Active Transport of Glutamate in Leishmania (Leishmania) amazonensis

Leishmania spp. are the causative agents of leishmaniasis, a complex of diseases with a broad spectrum of clinical manifestations. Leishmania (Leishmania) amazonensis is a main etiological agent of diffuse cutaneous leishmaniasis. Leishmania spp., as other trypanosomatids, possess a metabolism based significantly on the consumption of amino acids. However, the transport of amino acids in these organisms remains poorly understood with few exceptions. Glutamate transport is an important biological process in many organisms. In the present work, the transport of glutamate is characterized. This process is performed by a single kinetic system (K-m=0.59 +/- 0.04 mM, V-max=0.123 +/- 0.003 nmol/min per 20 x 10(6) cells) showing an energy of activation of 52.38 +/- 4.7 kJ/mol and was shown to be partially inhibited by analogues, such as glutamine, aspartate, alpha-ketoglutarate and oxaloacetate, methionine, and alanine. The transport activity was sensitive to the extracellular concentration of H+ but not to Na+ or K+. However, unlike other amino acid transporters presently characterized, the treatment with specific ionophores confirmed the participation of a K+, and not H+ membrane gradient in the transport process.

Growth, encystment and survival of Acanthamoeba castellanii grazing on different bacteria

Free-living amoebae of the genus Acanthamoeba are widely distributed in soil and water collections, where trophozoites (vegetative, multiplicative stages) feed mainly by phagocytosis and thus control bacterial populations in the environment. Here, we examined the growth, encystment and survival of Acanthamoeba castellanii receiving different bacteria (Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Bacillus subtilis, Bacillus megaterium, Micrococcus luteus, and Staphylococcus aureus) in nonnutrient saline. All bacteria assayed induced a dose-dependent proliferative response, in most cases maximized with a bacterial dose of 1 x 10(9) mL(-1); except for M. luteus, trophozoites grew better with viable than with heat-killed bacteria. In addition, Acanthamoeba growth was improved by adding bacteria on alternate days. Single-dose experiments indicated a temporal association between the growth of trophozoite and bacterial consumption, and higher consumption of M. luteus, E. coli and P. aeruginosa, bacterial species that allowed the highest trophozoite yields. Long-term Acanthamoeba-bacteria incubation revealed that encystment was significantly delayed by almost all the bacteria assayed (including S. aureus, which elicited a poor growth response) and that the presence of bacteria markedly increased cyst yield; final cyst recovery clearly depended on both the dose and the type of the bacterium given, being much higher with E. coli, M. luteus and P. aeruginosa.

Connective tissue mast cells are the target of formaldehyde to induce tracheal hyperresponsiveness in rats: Putative role of leukotriene B(4) and nitric oxide

Formaldehyde (FA) exposure induces upper airways irritation and respiratory abnormalities, but its mechanisms are not understood. Since mast cells are widely distributed in the airways, we hypothesized that FA might modify the airways reactivity by mechanism involving their activation. Tracheal rings of rats were incubated with Dulbecco`s modified medium culture containing FA (0.1 ppm) in 96-well plastic microplates in a humid atmosphere. After 30 min, 6 h, and 24-72 h, the rings were suspended in an organ bath and dose-response curve to methacholine (MCh) were determined. incubation with FA caused a transient tracheal hyperresponsiveness to MCh that was independent from tracheal epithelium integrity. Connective tissue mast cell depletion caused by compound 48/80 or mast cell activation by the allergic reaction, before exposure of tracheal rings to FA prevented the increased responsiveness to MCh. LTB(4) concentrations were increased in the culture medium of tracheas incubated with FA for 48 h, whereas the LTB(4)-receptor antagonist MK886 (1 mu M) added before FA exposure rendered the tracheal rings normoreactive to MCh. In addition, FA exposure did not cause hyperresponsiveness in tracheal segments incubated with L-arginine (1 mu M). We suggest that airway connective tissue mast cells constitute the target and may provide the increased LTB(4) generation as well as an elevated consumption of NO leading to tracheal hyperresponsiveness to MCh. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

The Crystal Complex of Phosphofructokinase-2 of Escherichia coli with Fructose-6-phosphate KINETIC AND STRUCTURAL ANALYSIS OF THE ALLOSTERIC ATP INHIBITION

Substrate inhibition by ATP is a regulatory feature of the phosphofructokinases isoenzymes from Escherichia coli (Pfk-1 and Pfk-2). Under gluconeogenic conditions, the loss of this regulation in Pfk-2 causes substrate cycling of fructose-6-phosphate (fructose-6-P) and futile consumption of ATP delaying growth. In the present work, we have broached the mechanism of ATP-induced inhibition of Pfk-2 from both structural and kinetic perspectives. The crystal structure of Pfk-2 in complex with fructose-6-P is reported to a resolution of 2 angstrom. The comparison of this structure with the previously reported inhibited form of the enzyme suggests a negative interplay between fructose-6-P binding and allosteric binding of MgATP. Initial velocity experiments show a linear increase of the apparent K(0.5) for fructose-6-P and a decrease in the apparent k(cat) as a function of MgATP concentration. These effects occur simultaneously with the induction of a sigmoidal kinetic behavior (n(H) of approximately 2). Differences and resemblances in the patterns of fructose-6-P binding and the mechanism of inhibition are discussed for Pfk-1 and Pfk-2, as an example of evolutionary convergence, because these enzymes do not share a common ancestor.

