Endometrial thickness following medical abortion is not predictive of subsequent surgical intervention.

OBJECTIVES: To evaluate the ability of endometrial thickness after medical abortion to predict the need for subsequent dilatation and curettage (D&C). METHODS: We pooled data from two multicenter medical abortion trials involving 2208 women who received mifepristone orally followed by misoprostol vaginally. Women returned for transvaginal ultrasonography approximately 7 days later. The endometrial thickness was measured if no gestational sac was present. Final status was confirmed by a phone interview at 5 weeks. The area under the receiver-operating characteristics (ROC) curve was calculated to assess the overall ability of endometrial thickness to predict the need for subsequent D&C. Endometrial thickness was dichotomized using threshold values at 5-mm increments from 10 to 30 mm. The sensitivity, specificity, negative predictive value and positive predictive value were calculated to evaluate the ability of each endometrial thickness threshold value to predict subsequent D&C. Multivariable regression analysis was performed to adjust endometrial thickness values for study, treatment group, and study site. RESULTS: At 7 days after misoprostol treatment, 1870 women (84.7%) had endometrial thickness assessed. Thirty of these women (1.6%) subsequently underwent D&C. The mean endometrial thickness was 14.5 mm for women who underwent D&C and 10.9 mm for those who did not (difference 3.5 mm (95% CI, 1.8-5.3 mm)). Endometrial thickness was poorly predictive of the need for D&C, with an area under the ROC curve of 0.65. All endometrial thickness thresholds had positive predictive values of 25% or less. The results were unchanged by adjustment of endometrial thickness values by multivariable modeling. CONCLUSIONS: Although endometrial thickness following successful expulsion of the gestational sac is thicker in women who will eventually require surgical intervention after medical abortion, endometrial thickness is not a clinically useful predictor of the subsequent need for D&C.

Endometrial thickness after misoprostol use for early pregnancy failure.

OBJECTIVES: To assess if there was any potential relationship between endometrial thickness and final treatment outcome in women successfully treated with misoprostol for a first trimester anembryonic gestation, embryonic demise or fetal demise. METHODS: Eighty women were treated with up to two doses of misoprostol 800 microg vaginally for early pregnancy failure. Subjects were scheduled to return 2 (range 1-4), 7 (range 5-9) and 14 (range 12-17) days after treatment. Transvaginal ultrasonography was performed at each follow-up visit. RESULTS: The median endometrial thickness at each of the follow-up visits for women who had expelled the gestational sac was 14 mm, 10 mm, and 7 mm, respectively. The endometrial thickness at the first follow-up visit exceeded 15 mm in 20 subjects (36%) and 30 mm in four subjects (7%). Only three women had a suction aspiration for bleeding after documented expulsion. The endometrial thickness for these women was 11, 13, and 14 mm at the first follow-up visit. CONCLUSIONS: There is no obvious relationship between increasing endometrial thickness and the need for surgical intervention in women treated with misoprostol for early pregnancy failure.

Interleukin-1 receptor antagonist as an adjunct in the treatment of Haemophilus influenzae otitis media in the chinchilla.

OBJECTIVES: This purpose of this study was to investigate the effect of blockade of the inflammatory cytokine pathway on experimentally induced otitis media in the chinchilla model. STUDY DESIGN: Pilot, randomized placebo-controlled trial. METHODS: Ampicillin-sensitive Haemophilus influenzae otitis media was induced in 45 adult chinchillas. The animals were randomly assigned to the following treatment groups: 1) transbullar injections (TBI) of interleukin-1 receptor antagonist (IL-1ra) and intramuscular ampicillin, 2) TBI of saline and intramuscular ampicillin, 3) TBI of IL-1ra and intramuscular sa-1 line or 4) TBI of saline and intramuscular saline. Blinded investigators measured resolution of otitis media by otomicroscopy, tympanogram, and culture results. RESULTS: Comparisons were made between the treatment groups to assess the ability of IL-1ra to assist with resolution of otitis media using exact two-group binomial tests with the StatXact statistical program. The group with TBI of IL-1ra and intramuscular ampicillin as a treatment demonstrated trends suggesting more rapid resolution of positive cultures and more rapid and complete return to normal results on tympanograms and otomicroscopic findings compared with the group treated with TBI of saline and intramuscular ampicillin. These trends did not achieve statistical significance with the relatively small sample sizes used in this pilot study. CONCLUSIONS: This investigation provides further evidence that the inflammatory cytokine cascade plays a significant role in the pathophysiology of otitis media and that modulation of this inflammatory pathway may provide novel and efficacious treatments for otitis media Further studies with larger groups of animals are warranted to determine whether the trends identified in this pilot study are reproducible and achieve statistical significance.

