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Effect of treatment with two intravaginal inserts on the uterine and vaginal microflora of early postpartum beef cows.

OBJECTIVE: To describe the bacterial species found within the uterus and vagina of early postpartum (10 to 20 d) beef cows treated for 14 d with (i) two intravaginal (CIDR-B) progesterone releasing inserts (C-P4; n=31); (ii) two identical but blank inserts (C-BL; n=15); (iii) untreated controls (CON; n=15). It was hypothesised that due to the locally immunosuppressive effects of progesterone on the uterus, the bacterial microflora of C-P4 would be altered by this treatment in contrast to CON and C-BL. PROCEDURE: Cows were enrolled at two intervals 7 d apart. Blood samples were collected at 0, 7 and 14 d after beginning treatments for subsequent progesterone assay. A triple guarded swabbing technique was used to collect bacteriological samples from the uterus of every cow on days 0, 7 and 14 following CIDR insertion. Swabs were also collected from the inserts and vagina of every cow on day 14. Due to the small sample sizes, only descriptive statistics were generated. RESULTS: Plasma progesterone levels were maintained at mid-luteal phase concentrations by the intravaginal progesterone releasing inserts (C-P4: 4.2 +/- 0.4 ng/mL at 7 d; 3.6 +/- 0.2 ng/mL at 14 d), although increased progesterone concentrations were found in 4/15 CON and 9/15 C-BL cows on day 14. Bacteria were isolated from 32/61 (52%) of all uterine samples collected at the time of insertion. Uterine and vaginal swabs from CON cows showed a marked reduction in isolates over time such that 14 d after insertion only 1/15 uterine swabs grew bacteria. In contrast, C-BL and C-P4 treated cows failed to show reductions in the number of uterine or vaginal isolates at 14 d after device insertion. Heavy growths of Pseudomonas aeruginosa and Actinomyces pyogenes were found on the intravaginal inserts from C-BL and C-P4 cows. Cows enrolled in the second week of the study that received intravaginal inserts (C-P4 + C-BL) were more likely to have Pseudomonas isolated from the uterus than those enrolled in week 1 (1/18 versus 14/28). CONCLUSIONS: The presence of two intravaginal inserts, regardless of hormone content, substantially altered the profile of uterine and vaginal bacteria in early postpartum beef cows. It was suspected that because of the early stage at insert application, the cervix had not involuted sufficiently to provide an adequate microbial barrier to the uterus. Pseudomonas aeruginosa was the predominant species isolated from both uterine and insert cultures after 14 d of treatment but may have been a contaminant, due to the greater proportion of cows infected with it that had been enrolled in the second week of the study.

Administration, Intravaginal↗

Interactions between cochlear implant electrode insertion depth and frequency-place mapping.

While new electrode designs allow deeper insertion and wider coverage in the cochlea, there is still considerable variation in the insertion depth of the electrode array among cochlear implant users. The present study measures speech recognition as a function of insertion depth, varying from a deep insertion of 10 electrodes at 28.8 mm to a shallow insertion of a single electrode at 7.2 mm, in four Med-El Combi 40+ users. Short insertion depths were simulated by inactivating apical electrodes. Speech recognition increased with deeper insertion, reaching an asymptotic level at 21.6 or 26.4 mm depending on the frequency-place map used. Başkent and Shannon [J. Acoust. Soc. Am. 116, 3130-3140 (2004)] showed that speech recognition by implant users was best when the acoustic input frequency was matched onto the cochlear location that normally processes that frequency range, minimizing the spectral distortions in the map. However, if an electrode array is not fully inserted into the cochlea, a matched map will result in the loss of considerable low-frequency information. The results show a strong interaction between the optimal frequency-place mapping and electrode insertion depth. Consistent with previous studies, frequency-place matching produced better speech recognition than compressing the full speech range onto the electrode array for full insertion ranges (20 to 25 mm from the round window). For shallower insertions (16.8 and 19.2 mm) a mild amount of frequency-place compression was better than truncating the frequency range to match the basal cochlear location. These results show that patients with shallow electrode insertions might benefit from a map that assigns a narrower frequency range than patients with full insertions.

