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The timing of norplant insertion and postpartum depression in teenagers.

BACKGROUND: This study tested the hypothesis that teenagers who have Norplant inserted during the puerperium report more depressive symptoms during the first postpartum year than their peers who do not receive Norplant. METHODS: We studied the prevalence of depressive symptoms in a group of 212 mothers aged 19 years less, in relation to the timing of Norplant insertion. The participants were divided into 3 groups: 100 (47%) had Norplant inserted during the puerperium (early Norplant users); 72 (34%) had Norplant inserted during the next 10 months (late Norplant users); and 40 (19%) used other contraceptives (40% oral contraceptives; 17% condoms; 43% nothing). Depressive symptoms were measured with the Center for Epidemiologic Studies - Depression Scale. Postpartum depression was defined as a scale score >/=16, 6-12 months after Norplant insertion or delivery. Variables examined as potential confounders were identified a priori from a review of the literature and controlled for in analysis of variance. RESULTS: At delivery, members of the 3 contraceptive groups did not differ significantly with regard to age, race, parity, educational, marital, or socioeconomic status. Late Norplant users were, however, more apt to have new boyfriends (p =. 03), to rate the support they received from the baby's father as poor (p =.004), and experience depression prior to Norplant insertion (p =.02). Contrary to the study hypothesis, late rather than early Norplant insertion was associated with postpartum depression. Multivariate analyses identified 3 independent predictors of the severity of depressive symptoms at follow-up (depression prior to Norplant insertion, a new boyfriend at delivery, and late Norplant insertion); R(2) = 41.3%. CONCLUSIONS: Contrary to the study hypothesis, puerperal Norplant insertion did not exacerbate postpartum depression. Delaying Norplant insertion may increase the risk of depression during the first postpartum year, particularly in teenagers with other psychosocial risk factors.

Adolescent↗

MTID: a database of Sleeping Beauty transposon insertions in mice.

The Sleeping Beauty (SB) transposon system provides the first random insertional mutagen available for germline genetic screens in mice. In preparation for a large scale project to create, map and manage up to 5000 SB insertions, we have developed the Mouse Transposon Insertion Database (MTID; http://mouse.ccgb.umn.edu/transposon/). Each insertion's genomic position, as well as the distance between the insertion and the nearest annotated gene, are determined by a sequence analysis pipeline. Users can search the database using a specified nucleotide or genetic map position to identify the nearest insertion. Mouse reports describe insertions carried, strain, genotype and dates of birth and death. Insertion reports describes chromosome, nucleotide and genetic map positions, as well as nearest gene data from Ensembl, NCBI and Celera. The flanking sequence used to map the insertion is also provided. Researchers will be able to identify insertions of interest and request mice or frozen sperm that carry the insertion.

Animals↗

Site preferences of insertional mutagenesis agents in Arabidopsis.

We have performed a comparative analysis of the insertion sites of engineered Arabidopsis (Arabidopsis thaliana) insertional mutagenesis vectors that are based on the maize (Zea mays) transposable elements and Agrobacterium T-DNA. The transposon-based agents show marked preference for high GC content, whereas the T-DNA-based agents show preference for low GC content regions. The transposon-based agents show a bias toward insertions near the translation start codons of genes, while the T-DNAs show a predilection for the putative transcriptional regulatory regions of genes. The transposon-based agents also have higher insertion site densities in exons than do the T-DNA insertions. These observations show that the transposon-based and T-DNA-based mutagenesis techniques could complement one another well, and neither alone is sufficient to achieve the goal of saturation mutagenesis in Arabidopsis. These results also suggest that transposon-based mutagenesis techniques may prove the most effective for obtaining gene disruptions and for generating gene traps, while T-DNA-based agents may be more effective for activation tagging and enhancer trapping. From the patterns of insertion site distributions, we have identified a set of nucleotide sequence motifs that are overrepresented at the transposon insertion sites. These motifs may play a role in the transposon insertion site preferences. These results could help biologists to study the mechanisms of insertions of the insertional mutagenesis agents and to design better strategies for genome-wide insertional mutagenesis.

Arabidopsis↗

Construction of an Enterococcus faecalis Tn917-mediated-gene-disruption library offers insight into Tn917 insertion patterns.

