The interplay of the N- and C-terminal domains of MCAK control microtubule depolymerization activity and spindle assembly

Spindle assembly and accurate chromosome segregation require the proper regulation of microtubule dynamics. MCAK, a Kinesin-13, catalytically depolymerizes microtubules, regulates physiological microtubule dynamics, and is the major catastrophe factor in egg extracts. Purified GFP-tagged MCAK domain mutants were assayed to address how the different MCAK domains contribute to in vitro microtubule depolymerization activity and physiological spindle assembly activity in egg extracts. Our biochemical results demonstrate that both the neck and the C-terminal domain are necessary for robust in vitro microtubule depolymerization activity. In particular, the neck is essential for microtubule end binding, and the C-terminal domain is essential for tight microtubule binding in the presence of excess tubulin heterodimer. Our physiological results illustrate that the N-terminal domain is essential for regulating microtubule dynamics, stimulating spindle bipolarity, and kinetochore targeting; whereas the C-terminal domain is necessary for robust microtubule depolymerization activity, limiting spindle bipolarity, and enhancing kinetochore targeting. Unexpectedly, robust MCAK microtubule (MT) depolymerization activity is not needed for sperm-induced spindle assembly. However, high activity is necessary for proper physiological MT dynamics as assayed by Ran-induced aster assembly. We propose that MCAK activity is spatially controlled by an interplay between the N- and C-terminal domains during spindle assembly.     

Effect of acetylcholine on the structuro-functional characteristics of bush-like interoceptors of the urinary bladder of the frog

In the preparation of the frog (Rana temporaria) isolated urinary bladder, it has been stated by morphological and physiological experiments that under an exogenic acetilcholine influence certain structural-functional changes take place in the bushy interoceptor. They are manifested as a sharp deterioration in coloration the receptor with methylene blue, the receptory plates decrease in their size, character of granuloformation changes in them. Simultaneously, an increased electrical activity is observed and, on the contrary, under the influence of atropine the impulse activity drops. It should be suggested that acetilcholine acts as an activator of metabolic and as a modulator of physiological processes in the receptor.     

Fluctuations in the enzymatic activity of the human endometrium

Cyclic fluctuations were studied in the activity of oxidoreductases playing a role in the major energy metabolic pathways, lysosomal and non-lysosomal hydrolases and some non-enzymatic cytochemical components demonstrable in different developmental physiological or pathophysiological phases of human endometrium. The total scope of the study involved 170 tissues and cytological specimens. The cytological material included microcurettings, aspirates, brush preparations and tissue prints. An evaluation of the usefulness of the application of enzyme cytochemistry to cytological material is included. The most important results were a cyclic fluctuation and a progestagenic controlled increase in the activity of many oxidoreductases, especially the NADPH regenerating enzymes of the pentose phosphate pathway, and of the NADP+ dependent isocitrate dehydrogenase. The histochemical evaluation of the activity of these NADP+ linked enzymes can therefore be recommended for the evaluation of the physiological status of the endometrial cells, especially in patients with infertility problems.     

Roles of DegP in prevention of protein misfolding in the periplasm upon overexpression of penicillin acylase in Escherichia coli

Enhancement of the production of soluble recombinant penicillin acylase in Escherichia coli via coexpression of a periplasmic protease/chaperone, DegP, was demonstrated. Coexpression of DegP resulted in a shift of in vivo penicillin acylase (PAC) synthesis flux from the nonproductive pathway to the productive one when pac was overexpressed. The number of inclusion bodies, which consist primarily of protein aggregates of PAC precursors in the periplasm, was highly reduced, and the specific PAC activity was highly increased. DegP was a heat shock protein induced in response to pac overexpression, suggesting that the protein could possibly suppress the physiological toxicity caused by pac overexpression. Coexpression of DegP(S210A), a DegP mutant without protease activity but retaining chaperone activity, could not suppress the physiological toxicity, suggesting that DegP protease activity was primarily responsible for the suppression, possibly by degradation of abnormal proteins when pac was overexpressed. However, a shortage of periplasmic protease activity was not the only reason for the deterioration in culture performance upon pac overexpression because coexpression of a DegP-homologous periplasmic protease, DegQ or DegS, could not suppress the physiological toxicity. The chaperone activity of DegP is proposed to be another possible factor contributing to the suppression.     