Adult↗

Gum elastic bougie-guided insertion of the ProSeal Laryngeal Mask Airway.

We tested the hypothesis that gum elastic-bougie-guided insertion of the ProSeal Laryngeal Mask Airway is more frequently successful than introducer tool guided insertion after failed digital insertion. One hundred anaesthetized patients (ASA 1-2, aged 18 to 80 years) were randomized for the second insertion attempt using either the gum elastic bougie-guided or introducer tool techniques. The bougie-guided technique involved priming the drain tube with the bougie, placing the bougie in the oesophagus using laryngoscope guidance, digital insertion along the palato-pharyngeal curve, and bougie removal. The introducer tool technique involved attaching the introducer tool, single-handed rotation along the palatopharyngeal curve, and introducer tool removal. Failed insertion was classified as (i) failed passage into the pharynx, (ii) malposition, or (iii) ineffective ventilation. Any blood staining was documented. Insertion was more frequently successful (50/50 vs 15/50, P=0.0002) and faster (35+/-17 s vs 54+/-45 s, mean+/-SD, P=0.006) with the bougie-guided technique. All failed insertions with the introducer tool technique were successful with the bougie-guided technique. The aetiology of failed insertion was similar for the digital and introducer tool techniques in 94% (33/35) of patients. There was no blood staining on the bougie, laryngoscope or introducer tool at removal, but blood staining was more common on the ProSeal Laryngeal Mask Airway with the introducer tool technique (9/50 vs 2/50, P=0.03). We conclude that the gum elastic bougie-guided insertion has a higher success rate and causes less trauma than the insertion tool insertion technique after failed digital insertion of the ProSeal Laryngeal Mask Airway.

Adolescent↗

Clinical results of modular polyethylene insert exchange with retention of total knee arthroplasty components.

BACKGROUND: Modular polyethylene inserts have enabled surgeons to perform an isolated tibial insert exchange while retaining well fixed components. The purpose of this study was to review the results of insert revision and to clarify the role of this option compared with that of revision total knee arthroplasty. METHODS: Fifty-six patients (sixty-three knees) were managed with revision of a tibial polyethylene insert and retention of well aligned and stable femoral and tibial components. The implants had been in situ for an average of fifty-nine months (range, two to 108 months) at the time of the insert exchange. The inserts that were removed at the time of exchange were evaluated with regard to wear of the articular surface according to the classification system of Hood et al. and with regard to undersurface wear according to the method described by Wasielewski et al. Forty-eight knees were followed for an average of 7.4 years (range, 3.0 to 12.2 years) after the insert exchange. Knees that did not require an additional operation were considered to have had a successful exchange. RESULTS: Seven of the forty-eight exchanges failed, at an average of fifty-four months, because of accelerated wear of the new insert. All seven knees required complete revision of all components. Of the twenty-two exchanges that were performed because of severe wear of the primary insert, six (27 percent) failed at an average of less than five years; thus, knees in which the exchange was performed because of advanced wear were more likely to fail again (p < 0.05). In addition, primary inserts that were removed from knees in which the exchange procedure subsequently failed had higher delamination scores than those that were removed from knees in which the exchange was successful (p < 0.05). Most of the primary inserts had substantial undersurface wear at the time of the exchange procedure. Metallosis (thirty knees) and osteolysis (nineteen knees) were unrelated to failure of the exchange. CONCLUSIONS: An isolated revision of the tibial polyethylene insert should not be performed when there is accelerated wear of the insert with severe delamination and grade-3 or 4 undersurface wear within ten years after the primary procedure. Because a variety of patient-related, implant-related, and technical factors influence polyethylene wear, the orthopaedist must consider multiple variables whenever contemplating a limited revision.

Arthroplasty, Replacement, Knee↗

Tibial interface wear in retrieved total knee components and correlations with modular insert motion.