Sequencing the insertion sites of 8,865 Tn917 insertions in Enterococcus faecalis strain OG1RF identified a hot spot in the replication terminus region corresponding to 6% of the genome where 65% of the transposons had inserted. In E. faecalis, Tn917 preferentially inserted at a 29-bp consensus sequence centered on TATAA, a 5-bp sequence that is duplicated during insertion. The regional insertion site preference at the chromosome terminus was not observed in another low-G+C gram-positive bacterium, Listeria monocytogenes, although the consensus insertion sequence was the same. The 8,865 Tn917 insertion sites sequenced in E. faecalis corresponded to only approximately 610 different open reading frames, far fewer than the predicted number of 2,400, assuming random insertion. There was no significant preference in orientation of the Tn917 insertions with either transcription or replication. Even though OG1RF has a smaller genome than strain V583 (2.8 Mb versus 3.2 Mb), the only E. faecalis strain whose sequence is in the public domain, over 10% of the Tn917 insertions appear to be in a OG1RF-specific sequence, suggesting that there are significant genomic differences among E. faecalis strains.

Bacterial Proteins↗

Immediate postabortal insertion of intrauterine devices.

BACKGROUND: Insertion of an intrauterine device (IUD) immediately after an abortion has several potential advantages. The woman is known not to be pregnant, a major concern for clinicians. For example, many clinicians refuse to insert an IUD in a woman who is not menstruating. After induced abortion, a woman's motivation to use contraception may be high. However, insertion of an IUD immediately after a pregnancy ends carries potential risks as well. For example, the risk of spontaneous expulsion may be increased due to recent cervical dilation. OBJECTIVES: To assess the safety and efficacy of IUD insertion immediately after spontaneous or induced abortion. SEARCH STRATEGY: We used MEDLINE, Popline, and EMBASE computer searches, supplemented by review articles and contacts with investigators. SELECTION CRITERIA: We sought all randomized controlled trials that had at least one treatment arm that involved IUD insertion immediately after an induced abortion or after curettage for spontaneous abortion. We identified 12 trials which described random assignment but excluded three from this review. Two of these revealed unethical research conduct, and one used alternate assignment to treatments. DATA COLLECTION AND ANALYSIS: We evaluated the methodological quality of each report and abstracted information onto a data collection form. We focused on gross discontinuation rates (single-decrement life table analysis) for accidental pregnancy, perforation, expulsion, and pelvic inflammatory disease. We entered the data into RevMan 3.1 for analysis of Peto odds ratios. MAIN RESULTS: In large multicenter trials, the TCu 220C device proved superior to either the Lippes Loop D or the Copper 7 IUDs for immediate postabortal insertion. In single-center trials, the Nova T IUD had a significantly higher discontinuation rate for pregnancy than did the Multiload 250 (OR 4.5; 95% CI 1.0-19.8), while the Nova T had a significantly lower discontinuation rate for pregnancy than did the TCu 200 (OR 0.3; 95% CI 0.1-0.9). The levonorgestrel-releasing device was more effective in preventing pregnancy than was the Nova T. Only one trial compared immediate vs. delayed insertion. In this trial, the performance of the Copper 7 IUD inserted immediately after abortion was inferior to that after interval insertion (remote from pregnancy), although the differences were not statistically significant. REVIEWERS' CONCLUSIONS: Insertion of an IUD immediately after abortion is both safe and practical. IUD expulsion rates appear higher than after interval insertions.

Abortion, Induced↗

Prenatal detection of velamentous insertion of the umbilical cord: a prospective color Doppler ultrasound study.