Relevant issues in revision canal-wall-down mastoidectomy.

The role of canal-wall-down (CWD) versus intact-canal-wall (ICW) mastoidectomy in the treatment of chronic otitis media with or without cholesteatoma remains a debated issue. Although past conventional wisdom has held that CWD mastoid surgery leads to a "safe" ear and is technically less demanding than the more controversial ICW mastoidectomy, this is often not the case. Although our preference is an ICW technique when possible, 47 of 109 (43%) mastoid procedures for chronic otitis media performed between January 1993 and June 1996 involved a CWD mastoid cavity. More than two thirds of these procedures (32 of 47) represented revision surgery, the most common indication being a poorly contoured, preexisting CWD mastoidectomy with persistent otorrhea. A dry, well-epithelialized ear was obtained in 90% of cases. Our preferred method of ossicular reconstruction (double cartilage block ossiculoplasty) is detailed, and hearing results according to American Academy of Otolaryngology Head and Neck Surgery guidelines are reported.

Antisense oligonucleotide inhibition of hepatitis C virus (HCV) gene expression in livers of mice infected with an HCV-vaccinia virus recombinant.

Hepatitis C virus (HCV) is the major cause of non-A, non-B hepatitis worldwide. Current treatments are not curative for most infected individuals, and there is an urgent need for both novel therapeutic agents and small-animal models which can be used to evaluate candidate drugs. A small-animal model of HCV gene expression was developed with recombinant vaccinia virus vectors. VHCV-IRES (internal ribosome entry site) is a recombinant vaccinia viral vector containing the HCV 5' nontranslated region (5'-NTR) and a portion of the HCV core coding region fused to the firefly luciferase gene. Intraperitoneal injection of VHCV-IRES produced high levels of luciferase activity in the livers of BALB/c mice. Antisense oligonucleotides complementary to the HCV 5'-NTR and translation initiation codon regions were then evaluated for their effects on the expression of these target HCV sequences in BALB/c mice infected with the vaccinia virus vector. Treatment of VHCV-IRES-infected mice with 20-base phosphorothioate oligonucleotides complementary to the sequence surrounding the HCV initiation codon (nucleotides 330 to 349) specifically reduced luciferase expression in the livers in a dose-dependent manner. Inhibition of HCV reporter gene expression in this small-animal model suggests that antisense oligonucleotides may provide a novel therapy for treatment of chronic HCV infection.

Doramectin efficacy against the kidney worm, Stephanurus dentatus in sows.

Sixteen sows passing Stephanurus dentatus eggs in their urine were bought on the local market and placed in individual pens with solid concrete floors in an open-sided bam. Water was supplied by nipple waterers and sows were fed individually 1.8 kg feed daily. First, urine samples were taken at dawn on days -8 and -7 and weights were taken on day 0. Sows were assigned to one of two groups on the basis of average urine egg counts and weights. Group T1 sows were each injected IM in the neck with sterile saline at the rate of 1.5 ml per 50 kg and group T2 sows were each injected IM with doramectin at the rate of 300 micrograms kg-1 on day 0. Urine samples were again taken on days 56 and 57 and the sows were necropsied on day 57. Urine of all doramectin treated sows were test negative for kidney work eggs on days 56 and 57 as was one control sow, whereas the average count for controls was 3762 eggs ml-1. No worms were found in doramectin treated sows and a total of 499 were found in the controls for an average of 62 per sow. The majority of worms were in the perirenal area and kidneys, a few were scattered in liver, lungs, abdominal muscles and peritoneal cavity. The efficacy of doramectin against Stephanurus dentatus in sows was 100% (P < 0.0001).

Doramectin efficacy against gastrointestinal nematodes in pigs.

