Pseudogene rescue: an adaptive mechanism of codon reassignment.

Alterations to the genetic code--codon reassignments--have occurred many times in life's history, despite the fact that genomes are coadapted to their genetic codes and therefore alterations are likely to be maladaptive. A potential mechanism for adaptive codon reassignment, which could trigger either a temporary period of codon ambiguity or a permanent genetic code change, is the reactivation of a pseudogene by a nonsense suppressor mutant transfer RNA. I examine the population genetics of each stage of this process and find that pseudogene rescue is plausible and also readily explains some features of extant variability in genetic codes.

Epigenomic plasticity within populations: its evolutionary significance and potential.

Epigenetics has progressed rapidly from an obscure quirk of heredity into a data-heavy 'omic' science. Our understanding of the molecular mechanisms of epigenomic regulation, and the extent of its importance in nature, are far from complete, but in spite of such drawbacks, population-level studies are extremely valuable: epigenomic regulation is involved in several processes central to evolutionary biology including phenotypic plasticity, evolvability and the mediation of intragenomic conflicts. The first studies of epigenomic variation within populations suggest high levels of phenotypically relevant variation, with the patterns of epigenetic regulation varying between individuals and genome regions as well as with environment. Epigenetic mechanisms appear to function primarily as genome defences, but result in the maintenance of plasticity together with a degree of buffering of developmental programmes; periodic breakdown of epigenetic buffering could potentially cause variation in rates of phenotypic evolution.

Tympanic temperature in confined beef cattle exposed to excessive heat load.

Angus crossbred yearling steers (n = 168) were used to evaluate effects on performance and tympanic temperature (TT) of feeding additional potassium and sodium to steers exposed to excessive heat load (maximum daily ambient temperature exceeded 32°C for three consecutive days) during seasonal summer conditions. Steers were assigned one of four treatments: (1) control; (2) potassium supplemented (diet containing 2.10% KHCO3); (3) sodium supplemented (diet containing 1.10% NaCl); or (4) potassium and sodium supplemented (diet containing 2.10% KHCO3 and 1.10% NaCl). Overall, additional KHCO3 at the 2% level or NaCl at the 1% level did not improve performance or heat stress tolerance with these diet formulations. However, the addition of KHCO3 did enhance water intake. Independent of treatment effects, TT of cattle displaying high, moderate, or low levels of stress suggest that cattle that do not adequately cool down at night are prone to achieving greater body temperatures during a subsequent hot day. Cattle that are prone to get hot but can cool at night can keep average tympanic temperatures at or near those of cattle that tend to consistently maintain lower peak and mean body temperatures. In addition, during cooler and moderately hot periods, cattle change TT in a stair-step or incremental pattern, while under hot conditions, average TT of group-fed cattle moves in conjunction with ambient conditions, indicating that thermoregulatory mechanisms are at or near maximum physiological capacity.

Replication of KIAA0350, IL2RA, RPL5 and CD58 as multiple sclerosis susceptibility genes in Australians.

A recent genome-wide association study (GWAS) conducted by the International Multiple Sclerosis Genetics Consortium (IMSGC) identified a number of putative MS susceptibility genes. Here we have performed a replication study in 1134 Australian MS cases and 1265 controls for 17 risk-associated single nucleotide polymorphisms (SNPs) reported by the IMSGC. Of 16 SNPs that passed quality control filters, four, each corresponding to a different non-human leukocyte antigen (HLA) gene, were associated with disease susceptibility: KIAA0350 (rs6498169) P=0.001, IL2RA (rs2104286) P=0.033, RPL5 (rs6604026) P=0.041 and CD58 (rs12044852) P=0.042. There was no association (P=0.58) between rs6897932 in the IL7R gene and the risk of MS. No interactions were detected between the replicated IMSGC SNPs and HLA-DRB1*15, gender, disease course, disease progression or age-at-onset. We used a novel Bayesian approach to estimate the extent to which our data increased or decreased evidence for association with the six most-associated IMSGC loci. These analyses indicated that even modest P-values, such as those reported here, can contribute markedly to the posterior probability of 'true' association in replication studies. In conclusion, these data provide support for the involvement of four non-HLA genes in the pathogenesis of MS, and combined with previous data, increase to genome-wide significance (P=3 x 10(-8)) evidence of an association between KIAA0350 and risk of disease.