OBJECTIVE: Velamentous insertion of the umbilical cord, with a reported incidence of 1% in singleton pregnancies, has been associated with several obstetric complications including fetal growth restriction, prematurity, congenital anomalies, low Apgar scores, fetal bleeding and retained placenta. The aim of this study was to determine the feasibility of identifying velamentous insertion of the umbilical cord during routine obstetric ultrasound. DESIGN: This was a prospective, cross-sectional ultrasound study in 832 unselected second- and third-trimester singleton pregnancies. Color Doppler ultrasound was routinely performed to identify the placental cord insertion site. The role of three-dimensional (3D) ultrasound in evaluating the placental cord insertion site was also studied in a subset of 50 pregnancies from this population. RESULTS: The placental cord insertion site was identified in 825/832 (99%) cases. Visualization was not achieved in seven third-trimester pregnancies with a posterior placenta. A velamentous insertion was suspected prenatally in eight cases, seven of which were confirmed after delivery as velamentous and one as markedly eccentric (battledore placenta). 3D ultrasound performed poorly at evaluating placental cord insertion site, being less efficient due to poor-quality resolution and far more time-consuming than the combined use of gray-scale and color Doppler ultrasound. CONCLUSIONS: Velamentous insertion of the umbilical cord can reliably be detected prenatally by gray-scale and color Doppler ultrasound. 3D imaging had limited value in the evaluation of the placental cord insertion site in our subset of patients. Systematic assessment of the placental cord insertion site at routine obstetric ultrasound has the potential of identifying pregnancies with velamentous insertion and, therefore, those at risk for obstetric complications including vasa previa.

Adult↗

The postnatal development of the insertions of the medial collateral ligament in the rat knee.

Bone soft tissue remodelling at the femoral and tibial insertions of the medial collateral ligament (MCL) of the rat knee was monitored at regular intervals from birth to 120 days of age in 40 Sprague Dawley rats. At birth the femoral insertion originated from the perichondrium of the epiphysis. By day 8 the perichondrium within the insertion had turned into fibrocartilage. Secondary ossification of the femoral epiphysis had progressed in the region near to the insertion site by day 15. The epiphyseal cartilage was entirely replaced by bone by day 40 except for the fibrocartilage within the insertion. After that stage, no qualitative change in zonal insertion characteristics was observed, but only increase in size and decrease in cellularity. At birth, the tibial ligament inserted onto the thin cortical bone of the metaphysis via periosteum. At day 8, osteoclasts started to resorb the thin cortical bone at the ligament insertion, thus forming a metaphyseal depression between days 10 and 20. From days 20 to 120, the insertion remained qualitatively unchanged, showing three zones, the ligament, periosteum, and metaphyseal trabecular bone. The deep periosteal layer showed osteoclastic activity in the proximal part and osteoblastic activity in the distal part. The migration-mechanism of the ligament insertion during growth seems to be caused by this growth-related osteoclastic resorption of the proximal metaphyseal bone and by simultaneous osteogenic activity, which successively cements the distal part of the ligament to bone. The persistence of the periosteal layer and the metaphyseal depression for up to 120 days may be regarded as a sign of continuing growth in this animal model. This is the first investigation showing that the formation of the metaphyseal depression is a purely postnatal event, and suggests that this process might be initiated by the change in mode of growth and joint biomechanics after birth, enabling ligament development and migration in a growing and increasingly loaded weight-bearing joint. The mainly resorptive process, which takes place during development of the tibial MCL insertion, may account for the tensile failure of this ligament that commonly occurs at this site during growth. The pronounced morphological differences between the chondral femoral and the periosteal tibial attachment of the adult MCL are apparently caused by the different postnatal development processes at epiphyses and metaphyses.

Animals↗

Co-administration of alfentanil-propofol improves laryngeal mask airway insertion compared to fentanyl-propofol.

PURPOSE: Insertion of the laryngeal mask airway (LMA) requires sufficient depth of anesthesia to relax the jaw and obtund airway reflexes. Recent studies suggest that the short-acting opioid alfentanil provides the best insertion conditions. We therefore compared the insertion conditions following co-administration of alfentanil-propofol with more commonly used fentanyl-propofol. METHODS: One hundred forty ASA I or II patients, age 18-81 yr, requiring minor surgery were recruited. They were randomized to receive either alfentanil (10 microg x kg(-1); n = 73) or fentanyl (1 microg x kg(-1); n = 67) with propofol (2.5 mg x kg(-1)) 90 sec prior to LMA (size 3 or 4) insertion. A six variable (mouth opening, ease of insertion, swallowing, coughing, movement and laryngospasm) three-point (nil / partial / total) score was used to assess insertion conditions. Duration of post-insertion apnea was recorded. Insertion conditions were compared using Chi-square for trends. RESULTS: The two groups were demographically similar. Mouth opening and ease of insertion were not improved with alfentanil co-administration. Alfentanil-propofol reduced the incidence of swallowing, gagging, movement and laryngospasm (P < 0.05), with 29% (alfentanil) compared to 45% (fentanyl) of patients responding (P = 0.05) to LMA insertion. Apnea [mean (SD)] following alfentanil lasted 154 (139) sec compared to 82 (61) sec following fentanyl (P = 0.001). CONCLUSION: Co-administration of alfentanil-propofol provided better insertion conditions than fentanyl-propofol, though apnea was prolonged by 72 sec.

