Disclosure of cardiac variants of uncertain significance results in an exome cohort.

This study examined the impact of disclosing subclassifications of genetic variants of uncertain significance (VUS) on behavioral intentions. We studied return of VUS results to 79 individuals with a cardiomyopathy-associated VUS, subclassified into VUS-high or VUS-low. Primary outcomes were perceived risk (absolute and comparative), perceived severity, perceived value of information, self-efficacy, decision regret, and behavioral intentions to share results and change behaviors. There was no significant difference between the 2 subclasses in overall behavioral intentions (t = 0.023, P = .982) and each of the individual items on the behavioral intentions scale; absolute (t = -1.138, P = .259) or comparative (t = -0.463, P = .645) risk perceptions; perceived value of information (t = 0.582, P = .563) and self-efficacy (t = -0.733, P = .466). Decision regret was significantly different (t = 2.148, P = .035), with VUS-low (mean = 17.24, SD = 16.08) reporting greater regret. Combining the subclasses, perceived value of information was the strongest predictor of behavioral intentions (ß = 0.524, P < .001). Participants generally understood the meaning of a genetic VUS result classification and reported satisfaction with result disclosure. No differences in behavioral intentions were found, but differences in decision regret suggest participants distinguish subclasses of VUS results. The perceived value of VUS may motivate recipients to pursue health-related behaviors.

Carcass orientation and drip time affect potential surface water carryover for broiler carcasses subjected to a post-chill water dip or spray1.

To estimate the potential for residual antimicrobial solution carryover, surface water accumulation and loss was measured on post-chill carcasses that were either dipped or sprayed with water. For all experiments, broilers were slaughtered, soft or hard scalded, defeathered, and eviscerated. Carcasses were immersion chilled, allowed to drip, and post-chill carcass weight (CW) recorded. For water dip treatment, carcasses were dipped for 0.5 min in water and hung by a wing (n = 33) or a leg (n = 30) and CW recorded at 0, 0.5, 1, 2, and 5 min post-dip. For water spray treatment, individual carcasses were hung by either the wings (n = 35) or legs (n = 34) from a shackle suspended from a scale. Water was sprayed at 80 psi and post-spray CW recorded. Initial water accumulation (0 min) for dipped carcasses was not significantly different (P > 0.05) for carcasses hung by the leg (101.0 g) or wing (108.8 g). Following the 5 min drip time, 31 g of water remained on the carcasses hung by the leg and only 10 g on carcasses hung by the wing (P < 0.05). When carcasses were sprayed with water, initial water accumulation (0 min) was 62 g for carcasses hung by the legs and 60 g for carcasses hung by the wings (P > 0.05). Following the 5 min drip time, 1 g or no water remained on the sprayed carcasses (P > 0.05). Carcasses that were dipped and hung by a leg for 5 min retained significantly more water (31 g) than carcasses that were dipped and hung by a wing (10 g) or sprayed carcasses hung either way (0.3 g) (P < 0.05). Post-chill water dip resulted in significantly higher initial carcass water accumulation than spraying (105 g vs. 61 g, P < 0.05). Carcass orientation during dripping only affected the amount of retained water for dipped carcasses. Dipped carcasses hung by a leg have the highest potential for residual carcass antimicrobial solution carryover and sprayed carcasses hung by either orientation have the lowest potential for residual antimicrobial solution carryover.

Effect of method of delivering nicarbazin to mallards on plasma 4,4'-dinitrocarbanilide levels and reproduction.

Nicarbazin (NCZ), a coccidiostat used in the poultry industry, has been developed as a contraceptive for resident Canada geese. We tested the efficacy of NCZ as a contraceptive using mallards (Anas platyrhynchos) as a model for Canada geese. Nicarbazin-treated corn was fed ad libitum for 14 d at 0, 750, 1,000, or 1,500 ppm. Plasma and egg levels of 4,4'-dinitrocarbanilide (DNC), the active anticoccidial component of NCZ, differed among treatment groups in a dose-response relationship, but plasma levels did not differ between sexes. Nicarbazin caused a decrease in egg weight, but there was no effect of NCZ on the numbers of eggs laid per female per day. Nicarbazin did not significantly impact bird health. An additional trial tested the effect of the method of NCZ delivery on plasma DNC levels. Mallards were given NCZ daily for 12 d either by gavage with a corn oil suspension, gavage with a water suspension, peroral administration of a capsule, or feeding 500 mg of NCZ/kg of pelleted feed ad libitum. The method of delivery significantly affected plasma DNC levels, with the highest levels in the corn oil suspension group and the lowest levels in the pelleted feed group. This is likely due to decreased availability of NCZ in a pellet compared with gavage with a suspension or capsule. Mallards receiving 34.2 mg of NCZ/kg of BW when fed cracked corn coated with NCZ daily for 14 d had higher plasma DNC levels than those obtained by liquid gavage, capsule, or pelleted NCZ feed. For maximum effect in the field, NCZ should be coated onto corn. A higher concentration of NCZ is needed in pelleted feed to obtain comparable plasma DNC levels to allow for the decreased absorption of DNC.