Functions of the activation loop in Csk protein-tyrosine kinase

Autophosphorylation in the activation loop is a common mechanism regulating the activities of protein-tyrosine kinases (PTKs). PTKs in the Csk family, Csk and Chk, are rare exceptions for lacking Tyr residues in this loop. We probed the function of this loop in Csk by extensive site-specific mutagenesis and kinetic studies using physiological and artificial substrates. These studies led to several surprising conclusions. First, specific residues in Csk activation loop had little discernable functions in phosphorylation of its physiological substrate Src, as Ala scanning and loop replacement mutations decreased Csk activity toward Src less than 40%. Second, some activation loop mutants, such as a single residue deletion or replacing all residues with Gly, exhibited 1-2% of wild type (wt) activity toward artificial substrates, but significantly higher activity toward Src. Third, introduction of a thrombin cleavage site to the activation loop also resulted in loss of 98% of wt activity for poly(E4Y) and loss of 95% of wt activity toward Src, but digestion with thrombin to cut the activation loop, resulted in full recovery of wt activity toward both substrates. This suggested that the catalytic machinery is fully functional without the activation loop, implying an inhibitory role by the activation loop as a regulatory structure. Fourth, Arg313, although universally conserved in protein kinases, and essential for the activity of other PTKs so far tested, is not important for Csk activity. These findings provide new perspectives for understanding autophosphorylation as a regulatory mechanism and imply key differences in Csk recognition of artificial and physiological substrates.     

The effect of the extremely weak field on the cultures of bacteria Escherichia coli and Staphylococcus aureus

The effect of extremely weak human electromagnetic field on the suspensions of Escherichia coli and Staphylococcus aureus in a physiological solution at 25 degrees C was studied. The results obtained indicate that the human field induces changes in the physical properties of liquid water in physiological solution and the induced structural and dynamic rearrangements of extracellular water are tranferred to bacterial cells and change their functional activity.     

论心理对生理的影响和作用

本文阐述了心理对生理的能动作用。在一定的条件下,人的心理可影响其生理功能,两者互为因果、相互影响。生理是心理活动的物质基础,心理是其生理的驱动力和标志。本文论述了精神、情绪、意念、信念、暗示等心理活动对生理的影响和作用及中医论心理对生理的作用,中医利用情志疗法的理论达到心理对生理趋向健康的作用。同时论证了肠道菌群通过心理间接影响生理功能,通过微生态制剂调节肠道菌群的微生态平衡,可改善患者的神经症状,使之心理活动达到最佳状态,进而使人的生理趋向健康。     

Criteria for the evaluation of the functional importance of endogenous analogues of pharmacological regulators

Criteria for the evaluation of the functional importance of endogenous analogues of pharmacological drugs are proposed. These include opposite changes in their content in opposite pathophysiological or physiological states, accompanied by corresponding changes in the functional activity of enzymes sensitive to changes in their level, and the regulation of target enzymes by physiological concentrations of such endogenous compounds. The applicability of these criteria has been demonstrated using tribulin, the endogenous family of inhibitors of monoamine oxidases.     

