Regulation of Leaf Senescence by Cytokinin, Sugars, and Light. Effects on NADH-Dependent Hydroxypyruvate Reductase

The aim of this study was to investigate the interactions between cytokinin, sugar repression, and light in the senescence-related decline in photosynthetic enzymes of leaves. In transgenic tobacco (Nicotiana tabacum) plants that induce the production of cytokinin in senescing tissue, the age-dependent decline in NADH-dependent hydroxypyruvate reductase (HPR), ribulose-1,5-bisphosphate carboxylase/oxygenase, and other enzymes involved in photosynthetic metabolism was delayed but not prevented. Glucose (Glc) and fructose contents increased with leaf age in wild-type tobacco and, to a greater extent, in transgenic tobacco. To study whether sugar accumulation in senescing leaves can counteract the effect of cytokinin on senescence, discs of wild-type leaves were incubated with Glc and cytokinin solutions. The photorespiratory enzyme HPR declined rapidly in the presence of 20 mM Glc, especially at very low photon flux density. Although HPR protein was increased in the presence of cytokinin, cytokinin did not prevent the Glc-dependent decline. Illumination at moderate photon flux density resulted in the rapid synthesis of HPR and partially prevented the negative effect of Glc. Similar results were obtained for the photosynthetic enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. It is concluded that sugars, cytokinin, and light interact during senescence by influencing the decline in proteins involved in photosynthetic metabolism

Analysis and preliminary design of optical sensors for propulsion control

A fiber-optic sensor concept screening study was performed. Twenty sensor subsystems were identified and evaluated. Two concepts selected for further study were the Fabry-Perot fiber-optic temperature sensor and the pulse-width-modulated phosphorescent temperature sensor. Various designs suitable for a Fabry-Perot temperature sensor to be used as a remote fiber-optic transducer were investigated. As a result, a particular design was selected and constructed. Tests on this device show that spectral peaks are produced from visible white light, and the change in wavelength of the spectral peaks produced by a change in temperature is consistent with theory and is 36 nm/C for the first order peak. A literature search to determine a suitable phosphor for implementing the pulse-width-modulated fiber optic temperature sensor was conducted. This search indicated that such a device could be made to function for temperatures up to approximately 200 C. Materials like ZnCdS and ZnSe activated with copper will be particularly applicable to temperature sensing in the cryogenic to room temperature region. While this sensing concept is probably not applicable to jet engines, the simplicity and potential reliability make the concept highly desirable for other applications

Regions of beta 2 and beta 4 responsible for differences between the steady state dose-response relationships of the alpha 3 beta 2 and alpha 3 beta 4 neuronal nicotinic receptors

We constructed chimeras of the rat beta 2 and beta 4 neuronal nicotinic subunits to locate the regions that contribute to differences between the acetylcholine (ACh) dose-response relationships of the alpha 3 beta 2 and alpha 3 beta 4 receptors. Expressed in Xenopus oocytes, the alpha 3 beta 2 receptor displays an EC50 for ACh approximately 20-fold less than the EC50 of the alpha 3 beta 4 receptor. The apparent Hill slope (n(app)) of alpha 3 beta 2 is near one whereas the alpha 3 beta 4 receptor displays an n(app) near two. Substitutions within the first 120 residues convert the EC50 for ACh from one wild-type value to the other. Exchanging just beta 2:104-120 for the corresponding region of beta 4 shifts the EC50 of ACh dose-response relationship in the expected direction but does not completely convert the EC50 of the dose- response relationship from one wild-type value to the other. However, substitutions in the beta 2:104-120 region do account for the relative sensitivity of the alpha 3 beta 2 receptor to cytisine, tetramethylammonium, and ACh. The expression of beta 4-like (strong) cooperativity requires an extensive region of beta 4 (beta 4:1-301). Relatively short beta 2 substitutions (beta 2:104-120) can reduce cooperativity to beta 2-like values. The results suggest that amino acids within the first 120 residues of beta 2 and the corresponding region of beta 4 contribute to an agonist binding site that bridges the alpha and beta subunits in neuronal nicotinic receptors

Stimulation of erythropoietin in renal insufficiency by hypobaric hypoxia

Patients with renal anaemia show inadequate levels of immunoreactive erythropoietin (Epo) related to the degree of anaemia. The purpose of our study is to compare the degree of stimulation of Epo by means of hypobaric hypoxia in normal controls and patients with renal anaemia. Baseline Epo concentrations were found to be 11.1±2.0 U/l in 10 healthy volunteers and 11.4±4.6 U/l in six patients with renal anaemia. After exposure to hypobaric hypoxia equivalent to 4560 m above sea level for a duration of 3.5 h, we observed a significant increase in serum Epo in healthy volunteers to 22.8±9.1 U/l (P<0.005), while there was no increase in patients with renal anaemia: 12.3±5.2 U/l (P<0.2). Our results show that in patients with renal anaemia serum Epo concentrations are comparable to those of normal controls, but inadequate in view of the concomitant degree of anaemia. Stimulation by acute hypobaric hypoxia was not possible in patients with renal insufficiency as opposed to normal controls. From these data it can be concluded that either Epo production is working at maximum capacity under baseline conditions, or an additional hybobaric stimulus is not able to influence a disturbed set point of the oxygen sensor regulating Epo synthesi