A green flow-based procedure for fluorimetric determination of acid-dissociable cyanide in natural waters exploiting multicommutation

A flow system designed with solenoid valves is proposed for determination of weak acid dissociable cyanide, based on the reaction with o-phthalaldehyde (OPA) and glycine yielding a highly fluorescent isoindole derivative. The proposed procedure minimizes the main drawbacks related to the reference batch procedure, based on reaction with barbituric acid and pyridine followed by spectrophotometric detection, i.e., use of toxic reagents, high reagent consumption and waste generation, low sampling rate, and poor sensitivity. Retention of the sample zone was exploited to increase the conversion rate of the analyte with minimized sample dispersion. Linear response (r=0.999) was observed for cyanide concentrations in the range 1-200 mu g L(-1), with a detection limit (99.7% confidence level) of 0.5 mu g L(-1)(19 nmol L(-1)). The sampling rate and coefficient of variation (n=10) were estimated as 22 measurements per hour and 1.4%, respectively. The results of determination of weak acid dissociable cyanide in natural water samples were in agreement with those achieved by the batch reference procedure at the 95% confidence level. Additionally to the improvement in the analytical features in comparison with those of the flow system with continuous reagent addition (sensitivity and sampling rate 90 and 83% higher, respectively), the consumption of OPA was 230-fold lower.

Thioxanthone Sensitized Photodegradation of Poly(alkyl methacrylate) Films

The thioxanthone-sensitized photodegradation of poly(alkyl methacrylate) films [alkyl = methyl, ethyl, butyl, and hexyl] was studied using near UV-vis light. The photooxidation process continued even after the total consumption of the sensitizer, possibly due to the excitation of the ketyl groups formed during the first stages of the process. The rate of oxidation, as well as the formation of hydroxy, peroxy, and ketyl groups was faster for polymers with larger ester groups. The decrease of the molecular weight of the degradated polymers was also larger for the hexyl substituted polymer. The side-chain size effect was attributed to the larger amount of secondary hydrogens available for abstraction by the triplet state of thioxanthone, present in the larger ester groups. The lower glass transition temperature of the hexyl substituted polymer allows a better diffusion of oxygen to the deeper layers of the films that also contributes to the faster photodegradation rate. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 115: 1283-1288, 2010

INVENTORY CONTROL SYSTEM : Optimization of production system and reliability

The main idea of this research to solve the problem of inventory management for the paper industry SPM PVT limited. The aim of this research was to find a methodology by which the inventory of raw material could be kept at minimum level by means of buffer stock level.The main objective then lies in finding the minimum level of buffer stock according to daily consumption of raw material, finding the Economic Order Quantity (EOQ) reorders point and how much order will be placed in a year to control the shortage of raw material.In this project, we discuss continuous review model (Deterministic EOQ models) that includes the probabilistic demand directly in the formulation. According to the formula, we see the reorder point and the order up to model. The problem was tackled mathematically as well as simulation modeling was used where mathematically tractable solution was not possible.The simulation modeling was done by Awesim software for developing the simulation network. This simulation network has the ability to predict the buffer stock level based on variable consumption of raw material and lead-time. The data collection for this simulation network is taken from the industrial engineering personnel and the departmental studies of the concerned factory. At the end, we find the optimum level of order quantity, reorder point and order days.

