Anatomy of a conformational change: hinged "lid" motion of the triosephosphate isomerase loop

Triosephosphate isomerase (TIM) is used as a model system for the study of how a localized conformational change in a protein structure is produced and related to enzyme reactivity. An 11-residue loop region moves more than 7 angstroms and closes over the active site when substrate binds. The loop acts like a "lid" in that it moves rigidly and is attached by two hinges to the remainder of the protein. The nature of the motion appears to be built into the loop by conserved residues; the hinge regions, in contrast, are not conserved. Results of molecular dynamics calculations confirm the structural analysis and suggest a possible ligand-induced mechanism for loop closure.     

施用竹炭对卷心菜叶绿素含量和荧光参数的影响

为探究人工施用竹炭对卷心菜叶绿素含量和荧光参数的影响,进而明确卷心菜的最适竹炭有机肥施用量,在田间试验条件下设置了无碳(CK)、中碳(MC)和高碳(SC)3个处理,对卷心菜莲座期和结球期的叶绿素含量及荧光参数进行测量研究.结果表明:(1)结球期的卷心菜叶绿素含量极显著高于莲座期(p0.01),且随着竹炭量的增加,两个时期的叶绿素含量变化趋势一致,从高到低依次为:MC,CK,SC.(2)结球期的卷心菜初始最小荧光Fo、初始最大荧光Fm、PSⅡ的最大光化学效率Fv/Fm,PSⅡ的潜在活性Fv/Fo、非光化学猝灭系数qN均低于莲座期,而PSⅡ实际光化学效率ΦPSⅡ、光化学猝灭系数qP高于莲座期,且随着竹炭量的增加,两个时期的这些参数均表现出一致的变化趋势,Fo和qN均呈"V"型变化,而Fm,Fv/Fm,Fv/Fo,ΦPSⅡ和qP均呈现倒"V"趋势,但对ΦPSⅡ和qP没有显著影响(p0.05).说明结球期的卷心菜光合作用高于莲座期;当竹炭量由CK增加到MC,有利于提高叶片叶绿素含量,增强光能利用率,减少植株热耗散,提高植株光合作用;但随着竹炭量进一步增加到SC,卷心菜叶片叶绿素含量降低,光能利用率减弱,热耗散增加,光合作用受到抑制,即中炭(MC)处理条件下,此卷心菜长势最优,表明中炭是该卷心菜生长的最适竹炭有机肥施用量.     

Hidden thermodynamics of mutant proteins: a molecular dynamics analysis

A molecular dynamics simulation method is used to determine the contributions of individual amino acid residues and solvent molecules to free energy changes in proteins. Its application to the hemoglobin interface mutant Asp G1(99) beta----Ala shows that some of the contributions to the difference in the free energy of cooperativity are as large as 60 kilocalories (kcal) per mole. Since the overall free energy change is only -5.5 kcal/mole (versus the experimental value of -3.4 kcal/mole), essential elements of the thermodynamics are hidden in the measured results. By exposing the individual contributions, the free energy simulation provides new insights into the origin of thermodynamic changes in mutant proteins and demonstrates the role of effects beyond those usually considered in structural analyses.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.     

PG99S智能型喷灌机的调试及维护

在蔬菜、花卉的工厂化育苗生产中,准确控制肥水的浓度和喷洒的均匀性显得尤为重要,而喷灌设备作为智能型连栋育苗温室的主要浇灌设备,被广泛应用于现代化育苗温室中。该设备自动作业,水量控制精准。本文就PG99S型智能喷灌机介绍其调试方法和故障维修。     