Protein kinase C modulates the activity of a cloned gamma-aminobutyric acid transporter expressed in Xenopus oocytes via regulated subcellular redistribution of the transporter

We report that activators and inhibitors of protein kinase C (PKC) and protein phosphatases regulate the activity of a cloned rat brain gamma- aminobutyric acid (GABA) transporter (GAT1) expressed in Xenopus oocytes. Four compounds known to activate PKC increased GABA uptake 2- 3.5-fold over basal control levels. Inhibition of PKC by bisindolylmaleimide reduced basal GABA uptake 80% and blocked the phorbol 12-myristate 13-acetate (PMA)-induced stimulation of transport. Okadaic acid, a protein phosphatase inhibitor, stimulated transport 2.5- fold; a 4-fold increase in GABA uptake occurred when oocytes were treated with cyclosporin A, a specific inhibitor of protein phosphatase 2B. Modulation resulted in changes to Vmax but not to Km and was influenced by the functional expression level of the transporter protein; as expression level increased, the ability to up-regulate transporter activity decreased. Down-regulation of transporter activity was independent of expression level. Modulation did not occur through phosphorylation of the three consensus PKC sites predicted by the primary protein sequence since their removal had no effect on the susceptibility of the transporter to modulation by PMA or bisindolylmaleimide. Subcellular fractionation of oocyte membranes demonstrated that under basal level conditions, the majority of GAT1 was targeted to a cytoplasmic compartment corresponding to the trans- Golgi or low density vesicles. Stimulation of PKC with PMA resulted in a translocation of transporters from this compartment to the plasma membrane. At higher expression levels of GAT1 protein, a larger portion of GAT1 was found on the plasma membrane during basal level conditions and treatment with bisindolylmaleimide resulted in removal of these transporters from the plasma membrane. At expression levels demonstrated to be resistant to modulation by PMA, PMA-treatment still resulted in translocation of transporters from the cytoplasm to the plasma membrane. Thus, the inability of PMA to increase uptake at high expression of the GAT1 protein is due to saturation at a step subsequent to translocation. These findings 1) demonstrate the presence of a novel regulated secretory pathway in oocytes and 2) suggest a modulatory mechanism for neurotransmitter transporters that could have significant effects upon synaptic function

Perfect design or practical study? A workshop on navigating the challenges of community based prevention research

Subject: There is a shared interest among public health researchers in tackling methodological issues surrounding community based research, and on moving beyond a focus on individual level change. As part of a project on community empowerment funded by the People’s Health Trust, we have conducted a feasibility study on quantitative and economic evaluation of complex community-based interventions. To understand different quantitative methods that can be used to evaluate community empowerment interventions, we have undertaken a methodological literature review that identified the following sets of challenges: Defining population of interest – interventions taking place at a community level are not specifically targeted at a well-defined group of individuals. Therefore it is challenging to even find those who are affected by an intervention. Diverse and un-prescribed effects – the effect of community empowerment interventions are likely broad, suggesting we need to measure multiple outcomes in order to detect change. This increases the likelihood of detecting spurious change and can require a lot of resource. Furthermore, in many cases these outcomes are not pre-defined by a programme (i.e. communities choose their own foci)

A discrete slug population model determined by egg production

Slugs are significant pests in agriculture (as well as a nuisance to gardeners), and it is therefore important to understand their population dynamics for the construction of efficient and effective control measures. Differential equation models of slug populations require the inclusion of large (variable) temporal delays, and strong seasonal forcing results in a non-autonomous system. This renders such models open to only a limited amount of rigorous analysis. In this paper, we derive a novel batch model based purely upon the quantity of eggs produced at different times of the year. This model is open to considerable reduction; from the resulting two variable discrete-time system it is possible to reconstruct the dynamics of the full population across the year and give conditions for extinction or global stability and persistence. Furthermore, the steady state temporal population distribution displays qualitatively different behavior with only small changes in the survival probability of slugs. The model demonstrates how small variations in the favorability of different years may result in widely different slug population fluctuations between consecutive years, and is in good agreement with field data

Optical temperature sensor utilizing birefringent crystals

A temperature sensor comprising an optical transducer member having an array of birefringent crystals. The length and, accordingly, the sensitivity to temperature change of successive birefringent crystals varies according to a particular relationship. The transducer is interconnected with a fiber optic transmission and detecting system. Respective optical output signals that are transmitted from the birefringent crystals via the fiber optic transmission system are detected and decoded so as to correspond to digits of a numbering system, whereby an accurate digital representation of temperature can ultimately be provided