Evaluation of nicarbazin as a potential waterfowl contraceptive using mallards as a model.

Contraception may provide a useful nonlethal management tool to reduce wild bird populations. We tested the efficacy of nicarbazin (NCZ) as a contraceptive for waterfowl and assessed health effects of NCZ, using domestic mallards (Anas platyrhynchos) as a model for Canada geese (Branta canadensis). Mallards were given gelatin capsules containing 0, 8.5, 17.0, or 33.75 mg of NCZ/kg of BW perorally once daily for 14 d. Fecal 4,4'-dinitrocarbanilide (DNC) and fluorescein were evaluated as potential markers of plasma and egg DNC levels. Plasma, egg, and fecal DNC levels differed among treatment groups in a dose response relationship. There were no significant effects on the numbers of eggs laid per female per day, proportion of fertile eggs, proportion of eggs hatching, or egg yolk mottling. Hatchability was 0.55 +/- 0.1 in the control group compared with 0.26 +/- 0.1 in the 33.75 mg/kg of BW group. Degeneration of the vitelline membrane was evident at all treatment levels; severity was dose-related and greater in the outer vitelline membrane than the inner vitelline membrane. No significant health effects were observed for birds treated with NCZ. The heterophil:lymphocyte ratio was elevated during the treatment and posttreatment periods in all groups, indicating birds were experiencing stress due to handling. Fecal DNC levels did not correlate well with plasma DNC levels, likely due to NCZ being administered as a bolus dose rather than being fed ad libitum. Fluorescein correlated well with plasma DNC levels during the treatment period and can therefore be used successfully as a noninvasive marker to determine the approximate amount of NCZ a bird is consuming. As a contraceptive, NCZ likely would have minimal adverse health effects on the target animal, although field studies with the species of interest need to be conducted. Further research using higher NCZ levels needs to be conducted to determine whether NCZ can inhibit reproduction in waterfowl.

Evaluation and significance of tetracycline stability in rabies vaccine baits.

Tetracycline is widely used as a biomarker for bait consumption by wildlife; tetracycline is incorporated into bones and teeth and can be detected by fluorescence microscopy several weeks postconsumption. During 2003, the United States Department of Agriculture distributed more than 10 million tetracycline-containing rabies-vaccine baits to control the spread of wildlife vectored rabies to humans, pets, and livestock. To estimate the percentage of target species consuming the baits, raccoons and skunks were collected in baited areas and teeth were analyzed for the presence of the biomarker. Several incidents of low biomarker detection rates prompted an investigation of the stability of the biomarker in the baits. Baits were collected at several points along the manufacturing and distribution chain. Baits were analyzed for free and polymer-bound tetracycline and the less active isomer epitetracycline. Results indicated that a portion of the tetracycline was converted to epitetracycline. Additionally, significant quantities of both compounds were trapped in the polymer, which is homogeneously distributed throughout the bait. The results of this study suggest that approximately 40% of the target quantity of tetracycline was unavailable for absorption. This situation could contribute to low biomarker detection rates and suggests that formulation modification should be considered.

Eureka! A surprising appearance after aspiration of a haemopneumothorax: treat the patient not the radiograph.

We describe the aspiration of a traumatic haemopneumothorax and an unexpected anteroposterior chest radiograph finding after the procedure. Chest aspiration is now routine emergency management for spontaneous pneumothorax. There have been no previous documented reports of this clinical scenario of radiological deterioration with clinical improvement after aspiration of a haemopneumothorax.

Discordant PKU phenotype in one family due to disparate genotypes and a novel mutation.

Classical phenylketonuria (PKU) and mild hyperphenylalaninaemia (MHP) are two ends of the broad diagnostic spectrum in phenylalanine hydroxylase (PAH) deficiency. We have analysed a family in which classical PKU, MHP and a normal phenotype occurred in family members with different mutations. Sequence analysis revealed three mutations segregating in the family. The individual with classical PKU had two previously reported deleterious mutations. A third novel mutation was identified in the other two individuals. This report demonstrates that when discordant phenotypes occur in a family, without protein loading or phenylalanine tolerance test, complete analysis of the PAH gene may be performed in order to support the diagnosis and assist in accurate genetic counselling and patient management.

Microsatellite variation in red-winged blackbirds (Agelaius phoeniceus).