Four controlled trials with growing pigs were performed to determine efficacy of doramectin against natural and induced populations of nematodes. In Trial 1 (T1), 20 pigs with natural infections were assigned to one of two like groups on the basis of weight, sex and worm egg counts. In Trial 2 (T2), 20 pigs with negative worm egg counts were assigned to one of two groups on the basis of weight and sex. Each pig was subsequently given (per os) 3000 Trichuris suis embryonated eggs; 2000 Ascaris suum embryonated eggs; 10000 Oesophagostomum spp. infective larvae and 10,000 Strongyloides ransomi infective larvae (SC injection). In Trial 3 (T3), 20 pigs with negative worm egg counts were assigned as in T2, and each pig was subsequently given (per os) 2000 A. suum embryonated eggs, 15000 Oesophagostomum quadrispinulatum infective larvae, and 2891 Hyostrongylus rubidus infective larvae. In Trial 4 (T4), 16 pigs with negative worm egg counts were each assigned to one of two groups as in T2 and were given (per os) 2670 T. suis embryonated eggs. On Day 0 of each trial, each pig of the control group was injected IM in the neck with sterile saline at the rate of 1.5 ml 50 kg-1. Each pig in the treated group of each trial was similarly injected with doramectin at the rate of 300 micrograms kg-1. All pigs were necropsied 14 or 15 days post-treatment and parasites recovered by standard parasitological procedures. Efficacies against natural infections were: A. suum, 100%; Oesophagostomum spp. 100%; H. rubidus, 99.4%; and Strongyloides ransomi, 99.9%. Efficacies against induced infections were: 4th stage A. suum, 100%; 4th stage O.dentatum, 99.9%; 4th stage O.quadrispinulatum, 97.1 and 99.6%; 4th stage H. rubidus, 100%; adult S. ransomi, 100%; adult Trichuris suis in mixed infection, 54.1%; and in pure infection, 95.3%.

Inhibition of human cytomegalovirus immediate-early gene expression by an antisense oligonucleotide complementary to immediate-early RNA.

ISIS 2922 is a phosphorothioate oligonucleotide that is complementary to human cytomegalovirus (CMV) immediate-early (IE) RNA and that exhibits potent and specific antiviral activity against CMV in cell culture assays. Specific assay systems were developed to separately characterize the antisense and nonantisense components of the antiviral activity mediated by ISIS 2922. In U373 cells transformed with cDNA encoding the CMV IE 55-kDa (IE55) protein, expression was inhibited at nanomolar concentrations comparable to effective concentrations in antiviral assays. The specificity of inhibition was demonstrated by using control oligonucleotides incorporating progressive base changes to destabilize oligonucleotide-RNA base pairing and by showing a lack of inhibition of the CMV IE72 product expressed from the same promoter. Inhibition of IE55 protein expression correlated with a reduction in mRNA levels consistent with an RNase H-mediated termination event. Studies with virus-infected cells demonstrated that antisense and nonantisense mechanisms contribute to the antiviral activity of ISIS 2922. Base complementarity to target RNA was important for optimal activity in antiviral assays, but base changes affecting parameters other than hybridization affinity also influenced antiviral activity. Sequence-independent inhibition of virus adsorption to host cells by phosphorothioate oligonucleotides was also observed at high concentrations. Therefore, at least three different mechanisms may contribute to the antiviral activity of ISIS 2922 in cell culture: antisense-mediated inhibition of target gene expression; nonantisense, sequence-dependent inhibition of virus replication; and sequence-independent inhibition of virus adsorption to host cells.

Antisense oligonucleotide inhibition of hepatitis C virus gene expression in transformed hepatocytes.

Genetic and biochemical studies have provided convincing evidence that the 5' noncoding region (5' NCR) of hepatitis C virus (HCV) is highly conserved among viral isolates worldwide and that translation of HCV is directed by an internal ribosome entry site (IRES) located within the 5' NCR. We have investigated inhibition of HCV gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon, and core protein coding sequences. Oligonucleotides were evaluated for activity after treatment of a human hepatocyte cell line expressing the HCV 5' NCR, core protein coding sequences, and the majority of the envelope gene (E1). More than 50 oligonucleotides were evaluated for inhibition of HCV RNA and protein expression. Two oligonucleotides, ISIS 6095, targeted to a stem-loop structure within the 5' NCR known to be important for IRES function, and ISIS 6547, targeted to sequences spanning the AUG used for initiation of HCV polyprotein translation, were found to be the most effective at inhibiting HCV gene expression. ISIS 6095 and 6547 caused concentration-dependent reductions in HCV RNA and protein levels, with 50% inhibitory concentrations of 0.1 to 0.2 microM. Reduction of RNA levels, and subsequently protein levels, by these phosphorothioate oligonucleotides was consistent with RNase H cleavage of RNA at the site of oligonucleotide hybridization. Chemically modified HCV antisense phosphodiester oligonucleotides were designed and evaluated for inhibition of core protein expression to identify oligonucleotides and HCV target sequences that do not require RNase H activity to inhibit expression. A uniformly modified 2'-methoxyethoxy phosphodiester antisense oligonucleotide complementary to the initiator AUG reduced HCV core protein levels as effectively as phosphorothioate oligonucleotide ISIS 6095 but without reducing HCV RNA levels. Results of our studies show that HCV gene expression is reduced by antisense oligonucleotides and demonstrate that it is feasible to design antisense oligonucleotide inhibitors of translation that do not require RNase H activation. The data demonstrate that chemically modified antisense oligonucleotides can be used as tools to identify important regulatory sequences and/or structures important for efficient translation of HCV.