Efficient targeted mutagenesis in Epichloë festucae using a split marker system.

A split-marker system for targeted gene deletion was developed for the model grass endophytic fungus Epichloë festucae. Compared to the conventional system that yields up to 25% homologous recombinants, the method resulted in 33-74% targeted deletions in E. festucae using as little as 1.5kb of targeting sequence.

Bone-seeking properties of Tc-99m carbonyl diphosphonic acid, dihydroxy-methylene diphosphonic acid and monohydroxy-methylene phosphoinic acid: concise communication.

Carbonyl diphosphonic acid, dihydroxy-methylene diphosphonic acid and monohydroxy-methylene diphosphonic acid were synthesized and labeled with Tc-99m. Their chemical structures were confirmed and their bone-seeking relationships were compared with those of Tc-99m MDP in experimental animals. MKonohydroxy-methylene diphosphonic acid was found to have higher bone affinity and faster blood clearance than MDP, whereas carabonyl diphosphonic acid and dihydroxy-methylene diphosphonic acid demonstrated lower bone affinity and slower blood clearance than MDP.

Effects from past solid waste disposal practices.

This paper reviews documented environmental effects experience from the disposal of solid waste materials in the U.S. Selected case histories are discussed that illustrate waste migration and its actual or potential effects on human or environmental health. Principal conclusions resulting from this review were: solid waste materials do migrate beyond the geometric confines of the initial placement location; environmental effects have been experienced from disposal of municipal, agricultural, and toxic chemical wastes; and utilization of presently known science and engineering principles in sitting and operating solid waste disposal facilities would make a significant improvement in the containment capability of shallow land disposal facilities.

The snoring spectrum: acoustic assessment of snoring sound intensity in 1,139 individuals undergoing polysomnography.

STUDY OBJECTIVES: To quantify the snoring sound intensity levels generated by individuals during polysomnographic testing and to examine the relationships between acoustic, polysomnographic, and clinical variables. DESIGN: The prospective acquisition of acoustic and polysomnographic data with a retrospective medical chart review. SETTING: A sleep laboratory at a primary care hospital. PARTICIPANTS: All 1,139 of the patients referred to the sleep laboratory for polysomnographic testing from 1980 to 1994. INTERVENTIONS: The acoustic measurement of snoring sound intensity during sleep concurrent with polysomnographic testing. MEASUREMENTS AND RESULTS: Four decibel levels were derived from snoring sound intensity recordings. L1, L5, and L10 are measures of the sound pressure measurement in decibels employing the A-weighting network that yields the response of the human ear exceeded, respectively, for 1, 5, and 10% of the test period. The Leq is a measure of the A-weighted average intensity of a fluctuating acoustic signal over the total test period. L10 levels above 55 dBA were exceeded by 12.3% of the patients. The average levels of snoring sound intensity were significantly higher for men than for women. The levels of snoring sound intensity were associated significantly with the following: polysomnographic testing results, including the respiratory disturbance index (RDI), sleep latency, and the percentage of slow-wave sleep; demographic factors, including gender and body mass; and clinical factors, including snoring history, hypersomnolence, and breathing stoppage. Men with a body mass index of > 30 and an average snoring sound intensity of > 38 dBA were 4.1 times more likely to have an RDI of > 10. CONCLUSIONS: Snoring sound intensity levels are related to a number of demographic, clinical, and polysomnographic test results. Snoring sound intensity is closely related to apnea/hypopnea during sleep. The noise generated by snoring can disturb or disrupt a snorer's sleep, as well as the sleep of a bed partner.

Optical light scatter imaging of cellular and sub-cellular morphology changes in stressed rat hippocampal slices.

Optical imaging, such as transmission imaging, is used to study brain tissue injury. Transmission imaging detects cellular swelling via an increase in light transmitted by tissue slices due to a decrease in scattering particle concentration. Transmission imaging cannot distinguish sub-cellular particle size changes from cellular swelling or shrinkage. We present an optical imaging method, based on Mie scatter theory, to detect changes in sub-cellular particle size and concentration. The system uses a modified inverted microscope and a 16-bit cooled CCD camera to image tissue light scatter at two angles. Dual-angle scatter ratio imaging successfully discriminated latex microsphere suspensions of differing sizes (0.6, 0.8, 1 and 2 microm) and concentrations. We applied scatter imaging to hippocampal slices treated with 100 microM N-methyl-D-aspartate (NMDA) to model excitotoxic injury or -40 mOsm hypotonic perfusion solution to cause edema injury. We detected light scatter decreases similar to transmission imaging in the CA1 region of the hippocampus for both treatments. Using our system, we could distinguish between NMDA and hypotonic treatments on the basis of statistically significant (P<0.0003) differences in the scatter ratio measured in CA1. Scatter imaging should be useful in studying tissue injuries or activity resulting in brain tissue swelling as well as morphological changes in sub-cellular organelles such as mitochondrial swelling.

