A novel model predictive control for a piecewise affine class of hybrid system with repetitive disturbance.

The present investigation addresses an innovative method based on explicit form of the model predictive control (EMPC) for a constrained Piecewise affine (PWA) class of hybrid systems, considering repetitive disturbance. This model of hybrid systems is investigated due to the fact that PWA modeling structure can approximate nonlinear systems via various operating points, and also because the simulation of PWA models are easy. With EMPC, the problem of optimization is solved in an offline way only once. Unlike conventional EMPC, the process information of the past and the data which are predicted are applied in the proposed strategy. This is the first time that in this study, the investigators adopt an approach in which these predicted data are weighted by another optimization problem (OP) and this weighted predicted sequence along with the past information of the process as an updating control input formula. In fact, two separate OPs are solved simultaneously at each step of proposed EMPC. The first one is linked with calculating the control input from the constrained cost function of EMPC algorithm and the second one concerns finding the optimal weighting factors in order to minimize the error signal, i.e. the difference between the reference path and the output signal at each optimization step of EMPC strategy. The precision of the proposed method is extremely dependent on the accuracy of the process model, so iterative learning control (ILC) algorithm is applied to protecting the process model against the periodic disturbances. These mathematical analyses are proven and validated by simulation results.

Molecular cloning of chitinase 33 (chit33) gene from Trichoderma atroviride / Clonagem molecular do gene quitinase 33 (chit33) em Trichoderma atroviride

In this study Trichoderma atroviride was selected as over producer of chitinase enzyme among 30 different isolates of Trichoderma sp. on the basis of chitinase specific activity. From this isolate the genomic and cDNA clones encoding chit33 have been isolated and sequenced. Comparison of genomic and cDNA sequences for defining gene structure indicates that this gene contains three short introns and also an open reading frame coding for a protein of 321 amino acids. The deduced amino acid sequence includes a 19 aa putative signal peptide. Homology between this sequence and other reported Trichoderma Chit33 proteins are discussed. The coding sequence of chit33 gene was cloned in pEt26b(+) expression vector and expressed in E. coli.

Neste estudo Trichoderma atroviride foi escolhido como superprodutor da enzima quitinase dentre 30 isolados de Trichoderma sp. com base na atividade específica de quitinase. Clones de cDNA e genômico codificando chit33 foram obtidos deste isolado e seqüenciados. A comparação das seqüências genômica e de cDNA para definir a estrutura do gene indicou que este contém três pequenos introns e uma fase aberta de leitura codificando uma proteína de 321 aminoácidos. A seqüência de aminoácidos deduzida inclui um possível peptídio sinal de 19 aminoácidos. Homologia entre esta seqüência e outras proteínas Chit33 descritas de Trichoderma é discutida. A seqüência codificadora do gene chit33 foi clonada no vetor de expressão pET26b(+) e expressa em E. coli.

Molecular cloning of chitinase 33 (chit33) gene from Trichoderma atroviride.

In this study Trichoderma atroviride was selected as over producer of chitinase enzyme among 30 different isolates of Trichoderma sp. on the basis of chitinase specific activity. From this isolate the genomic and cDNA clones encoding chit33 have been isolated and sequenced. Comparison of genomic and cDNA sequences for defining gene structure indicates that this gene contains three short introns and also an open reading frame coding for a protein of 321 amino acids. The deduced amino acid sequence includes a 19 aa putative signal peptide. Homology between this sequence and other reported Trichoderma Chit33 proteins are discussed. The coding sequence of chit33 gene was cloned in pEt26b(+) expression vector and expressed in E. coli.

Effect of plant growth regulators on in vitro biological control of Fusarium oxysporum by Trichoderma harzianum (T8).

In this study the effect of two plant growth regulators (indolacetic acid, IAA and gibberellic acid, GA3) and also Trichoderma harzianum (T8) on the phytopathogen fungus Fusarium oxysporium (F15) was investigated. IAA and GA3 with 15 and 30 ppm concentration have no significant effect on T. harzianum (T8) growth. The biocontrol activity of T. harzianum on F. oxysporum was slightly decreased by the presence of IAA and/or GA3. Addition of 40 ppm of GA3 to the culture medium of F. oxsporum increased polygalacturonase activity about 100%. A strong increasing effect on chitinase activity (60%) by T. harzianum (T8) was observed in the presence of phytopathogenic fungus F. oxysporum, but 40 ppm IAA and/or GA3 decreased about 47% of chitinase activity of T. harzianum.

Estradiol increases delayed, N-methyl-D-aspartate receptor-mediated excitation in the hippocampal CA1 region.