Adjustment of metabolite composition in the haemolymph to seasonal variations in the land snail <Emphasis Type="Italic">Helix pomatia</Emphasis>

In temperate regions, land snails are subjected to subzero temperatures in winter and hot temperatures often associated to drought in summer. The response to these environmental factors is usually a state of inactivity, hibernation and aestivation, respectively, in a temperature and humidity buffered refuge, accompanied by physiological adjustments to resist cold or heat stress. We investigated how environmental factors in the microhabitat and body condition influence the metabolite composition of haemolymph of the endangered species Helix pomatia. We used UPLC and GC–MS techniques and analyzed annual biochemical variations in a multivariate model. Hibernation and activity months differed in metabolite composition. Snails used photoperiod as cue for seasonal climatic variations to initiate a physiological state and were also highly sensitive to temperature variations, therefore constantly adjusting their physiological processes. Galactose levels gave evidence for the persistence of metabolic activity with energy expenditure during hibernation and for high reproductive activity in June. Triglycerides accumulated prior to hibernation might act as cryoprotectants or energy reserves. During the last month of hibernation snails activated physiological processes related to arousal. During activity, protein metabolism was reflected by high amino acid level. An exceptional aestivation period was observed in April giving evidence for heat stress responses, like the protection of cells from dehydration by polyols and saccharides, the membrane stabilization by cholesterol and enhanced metabolism using the anaerobic succinic acid pathway to sustain costly stress responses. In conclusion, physiological adjustments to environmental variations in Helix pomatia involve water loss regulation, cryoprotectant or heatprotectant accumulation.     

A quantitative genetic study for elucidating the contribution of glutamine synthetase,glutamate dehydrogenase and other nitrogen-related physiological traits to the agronomic performance of common wheat

To better understand the genetic variability for nitrogen use efficiency in winter wheat is a necessity in the frame of the present economic and ecological context. The objective of this work was to investigate the role of the enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH), and other nitrogen (N)-related physiological traits in the control of agronomic performance in wheat. A quantitative genetics approach was developed using the Arche × Récital population of doubled haploid lines grown for 3 years in the field. GS and GDH activities, ammonium, amino acid and protein contents were measured at different stages of plant development in different organs after flowering. Significant genotypic effects were observed for all measured physiological and agronomical traits. Heading date was negatively correlated with ammonium, amino acid, protein contents and GS activity in the flag leaf lamina. Grain protein content was positively correlated with both ammonium and amino acid content, and to a lesser extent with soluble protein content and GS activity. A total of 148 quantitative trait loci (QTLs) were detected, 104 QTLs for physiological traits and 44 QTLs for agronomic traits. Twenty-six QTLs were detected for GDH activity spread over 13 chromosomes and 25 QTLs for GS activity spread over 12 chromosomes. We found only a co-localization between a QTL for GS activity and GSe, a structural gene encoding cytosolic GS on chromosome 4B. A coincidence between a QTL for GDH activity and a gene encoding GDH was also found on chromosome 2B. QTL regions combining both physiological and agronomical QTLs were mainly identified on linkage groups 2A, 2B, 2D, 5A, 5B and 5D. This approach allowed us to propose possible functions of physiological traits to explain the variation observed for agronomic traits including yield and its components. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Physiological relaxation induced by horticultural activity: transplanting work using flowering plants

Background

Despite increasing attention and a growing volume of research data, little physiological evidence is available on the benefits of horticultural activity and the different effects on individuals. Therefore, the aim of the present study was to investigate the physiological effects of horticultural activity and to examine how differences in personality alter these effects.

Results

The effects of transplanting real flowers (horticultural activity) and handling artificial flowers (control activity) on human physiological activity were compared. On the first day, eight participants engaged in horticultural activity and another eight in the control activity. On the second day, participants switched roles. Participants’ physiological conditions during each activity were assessed by measuring the heart rate and heart rate variability (HRV). Psychological responses, which were measured using a semantic differential rating scale, showed that the horticultural activity promoted comfortable, soothed, and natural feelings, compared to the control activity. Analysis of physiological responses using two-way repeated measures analysis of variance (ANOVA) revealed that sympathetic nervous activity significantly decreased in the late time period (11 to 15 minutes) of horticultural activity only in the type A group.

Conclusions

This study supports the fact that the horticultural activity can enhance psychological and physiological relaxation effects, although these physiological effects can differ among individuals with different personalities.     