Territorial male red-winged blackbirds from five locations in the United States and Canada were genotyped using a suite of six microsatellite loci. Each population possessed unique alleles, but numbers of alleles per locus (range = 7.3-8.8) and expected multilocus heterozygosities (range = 0.76-0.80) were similar in all populations. Significant overall allele frequency differences were detected between some population pairs, and some pairwise Fst values were significant (but small). However, Fst among populations, although significant, was also small (0.009). Despite revealing low levels of population structure, the high multilocus polymorphism indicates these loci will be valuable in the genetic analysis of behavior and reproductive strategies in this species.

Alcohol related falls: an interesting pattern of injuries.

OBJECTIVE: To discover if there is a significant difference in the pattern and severity of injury sustained during falls in patients who have consumed alcohol and those who have not. To determine how pattern and severity of injury correlates with blood alcohol concentration. METHOD: A prospective quasi-randomised controlled study between November 2001 and July 2002. All healthy adults between 16 and 60 years who had fallen from standing height were included. A systematic history and examination permitted calculation of injury severity scores as per abbreviated injury scale update 1998. Blood alcohol concentrations were obtained from intoxicated patients with consent. RESULTS: 351 healthy adult patients were included in the study, there were 238 in the no alcohol group, 113 had consumed alcohol and blood alcohol intake were obtained for 47. The alcohol group had a higher incidence of head injuries (46 (48%) versus 22 (9%)) with a lower incidence of limb injuries (39 (39%) versus 183 (76%)) than the no alcohol group. There was a significant difference in the pattern of injury between the alcohol and no alcohol groups (chi(2), p<0.001) and there was a significant difference in the injury severity scores (p<0.001, Z = -2.5). In the alcohol group severity and pattern correlated with alcohol concentration at the time of injury. Patients with an alcohol concentration<2 g/l had mostly soft tissue limb injuries (58%), 2-2.5 mostly significant limb fractures (55%), and >2.5 mostly significant head injuries (90%). CONCLUSIONS: Alcohol related falls are more often associated with severe craniofacial injury. The severity of both limb and head injury is greater and correlates directly with blood alcohol concentration.

Effectiveness of twenty, twenty-five diazacholesterol, avian gonadotropin-releasing hormone, and chicken riboflavin carrier protein for inhibiting reproduction in Coturnix quail.

Contraception may provide a useful nonlethal management tool when it is desirable to reduce populations of birds. We tested the efficacy of 20,25 diazacholesterol, and immunization with avian gonadotropin-releasing hormone (AGnRH-I) and chicken riboflavin carrier protein (cRCP) as contraceptives and investigated their modes of action in Coturnix quail (Coturnix coturnix japonica). Females that were paired with males treated with 20,25 diazacholesterol produced lower percentages of eggs that were fertile and hatched. Females treated with 20,25 diazacholesterol and paired with control males laid fewer eggs, and lower percentages of their eggs were fertile and hatched. Treatment with 20,25 diazacholesterol reduced testosterone levels in males and progesterone levels in females. Nonesterified cholesterol levels were reduced, whereas desmosterol levels increased in birds treated with 20,25 diazacholesterol. Treatment with AGnRH-I and cRCP immunocontraceptive vaccines did not decrease average egg production and hatchability or hormone levels, but this failure might have been due to the vaccination protocol. If registered, wildlife managers may be able to use 20,25 diazacholesterol when other methods, such as lethal control, are undesirable for reducing damage caused by specific breeding behaviors such as the building of nests.

Risk assessment of an acetaminophen baiting program for chemical control of brown tree snakes on Guam: evaluation of baits, snake residues, and potential primary and secondary hazards.

The brown tree snake (Boiga irregularis) is a significant ecological, agricultural, and economic pest on Guam. Acetaminophen has recently been identified as a promising snake toxicant. Subsequent experimentation has shown that acetaminophen-mouse baits are readily consumed by and acutely toxic to browntree snakes. Before implementing an island-wide acetaminophen-mouse baiting program for the reducton of brown tree snake populations,the potential risks to nontarget wildlife must be evaluated. Quantification of nontarget hazards by comparing potential exposure levels to toxicity values suggested a significant level of concern for rodents, cats, pigs, and birds. For these species, subsequent calculations and field and laboratory experiments, which quantified acetaminophen consumption under field conditions, indicated that acetaminophen consumption was minimal. These results indicate that the advantages of using acetaminophen to reduce brown tree snake populations on Guam outweigh the minimal risks to nontarget feral and wildlife species.

Novel field sampling procedure for the determination of methiocarb residues in surface waters from rice fields.