Hippocampal functions, e.g. synaptic plasticity and hippocampal-dependent behavior, are influenced by the circulating levels of ovarian steroids in adult, female rats. The mechanisms underlying this estradiol-dependent modulation, however, are poorly understood. One possibility is that estradiol alters N-methyl-D-aspartate (NMDA)-receptor functioning in the hippocampus. Here, using the in vitro hippocampal slice preparation, we evaluate estradiol-dependent changes in the NMDA receptor- and the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor-mediated components of excitatory postsynaptic potentials (EPSPs) evoked in CA1 by Schaffer collateral test stimulation. Using established experimental conditions [J Neurosci 17 (1997) 1848], we replicate the observation that estradiol pretreatment of ovariectomized rats increases a pharmacologically isolated NMDA receptor-mediated EPSP evoked by Schaffer collateral stimulation. However, using different conditions that optimize study of this evoked response, the estradiol-dependent increase in the monosynaptic NMDA receptor-mediated EPSP is eliminated. Low-intensity test stimulation of the Schaffer collaterals in this optimized medium reveals a novel, late NMDA receptor-mediated EPSP in CA1 from estradiol-pretreated rats. The mechanism(s) underlying this estradiol-dependent increase in a late, NMDA receptor-mediated EPSP is not known, but enhanced CA1-CA1 excitatory circuitry and glutamate spillover could contribute to this response. We conclude that estradiol pretreatment enhances NMDA receptor function in the female hippocampus by increasing not the monosynaptic, but rather a late NMDA receptor-mediated response. Variations in the magnitude of this late response may well contribute to ovarian steroid-dependent modulation of hippocampal synaptic plasticity.

Nicotine and its interaction with beta-amyloid protein: a short review.

Two features of Alzheimer's disease (AD) are beta-amyloid protein (betaAP) deposition and a severe cholinergic deficit. beta-Amyloid protein is a 39- to 43-amino acid transmembrane fragment of a larger precursor molecule, amyloid precursor protein. It is a major constituent of senile plaque, a neuropathologic hallmark of AD, and has been shown to be neurotoxic in vivo and in vitro. The cholinergic neurotransmission system is seen as the primary target of AD. However, other systems are also found to show functional deficit. An association between cholinergic deficit and betaAP is suggested by a negative correlation between cigarette smoking and AD. Evidence hitherto suggests that betaAP causes neuronal death possibly via apoptosis by disrupting calcium homeostasis, which may involve direct activation or enhancement of ligand-gated or voltage-dependent calcium channels. Selective second messengers such as protein kinases are triggered that signal neuronal death. Nicotine or acetylcholinesterase inhibitors can partially prevent the neurotoxicity of betaAP in vivo and in vitro. However, the exact mechanism by which nicotine provides its protective effects is not fully understood, but clearly there are protective roles for nicotine. Here, some aspects of betaAP neurotoxicity and nicotinic intervention as a protective agent are discussed.

Estradiol modulates long-term synaptic depression in female rat hippocampus.

Fluctuating estradiol levels in the adult, female rat modify the anatomical and functional organization of the hippocampal CA1 region. When systemic levels of estradiol are low, e.g., on estrus or in ovariectomized (OVX) rats, long-term synaptic potentiation is difficult to induce in vivo. However, little is known about the role of this ovarian hormone in long-term synaptic depression. Using multiple conditioning paradigms, we assess the magnitude of long-term depression (LTD) at CA3-CA1 synapses in vitro from adult, ovariectomized rats as a function of systemic estradiol replacement. In hippocampal slices from control OVX rats with low levels of estradiol, a low-frequency (2 Hz), asynchronous conditioning stimulation protocol does not produce LTD at 1 h postconditioning. However, this same protocol induces robust LTD in slices from estradiol-treated OVX rats. When the conditioning frequency is increased to 4 Hz, slices from both groups of rats show robust LTD in vitro. At an even higher conditioning frequency (10 Hz), the 2-Hz-based observations are reversed; no consistent changes in synaptic transmission are observed in slices from estradiol-treated OVX rats, but those from control rats (OVX + oil) show robust LTD. Thus estradiol reduces the frequency threshold for LTD induction at the CA3-CA1 synapses. Further, regardless of the conditioning frequency employed, where robust LTD is seen, its induction depends on normally functioning N-methyl-D-aspartate (NMDA) receptors during conditioning. The shift in conditioning frequency needed to elicit LTD is consistent with a decrease in NMDA receptor activation with decreasing estradiol levels.

Serological and biochemical characteristics of virulence plasmid of Yersinia enterocolitica isolates from chicken in the Islamic Republic of Iran.

