Mechanism of Na+/H+ antiporting

Na+/H+ antiporters are central to cellular salt and pH homeostasis. The structure of Escherichia coli NhaA was recently determined, but its mechanisms of transport and pH regulation remain elusive. We performed molecular dynamics simulations of NhaA that, with existing experimental data, enabled us to propose an atomically detailed model of antiporter function. Three conserved aspartates are key to our proposed mechanism: Asp164 (D164) is the Na+-binding site, D163 controls the alternating accessibility of this binding site to the cytoplasm or periplasm, and D133 is crucial for pH regulation. Consistent with experimental stoichiometry, two protons are required to transport a single Na+ ion: D163 protonates to reveal the Na+-binding site to the periplasm, and subsequent protonation of D164 releases Na+. Additional mutagenesis experiments further validated the model.     

Oral glucose leads to a differential response in glucose,insulin, and GLP-1 in lean versus obese cats

The response to oral glucose was examined in 10 obese and 9 lean age-matched, neutered cats. In all cats, oral administration of 2 g/kg glucose was followed by a prompt increase in glucose, insulin, and glucagon-like peptide (GLP)-1. There were significant differences between lean and obese cats in the areas under the curve for glucose, insulin, and GLP-1. However, the responses were variable, and a clear distinction between individual lean and obese cats was not possible. Therefore, this test cannot be recommended as a routine test to examine insulin resistance in individual cats as it is used in people. A further disadvantage for routine use is also the fact that this test requires gastric tubing for the correct administration of the glucose and associated tranquilization to minimize stress and that it was associated with development of diarrhea in 25% of the cats. GLP-1 concentrations were much lower in obese than lean cats. The low GLP-1 concentrations in obese cats might indicate a contribution of GLP-1 to the lower insulin sensitivity of obese cats, but this hypothesis needs to be further investigated.     

A comparison of two combinations of xylazine-ketamine administered intramuscularly to alpacas and of reversal with tolazoline

ObjectiveTo evaluate the anesthetic and cardiorespiratory effects of two doses of intramuscular (IM) xylazine/ketamine in alpacas, and to determine if tolazoline would reduce the anesthetic recovery time.Study designProspective randomized crossover study.AnimalsSix castrated male alpacas.MethodsEach alpaca received a low dose (LD) (0.8 mg kg⁻¹ xylazine and 8 mg kg⁻¹ ketamine IM) and high dose (HD) (1.2 mg kg⁻¹ xylazine and 12 mg kg⁻¹ ketamine IM) with a minimum of one week between trials. Time to sedation, duration of lateral recumbency and analgesia, pulse rate, respiratory rate, hemoglobin oxygen saturation, arterial blood pressure, blood-gases, and the electrocardiogram were monitored and recorded during anesthesia. With each treatment three alpacas were randomly selected to receive tolazoline (2 mg kg⁻¹ IM) after 30 minutes of lateral recumbency.ResultsOnset of sedation, lateral recumbency and analgesia was rapid with both treatments. The HD was able to provide ≥30 minutes of anesthesia in five of six alpacas. The LD provided ≥30 minutes of anesthesia in three of six alpacas. Respiratory depression and hypoxemia occurred with the HD treatment during the first 10 minutes of lateral recumbency: two animals were severely hypoxemic and received nasal oxygen for 5 minutes. Heart rate decreased, but there were no significant changes in arterial blood pressure. Tolazoline significantly shortened the duration of recumbency with the HD.ConclusionsThe HD provided more consistent clinical effects in alpacas than the LD. Intramuscular tolazoline shortened the duration of lateral recumbency in alpacas anesthetized with the HD combination.Clinical relevanceBoth doses of the combination were effective in providing restraint in alpacas and the duration of restraint was dose dependent. Supplemental oxygen should be available if using the HD and IM administration of tolazoline will shorten the recovery time.     

Use of 25-hydroxyvitamin D3 and dietary calcium to improve tenderness of beef from the round of beef cows

The objective of this trial was to determine how 25-hydroxyvitamin D(3) (25-OH D(3)) supplementation, altering supplemental dietary calcium, or their combination influence postmortem biochemical and tenderness changes in muscles from the round of mature cows. Twenty-seven Angus cows (3 to 7 yr old) were allotted randomly to 9 pens with 3 cows per pen. Treatments were arranged in a 3 x 3 factorial design with 3 dosages of 25-OH D(3) (0, 250, or 500 mg of 25-OH D(3) administered as a 1-time oral bolus 7 d before slaughter) and 3 percentages of supplemental limestone (0.5, 0.75, and 1.0%) replenished in the diet for 3 d before slaughter and after a 2-wk limestone withdrawal. Plasma samples were obtained during the feeding period. Upon slaughter, adductor, gracilus, pectineus, sartorius, semimembranosus, vastus intermedius, and vastus lateralis muscles were obtained and aged for 1, 3, or 7 d. Calcium concentrations were increased in plasma when 250 or 500 mg of 25-OH D(3) were administered (P 相似文献   

Food aversion learning: ability of lambs to distinguish safe from harmful foods

We studied the ability of lambs to select safe foods in the presence of harmful foods. In a series of feeding experiments, 3- to 6-mo-old lambs were offered a choice between 1) a familiar, safe and a novel, harmful food, 2) a novel, safe and a novel, harmful food and 3) a familiar, harmful and a novel, safe food. All harmful foods were palatable feeds that had been treated with lithium chloride (LiCl), a non-lethal gastrointestinal poison. When lambs ingested a meal composed of a novel food containing LiCl and a familiar, safe food, lambs subsequently avoided the novel food. Lambs avoided the novel food even when the familiar food contained LiCl. The response of lambs varied when lambs were given a choice between a novel, safe food and a novel food containing LiCl. Lambs either avoided the novel food containing LiCl and ingested the safe food or they limited their intake of both novel foods. Their response was dependent on the novelty of the food containing LiCl. When a novel, palatable food contained 2% LiCl, lambs always ate some of the food, even after experiencing illness from ingesting it. Lambs experienced with foods containing LiCl displayed greater food neophobia than lambs naive to LiCl-treated foods. Thus, novelty was the major criterion that lambs used to associate foods with gastrointestinal illness.     