BACKGROUND: Wear occurring at the interface between the polyethylene insert and metal baseplate of a modular tibial component has become an increasingly common finding at the time of revision total knee arthroplasty. Although this so-called backside wear on retrieved polyethylene inserts has been evaluated in prior studies, wear on retrieved metal baseplates has not been described, to our knowledge. The purposes of the present study were to characterize backside wear on retrieved polyethylene inserts and on the mating surfaces of their corresponding baseplates and to investigate if there is a relationship between backside wear and relative motion of the modular elements. METHODS: Twenty-nine retrieved modular tibial components of twelve fixed-bearing designs were analyzed in vitro with regard to backside wear and relative motion between the polyethylene insert and the metal baseplate. We graded the backside of each polyethylene insert and the mating surface of the metal baseplate for wear with use of a scoring system that consisted of three modes of wear and three levels of severity of wear. Relative motion between the insert and the baseplate was measured in the transverse plane with use of a mechanical testing machine. These measurements were used to compute the insert motion index, which served to quantify unrestricted motion of the insert with respect to the baseplate. RESULTS: The mean insert motion index for the tibial components was 416 micro m (range, 104 micro m to 760 micro m). On a wear-grading scale ranging from 0 to 54 (with 0 indicating no wear), the mean backside wear score was 30 (range, 12 to 48) for the inserts and 28 (range, 7 to 51) for the baseplates. Insert motion was positively correlated with backside polyethylene wear (p = 0.003) and baseplate wear (p < 0.001). Baseplate wear was strongly correlated with backside polyethylene wear (p < 0.001). CONCLUSIONS: Backside wear was correlated with the relative motion between the polyethylene insert and the metal baseplate. New locking mechanism designs directed toward better methods of securing the polyethylene insert to the tibial tray are needed to minimize the generation of particulate wear debris at the modular interface.

Adult↗

Ovulation and estrus characteristics in crossbred Brahman heifers treated with an intravaginal progesterone-releasing insert in combination with prostaglandin F2alpha and estradiol benzoate.

Crossbred Brahman heifers (n = 60) were studied to determine the effect of a 7-d intravaginal progesterone-releasing insert (INSERT) in combination with PG (Lutalyse; 25 mg i.m.) and estradiol benzoate (EB; .5 mg i.m.) on time of ovulation and estrous behavior. In Phase I, heifers at unknown stages of the estrous cycle were assigned by BW and body condition score to one of the three treatments on d 0: 1) INSERT for 7 d and PG on d 7 (CONTROL; n = 10); 2) INSERT for 7 d, PG on d 7, and EB 24 h after INSERT removal (EB24; n = 10); or 3) INSERT for 7 d, PG on d 7, and EB 48 h after INSERT removal (EB48; n = 10). Blood samples were collected every 8 h after INSERT removal. Also, blood sampling and ultrasonography began 8 h after the onset of estrus, determined with HeatWatch devices, and every 4 h thereafter to detect ovulation. In Phase II, Phase-I treatments (n = 10/treatments) were replicated, but only behavioral estrus data were collected to minimize handling of heifers. Frequent handling of heifers did not influence (P > .1) the interval from INSERT removal to the onset of HeatWatch and visual estrus and duration of estrus, so behavioral estrus data were combined for Phases I and II. Interval from INSERT removal to HeatWatch estrus was decreased (P < .05) in EB24 (45.5 h) vs EB48 (55.9 h) and CONTROL (59.2 h). Interval from INSERT removal to ovulation differed (P < .04) between CONTROL, EB24, and EB48 (93.5, 74.5, and 78.9 h, respectively). Ovulatory follicle size was similar (P > .1) between CONTROL, EB24, and EB48 (14.4, 12.5, and 14.1 mm, respectively). Duration of estrus was similar for CONTROL, EB24, and EB48 (14.0, 15.1, and 17.6 h, respectively). No difference (P > . 1) was observed in number of mounts received between CONTROL, EB24, and EB48 (28.0, 25.7, and 39.4, respectively), but number of mounts received increased in Phase II vs Phase I (40.0 and 22.2, respectively; P < .05). In conclusion, EB hastened the interval from INSERT removal to ovulation without altering duration of estrus or number of mounts received. Frequent handling of heifers did not affect interval to first mount received after INSERT removal or duration of estrus, but it decreased the total number of mounts received.

Administration, Intravaginal↗

Appropriately trained nurses are competent at inserting intrauterine devices: an audit of clinical practice.