Cytopathological and histopathological diagnosis of canine splenic disorders

Objectives To determine (1) the common types of canine splenic disorders, and the breeds affected, that are diagnosed by cytopathological and histopathological examination in Sydney, Australia and (2) the accuracy of cytopathological examination compared with histopathological examination for the diagnosis of canine splenic disorders. Design 69 cytopathological and 51 histopathological diagnoses of canine splenic disorders presented to the Veterinary Pathology Diagnostic Services, The University of Sydney during 2006 and 2007 were tabulated and analysed; 17 cases examined both cytopathologically and histopathologically during 2001–07 were also analysed. Results The most common cytopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (29%), followed by no abnormalities detectable (28%), malignant neoplasms (20%), equivocal diagnoses (20%) and inflammatory disorders (3%). The most common breeds were Kelpie crosses and mixed breeds. The most common histopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (49%), followed by malignant neoplasms (43%) and inflammatory disorders (8%). The most common breeds were German Shepherd Dogs, Boxers and Maltese Terriers. Cytopathological and histopathological diagnoses were in complete agreement in 59% of cases, partial agreement in 29% and disagreement in 12%. Conclusion Benign disorders of growth, vascular disturbances and necrosis were the most commonly diagnosed canine splenic disorders, both cytopathologically and histopathologically. Kelpie crosses presented most frequently for cytopathological examination. German Shepherd Dogs were the most common breed diagnosed histopathologically with haemangiosarcoma. Although cytopathological and histopathological splenic examinations are complementary for diagnosis, this study has shown a high correlation for complete and partial agreement between the two.     

Topical anaesthesia alleviates short-term pain of castration and tail docking in lambs

Objective To investigate the effect of a topical anaesthetic formulation on pain alleviation, wound healing and systemic levels of local anaesthetic actives in lambs undergoing castration and tail docking. Design Three placebo-controlled and/or randomised experiments were conducted using three groups of Merino lambs (n = 62, 68 and 19) undergoing routine castration and tail docking. Procedure Surgical castration, with either surgical or hot-iron tail docking, was performed with and without the application of topical anaesthetic (Tri-Solfen®) or placebo. The effects of this procedure were compared with those of rubber ring castration and tail docking, and of the handled but unmarked controls. Wound pain was assessed using calibrated Von-Frey monofilaments over a 4-h period, pain-related behaviour was assessed over 5 h, wound healing was assessed at 14 and 28 days, and the plasma levels of lignocaine and bupivacaine were determined. Results Rapid and up to 4 h primary hyperalgesia developed following surgical castration and tail docking in the untreated and placebo-treated lambs. It was absent in the castration wounds, and significantly reduced in the tail-docking wounds, of the treated lambs. Hot-iron docking was associated with mild and transient secondary hyperalgesia, which was abolished by the topical anaesthesia. There was a significant reduction in pain-related behaviours in treated lambs, which were not significantly different in their behaviour to the sham-operation handled controls. Plasma lignocaine and bupivacaine levels were below the toxic thresholds in all tested lambs. Conclusion Topical anaesthesia alleviates wound pain and significantly reduces pain-related behaviours in lambs undergoing surgical castration plus surgical or hot-iron tail docking, without a negative effect on wound healing or a risk of systemic toxicity.     

Classification of guppies’ (Poecilia reticulata) gender by computer vision

Labor costs of guppy growers and breeders are largely those of manual sorting (by strain, quality and gender) and counting fish. In most farms, female and male fish are grown together and sold either separately or together. Sorting fish according to gender is important for marketing as well as for breeding programs, so that a device for sorting and counting fish can potentially reduce production costs and improve quality.

A project aiming to develop sorting and counting technologies for ornamental fish growers included development and testing of image-processing algorithms for sorting guppy fish (Poecilia reticulata) by gender. The algorithms are derived from shape and color differences between female and male guppies. An algorithm for the determination of landmarks on fish contours was developed and found to be accurate in accordance with human judgment, enabling extraction of specific shape and color features of the tail and the body.

The algorithms were applied to three sets of images of guppies of the “Red-Blond” strain. Gender identification accuracy was approximately 90% using shape features, approximately 96% using color features and was slightly improved when both color and shape features were used.

Some of the components used are essential for future development of a computer vision based system for sorting and grading ornamental fish by strain and quality.