Use of Photovoltaic on an E-bike? : A Feasibility Study

In recent years the number of bicycles with e-motors has been increased steadily. Within the pedelec – bikes where an e-motor supports the pedaling – a special group of transportation bikes has developed. These bikes have storage boxes in addition to the basic parts of a bike. Due to the space available on top of those boxes it is possible to install a PV system to generate electricity which could be used to recharge the battery of the pedelec. Such a system would lead to grid independent charging of the battery and to the possibility of an increased range of motor support. The feasibility of such a PV system is investigated for a three wheeled pedelec delivered by the company BABBOE NORDIC.The measured data of the electricity generation of this mobile system is compared to the possible electricity generation of a stationary system.To measure the consumption of the pedelec different tracks are covered, and the energy which is necessary to recharge the bike battery is measured using an energy logger. This recharge energy is used as an indirect measure of the electricity consumption. A PV prototype system is installed on the bike. It is a simple PV stand alone system consisting of PV panel, charge controller with MPP tracker and a solar battery. This system has the task to generate as much electricity as possible. The produced PV current and voltage aremeasured and documented using a data logger. Afterwards the average PV power is calculated. To compare the produced electricity of the on-bike system to that of a stationary system, the irradiance on the latter is measured simultaneously. Due to partial shadings on the on-bike PV panel, which are caused by the driver and some other bike parts, the average power output during riding the bike is very low. It is too low to support the motor directly. In case of a similar installation as the PV prototype system and the intention always to park the bike on a sunny spot an on-bike system could generate electricity to at least partly recharge a bike battery during one day. The stationary PV system using the same PV panel could have produced between 1.25 and 8.1 times as much as the on-bike PV system. Even though the investigation is done for a very specific case it can be concluded that anon-bike PV system, using similar components as in the investigation, is not feasible to recharge the battery of a pedelec in an appropriate manner. The biggest barrier is that partial shadings on the PV panel, which can be hardly avoided during operation and parking, result in a significant reduction of generated electricity. Also the installation of the on-bike PV system would lead to increased weight of the whole bike and the need for space which is reducing the storage capacity. To use solar energy for recharging a bike battery an indirect way is giving better results. In this case a stationary PV stand alone system is used which is located in a sunny spot without shadings and adjusted to use the maximum available solar energy. The battery of the bike is charged using the corresponding charger and an inverter which provides AC power using the captured solar energy.

Attitudes Towards and Uses of the Japanese Adverbzenzen by Swedish Learners of Japanese

The word zenzen is an adverb that is used frequently in daily conversational Japanese. From the Meiji period (1868-1912) until the early Showa period (1924-1989) the word was used together with both affirmative and negative words to form expressions. In the early Showa period the grammatical rules in education changed so that the only acceptable use was together with a negative word. From the 1990’s onward, the use together with an affirmative word has made a comeback especially among younger Japanese people. However even though the usage together with an affirmative word has made a comeback and was considered normal once in history, in today’s society it is still considered as slang and thus not recommended usage in formal situations. Foreign language learners however, tend not to learn a language only by textbooks but also by imitating the language of native Japanese speakers and Japanese popular culture. This may lead to a confusion regarding what words are acceptable to use in conversations. Therefore in this study, an online survey that examines the usage and attitudes regarding the word zenzen aimed at Japanese language learners at Swedish universities was conducted. The results of the survey showed that although a majority of the learners showed a good understanding of the usage, more than half of the learners displayed a feeling of confusion regarding the usage of the word. The gender comparison regarding the usage showed no major differences. Having lived in Japan, having Japanese friends whom you speak Japanese with regularly and length of Japanese study was associated with an increased understanding of the usage. Regular consumption of Japanese popular culture, however, was not associated with an increased understanding of the usage. A literature analysis was also conducted to examine the attitudes regarding the usage of zenzen in a variety of books with topics including business language and books aimed at Japanese language teachers. The results showed that zenzen used together with a negative word was considered as the norm while zenzen used together with an affirmative word was not recommended to be used in formal situations. When recommending proper usage of the word zenzen together with an affirmative word to foreign learners of Japanese, hijou-ni and totemo was seen as better alternatives to zenzen in a formal situation.

Outdoor performance of forest seedlings pre-cultivated under artificial lights : effects of the light spectra used for pre-cultivation on the future establishment and development

Forest nurseries are essential for producing good quality seedlings, thus being a key element in the reforestation process. With increasing climate change awareness, nursery managers are looking for new tools that can help reduce the effects of their operations on the environment. The ZEPHYR project, funded by the European Commission under the Seventh Framework Programme (FP7), has the objective of finding new alternatives for nurseries by developing innovative zero-impact technologies for forest plant production. Due to their direct relationship to the energy consumption of the nurseries, one of the main elements addressed are the grow lights used for the pre-cultivation. New LED luminaires with a light spectrum tailored to the seedlings’ needs are being studied and compared against the traditional fluorescent lamps. Seedlings of Picea abies and Pinus sylvestris were grown under five different light spectra (one fluorescent and 4 LED) during 5 weeks with a photoperiod of 16 hours at 100 μmol∙m-2∙s-1 and 60% humidity. In order to evaluate if these seedlings were able cope with real field stress conditions, a forest field trial was also designed. The terrain chosen was a typical planting site in mid-Sweden after clear-cutting. Two vegetation periods after the outplanting, the seedlings that were pre-cultivated under the LED lamps have performed at least as well as those that were grown under fluorescent lights. These results show that there is a good  potential for lightning substitution in forestry nurseries.