The role of cytokines in the behavioral responses to endotoxin and influenza virus infection in mice: effects of acute and chronic administration of the interleukin-1-receptor antagonist (IL-1ra).

Following infection with influenza virus, animals display decreased locomotor activity and feeding behavior and loss of body weight. It has been suggested that these effects may be mediated by cytokines, such as interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha), induced by the infection. To assess the potential role of IL-1, we tested the ability of a naturally occurring IL-1-receptor antagonist (IL-1ra) to antagonize the changes in feeding behavior induced by IL-1, endotoxin (lipopolysaccharide, LPS), and infection with influenza virus. Feeding behavior was assessed by measuring the daily intake of food pellets and sweetened milk in a 30-min period. Acute injection of IL-1 beta decreased milk intake, but mouse IL-6 and mouse TNF-alpha did not. However, TNF-alpha decreased food pellet intake slightly, especially when it was injected at the beginning of the dark phase. The reductions in milk intake induced by mouse IL-1 beta were largely prevented by IL-1ra pretreatment (100 micrograms/mouse i.p.). The LPS-induced reductions in milk intake were attenuated, but not blocked, by IL-1ra treatment (300 micrograms/mouse). LPS still induced significant decrements in the presence of the antagonist. In influenza virus-infected mice, IL-1ra was administered either by repeated subcutaneous (s.c.) injections, or by continuous s.c. infusion from osmotic minipumps. These IL-1ra treatments produced small, but statistically significant, attenuations of the depression in milk and food pellet intake in the virus-infected mice. In several experiments, IL-1ra treatment increased the survival of influenza virus-infected mice. Thus the attenuation of the hypophagia may have been caused by this IL-1ra-induced increase in survival. The results suggest that IL-1 contributes to sickness behavior induced by LPS and influenza virus infection, but it is not the only factor involved.

A biomechanical study of torque and accuracy of halo pin insertional devices.

The purpose of the present study was to assess the accuracy and repeatability of the torque achieved with a variety of devices used for the insertion of halo pins. We found that equivalent devices from the same manufacturer did not reliably achieve the same torque, that the same device used by different surgeons did not reliably achieve the same torque, and that the devices achieved varying levels of accuracy and repeatability. The proportion of trials in which the achieved torque was within +/- 10 per cent of the intended torque averaged 64 per cent (range, less than 1 to 100 per cent). Our data suggest that the insertion of halo pins with use of most currently available torque-setting devices is an inherently inaccurate process.

Muscarinic signalling in submandibular salivary acinar cells of ageing rats.

Muscarinic cholinergic stimulation of submandibular acinar cells results in the activation of Ca(2+)-dependent ion-transport pathways responsible for the secretion of primary saliva. Decreased saliva production is common among elderly people and may compromise oral health with implications for systemic health, nutrition, and quality of life. The density and affinity of muscarinic receptors in the submandibular gland of rats and the Ca2+ responses to stimulation of these receptors in the acinar cells were examined. An increase in the number of receptors and increases in the affinities of the receptors were found as the rats age from 7 weeks to 11 months. However, the coupling of the receptors to the intracellular Ca2+ signals in acinar cell clusters was substantially reduced in the older animals.

The Maine State Patient Education Forum: networking system for a rural area.

Patient education councils are growing and prospering. The Maine State Patient Education Forum is one such council which has gone through a continuing developmental process to meet needs of patient educators in Maine. Involving many organizations and individuals who do patient education, having a clear direction and a common locus, scheduling regular meetings, and participating in national networking have all been ways to vitalize and strengthen the council. Maine as a rural state has found the forum to be a dynamic model to motivate patient educators to maintain flexibility and acquire new concepts so they can continue to provide high quality patient education services.

Gambling behavior of Louisiana students in grades 6 through 12.

