the carrier import pathway contain no cleavable extension, displayed high-level stimulation of insertion into the inner membrane by addition of the intermembrane space fraction. Addition of the N-terminal cleavable extension from carrier proteins to TIM23 enhanced insertion of TIM23 into the inner membrane even in the absence of the soluble intermembrane space fraction. Together, these results demonstrate that the cleavable N-terminal extensions present on carrier proteins from plants are required for efficient insertion into the inner mitochondrial membrane, and that they can stimulate insertion of any carrier-like protein into the inner membrane.

Amino Acid Sequence↗
PubMed2006

Spatiotemporal pH dynamics following insertion of neural microelectrode arrays.

Insertion trauma is a critical issue when assessing intracortical electrophysiological and neurochemical recordings. Previous reports document a wide variety of insertion techniques with speeds ranging from 10 microm/s to 10 m/s. We hypothesize that insertion speed has an effect on tissue trauma induced by implantation of a neural probe. In order to monitor the neural interface during and after probe insertion, we have developed a silicon-substrate array with hydrous iridium oxide microelectrodes for potentiometric recording of extracellular pH (pH(e)), a measure of brain homeostasis. Microelectrode sites were sensitive to pH in the super-Nernstian range (-85.9 mV/pH unit) and selective over other analytes including ascorbic acid, Na(+), K(+), Ca(2+), and Mg(2+). Following insertion, arrays recorded either triphasic or biphasic pH(e) responses, with a greater degree of prolonged acidosis for insertions at 50 microm/s than at 0.5 mm/s or 1.0 mm/s (p<0.05). Spatiotemporal analysis of the recordings also revealed micro-scale variability in the pH(e) response along the array, even when using the same insertion technique. Implants with more intense acidosis were often associated histologically with blood along the probe tract. The potentiometric microsensor array has implications not only as a useful tool to measure extracellular pH, but also as a feedback tool for delivery of pharmacological agents to treat surgical brain trauma.

Acidosis↗
PubMed2004

Quality of permanent prostate implants using automated delivery with seedSelectron versus manual insertion of RAPID Strands.

BACKGROUND AND PURPOSE: To compare the quality of manually inserted RAPID Strand implants with automatically inserted selectSeed implants using volumetric and dosimetric parameters. PATIENTS AND METHODS: Patients with T1 to T2 prostate carcinoma were treated with brachytherapy. The (125)I seeds were implanted in the prostate in three different ways: manual insertion of RAPID Strands (R); insertion of selectSeeds using the seedSelectron (S); a combination of both techniques: manual insertion of RAPID Strands in the left half of the prostate and insertion of selectSeeds with the seedSelectron in the right half of the prostate (RS). The comparison is based on implant and target specific parameters. The implant specific parameters, V(100), homogeneity index (HI), and natural dose ratio (NDR), were determined at the time of implantation and four weeks later. MR images taken four weeks after the implantation were used for the calculation of the target specific parameters: D(90), HI, external index (EI), and conformation number (CN). RESULTS: We found no significant difference between the groups of implants (R, S, RS) for the implant specific parameters V(100), HI, and NDR at t(0) and neither at t(4w). For each group, the V(100) values decreased significantly with time between t(0) and t(4w). The target specific parameters D(90), HI, EI and CN were not significantly different between the groups. For the group of patients with both RAPID Strands and selectSeeds, we found a significant difference in D(90) between both halves of the prostate. CONCLUSIONS: The dosimetry parameters of a newly introduced implant technique using an automatic seed afterloader were not significantly different from the parameters of a manual insertion technique using RAPID Strands. Since either technique has its advantages and disadvantages regarding seed migration, physics quality assurance, efficiency, logistics, and ease of use, it was decided to use both techniques and to continue evaluations.

Brachytherapy↗
PubMed2004

Influence of a 12-nt insertion present in the 3' untranslated region of classical swine fever virus HCLV strain genome on RNA synthesis.