OBJECTIVE: Very few nurses in the UK insert intrauterine devices (IUDs). Within King's Healthcare, nurses with the advanced family planning qualification (ENB AO8), who have also undertaken further training in IUD insertion, have done so for many years. This is a controversial issue and our aim was to evaluate whether women who had IUDs inserted by specialist nurses had significantly more problems compared with those inserted by doctors. STUDY DESIGN: This was a prospective, ongoing, cohort study of all IUD insertions in hospital-based family planning clinics over a 3-month period. A medical, gynecological, obstetric and contraceptive history was taken, paying particular regard to the reason for requesting an IUD. Clinical assessment, bimanual examination findings and any difficulties with insertion were noted at the time. Any postinsertion problems were noted at the follow-up visit. RESULTS: Fifty IUDs were inserted, 28 by doctors and 22 by nurses. Sixteen were for postcoital contraception and five were routine reinsertions alter removal of an existing IUD. Eleven nulliparous women had insertions (six by doctors and five by nurses). The specialist nurses did not have to consult the doctor with any problems or difficulties with insertion. Twenty-nine clients attended for their follow-up appointment at 6 weeks. Three IUDs required early removal due to unacceptable bleeding and pain--all three had been inserted by doctors. Two postcoital IUDs were removed following menstruation. Of the remaining patients, eight were noted to have minor problems (divided equally between doctor and nurse insertions). CONCLUSION: This study provides favorable evidence that adequately trained nurses are proficient and safe at IUD insertions, regardless of the woman's parity. IUDs inserted by nurses are also likely to have additional benefits of cost-effectiveness and increased client and nurse satisfaction. Follow-up is ongoing.

Adolescent↗

[Comparative study of two methods for laryngeal mask insertion].

OBJECTIVE: To compare two ways of inserting laryngeal airway masks: uninflated and partially inflated to 75% of the volume, as recommended by manufacturers. PATIENTS AND METHOD: We studied 60 ASA I-II patients scheduled for outpatient surgery under general anesthesia with numbers 3 or 4 laryngeal masks, after having obtained informed consent from the patients (or parents in the case of minors). The patients were randomly assigned to two groups. In group A the masks were inserted inflated to 75% of volume as recommended by manufacturers, whereas in group B deflated masks were inserted as described by Brain. Anesthesia was standardized for all patients. One patient was withdrawn from the study when a technical error was detected. We recorded the presence of criteria predictive of difficult airway management, systolic and diastolic blood pressures, heart rate at four times (baseline, before and after induction and after insertion of the mask), number of insertion tries, final mask volume needed for adequate ventilation, need for an additional dose of the hypnotic drug and complications. RESULTS: No statistically significant differences between the two insertion methods were found. Difficult airway management criteria were unrelated to difficulty of mask insertion. Correct insertion of an uninflated mask proved impossible in one patient, in whom we were then able to insert an inflated mask. CONCLUSION: No differences were found between inflated insertion and Brain's uninflated insertion technique. We believe that inflated mask insertion might be useful when uninflated insertion proves impossible.

Adult↗

Prophylactic inferior vena cava filter insertion for trauma: intensive care unit versus operating room.

The frequency of insertion of prophylactic inferior vena cava filters (IVCF) among traumatized patients has increased nationally. That has placed a substantial operational and economic burden upon trauma centers. The purpose of this study was to compare and contrast successful implantation, morbidity, and cost-effectiveness of prophylactic IVCF insertion in a surgical-trauma intensive care unit (STICU) versus an operating room (OR). A retrospective chart review was conducted of all trauma patients who received a prophylactic IVCF at an urban Level I trauma center between January 1999 and December 2003. Data were collected to identify patient demographics, indications, anatomical site of insertion, hospital location of insertion, hospital days before insertion, and complications associated with insertion. One hundred thirty-four patients underwent prophylactic IVCF during the study period: seventy-eight (58%) in the OR and fifty-six (42%) in the STICU. The average age of patients for the OR and STICU groups were 38.6 years and 39.6 years, respectively. The average number of days to IVCF insertion was 6.5 days and 7.0 days in the OR and STICU groups, respectively. Indications for IVCF among patients who had placement in the OR were orthopedic injury (60%), spinal cord injury (25%), and head injury (15%). Indications for IVCF among patients who had placement in the STICU were head injury (38%), orthopedic injuries (34%), and spinal cord injury (25%). Three (3.8%) patients in the OR group and two (3.6%) patients in the STICU group required a change of anatomic insertion site from the femoral to the internal jugular vein. There were two (2.6%) complications associated with IVCF insertion in the OR and two (3.5%) complications associated with IVCF insertion in the STICU (P > 0.05). Insertion of IVCF in the STICU decreased patient-cost by an average of dollar 1636 per patient. Prophylactic IVCF insertion in an STICU is cost-effective and can be performed with similar success and complication rates to IVCF insertion in an OR.