The Lunar Laser Ranging Experiment: Accurate ranges have given a large improvement in the lunar orbit and new selenophysical information

The lunar ranging measurements now being made at the McDonald Observatory have an accuracy of 1 nsec in round-trip travel time. This corresponds to 15 cm in the one-way distance. The use of lasers with pulse-lengths of less than 1 nsec is expected to give an accuracy of 2 to 3 cm in the next few years. A new station is under construction in Hawaii, and additional stations in other countries are either in operation or under development. It is hoped that these stations will form the basis for a worldwide network to determine polar motion and earth rotation on a regular basis, and will assist in providing information about movement of the tectonic plates making up the earth's surface. Several mobile lunar ranging stations with telescopes having diameters of 1.0 m or less could, in the future, greatly extend the information obtainable about motions within and between the tectonic plates. The data obtained so far by the McDonald Observatory have been used to generate a new lunar ephemeris based on direct numerical integration of the equations of motion for the moon and planets. With this ephemeris, the range to the three Apollo retro-reflectors can be fit to an accuracy of 5 m by adjusting the differences in moments of inertia of the moon about its principal axes, the selenocentric coordinates of the reflectors, and the McDonald longitude. The accuracy of fitting the results is limited currently by errors of the order of an arc second in the angular orientation of the moon, as derived from the best available theory of how the moon rotates in response to the torques acting on it. Both a new calculation of the moon's orientation as a function of time based on direct numerical integration of the torque equations and a new analytic theory of the moon's orientation are expected to be available soon, and to improve considerably the accuracy of fitting the data. The accuracy already achieved routinely in lunar laser ranging represents a hundredfold improvement over any previously available knowledge of the distance to points on the lunar surface. Already, extremely complex structure has been observed in the lunar rotation and significant improvement has been achieved in our knowledge of lunar orbit. The selenocentric coordinates of the retroreflectors give improved reference points for use in lunar mapping, and new information on the lunar mass distribution has been obtained. Beyond the applications discussed in this article, however, the history of science shows many cases of previously unknown, phenomena discovered as a consequence of major improvements in the accuracy of measurements. It will be interesting to see whether this once again proves the case as we acquire an extended series of lunar distance observations with decimetric and then centimetric accuracy.     

A role for interaction of the RNA polymerase flap domain with the sigma subunit in promoter recognition

In bacteria, promoter recognition depends on the RNA polymerase sigma subunit, which combines with the catalytically proficient RNA polymerase core to form the holoenzyme. The major class of bacterial promoters is defined by two conserved elements (the -10 and -35 elements, which are 10 and 35 nucleotides upstream of the initiation point, respectively) that are contacted by sigma in the holoenzyme. We show that recognition of promoters of this class depends on the "flexible flap" domain of the RNA polymerase beta subunit. The flap interacts with conserved region 4 of sigma and triggers a conformational change that moves region 4 into the correct position for interaction with the -35 element. Because the flexible flap is evolutionarily conserved, this domain may facilitate promoter recognition by specificity factors in eukaryotes as well.     

Genetic variation in dieback resistance in Fraxinus excelsior confirmed by progeny inoculation assay

Ash dieback caused by the pathogenic fungus Hymenoscyphus fraxineus [previously known as H. pseudoalbidus (sexual stage) and Chalara fraxinea (asexual stage)] is a widespread problem in Europe. Here, we assess crown damage from natural infection and necrosis development following artificial controlled inoculations on full‐sib and half‐sib progeny from Danish Fraxinus excelsior clones with contrasting and well‐characterized levels of susceptibility to the disease. The inoculation assay was performed on a total of 123 offspring, and necrosis development monitored over two years. The offspring from low susceptible mother clones developed smaller necroses when compared to offspring from susceptible clones. Their crown damage due to natural infections was also significantly less. The correlation coefficient between average crown damages of mother clones and the average of their progeny was 0.85 (natural infections), while the correlation between crown damage of mother clones and the average necrosis development in their progeny after controlled inoculation was 0.73. The correlation between resistance of parent trees and crown damage/necrosis development on their offspring confirms the presence of heritable resistance and indicates that a bioassay based on controlled inoculations has the potential of becoming a fast and cost‐effective tool for estimation of dieback susceptibility in breeding programmes for resistance in ash trees.     

Effect of cortisol on luteinizing hormone release during sexual behavior in the boar.

The effect of inhibiting the rise in cortisol concentrations that occurs at copulation upon luteinizing hormone release was studied in seven adult boars. Plasma samples were collected for assay of luteinizing hormone, testosterone and cortisol on a control day and before, during and after exposure to an estrous sow. The area under the curve was used to evaluate hormone production and treatment effects were evaluated by a paired Student's t-test. The 11 beta-hydroxylase inhibitor, metyrapone, was used to suppress glucocorticoid hormone production. Cortisol concentrations increased significantly (p less than 0.05) after breeding compared to values on the control day while treatment with metyrapone prior to breeding prevented the cortisol increase (p greater than 0.05). Although luteinizing hormone production increased significantly after copulation in both breeding experiments, metyrapone pretreatment resulted in a reduction of luteinizing hormone secretion (p less than 0.05). Testosterone production was also reduced in boars pretreated with metyrapone. The results suggest that the increased levels of cortisol occurring at copulation may enhance luteinizing hormone release in boars.