The function of a 12-nt insertion 'CTTTTTTCTTTT' in the 3' untranslated region (3'UTR) of the HCLV strain, a vaccine strain derived from the Shimen strain of classical swine fever virus (CSFV), was examined in vitro. RNA synthesis increased when the 12-nt insertion was deleted from the 3'UTR of the HCLV strain. RNA synthesis also decreased when the 12-nt insertion was introduced into the 3'UTR of the CSFV Shimen, virulent strain. Therefore, the 12-nt insertion present in the 3'UTR of the HCLV strain may be a marker for avirulence. It was also found that the predicted secondary structure of the 3'UTR of the HCLV strain was more stable when the 12-nt insertion was deleted, and that the 3'UTR of Shimen strain was more unstable when the 12-nt insertion was introduced. Replacement of the 12-nt insertion 'CTTTTTTCTTTT' by another 12-nt fragment 'ATTATTATTTAT' in the 3'UTR of HCLV strain facilitated RNA synthesis and stabilized the predicted secondary structure, which was also observed in the 3'UTR of the Shimen strain.

3' Untranslated Regions↗
PubMed2004

Intracellular assembly and sorting of intermediate filament proteins: role of the 42 amino acid lamin insert.

Nuclear and cytoplasmic intermediate filament (IF) proteins segregate into two independent cellular networks by mechanisms that are poorly understood. We examined the role of a 42 amino acid (aa) insert unique to vertebrate lamin rod domains in the coassembly of nuclear and cytoplasmic IF proteins by overexpressing chimeric IF proteins in human SW13+ and SW13- cells, which contain and lack endogenous cytoplasmic IF proteins, respectively. The chimeric IF proteins consisted of the rod domain of human nuclear lamin A/C protein fused to the amino and carboxyl-terminal domains of the mouse neurofilament light subunit (NF-L), which contained or lacked the 42 aa insert. Immunofluorescence microscopy was used to follow assembly and targeting of the proteins in cells. Chimeric proteins that lacked the 42 aa insert colocalized with vimentin, whereas those that contained the 42 aa insert did not. When overexpressed in SW13- cells, chimeric proteins containing the 42 aa formed very short or broken cytoplasmic filaments, whereas chimeric proteins that lacked the insert assembled efficiently into long, stable cytoplasmic filaments. To examine the roles of other structural motifs in intracellular targeting, we added two additional sequences to the chimera, a nuclear localization signal (NLS) and a CAAX motif, which are found in nuclear IF proteins. Addition of an NLS alone or an NLS in combination with the CAAX motif to the chimera with the 42 aa insert resulted in cagelike filament that assembled close to the nuclear envelope and nuclear lamina-like targeting, respectively. Our results suggest that the rod domains of eukaryotic nuclear and cytoplasmic IF proteins, which are related to each other, are still compatible upon deletion of the 42 aa insert of coassembly. In addition, NF-L end domains can substitute for the corresponding lamin domains in nuclear lamina targeting.

Amino Acid Sequence↗
PubMed2000

A treatment algorithm for pneumothoraces complicating central venous catheter insertion.

BACKGROUND: We investigated the role of observation or insertion of a small French pigtail catheter with Heimlich valve as alternative management to a tube thoracostomy for iatrogenic pneumothorax complicating central venous catheter (CVC) insertion. METHODS: A retrospective review of 9,637 consecutive patients who had had subclavian CVCs inserted on an outpatient basis identified 100 patients with pneumothoraces. Treatment consisted of (1) observation, (2) outpatient insertion of a Heimlich valve, or (3) inpatient tube thoracostomy. RESULTS: The median pneumothorax size was 10% (range 1% to 100%). Fifty-eight patients had observation as initial treatment, and this strategy was successful in 35 (60%). Thirty-four patients were treated initially with Heimlich valves, and this strategy was successful in 29 (85%). Tube thoracostomy as initial therapy was successful in 7 (88%) of 8 patients. Patients in who initial treatment failed were treated with insertion of a Heimlich valve or tube thoracostomy. CONCLUSION: In appropriately selected patients, pneumothorax after insertion of a subclavian CVC can be successfully managed in the outpatient setting with observation. Patients in whom observation fails can be treated with insertion of a Heimlich valve. Tube thoracostomy can be reserved for refractory PTX or emergent situations.