Adolescent↗

Glottic insertion of the ProSeal LMA occurs in 6% of cases: a review of 627 patients.

PURPOSE: Glottic insertion of the ProSeal Laryngeal Mask Airway (PLMA) has received little attention in the anesthesiology literature. We investigated the incidence and depth of insertion associated with this important cause for a failed insertion attempt with the PLMA. METHODS: With Institutional Review Board approval, we reviewed 15-months' use of the PLMA. Diagnosis of glottic insertion involved a test with children's bubble solution placed on the drain tube port, as well as a fibreoptic examination of the airway of patients experiencing airway obstruction. Patients were anesthetized and paralyzed and the PLMA was inserted deflated with the fingertip method (women size 4, men size 5). The cuff was inflated and a soap membrane established on the drain tube port. Glottic insertion was diagnosed by applying fingertip pressure to the patient's chest wall and observing pulmonary exhalation via the drain tube and bubble formation. The PLMA was then removed and reinserted without further assessment. For all patients, we used a fibrescope to determine the cause of unexplained airway obstruction after the PLMA was considered successfully inserted. RESULTS: There were 627 patients (391 women, 236 men). We diagnosed glottic insertion in 38/627 (6.1%) patients, 37 by the soap membrane test and one with airway obstruction and direct fibreoptic visualization of malposition. Following glottic insertion, successful reinsertion of the PLMA behind the larynx was always associated with greater depth of insertion by an average 2.0 cm. CONCLUSION: Glottic insertion can be easily and quickly diagnosed and our results suggest the incidence and importance of malposition are under-reported in the literature.

Adolescent↗

Characterization of an internal permissive site in the cholera toxin B-subunit and insertion of epitopes from human immunodeficiency virus-1, hepatitis B virus and enterotoxigenic Escherichia coli.

We previously described the construction of novel hybrid proteins based on the B-subunit of cholera toxin (CTB) [Bäckström et al., Gene 149 (1994) 211-217], in which a neutralizing B-cell epitope from the third variable (V3) loop in the envelope glycoprotein gp120 from human immunodeficiency virus type 1 (HIV-1) was inserted within a surface-exposed region between amino acids (aa) 55 and 64. The resulting protein retained properties of native CTB and could induce strong anti-CTB antibody (Ab) responses, but the inserted gp120 epitope was only modestly immunogenic. In this study, the potential use of this internal permissive site in CTB for the insertion of heterologous epitopes has been further investigated. Six additional plasmids were constructed encoding HIV::CTB hybrid proteins with ten to fourteen aa from the V3 loop of gp120 genetically inserted at different positions between aa 52 and 65, with deletions of different CTB aa. Plasmids encoding proteins with peptides inserted between aa 53 and 64 in CTB gave rise to stable proteins which reacted with CTB-specific monoclonal antibodies (mAb) and bound to GM1 gangliosides (GM1), indicating that insertions between these positions do not drastically alter the conformation or the receptor-binding properties of native CTB. Plasmids were also constructed encoding CTB hybrid proteins which had either an 11-aa peptide from hepatitis B virus (HBV) pre-S(2) or one of two peptides related to the heat-stable toxin (STa) of enterotoxigenic Escherichia coli inserted between aa 55 and 64 of CTB. This resulted in the production of HBV::CTB or ST::CTB hybrid proteins and illustrated that the internal permissive site can be used for insertion of peptides of varying aa composition. The reactivity of the inserted epitopes with epitope-specific mAb in GM1-ELISA and immunoblots varied greatly between hybrid proteins and depended on the position in CTB and the aa composition of the inserted peptides. Despite these differences, all the HIV::CTB, ST::CTB and HBV::CTB hybrid proteins could induce low, but significant, levels of serum Ab in mice against gp120, STa or pre-S(2), in addition to strong serum Ab responses against CTB. The Ab response against the internally inserted gp120 peptide was similar to that against the same peptide fused to the N-terminus of CTB, indicating that internally placed peptides had similar immunogenicity to the same peptides added terminally.