Adult↗

High throughput T-DNA insertion mutagenesis in rice: a first step towards in silico reverse genetics.

A library of 29,482 T-DNA enhancer trap lines has been generated in rice cv. Nipponbare. The regions flanking the T-DNA left border from the first 12,707 primary transformants were systematically isolated by adapter anchor PCR and sequenced. A survey of the 7480 genomic sequences larger than 30 bp (average length 250 bp), representing 56.4% of the total readable sequences and matching the rice bacterial artificial chromosome/phage artificial chromosome (BAC/PAC) sequences assembled in pseudomolecules allowed the assigning of 6645 (88.8%) T-DNA insertion sites to at least one position in the rice genome of cv. Nipponbare. T-DNA insertions appear to be rather randomly distributed over the 12 rice chromosomes, with a slightly higher insertion frequency in chromosomes 1, 2, 3 and 6. The distribution of 723 independent T-DNA insertions along the chromosome 1 pseudomolecule did not differ significantly from that of the predicted coding sequences in exhibiting a lower insertion density around the centromere region and a higher density in the subtelomeric regions where the gene density is higher. Further establishment of density graphs of T-DNA inserts along the recently released 12 rice pseudomolecules confirmed this non-uniform chromosome distribution. T-DNA appeared less prone to hot spots and cold spots of integration when compared with those revealed by a concurrent assignment of the Tos17 retrotransposon flanking sequences deposited in the National Center for Biotechnology Information (NCBI). T-DNA inserts rarely integrated into repetitive sequences. Based on the predicted gene annotation of chromosome 1, preferential insertion within the first 250 bp from the putative ATG start codon has been observed. Using 4 kb of sequences surrounding the insertion points, 62% of the sequences showed significant similarity to gene encoding known proteins (E-value < 1.00 e(-05)). To illustrate the in silico reverse genetic approach, identification of 83 T-DNA insertions within genes coding for transcription factors (TF) is presented. Based both on the estimated number of members of several large TF gene families (e.g. Myb, WRKY, HD-ZIP, Zinc-finger) and on the frequency of insertions in chromosome 1 predicted genes, we could extrapolate that 7-10% of the rice gene complement is already tagged by T-DNA insertion in the 6116 independent transformant population. This large resource is of high significance while assisting studies unravelling gene function in rice and cereals, notably through in silico reverse genetics.

Base Sequence↗

Structural and energetic differences between insertions and substitutions in staphylococcal nuclease.

In a previous study, the small protein staphylococcal nuclease was shown to readily accommodate single alanine and glycine insertions, with average losses in stability comparable to substitutions at the same sites (PROT. 7:299-305, 1990). To more fully explore this unexpected adaptability to changes in residue spacing, 2 double amino acid insertions (alanyl-glycine, glycyl-glycine) and 3 additional single amino acid insertions with dissimilar side chains (proline, leucine, and glutamine) were constructed at 10 of the sites previously studied. At 8 of these sites, the type of amino acid side chain on the inserted residue significantly influenced the stability of the mutant protein. However, at 9 of the 10 sites, the double insertions were found to be no more destabilizing than the single alanine or glycine insertions. In contrast, double substitution mutations of staphylococcal nuclease, which replace two adjacent residues with alanine, do not show this striking degree of non-additivity. A comparison of the effects of single glutamine and single glycine insertions with alanyl-glycine insertions indicates that insertion of alanine into the peptide backbone is, on average, less destabilizing than appending the equivalent atoms onto the side chain of a glycine insertion. To explain their very different energetic effects, we propose that, unlike most substitutions, the inserted residue(s) must induce lateral displacements of the polypeptide chain, forcing the folded conformation away from that of wild type. The resulting obligatory shifts in the positioning of residues flanking the insertion generate a large number of degrees of freedom around which the mutant structure can relax.(ABSTRACT TRUNCATED AT 250 WORDS)