OBJECTIVES: The prevalence of problem and gambling behavior, the average age of onset of gambling behavior, and the co-occurrence of gambling disorder with substance use were determined in the Louisiana student population grades 6 through 12. METHODS: A stratified randomized sample of 12,066 students in Louisiana schools during the 1996-1997 school year was surveyed about gambling behavior using the South Oaks Gambling Screen--Revised for Adolescents (SOGS-RA). RESULTS: Fourteen percent of the students never gambled, 70.1 percent gambled without problems, 10.1 percent indicated problem gambling in the past year (level 2 according to the SOGS-RA), and 5.8 percent indicated pathological gambling behavior in the past year (level 3). Weekly or more frequent lottery play was reported by 16.5 percent. The average age of onset of gambling behavior was 11.2 years. Fifty-nine percent of the students with problem and pathological gambling behavior reported frequent alcohol and illicit drug use. CONCLUSIONS: A significant minority of Louisiana students in grades 6 through 12-15.9 percent--acknowledged gambling-related symptoms and life problems. The association of problem and pathological gambling with use of alcohol, tobacco, and marijuana provides preliminary support for the inclusion of gambling among other adolescent risk behaviors.

Evaluation of vitamin K3 feed additive for prevention of sweet clover disease.

Sweet clover poisoning in cattle is caused by an anticoagulant (dicumarol) that is formed in moldy sweet clover hay. Previous experiments with vitamin K3 and vitamin K1 in therapy trials indicated that vitamin K1 was effective in reducing prothrombin times but vitamin K3 was not. As a possible alternative in the use of toxic sweet clover hays, vitamin K3 was evaluated to see if it would prevent hemorrhagic crises when fed to cattle consuming toxic sweet clover hay. Vitamin K3 levels of 0, 0.45, 4.5, 11, and 45 mg/kg body weight/day were fed to 173-235-kg steers consuming toxic (40-50 ppm dicumarol) sweet clover. The 45-mg K3/kg/day supplement was not palatable and had to be discontinued. The 0.45, 4.5, and 11-mg K3/kg/day supplements did not significantly reduce the prothrombin times as compared to the 0-mg K3/kg/day group.

Effect of ammonium carbamate on nutritive and preservative characteristics of high-moisture coastal bermudagrass hay.

Two studies evaluated the effects of ammonium carbamate (AC) on preservation and digestibility of high-moisture (HM) Coastal bermudagrass (Cynodon dactylon) hay. A 3 X 7 factorial arrangement of treatments in a completely random design was used to estimate the efficacy of AC in laboratory conditions. Treatments were 25, 30, or 35% moisture (M) forage treated with levels (L) of 0, 1.14, 2.28, 3.42, 4.56, 5.70, or 6.84% AC. Nitrogen in forage increased (P less than .01) linearly due to M and L. There was a linear decrease (P less than .01) in ADF N due to M. Neutral detergent fiber decreased (P less than .01) linearly as M increased and increased (P less than .01) linearly with increased L. Acid detergent fiber decreased (P less than .01) linearly due to L. There was an M X L interaction (P less than .01) for hemicellulose (HC) concentrations and total aerobic fungal counts. The digestibility of HM hay treated with a product (ACNH) containing 57% AC was compared to that of untreated hay (UH) and UH plus urea (UHU) when fed to lambs (four lambs per treatment). Digestibility of NDF and hemicellulose (P less than .01) and ADF (P less than .05) was increased for ACNH forage compared with UH or UHU. Ammonium carbamate seems to be beneficial for preserving HM forage, and an admixture containing ammonium carbamate is effective for increasing the digestibility of Coastal bermudagrass hay.

Effect of suckling on pituitary responsiveness to gonadotropin-releasing hormone throughout the early postpartum period of beef cows.