Amino Acid Sequence↗

Characterization of a nondeleterious L1 insertion in an intron of the human factor VIII gene and further evidence of open reading frames in functional L1 elements.

We have characterized an insertional event in IVS-10 of the factor VIII gene in a pedigree containing a hemophilia A patient (JH-25). The inserted DNA is a 5' truncated L1 element that is 681 bp long followed by a 3'66-bp poly(A) tract. The L1 element is inserted 154 bp 5' to the start of exon 11 and is flanked by a 13- to 17-bp target site duplication. The L1 insertion is present in four generations of the patient's family. The maternal grandfather who carries the insertion does not have hemophilia A, indicating that the insertion is not the cause of hemophilia A in the patient. We have sequenced this insertion and two previously reported de novo L1 insertions in the factor VIII gene in patients JH-27 (3785 bp) and JH-28 (2132 bp). The three nucleotide sequences differ by 0.2-0.8%. All three of these L1 insertions have open reading frames (ORFs) (1192, 642, and 157 aa) and the three derived amino acid sequences are 98-99% identical. The previously reported sequence similarity between L1 3' ORFs and the polymerase domain of reverse transcriptases is maintained in the ORFs of the JH-27 and JH-28 L1 insertions. The presence of open reading frames and the close sequence similarity of these recently inserted L1 elements provide indirect evidence for the existence of a set of functional L1 elements that encode one or more proteins necessary for their retrotransposition.

Base Sequence↗

Parameters affecting the drug release from in situ gelling nasal inserts.

The purpose of the study was to investigate the influence of physicochemical drug properties, drug loading, and composition of the release medium on the drug release from in situ gelling nasal inserts. Sponge-like nasal inserts of carrageenan and HPMC K15M with the model drugs oxymetazoline HCl, diprophyllin, and acetaminophen (APAP) were prepared by lyophilization. Drug release studies at different drug loadings were performed in various release media. Raman analysis, DSC, and SEM were conducted to analyze the physical state of the drugs in the inserts. All drugs were dissolved in the solid HPMC inserts and were released at similar rates at all investigated loadings except for the least soluble APAP. APAP concentrations in the hydrating HPMC K15M inserts in excess of its solubility limit resulted in reduced relative release rates at higher drug loadings. Drug-polymer interactions (formation of less soluble drug-polymer salts) resulted in a slower release of oxymetazoline HCl from carrageenan inserts than from HPMC K15M inserts. Changes in the composition of the release medium affected the water uptake of carrageenan but not of HPMC K15M inserts. Oxymetazoline release from carrageenan inserts increased with higher Na+-content of the medium because of ion exchange and at low (pH 2) as well as at high pH (pH 10). The osmolality of the release medium had no effect. The solubility of the drug, its physical state in the polymer matrix, and drug-polymer interactions governed the drug release from nasal inserts. The release from inserts prepared with oppositely charged polymers and drugs was influenced by electrostatic drug-polymer interactions and by the composition of the release medium.

Administration, Intranasal↗

Ultrasound: an effective method for localization of the echogenic Essure sterilization micro-insert: correlation with radiologic evaluations.

STUDY OBJECTIVE: To examine the reliability and practicality of performing office-based transvaginal ultrasound for determining the ease of locating the Essure hysteroscopic sterilization micro-insert and compare its usefulness against established radiologic evaluations. DESIGN: Prospective single-center, single-arm, clinical study (Canadian Task Force classification xx). SETTING: Hospital-based clinical research center. PATIENTS: One hundred forty-five women of reproductive age and proven fertility. INTERVENTION: Thirty-seven women who underwent the Essure hysteroscopic method of sterilization had routine radiologic and transvaginal ultrasound assessments for determining the retention of these micro-inserts from 3 months to 2 years after placement. An additional 108 women had ultrasound assessment as the only means of micro-insert localization 3 months after placement. MEASUREMENTS AND MAIN RESULTS: The 145 women (100%) who underwent an ultrasound assessment at a 3-month, posthysteroscopic-sterilization office visit had their micro-inserts readily identified and localized to the uterotubal area due to the micro-inserts' dense echogenic properties. For the 37 women who had both assessments, the ultrasound findings correlated with pelvic radiograph and hysterosalpingogram assessments of micro-insert location in all instances. In addition, the ultrasound evaluations provided additional information about the micro-inserts relative position to the surrounding, less-echogenic soft tissue structures of the upper uterotubal area. In the 37 women who had serial ultrasound and radiologic evaluations performed for up to 2 years after micro-insert placement, ultrasound was found to be equally effective in identifying the location of the micro-inserts and indicted that their position remained identifiable and stable over time. CONCLUSION: A single transvaginal ultrasound in-office examination, performed 3 months after hysteroscopic micro-insert placement, was found to be a simple, reliable, and convenient method of assessing micro-insert location, when compared with radiologic assessments.