Pathogenic Yersinia enterocolitica harbour plasmid that is essential for virulence. We studied the characteristics of virulence plasmid using serological, biochemical and bioassay tests in Y. enterocolitica isolates of chicken using plasmid curing. Plasmid-cured isogenic derivatives (2029c and 2150c) were obtained from two isolates of Y. enterocolitica (RTCC 2029 and RTCC 2150). The results demonstrated that plasmid-bearing isolates (2029 and 2150) were human-serum-resistant when grown at 37 degrees C, but were sensitive when grown at 25 degrees C, whereas plasmid-cured isolates (2029c and 2150c) were sensitive when grown at both temperatures. Also autoagglutination, calcium-dependency tests and experimental infection in mice demonstrated that these phenotypes were associated with the virulence plasmid.

Serological and biochemical characteristics of virulence plasmid of Yersinia enterocolitica isolates from chicken in the Islamic Republic of Iran

Pathogenic Yersinia enterocolitica harbour plasmid that is essential for virulence. We studied the characteristics of virulence plasmid using serological, biochemical and bioassay tests in Y. enterocolitica isolates of chicken using plasmid curing. Plasmid-cured isogenic derivatives [2029c and 2150c] were obtained from two isolates of Y. enterocolitica [RTCC 2029 and RTCC 2150]. The results demonstrated that plasmid-bearing isolates [2029 and 2150] were human-serum-resistant when grown at 37 ّC, but were sensitive when grown at 25 ّC, whereas plasmid-cured isolates [2029c and 2150c] were sensitive when grown at both temperatures. Also autoagglutination, calcium-dependency tests and experimental infection in mice demonstrated that these phenotypes were associated with the virulence plasmid

Nicotine modulates the neurotoxic effect of beta-amyloid protein(25-35)) in hippocampal cultures.

Two major features of Alzheimer's disease (AD) are beta-amyloid protein (beta AP) deposition and a severe cholinergic deficit. An association between the two is suggested by the negative correlation found between cigarette smoking and AD. We sought to investigate this further by examining the effects of acute and chronic nicotine exposure on beta AP-induced neuronal loss in rat hippocampal cultures. Nicotine was found to attenuate the neurotoxicity of higher concentrations of beta AP(25-35), an effect which was enhanced by longer nicotine pretreatment and significantly inhibited by the nicotine receptor antagonist mecamylamine. Our results suggest that nicotine partially protects against the neurotoxic actions of beta AP(25-35) via a receptor-mediated pathway.

The histamine H1 receptor in GT1-7 neuronal cells is regulated by calcium influx and KN-62, a putative inhibitor of calcium/calmodulin protein kinase II.

1. In GT1-7 cells, histamine stimulated the initial [Ca2+]i transient in a dose-dependent manner with a best-fit EC50 value of 4.2 +/- 4.2 microM (mean +/- s.e.mean, n = 4) and a best-fit maximal effect of 138 +/- 56 nM (n = 4) increase above basal calcium levels. 2. Pretreatment of cells with 30 microM histamine for 30 min desensitized the population mean peak calcium signal by 53% to 75 +/- 9 nM, (n = 3, P < 0.04). Analysis of the individual cells revealed that 39 +/- 7% (n = 94 cells from 8 experiments) of pretreated cells exhibited desensitized histamine-stimulated [Ca2+]i transients of < or = 1 standard deviation below the control cells mean calcium transient level. 3. The desensitization induced by histamine was prevented (P < 0.01) by KN-62 (10 microM), a putative inhibitor of the calcium/calmodulin-dependent protein kinase II (CaMKII). KN-62 (10 microM) alone did not induce [Ca2+]i mobilization, nor did it antagonize the histamine-stimulated [Ca2+]i signal. In addition, KN-62 did not appear to have its effect by hastening the rate of recovery from desensitization. 4. Histamine pretreatment in nominal (zero calcium + 0.2 mM EGTA) or in low (0.3 mM) extracellular calcium did not induce histamine receptor desensitization, supporting a role for extracellular calcium in the homologous H1 receptor desensitization process. 5. Histamine (30 microM) stimulated at least four different types of [Ca2+]i signals in GT1-7 cells. The majority (61%) were of single spikes with the remaining cells showing some form of calcium oscillatory behaviour. The proportion of GT1-7 cells showing histamine-induced calcium oscillations was histamine concentration-dependent and significantly reduced after acute desensitization. KN-62, when present during histamine pretreatment, prevented this fall in calcium oscillation. Under the conditions of nominal or 0.3 mM extracellular calcium the proportion of cells exhibiting histamine-stimulated calcium oscillations was not significantly different from the controls. 6. Bradykinin stimulated a [Ca2+]i transient in GT1-7 cells with a population mean peak response of 147 +/- 8 nM (n = 5) over basal levels. The bradykinin-induced [Ca2+]i signal was without any calcium oscillatory activity. Histamine pretreatment caused the heterologous desensitization of the bradykinin [Ca2+]i signal (44% reduction, P < 0.007), which was unaffected by KN-62. 7. The results presented here suggest that the histamine-mediated homologous H1 receptor desensitization process involves extracellular calcium and can be blocked by KN-62, a putative inhibitor of CaMKII. In contrast, KN-62 does not appear to prevent the histamine-mediated heterologous desensitization cascade. These findings suggest fundamental differences in the mechanisms underlying homologous and heterologous H1 receptor desensitization pathways in GT1-7 neuronal cells.