Alanine↗

The size distribution of insertions and deletions in human and rodent pseudogenes suggests the logarithmic gap penalty for sequence alignment.

The size distributions of deletions, insertions, and indels (i.e., insertions or deletions) were studied, using 78 human processed pseudogenes and other published data sets. The following results were obtained: (1) Deletions occur more frequently than do insertions in sequence evolution; none of the pseudogenes studied shows significantly more insertions than deletions. (2) Empirically, the size distributions of deletions, insertions, and indels can be described well by a power law, i.e., fk = Ck-b, where fk is the frequency of deletion, insertion, or indel with gap length k, b is the power parameter, and C is the normalization factor. (3) The estimates of b for deletions and insertions from the same data set are approximately equal to each other, indicating that the size distributions for deletions and insertions are approximately identical. (4) The variation in the estimates of b among various data sets is small, indicating that the effect of local structure exists but only plays a secondary role in the size distribution of deletions and insertions. (5) The linear gap penalty, which is most commonly used in sequence alignment, is not supported by our analysis; rather, the power law for the size distribution of indels suggests that an appropriate gap penalty is wk = a + b ln k, where a is the gap creation cost and blnk is the gap extension cost. (6) The higher frequency of deletion over insertion suggests that the gap creation cost of insertion (ai) should be larger than that of deletion (ad); that is, ai - ad = ln R, where R is the frequency ratio of deletions to insertions.

Animals↗

Expression of ribosomal DNA insertions in Drosophila melanogaster.

Approximately half of the ribosomal genes on the X chromosome of Drosophila melanogaster are interrupted by an insertion of type 1. Nuclear RNA from D. melanogaster embryos was transferred to DBM paper and hybridized with cloned type 1 insertion sequences. With a DNA fragment derived specifically from large insertions, transcripts were detected between 5 and 10 kb. These insertion transcripts represent less than one RNA molecule per nucleus, which is more than three orders of magnitude below the concentration of nascent rRNA chains, as determined by kinetics of hybridization. With a DNA fragment derived from the right end of large insertions which is also complementary to short insertions, more discrete RNA bands appeared with sizes between 1 and 8.5 kb, representing altogether about 13 RNA molecules per nucleus. Insertion transcripts large enough to be potential precursors to 28S rRNA represent less than one molecule per nucleus. It was shown by sandwich hybridization that at least some of the insertion transcripts are derived from rDNA. No significant difference was found between insertion transcripts in RNA extracted from ovaries, embryos, larvae, pupae or adult flies. Unless a mechanism other than splicing is involved, ribosomal genes with insertions cannot contribute significantly to the synthesis of 28S rRNA. A cytoplasmic RNA approximately 1 kb long, which is complementary to a short insertion and to ribosomal gene sequences flanking both sides of the insertion, was found. The abundance of this short unspliced RNA is about 50 molecules per embryo cell.

Animals↗

Insertion-site anatomy of the human menisci: gross, arthroscopic, and topographical anatomy as a basis for meniscal transplantation.