In vitro evaluation of phosphorothioate oligonucleotides targeted to the E2 mRNA of papillomavirus: potential treatment for genital warts.

Papillomaviruses induce benign proliferative lesions, such as genital warts, in humans. The E2 gene product is thought to play a major role in the regulation of viral transcription and DNA replication and may represent a rational target for an antisense oligonucleotide drug action. Phosphorothioate oligonucleotides complementary to E2 mRNAs were synthesized and tested in a series of in vitro bovine papillomavirus (BPV) and human papillomavirus (HPV) models for the ability to inhibit E2 transactivation and virus-induced focus formation. The most active BPV-specific compounds were complementary to the mRNA cap region (ISIS 1751), the translation initiation region for the full-length E2 transactivator (ISIS 1753), and the translation initiation region for the E2 transrepressor mRNA (ISIS 1755). ISIS 1751 and ISIS 1753 were found to reduce E2-dependent transactivation and viral focus formation in a sequence-specific and concentration-dependent manner. ISIS 1755 increased E2 transactivation in a dose-dependent manner but had no effect on focus formation. Oligonucleotides with a chain length of 20 residues had optimal activity in the E2 transactivation assay. On the basis of the above observations, ISIS 2105, a 20-residue phosphorothioate oligonucleotide targeted to the translation initiation of both HPV type 6 (HPV-6) and HPV-11 E2 mRNA, was designed and shown to inhibit E2-dependent transactivation by HPV-11 E2 expressed from a surrogate promoter. These observations support the rationale of E2 as a target for antiviral therapy against papillomavirus infections and specifically identify ISIS 2105 as a candidate antisense oligonucleotide for the treatment of genital warts induced by HPV-6 and HPV-11.

Performance of pigs with mixed nematode infections before and after ivermectin treatment.

Ninety crossbred parasitized pigs were used in two controlled experiments to compare performance before and after ivermectin treatment. Four groups of 15 pigs were sequentially infected with 2000 Ascaris suum, 10,000 Oesophagostomum spp. and 10,000 Strongyloides ransomi beginning at average pig weights of either 38 kg (Experiment I) or 14 kg (Experiment II). Two infected groups of pigs were treated with ivermectin on Day 34 (Experiment I) or 37 (Experiment II) of the experiments and all pigs were fed to slaughter weights of approximately 100 kg. Average daily gain (ADG) of heavier ivermectin-treated pigs was greater than that of infected non-treated pigs (P less than 0.03) and feed conversion was numerically greater by 5% (P greater than 0.10). ADG was not different between lighter treated or non-treated pigs, but feed conversion was numerically 4% greater in ivermectin-treated compared to non-treated pigs (P greater than 0.10). No differences existed in carcass dressing percentage among the three groups of heavier pigs, but among the lighter groups ivermectin-treated pigs had a higher dressing percentage (P less than 0.03) than infected non-treated or control pigs. Results of the calculation of production costs based on combined data from the two experiments for the post-treatment period, including feed, drug and maintenance costs, indicate that the production cost for ivermectin-treated pigs was $1.53 per pig less than that for infected non-treated pigs when the cost for each group to attain control pig weights is compared with the actual cost for control pigs.

Effect of diaphragmatic respiration with end-tidal CO2 biofeedback on respiration, EEG, and seizure frequency in idiopathic epilepsy.

Breathing rate (RR), end-tidal percent CO2, and EEG were obtained in three groups: psychiatric referral subjects presenting with anxiety, panic phobia, depression and migraine; a group of idiopathic seizure sufferers; and a group of asymptomatic controls. Virtually all the noncontrol subjects were found to show moderate to severe hyperventilation and the accompanying EEG dysrhythmia. The seizure group subjects were taught diaphragmatic respiration with end-tidal percent CO2 biofeedback. The training normalized their respiration and altered their EEGs and seizure frequency.

Excess crude protein for nongravid gilts.