Neurotoxic mechanisms of transactivating protein Tat of Maedi-Visna virus.

Infection by lentiviruses such as human immunodeficiency virus (HIV) and Maedi-Visna virus (MVV) is associated with neurodegenerative disorders. We have investigated the neurotoxic mechanisms of a synthetic peptide of transactivating protein tat of MVV in striatal neuronal cultures. Tat peptide (but not control peptide) caused neuronal death, without affecting glial viability, in a time- and dose-dependent manner. Significant neuronal death was not observed until 6-8 h after tat peptide application (2.35-2350 nM), whereas half maximal and maximal cell death was observed after 12 and 24 h respectively. Tat peptide neurotoxicity could be partially inhibited by blockade of either N-methyl-D-aspartate (NMDA)- or non-NMDA receptors, suggesting that excessive neuroexcitation by glutamate or its analogues may contribute to tat-neurotoxicity. Furthermore, when both these glutamate receptor subtypes were blocked simultaneously, an increased degree of neuroprotection was observed. Finally, tat peptide toxicity was also reduced by blockade of L-type calcium channels. Calcium imaging revealed that intracellular calcium increases slowly upon tat application, predominantly due to entry of extracellular calcium. These results indicate that cellular calcium entry through voltage-gated calcium channels following activation of both NMDA and non-NMDA receptors, and subsequent accumulation of intracellular calcium may contribute to the neuronal death induced by tat protein.

Receptor-mediated desensitisation of histamine H1 receptor-stimulated inositol phosphate production and calcium mobilisation in GT1-7 neuronal cells is independent of protein kinase C.

GT1-7 cells, a clonal line derived from specific tumours of gonadotropin-releasing hormone-secreting neurons from mouse hypothalamus, were used as a model system to investigate the cellular mechanisms underlying the histamine H1 receptor-mediated desensitisation. GT1-7 cells contain H1 receptors, acute stimulation of which leads to the desensitisation of histamine-mediated calcium mobilisation and is manifest as a concurrent reduction in both the magnitude of the calcium transient and of the sustained phase. Acute pretreatment of the cells with the phorbol ester, phorbol 12-myristate 13-acetate, can also ablate the histamine-stimulated calcium mobilisation. In addition, acute H1-receptor stimulation and acute phorbol ester treatment result in the attenuation of histamine-mediated inositol phosphate production. Receptor desensitisation resulting from acute stimulation with histamine is not affected by inhibiting protein kinase C (PKC) activity with Ro 31-7549 or staurosporine. In contrast, the desensitisation of H1-receptor responses induced by direct activation of protein kinase C is preventable by PKC inhibitors. Thus, these results imply that a PKC-dependent mechanism and PKC-independent mechanism are involved in the H1-receptor desensitisation cascade in GT1-7 cells and do not support the involvement of PKC in the receptor-mediated desensitisation of H1 receptor-stimulated calcium and inositol phosphate responses.

An immortalised murine hypothalamic neuronal cell, GT1-7, expresses functional histamine H1 receptors.

Histamine, acting via H1 receptors, dose-dependently stimulated [3H]inositol phosphate production in GT1-7 neuronal cells. GT1-7 cells also responded to Substance P but not to other neuroactive drugs tested. Acute histamine pretreatment desensitised the histamine-induced response, resulting in a reduction in the maximal response and a slower time-course of [3H]-inositol phosphate production. The desensitisation phenomenon was reversible, with full recovery by 2 h.

A role for the neural cell adhesion molecule in a late, consolidating phase of glycoprotein synthesis six hours following passive avoidance training of the young chick.