Methiocarb was extracted from surface water samples collected at experimental rice field sites in Louisiana and Texas. The sampling system consisted of a single-stage 90-mm Empore extraction disk unit equipped with a battery-powered vacuum pump. After extraction, the C-18 extraction disks were stored in an inert atmosphere at -10 degrees C and shipped overnight to the laboratory. The disks were extracted with methanol and the extracts analyzed by reversed-phase high-performance liquid chromatography with a methanol/water mobile phase. Methiocarb was detected by ultraviolet absorption at 223 nm and quantified with the use of calibration standards. Recoveries from control surface water samples fortified at 5.0, 10, 50, and 100 ng/mL methiocarb averaged 92 +/- 7%. A method limit of detection for methiocarb in rice field surface water was estimated to be 0.23 ng/mL at 223 nm.

Determination of cabergoline by electrospray ionization tandem mass spectrometry: picogram detection via column focusing sample introduction.

An electrospray ionization tandem mass spectrometric method was developed for low-picogram detection of an ergot alkaloid, cabergoline, in coyote plasma extracts. Cabergoline is under investigation as an abortifacient in canid species. Central to the successful development of this method was the ability to introduce relatively large sample volumes into the mass spectrometer. This was achieved by focusing the analyte on a conventional high-performance liquid chromatography guard column prior to elution into the spectrometer. Volumes up to at least 900 microL could be injected onto the guard column using a 100% aqueous mobile phase. Cabergoline retained on the column was eluted as a discreet band into the mass spectrometer by the rapid addition of methanol (30%) to the mobile phase. As compared to flow injection sample introduction, the ability to inject larger sample volumes led to a greatly lowered detection limit. Using this technique and a modification of a previously reported extraction procedure, cabergoline could be determined in coyote plasma at concentrations as low as 9 pg of cabergoline/mL of plasma.

Determination of 4,4'-dinitrocarbanilide (DNC), the active component of the antifertility agent nicarbazin, in chicken, duck, and goose plasma.

4,4'-Dinitrocarbanilide (DNC) was extracted from chicken, duck, and goose plasma and isolated by reversed-phase high-performance liquid chromatography. DNC was detected by ultraviolet absorbance at 347 nm and quantified by comparison to a calibration standard. Recovery data were determined by analyzing DNC-fortified control plasma. The mean recovery of DNC in fortified chicken plasma samples was 99.7 +/- 1.9% for 0.18 and 9.1 ppm DNC, and in fortified duck and goose plasma samples was 99.5 +/- 4.9% and 101.4 +/- 4.5%, respectively, for 0.18, 9.1, and 18 ppm DNC.

Analysis of pesticide gas cartridges.

Charcoal and sodium nitrate, the active ingredients in pesticide gas cartridges, are quantified via carbon analysis and ion chromatography, respectively. Linearity was excellent (R(2) > 0.995) over a range consisting of 50-150% of the target concentration for both ingredients. The coefficient of variation for the replicate analyses of gas cartridges over multiple days was <4% for both analytes. Using the results from the analysis of two batches of gas cartridges, theoretical populations were modeled and used to determine practical sampling strategies to support a quality control program for a gas cartridge manufacturing operation. This modeling indicates that the analysis of three cartridges from each of five different lots would produce mean values for both active ingredients that are within 5% of the true mean >99% of the time.

Solid phase extraction gas chromatography/electron capture detector method for the determination of organochlorine pesticides in wildlife whole blood.

A gas chromatographic method for the analysis of 10 organochlorine pesticides in 0.5 mL of whole blood is described. Sample preparation involved an ethyl ether and hexane extraction, followed by a silica solid phase extraction cleanup. The pesticides are quantified by gas chromatography/electron capture detection. Method limits of detection ranged from 1.1 to 5.2 microg/L. The mean and standard deviation for the recovery of 10 pesticides was 97.9 +/- 5.5%. Recoveries from whole blood were comparable to recoveries from plasma. This indicates that the preparation of plasma is unnecessary for the quantification of organochlorine pesticides in blood. This approach is particularly useful as a nonlethal approach for monitoring pesticide contamination in small animals for which the volume of blood is limiting.

Determination of DiazaCon in quail feed and quail serum by ion pair reversed-phase chromatography.

Liquid chromatographic (LC) methods were developed for quantitating the potential avian contraceptive DiazaCon in quail feed and serum. DiazaCon was extracted from ground quail feed with basic n-butyl chloride. The n-butyl chloride extract was evaporated to dryness. The DiazaCon residues were dissolved in an aqueous methanolic ion pairing solution and quantitated by LC at 206 nm. Avian sera was combined with an equal volume of a pH 4 aqueous solution of ion pairing reagent and filtered to remove interfering proteins. DiazaCon was quantitated by LC. Mean recoveries for 500 and 2000 ppm fortified feed were 89.1 and 91.0%, respectively. The mean recovery for sera fortified at 5 levels ranging from 35 to 2000 ppm was 84.9%. Method limits of detection were approximately 14 and 13 ppm for feed and sera, respectively.