Twenty-five nongravid crossbred gilts (avg initial wt, 126 kg) were placed on either a high (38%) or a low (13%) crude protein (CP) diet and fed either at the rate of 1.82 kg/d or had ad libitum access to feed. In addition, a fifth group was pair-fed the 13% CP diet to the average intake of the gilts fed high CP ad libitum. The experimental period lasted 30 d. Corn-soybean meal diets were used and CP levels were varied by altering the corn:soybean meal ratio. Gain and gain/feed were reduced (P less than .01) in gilts fed 1.82 kg/d compared with the gilts fed ad libitum or pair-fed gilts. Gain (P less than .03) and feed intake (P less than .01) of gilts with ad libitum access to the 13% CP diet were higher than those of gilts with ad libitum access to the 38% CP diet. Gain/feed was not different (P greater than .10) between the two groups, however. Rate of gain and feed efficiency of gilts pair-fed the 13% CP diet were similar (P greater than .10) to those of gilts with ad libitum access to the 38% CP diet. Plasma total free amino acids, NH3 and total protein were not (P greater than .10) affected by treatment. Plasma urea-N and urinary total N, urea-N and orotic acid were increased (P less than .01) in gilts fed the high CP diet regardless of feed intake level. However, urinary NH3 was higher (P less than .01) in gilts fed the low-protein diet. These results indicate that excess dietary CP for nongravid gilts decreases gain and feed intake and has no effect on efficiency of feed utilization, but it increases plasma urea-N and urinary total N, urea-N and orotic acid.

Effect of ammonium chloride ingestion on Eimeria acervulina-infected chicks fed excess copper.

Four experiments were conducted with broiler chicks to investigate the effect of dietary NH4Cl on gain, feed efficiency, duodenal pH, and liver Cu concentration of Eimeria acervulina-infected chicks. Experimental coccidial infection reduced chick gain, feed efficiency, and duodenal pH, but it increased liver Cu concentration in chicks fed excess Cu. Ammonium chloride had no effect on gain, feed efficiency, liver Cu concentration, or on duodenal pH.

Effect of dietary buffer additions on gain, efficiency, duodenal pH, and copper concentration in liver of Eimeria acervulina-infected chicks.

Three experiments were conducted with broiler chicks to investigate the effect of dietary additions of NaHCO3(1%), A1(OH)3(.5%), kaolin(1%), A1(OH)2NaCO3(.23%), CaCO3(.37%), and MgO(1%) on gain, efficiency, duodenal pH, and liver Cu concentration of Eimeria acervulina-infected chicks. Experimental coccidial infection consistently reduced chick gain, efficiency, and duodenal pH, but it increased liver Cu concentration of chicks fed excess Cu. Sodium bicarbonate addition improved chick gain and efficiency slightly, whereas the MgO addition reduced these performance criteria. Sodium bicarbonate improved gain more in coccidiosis-infected chicks than in uninfected chicks, but it failed to alleviate, to any extent, the coccidiosis-induced liver Cu increase of the duodenal pH decrease.

Excess supplemental choline for swine.

Seven experiments were conducted with 280 crossbred pigs to investigate the effect of excess choline on rate and efficiency of gain of weanling, growing and finishing swine Choline additions were made to a conventional corn-soybean meal diet. Daily gain of weanling pigs was reduced slightly by 6,000 ppm excess choline compared with 0, 500, 1,000, 2,000 or 4,000 ppm excess choline. Excess supplemental choline (2,000 ppm) fed throughout the weanling, growing and finishing (121 to 126 d) phases of growth reduced (P less than .08) daily gain but it did not affect (P greater than .10) feed utilization. The 2,000-ppm choline addition, however, did not affect (P less than .10) pig gain when fed only during the growing and finishing stages of growth (68 to 86 d). Excess choline should be avoided in swine diets if maximum rate of gain is to be achieved.

Behavioral control of intractable idiopathic seizures: I. Self-regulation of end-tidal carbon dioxide.

Eleven women and seven men with moderate to severe chronic hyperventilation and idiopathic seizures refractory to therapeutic serum levels of anticonvulsant medication were given diaphragmatic respiration training with percent end-tidal CO2 biofeedback. The training had a rapid correcting effect on their respiration, making it comparable to that of 18 asymptomatic control subjects. Ten of the seizure-group subjects were in the study at least 7 months and following treatment, 8 showed EEG power spectrum "normalization", restoration of cardio-respiratory synchrony (RSA), and their seizure frequency and severity were significantly reduced.