We have investigated the effect of intracranial injections of the amnestic anti-metabolite, 2-deoxygalactose, and antibodies to the neural cell adhesion molecule on retention of a one-trial passive avoidance task in chicks. Groups of chicks received bilateral intracranial injections of 10 mumol/hemisphere 2-deoxygalactose or 10 microliters/hemisphere anti-neural cell adhesion molecule and were tested 24 h following training. 2-Deoxygalactose injections were amnestic when administered at a previously established time (30 min pre-training). Here we show that the agent is also amnestic when injected within a second time window occurring specifically 6-8 h after training. Administration of 2-deoxygalactose between 2 and 6 h or after 8 h post-training was without effect on retention tested 24 h following training. Anti-neural cell adhesion molecule injections were amnestic only when performed at a time which coincided with the second phase of 2-deoxygalactose susceptibility. Further experiments demonstrated that the neural cell adhesion molecule is one of the molecules into which 2-deoxygalactose is incorporated. Additionally, we investigated the extent of diffusion of 2-deoxygalactose and anti-neural cell adhesion molecule following their injection, with respect to their residence in forebrain loci known to be involved in the memory for passive avoidance. We interpret these data as indicating that two waves of glycoprotein synthesis are necessary for the establishment of long-term memory for the experience of passive avoidance training. The evidence is discussed in the context of earlier results indicating that the two waves involve different glycoprotein species and, possibly, different forebrain regions. We speculate that the late phase of glycoprotein synthesis coincides with, and is required for, modulation of cell-cell adhesion processes, reflecting the selection and stabilization of synapses which maintain an enduring representation of long-term memory.

Desensitization of histamine H1 receptor-mediated inositol phosphate production in HeLa cells.

1. Histamine stimulated the accumulation of total [3H]-inositol phosphates (IPn) in control HeLa cells with an EC50 of 3.7 +/- 0.7 microM in the presence of 10 mM LiCl. The maximum response to histamine after 15 min incubation was 43 +/- 5% over basal accumulation and occurred at a concentration of 1 mM histamine. 2. The histamine-induced IPn production in HeLa cells was confirmed as H1 receptor-mediated, since the H1 antagonist mepyramine (10(-6) M) inhibited the histamine response (10(-4) M) by 83 +/- 7%, whereas the H2 antagonist, ranitidine (10(-4) M), and H3 antagonist, thioperamide (10(-6) M), were ineffective. 3. Histamine (10(-4) M) pretreatment of HeLa cells for 30 min desensitized the subsequent histamine-induced IPn accumulation. The desensitized cells accumulated IPn in response to histamine with an EC50 of 1.7 +/- 0.7 microM after 15 min incubation. The maximum histamine-induced IPn accumulation at 10(-4) M was 19 +/- 5% over basal and was significantly lower (P < 0.03) than the maximum response in control cells. 4. The desensitization of histamine-induced IPn accumulation was time-dependent and, at a desensitizing histamine concentration of 10(-4) M, the half-maximal attenuation occurred after approximately 9 min and maximum desensitization was achieved by 15-20 min. The desensitization of the IPn accumulation was a reversible phenomenon and full recovery of the response occurred 150 min after the removal of the desensitizing histamine-containing medium. The half-time for the recovery of the histamine-induced response was estimated at 120 min. 5. Bradykinin stimulated IPn, accumulation in HeLa cells, and the ECm in control cells of 1.9 +/- 0.2 nM was not significantly different from the EC50 value from histamine-pretreated cells of 1.6 +/- 0.9 nM. The bradykinin response at 1 microM was 194 +/- 48% over basal IPn accumulation in control cells and this value was significantly different (P <0.04) from the 1 microM bradykinin-induced IPn accumulation in histamine pretreated HeLa cells of 143 +/- 38% over basal.6. NaF stimulated IP,, accumulation in control HeLa cells in a dose-related manner, with the maximum effect occurring at 15-20 mM. The EC50 value for NaF-stimulated IPn accumulation in control cells was 10.5 +/- 1.1 mm and the maximum response was 136 +/- 41% over basal after 20 min incubation. In histamine desensitized HeLa cells the EC50 value for NaF was 12.3 +/- 0.4 mM after 20 min stimulation,which was not significantly different from the value obtained in control cells. The maximum NaF stimulated IPn formation in desensitized cells of 68 +/- 23% over basal occurred at 15 -20 mM and was significantly lower (P<0.01) than that obtained in control cells.7. We show here that the acute histamine pretreatment of HeLa cells results in the desensitization of histamine H1 receptor-mediated IPn production. The desensitization was not restricted to the H1 receptor-mediated signal transduction pathway, but also includes both the bradykinin- and NaF mediated responses, supporting a heterologous desensitization mechanism. Our results are consistent with the site of attenuation being at or distal to the G-protein and the underlying mechanism being a slowed time-course for the production of inositol phosphates.