The objective of this study was to compare pituitary responsiveness to gonadotropin-releasing hormone (GnRH) throughout the early postpartum period (PP) in beef cows suckling in single calf vs cows which had calves removed at birth. The experiment utilized 48 cows in a 2 X 4 factorial. Main effects studied were suckling status (suckled or nonsuckled) and days PP (d 3, 10, 20 or 30). Covariates studied were pre-treatment concentrations of progesterone (P), estradiol-17 beta (E), luteinizing hormone (LH) and follicle stimulating hormone (FSH). Suckled (S) cows nursed one calf, while nonsuckled (NS) cows had calves removed at birth. Treatment was a single im injection of 100 micrograms GnRH dissolved in saline. Pre-GnRH plasma concentrations of E, P, LH and FSH and post-treatment plasma concentrations of LH and FSH were determined at 15 min intervals (-30 to 360 min) in blood samples collected via jugular cannulae. Post-treatment plasma concentrations of P were determined at 6 (n = 23) or 8 (n = 23) days after treatment to monitor ovarian response. Concentrations of E, LH and FSH before GnRH injection did not differ (P greater than .1) between S and NS cows at any PP stage and data were pooled for statistical analyses. Pre-treatment LH, but not FSH, increased (r = .43; P less than .002 (with days PP and concentrations of LH were greater (P less than .05) at 30 d than at 3 d PP. Pre-treatment P was greater (P less than .05) in NS than in S cows on d 30 PP and P was correlated (r = .51; P less than .01) with days PP in NS cows. However, the frequency of cows exhibiting plasma P greater than or equal to 1 ng/ml prior to GnRH was low on all days studied and was similar for both groups. LH release after GnRH was correlated with increasing days PP in both S (r = .73; P less than .001) and NS (r = .61; P less than .002) cows, Releasable FSH was not related to days PP in either group. Mean peak LH and total LH released were greater (P less than .025 and P less than .05), respectively) in S than in NS cows at 30 d PP. Mean peak FSH and total FSH released were greater (P less than .05 and P less than .07, respectively) at 10 and 30 d PP in S cows than in NS cows. Pre-treatment E, but no P, LH or FSH was positively related (P less than .01) to GnRH-mediated LH release in both groups. When pre-treatment E was partitioned into separated regressions for S (ES) and NS (ENS) cows, increased pituitary LH response in S cows on d 30 appeared to be due in part to a greater sensitivity of S cows to E modulation. We found no evidence to suggest that pituitary responsiveness per se is decreased in beef cows suckling a single calf.

Weight gain and reproductive performance of spring-born beef heifer calves intraruminally administered oxfendazole.

The effect of oxfendazole (Synanthic) on weight gain and reproductive performance of spring-born heifer calves was evaluated at five locations (IL, IN, MI, MO, ND). Three hundred thirty-eight mixed-breed beef heifers (293.4 kg) blocked by weight and age were randomly allotted to one of two treatments. Treatment 1 heifers were dewormed via intraruminal injection of oxfendazole. Treatment 2 heifers served as a control. Heifers were comingled during the winter phase in semi-confinement during the period from late January to late May (winter phase). Heifers were sorted by treatment at the beginning of the grazing season (approximately May 24, 1991) and remained separate until the end of the study (approximately July 31, 1991; summer phase). Dewormed heifers received oxfendazole (4.5 mg/kg BW) in late January and again 28 and 56 d after the beginning of the grazing season. Fecal samples were taken every 21 d from January through September and assayed for gastrointestinal parasite eggs. Fecal egg counts were similar across treatments at the beginning of the trial (P = .34). Deworming decreased fecal egg counts throughout the trial. Twenty-eight days after initial deworming a significant treatment effect on fecal egg counts was observed (P < .01). Winter ADG was significantly higher (P < .01) for dewormed heifers than for controls (.60 vs .52 kg, respectively). Summer ADG was not different between treatments. Dewormed heifers were 7.4 kg heavier than controls (P < .05) at the end of the trial. Age of puberty was not affected by treatment (P = .64). First-service conception and final pregnancy rates were not different between treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Occurrence and range of dicumarol concentrations in sweet clover.

The dicumarol concentration in 272 cured sweet clover samples from 4 counties in North Dakota was determined. Dicumarol concentrations ranged from 0 to 164.7 mg/kg of sweet clover, with 64.6% of the 272 samples containing less than 10 mg/kg. Round bales were significantly (P less than 0.05) higher in dicumarol concentration (mean of 22.9 +/- 3.10 mg/kg) than were stacks or silage (means of 1.8 +/- -6.3 mg/kg and 0.6 +/- 2.1 mg/kg, respectively). The outer section of core samples taken from round bales contained significantly (P less than 0.0021) higher concentrations of dicumarol than did the inner section of the core, with means of 30.8 mg/kg and 16.6 mg/kg, respectively. A significant (P less than 0.0001) correlation existed between crude protein and dicumarol values (r = 0.44).