抗肿瘤海绵放线菌的分离及菌株HA01184的鉴定

目的：对采自海南、湛江等海域的海绵样品进行放线菌选择性分离,采用其发酵液进行抗肿瘤活性筛选,并对活性较好的菌株进行鉴定。方法：用含50μg／mL重铬酸钾为抑制剂的海水高氏一号合成培养基分离培养海绵放线菌;以MTT法进行菌株的抗肿瘤活性筛选;通过培养特征、形态特征、生理生化特征、16S rDNA序列测定及系统发育分析,对菌株HA01184进行鉴定。结果与结论：海水高氏一号合成培养基用于海绵放线菌分离培养具有很好的选择性,从海绵样品中共分离得到放线菌165株,细胞毒活性达80％以上的阳性菌株有10株,其中菌株HA01184的发酵液细胞毒活性为90％。结合形态观察、生理生化特征和16S rDNA序列比对分析,将HA01184归于链霉菌属,可能是来自海洋环境的一个潜在新种。     

Lactoferrin Is an Allosteric Enhancer of the Proteolytic Activity of Cathepsin G

Protease-mediated degradation of proteins is critical in a plethora of physiological processes. Neutrophils secrete serine proteases including cathepsin G (CatG), neutrophile elastase (NE), and proteinase 3 (PR3) together with lactoferrin (LF) as a first cellular immune response against pathogens. Here, we demonstrate that LF increases the catalytic activity of CatG at physiological concentration, with its highest enhancing capacity under acidic (pH 5.0) conditions, and broadens the substrate selectivity of CatG. On a functional level, the enzymatic activity of CatG was increased in the presence of LF in granulocyte-derived supernatant. Furthermore, LF enhanced CatG-induced activation of platelets as determined by cell surface expression of CD62P. Consequently, LF-mediated enhancement of CatG activity might promote innate immunity during acute inflammation.     

Variation in ecosystem carbon dynamics of saltwater marshes in the northern Gulf of Mexico

Wetlands are highly productive ecosystems that can sequester large quantities of carbon. However, variation in physiological potential can alter their capacity to sequester carbon. To examine variation in ecosystem physiological capacity, we established three coastal marsh sites in Mississippi and Alabama spanning a productivity gradient. Over 1 year, we measured ecophysiological activity and spectral indices in two vegetation zones within each marsh to develop a better understanding of variation in ecosystem responses and health. Gross ecosystem exchange of carbon and ecosystem respiration rates (R_eco) differed significantly among sites, with the highest activity at Grand Bay, Mississippi and Point Aux Pines, Alabama and lower ecophysiological activity at Dauphin Island, Alabama. Net ecosystem exchange was similar for all three study areas because greater carbon assimilation was negated by higher levels of respiration. Spectral indices and leaf area were significantly different by marsh vegetation zone, suggesting that alterations in species composition and plant productivity can have important implications for carbon sequestration. While limited to 1 year, this study establishes a foundation by which to evaluate future research conducted over greater temporal and spatial scales, thereby enhancing our understanding of marsh physiological activity.     

Functional morphology of the adrenal cortex during the physiological rhythm of sex steroid synthesis]

A method of a quantitative histenzymological analysis was used for determination of the extent of participation of various zones of the adrenal cortex in provision of physiological rhythm of the steroid synthesis. The activity of 3beta-OH-steroid dehydrogenase, glucoso-6-phosphat dehydrogenase, NAD- and NADP-diaphorases, acid and alkaline phosphatases and non-specific esterase was investigated. The data of a histoenzymotological study were compared with the result of biochemical analysis of corticosteroids in the peripheral blood. Under physiological conditions intensification of steroidogenesis was realized by mobilization of individual groups of cells of the adrenal gland, not its whole parenchyma. Physiological rhythm of steroid synthesis was provided by an integrated function of all the zones; however, by duration and quantitative expression of the secretory activity a structural heterogeneity was revealed in the adrenal cortex.     