A cadaveric study was performed to determine the insertion-site anatomy of the human menisci, their topographical relationships to adjacent intra-articular structures, and which arthroscopic portal provides for optimal visualization of each insertion site. Fifteen fresh-frozen cadaver knees were studied (ages 48 to 63 years). Ten knees underwent arthroscopy using four standard arthroscopic portals. Visualization and placement of an arthroscopic guide over each meniscal horn insertion site was attempted through the four arthroscopic portals. Guide wires were drilled to mark horn insertions followed by a gross dissection to evaluate accuracy of the guide wire gross dissection to evaluate accuracy of the guide wire placement and to isolate meniscal horn insertion sites. Insertion sites were outlined and evaluated for size and topographical relationships to other intra-articular structures. Five additional knees were dissected free of all soft tissues except the tibial insertions of the meniscal roots and anterior cruciate ligament/posterior cruciate ligament. Each tibia was mounted in a jig and a digitizing system was used to record coordinates of points along the outline of each bony meniscal horn insertion site, the ACL tibial insertion, and the articular surface of each tibial plateau. The x, y, z coordinates for each point were calculated and loaded into a computer program allowing for surface area determination and computer-generated topographical maps to assess relative position of each specific insertion site. Placement of the arthroscope in the anterolateral portal allows optimal visualization and guide wire placement for both lateral meniscal horn insertion sites. Medial meniscal anterior and posterior horn insertion sites are best visualized with the arthroscope in the anteromedial and posteromedial portals respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Arthroscopy↗

Domain insertions in protein structures.

Domains are the structural, functional or evolutionary units of proteins. Proteins can comprise a single domain or a combination of domains. In multi-domain proteins, the domains almost always occur end-to-end, i.e., one domain follows the C-terminal end of another domain. However, there are exceptions to this common pattern, where multi-domain proteins are formed by insertion of one domain (insert) into another domain (parent). Here, we provide a quantitative description of known insertions in the Protein Data Bank (PDB). We found that 9% of domain combinations observed in non-redundant PDB are insertions. Although 90% of all insertions involve only one insert, proteins can clearly have multiple (nested, two-domain and three-domain) inserts. We also observed correlations between the structure and function of a domain and its tendency to be found as a parent or an insert. There is a bias in insert position towards the C terminus of parents. We observed that the atomic distance between the N and C terminus of an insert is significantly smaller when compared to the N-to-C distance in a parent context or a single domain context. Insertions are found always to occur in loop regions of parent domains. Our observations regarding the relationship between domain insertions and the structure, function and evolution of proteins have implications for protein engineering.

Amino Acid Sequence↗

Pedicle screw fixation strength: pullout versus insertional torque.

BACKGROUND CONTEXT: Researchers studying early pedicle screw designs have suggested that pullout strength and insertional torque are correlated. For surgeons using pedicle screws, insertional torque is widely believed to be a good predictor of pullout strength and initial stability of the screw and construct. How appropriate is this assumption when applied to new screw and thread designs? PURPOSE: This study investigated the correlation between insertional torque and pullout strength of three different pedicle screw designs, with different insertional torque characteristics. We hypothesized that a significant increase in insertional torque would indicate a commensurate increase in pullout strength. STUDY DESIGN: Biomechanical analysis of instrumented vertebral specimens. METHODS: Calf lumbar vertebra were prepared and instrumented with one of three pedicle screws. Pilot hole preparation was standardized and coaxial orientation was confirmed by direct inspection. Screws did not penetrate the pedicle cortex or abut or penetrate the anterior vertebral cortex. Any specimen with pedicle wall breach was discarded. The pedicles were instrumented with one of three screws: 1) 7.5 x 40 mm conical, asymmetric progressive thread (Xia; Stryker Spine, Allendale, NJ), 2) 7.5 x 40 mm conical with traditional V-shaped thread (Osteonics, Stryker Spine, Allendale, NJ)) or 3) 6.5 x 40 mm cylindrical with V thread (Osteonics, cylindrical). Paired testing allowed individual screws to be directly compared with a contralateral "control." Insertional torque and peak torque values were recorded for each rotation up to full insertion. Pullout testing was conducted at a rate of 1 mm/minute. Load-displacement data were recorded at 20 Hz. Stiffness was considered the slope of the most linear part of the curve before the yield point. RESULTS: Peak loads for 7.5 conical Xia screws measured 1,783+/-589.1 N compared with 1,943.0+/-625.8 N for 7.5 conical Osteonics screws and 1,641.0+/-356.7 N for 6.5 cylindrical Osteonics screws. The peak insertional torque values were 6.7+/-1.9 Nm (158% greater than control), 4.5+/-1.1 Nm (73% greater than control) and 2.6+/-0.7 Nm, respectively. Insertional torques for Xia screws were significantly greater than conical (p=.001) and cylindrical Osteonics screws (p<.0001), and insertional torques for Osteonics conical screws were significantly greater than those of cylindrical screws (p<.0001). Although pullout loads for the conical Osteonics screws were consistently higher than either the Xia or Osteonics cylindrical screws, the differences were not significant (p>.05). There was no significant correlation between pullout strength and insertional torque (p>.05). CONCLUSIONS: This unexpected result is best explained by the progressively narrowing flutes of the Xia screw, which compact the trabeculae into a smaller volume as the screw nears full insertion. The trapezoidal threads also increase contact with the cortical surface area and compress trabeculae toward the cortex, thus creating greater friction and higher torque values. This increase in torque did not translate into a commensurate increase in pullout strength, where trabeculae fail in shear.