Ambulatory Surgical Procedures↗

Declines in stimulated striatal dopamine release over the first 32 h following microdialysis probe insertion: generalization across releasing mechanisms.

In a recent paper [R.R. Holson, J.F. Bowyer, P. Clausing, B. Gough, Methamphetamine-stimulated striatal dopamine release declines rapidly over time following microdialysis probe insertion, Brain Res. 739 (1996) 301-307] we reported that methamphetamine-stimulated striatal dopamine release declined rapidly over the first eight hours following microdialysis probe insertion. This decline was strictly a function of time post-probe implantation, and not due to tolerance or desensitization. To further examine this phenomenon, we subjected rats to three brief pulses of several DA-releasing compounds at 2, 4 and 6 h post-probe insertion, and compared these results to those caused by a single pulse 6 h post-insertion, or in some cases to pulses given more than 24 h post-insertion. We found that when buproprion, a dopamine reuptake blocker, was infused briefly into the striatum via the microdialysis probe, there was a pronounced drop in the amount of dopamine released at 6 h vs. 2 h post-insertion; this drop was not due to repeated exposure, since dopamine release at 6 h post-insertion was the same for a single pulse, or when preceded by two earlier pulses. Twenty-four hours later, buproprion-stimulated dopamine release was still lower, but did not appear to drop further thereafter. Potassium-stimulated dopamine release, on the other hand, dropped rapidly over the first 8 h post-insertion, and this decline continued throughout the 24-32 h interval post-insertion. Similarly, a single i.p. injection of 0.5 mg/kg haloperidol released three times as much dopamine when given two compared to six hours post-implantation. Both bupropion- and potassium-stimulated dopamine release were accompanied by declines in extracellular DOPAC concentrations, and these declines were the same 2 or 26 h post-insertion. In contrast, haloperidol exposure increased extracellular DOPAC, and this haloperidol-stimulated DOPAC increase was also greatly attenuated at 6 compared to 2 h post-insertion. We conclude that there is a general decline over time post-probe implantation in the ability of the striatal dopamine system to release dopamine, and perhaps to increase dopamine synthesis, in response to pharmacological challenges.

3,4-Dihydroxyphenylacetic Acid↗

Mucoadhesive ocular insert based on thiolated poly(acrylic acid): development and in vivo evaluation in humans.

The aim of the study was to develop a mucoadhesive ocular insert for the controlled delivery of ophthalmic drugs and to evaluate its efficacy in vivo. The inserts tested were based either on unmodified or thiolated poly(acrylic acid). Water uptake and swelling behavior of the inserts as well as the drug release rates of the model drugs fluorescein and two diclofenac salts with different solubility properties were evaluated in vitro. Fluorescein was used as fluorescent tracer to study the drug release from the insert in humans. The mean fluorescein concentration in the cornea/tearfilm compartment as a function of time was determined after application of aqueous eye drops and inserts composed of unmodified and of thiolated poly(acrylic acid). The acceptability of the inserts by the volunteers was also evaluated. Inserts based on thiolated poly(acrylic acid) were not soluble and had good cohesive properties. A controlled release was achieved for the incorporated model drugs. The in vivo study showed that inserts based on thiolated poly(acrylic acid) provide a fluorescein concentration on the eye surface for more than 8 h, whereas the fluorescein concentration rapidly decreased after application of aqueous eye drops or inserts based on unmodified poly(acrylic acid). Moreover, these inserts were well accepted by the volunteers. The present study indicates that ocular inserts based on thiolated poly(acrylic acid) are promising new solid devices for ocular drug delivery.