The effect of quercetin on the phosphorylation activity of the Rous sarcoma virus transforming gene product in vitro and in vivo

The phosphotransferase activity of the Rous sarcoma virus src gene product, pp60src, was inhibited both in vitro and in vivo by the bioflavonoid quercetin. The Ki for the inhibitory effect was in the range of 6-11 microM under conditions in vitro. The inhibitory effect of quercetin was competitive towards the nucleotides ATP and GTP as substrates for pp60src and was non-competitive towards alpha-casein as the protein substrate of this kinase activity. In contrast, studies in vitro of the phosphotransferase activity of the catalytic subunit of the cAMP-dependent protein kinase showed that this flavonoid did not inhibit the phosphorylation of physiological substrates of this enzyme. In cultured cells the half-maximal inhibition of tyrosine phosphorylation of pp60src as well as the phosphorylation of the Mr = 34000 protein, a physiological substrate of pp60src, was in the range 0.06-0.08 mM.     

Long-term changes in the physiologic and biochemical status of the body through immunization with endogenous regulators

Immunization with protein and oligopeptide, and low-molecular weight nonpeptide compounds demonstrated a possibility for selective decrease in the level (activity) of several endogenous compounds, and consequently, for a significant prolonged changes is physiological functions connected with these compounds. The method opens possibilities for directed regulation of physiological status of organism, correction of pathological changes, and creation of relatively stable functional systems with new properties.     

干旱胁迫对不同烤烟品种根系生长和生理特性的影响

以抗旱型烤烟品种‘农大202'及一般型烤烟品种‘NC89'和‘K326'为材料,采用盆栽试验研究了干旱胁迫对根系生长和生理特性的影响,以明确各烟草品种的抗旱能力及其与根系生长和生理特性的关系.结果显示:严重干旱胁迫之后各烤烟品种根系鲜重、干重上升,而根系活力、根系吸收面积以及根系SOD和POD活性等根系生理指标则呈下降趋势.在干旱胁迫条件下,‘农大202'的根系总吸收面积、活跃吸收面积、根系活力、SOD活性和POD活性均显著高于两对照品种‘NC89'和‘K326',但其根系鲜重和干重等生物量并不具有太大的优势.研究表明,烟草的根系生长和生理特性对环境水分条件的响应存在明显的基因型差异;在干旱胁迫条件下, ‘农大202'根系能保持较高总吸收面积、活跃吸收面积、根系活力和保护酶活性,是其具有较强抗旱性的生理基础.     

Activation of trout gill AMP deaminase by an endogenous proteinase--I. Effects on the regulatory properties of the enzyme

Some regulatory properties of trout gill AMP deaminase were determined in crude extracts, before or after modification of the enzyme by the endogenous proteinase. After proteolysis, the optimal concentrations for activation by sodium and potassium were shifted from 10 to 75 mM, resulting in a large increase of enzyme activity near the physiological potassium concentration. This activation was shown to be the consequence of a much lower sensitivity of AMP deaminase to inhibition by increasing ionic strength. The modified enzyme was also less sensitive to modifications of pH and to inhibition by physiological concentrations of inorganic phosphate. When all these modifications were considered, limited proteolysis of gill AMP deaminase resulted in a 40 times increase of enzyme activity under in vivo conditions.     

Cephalic vasomotor feedback in the modification of migraine headache

The effect of cephalic vasomotor response (CVMR) and frontalis electromyographic (EMG) feedback on control of temporal arterial vasoconstriction and frontalis muscle activity in migraine and muscle contraction headache patients was investigated. A single subject multiple baseline design (across subjects and responses) was introduced to evaluate (1) patterning in the two physiological systems and (2) the effects of CVMR and EMG feedback on headache activity. The data indicated that (a) all four patients demonstrated an ability to control CVMR activity during CVMR feedback and EMG during EMG feedback, (b) idiosyncratic patterns of physiological activity emerge during feedback training, and (c) learned control of the pain mechanism for muscle contraction and migraine headaches was related to reduced frequency and duration of these headaches.Portions of this paper were presented at the Ninth Annual Convention of the Association for Advancement of Behavior Therapy, San Francisco, 1975.