Animals↗

Catalyzed insertion of proteins into phospholipid membranes: specificity of the process.

The process of insertion of intrinsic proteins into phospholipid membranes conjures up the thought of enormous energy barriers but is a routine occurrence in cells. Proteinaceous complexes responsible for protein targeting/translocation/insertion into membranes have been studied intensively. However, the mitochondrial voltage-dependent anion channel (VDAC), can insert into phospholipid membranes by an auto-catalytic process called "auto-directed insertion." This process results in an oriented insertion of VDAC channels and an increase in insertion rate per unit area of 10 orders of magnitude. Here we report that VDAC catalyzes the insertion of PorA/C1 and KcsA by increasing their calculated insertion rate per unit area by 9 orders of magnitude with no detectable effect on the insertion of alpha-hemolysin. This was measured as a reduction in the delay before the first insertion of these proteins. Gramicidin and PorA/C1 accelerate the calculated insertion rate per unit area of VDAC by 8 and 9 orders of magnitude, respectively. Only PorA/C1 increases the overall rate of VDAC insertion (50-fold) over the self-catalyzed rate. Our results indicate that catalyzed insertion of proteins into phospholipid membranes does not arise simply from disturbance of the phospholipid membrane because it shows strong specificity.

Bacterial Proteins↗

Follicular growth, estrus and pregnancy after fixed-time insemination in beef cows treated with intravaginal progesterone inserts and estradiol benzoate.

An experiment was performed to compare the effects of 3 short-term treatments with progesterone and estradiol benzoate (EB) on follicular growth, synchrony of estrus and pregnancy rate after fixed-time insemination in lactating postpartum beef cows. In Treatment 1 (n = 46), each cow received a progesterone-containing intravaginal insert for 7 d with injection of EB (2 mg, i.m.) at the time of device insertion. In Treatment 2 (n = 46), the insert was used for only 5 d with injection of EB (2 mg, i.m.) at the time of insertion. Cows in Treatment 3 (n = 47) received an insert for 5 d with no EB at the time of insertion. Each cow in the 3 groups received PGF2 alpha (25 mg, i.m.) at the time of insert removal, followed by EB (1 mg, i.m.) 30 h later. The cows were then inseminated 28 to 30 h after treatment with EB (58 to 60 h after insert removal). Treatment with 2 mg EB terminated the growth of the largest ovarian follicle (> 5 mm in diameter) at device insertion in 16/16 and 14/15 cows in Treatments 1 and 2, respectively. Estrus was detected within an 8-h target period (48 to 56 h after insert removal) in 93, 87 and 81% of cows in Treatments 1, 2 and 3, respectively (P > 0.05). Pregnancy rates at 39 d post insemination were 60, 50 and 51% for Treatments 1, 2 and 3, respectively (P > 0.05). The pregnancy rates did not differ between cows that were anovulatory or those that had ovulated before the initiation of treatments (54%), or among cows that were 28 to 40, 41 to 60 or > 60 days post partum at insemination (43, 59 and 54%, respectively). Treatment with progesterone inserts for 5 or 7 d, PGF2 alpha at the time of insert removal and 1 mg EB 30 h later induced the high degree of synchrony of estrus and ovulation necessary for fixed-time insemination.

Administration, Intravaginal↗