Ambulatory Care↗
PubMed1999

Duplex ultrasound insertion of inferior vena cava filters in multitrauma patients.

BACKGROUND: Techniques for placement of inferior vena cava (IVC) filters have undergone continued evolution from open surgical exposure of the venous insertion site to percutaneous insertion in most cases today. However, the required transport either to an operating room or interventional suite can be complex and potentially hazardous for the multiply injured trauma patient who may require ventilator support, controlled intravenous infusions, or skeletal immobilization. Increased experience with color-flow duplex scanning for routine IVC imaging and portability of ultrasound equipment have suggested the usefulness of duplex-guided IVC filter insertion (DGFI) in critically ill trauma and intensive care unit (ICU) patients. METHODS: A total of 25 multitrauma/ICU patients were considered for DGIF. Screening color-flow duplex scans were performed on all patients, and obesity or bowel gas prevented ultrasound imaging in 2 cases, leaving 23 patients suitable for DGFI. In each case, the IVC was imaged in the transverse and longitudinal planes. The right renal artery was identified as it passed posterior to the IVC and was used as a landmark of the infrarenal segment of the IVC. All procedures were performed at the bedside in a monitored ICU setting using percutaneous placement of titanium Greenfield filters. Duplex scanning after insertion was used to document proper placement, and circumferential engagement of the filter struts in the IVC wall. An abdominal radiograph was also obtained in each case to confirm proper filter location. Duplex ultrasound imaging was repeated within 1 week of insertion to assess IVC and insertion site patency. RESULTS: DGFI was successful in all cases. The filter was deployed at a suprarenal level in one case, as was recognized at the time of postprocedural scanning. Three patients died as a result of their injuries but there were no pulmonary embolism deaths. Repeat duplex scanning was obtained in 17 patients, and revealed no case of IVC or insertion site thrombosis. CONCLUSIONS: Vena caval interruption can be safely performed under ultrasound guidance in a monitored, ICU environment. In selected multiply injured trauma patients, this will reduce the risk, complexity and cost of transport for these critically ill patients. DGFI also reduces procedural costs compared with an operating room or interventional suite, and eliminates intravenous contrast exposure. Preprocedural scanning is essential to identify patients suitable for DGFI, and careful attention must be paid to the known ultrasonographic anatomical landmarks.

Adult↗
PubMed2001

Spontaneous insertion of gene 9 minor coat protein of bacteriophage M13 in model membranes.

Gene 9 minor coat protein from bacteriophage M13 is known to be located in the inner membrane after phage infection of Escherichia coli. The way of insertion of this small protein (32 amino acids) into membranes is still unknown. Here we show that the protein is able to insert in monolayers. The limiting surface pressure of 35 mN/m for 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphoglycerol lipid systems indicates that this spontaneous insertion can also occur in vivo. By carboxyfluorescein leakage experiments of vesicles it is demonstrated that protein monomers, or at least small aggregates, are more effective in releasing carboxyfluorescein than highly aggregated protein. The final orientation of the protein in the bilayer after insertion was addressed by proteinase K digestion, thereby making use of the unique C-terminal location of the antigenic binding site. After insertion the C-terminus is still available for the enzymatic digestion, while the N-terminus is not. This leads to the overall conclusion that the protein is able to insert spontaneously into membranes without the need of any machinery or transmembrane gradient, with the positively charged C-terminus remaining on the outside. The orientation after insertion of gene 9 protein is in agreement with the 'positive inside rule'.

Bacteriophage M13↗