Acrylic Resins↗

Cellular insertions in the NS2-3 genome region of cytopathic bovine viral diarrhoea virus (BVDV) isolates.

When compared to noncytopathic (ncp) bovine viral diarrhoea virus (BVDV), some cytopathic (cp) BVDV contain additional sequences in the NS2-3 genomic region. One of these insertions, which is 270 nucleotides long and of host origin (cINS), was first described for strain NADL. To find out how frequently this type of insertion occurs in other cp BVDV, 32 cp BVDV field isolates and the BVDV reference cp strain Indiana were screened using RT-PCR which detected cINS in NADL. For most cp viruses an RT-PCR product of 402bp indicated the presence of NS2-3 genes without insertions. In addition, one or two DNA fragments, around 600-850bp in size, were amplified from the genomes of 13 cp viruses indicating the presence of insertions. Sequencing of the PCR products, i.e. 402bp DNA fragment (with no insertion) and longer fragments (with insertion) revealed the location of the insertions in the NS2-3 coding region of eight cp BVDV genomes. All of the insertions were confirmed to be of the cINS type and were located in a very similar position to that found previously in the NADL genome. They were in the same reading frame as the viral polypeptide and they encoded 90-140 amino acids. The 5' and 3' ends of the insertions were different in most of the cp isolates studied. Interestingly, a 14-amino-acid stretch at the 5'-end of the insertion in the cp 5569 isolate as well as 15 amino acids at the 3'-end of the insertion in the cp 5.19516 isolate were not homologous to the cINS sequence. No significant matches for these stretches were found in the EMBL and Swissprot databases.

Amino Acid Sequence↗

Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation.

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.

Administration, Intravaginal↗

Laryngeal mask insertion during target-controlled infusion of propofol.

STUDY OBJECTIVE: To compare the Laryngeal Mask Airway (LMA; The Laryngeal Mask Airway Co., Ltd., Nicosia, Cyprus) insertion conditions produced by 6 and 8 microg/mL of target plasma concentrations (Cpt) during the induction of anesthesia with target-controlled infusion (TCI) of propofol. DESIGN: Randomized, prospective, single-blind, clinical study. SETTING: University hospital. PATIENTS: 44 ASA physical status I and II patients, 16 to 54 years of age, weighing between 45 and 100 kg, undergoing minor surgery in which the use of LMA was indicated. INTERVENTIONS: Patients were randomly divided into two groups (1 and 2) of 22 to compare the effects of different propofol concentrations. Three minutes after intravenous (IV) injection of midazolam 0.04 mg/kg, group 1 and 2 received TCI of propofol with 6 and 8 microg/mL of Cpt, respectively. LMA was inserted when the effect-site concentration (EC) reached 2.5 microg/mL, which was displayed on the infusion pump. MEASUREMENTS: The LMA insertion conditions (mouth opening, gagging, coughing, head or limb movement, laryngospasm, overall ease of insertion) were assessed, and hemodynamic responses were evaluated until 3 minutes after LMA insertion. Total dose of propofol, EC, and elapsed time since the start of TCI were recorded at five times: at the loss of consciousness and eyelash reflex, at 2.5 microg/mL of EC, and immediately, 1 minute, and 3 minutes after the insertion of LMA. MAIN RESULTS: There was no significant difference between the two groups in insertion conditions, despite the significantly larger total dose and shorter elapsed time (2.6 +/- 0.08 mg/kg and 109 +/- 5.0 s) in Group 2 than those (2.1 +/- 0.02 mg/kg and 140 +/- 4.1 s) in Group 1 at 2.5 microg/mL of EC (p < 0.05). Systolic and diastolic blood pressure decreased and heart rate increased significantly throughout the study period in both groups (p < 0.05). But there was a significant decrease in arterial pressure in Group 2 compared with Group 1 1 and 3 minutes after the insertion (p < 0.05). CONCLUSIONS: Induction with 8 microg/mL of Cpt, compared with 6 microg/mL, allowed earlier LMA insertion but, could not improve the conditions for LMA insertion and required more careful attention to the decrease in blood pressure after LMA insertion.

Adolescent↗