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PubMed2002

Anticonvulsant valproate reduces seizure-susceptibility in mutant Drosophila.

Despite the frequency of seizure disorders in the human population, the genetic basis for these defects remains largely unclear. Currently, only a fraction of the epilepsies can be linked conclusively to a genetic determinant. In addition, a significant number of epileptics do not respond to the current anticonvulsant therapies. We have turned to Drosophila as a model to address these problems and have identified genetic mutants that are more sensitive to seizures, bang-sensitive (BS) mutants, such as slamdance (sda), bangsenseless (bss) and easily shocked (eas), as well as mutants that are resistant to seizures, such as paralytic, maleless(napts), shaking-B(2) and Shaker. Here, we have developed a new method for evaluating compounds with anticonvulsant activity. The methodology uses Drosophila BS mutants to assay the ability of compounds to suppress the seizure susceptible phenotype normally seen in the BS mutants. To test the effectiveness of this method, two BS mutant strains were administered the anticonvulsant valproate and in both cases the drug was able to suppress seizures. The Drosophila system provides a potentially powerful way of developing and testing new drugs with anticonvulsant properties.

Action Potentials↗
PubMed1986

Identification of a alpha-NeuAc-(2----3)-beta-D-galactopyranosyl N-acetyl-beta-D-galactosaminyltransferase in human kidney.

Microsomal preparations from human kidney were found to contain enzymic activity capable to transfer N-acetylgalactosamine from UDP-N-acetylgalactosamine to native bovine fetuin. The acceptor structures on the fetuin molecules were identified as N- as well as O-linked glycans with a markedly higher incorporation into the N-linked carbohydrate chains. Analysis of the alkali-labile transferase products by thin-layer chromatography indicated that the enzyme is able to synthesize structures having mobilities identical with those found on glycophorin from Cad erythrocytes. Mild acid treatment and enzymic hydrolysis with N-acetylhexosaminidase from jack beans of the N-linked transferase products suggested that beta-D-GalpNAc-(1----4)-[alpha-NeuAc-(2----3)]-beta-D-Galp-(1----s tructures were formed by the enzymic reaction on both N- and O-linked acceptors. The enzyme might, therefore, be involved in the biosynthesis of Sda (and Cad) antigenic structures. By use of various oligosaccharides, glycopeptides, and glycolipids having well characterized carbohydrate sequences, the acceptor-substrate specificity of the N-acetylgalactosaminyltransferase was determined. The enzyme generally recognized alpha-NeuAc-(2----3)-beta-D-Gal groups as acceptors, but in a certain conformation. Thus, tri- and tetra-saccharide alditols, native human glycophorin A, and GM3 were not acceptor substrates although they carry the potential disaccharide acceptor unit. When these structures were presented as sialyl-(2----3)-lactose or as a tryptic peptide from glycophorin A, they were shown to be rather good acceptor substrates for the N-acetyl-beta-D-galactosaminyltransferase from human kidney.

Animals↗
PubMed1986

Ascorbate regeneration in bovine ocular tissues by NADH-dependent semidehydroascorbate reductase.

Despite its fast autoxidation in vitro, ascorbate remains in its reduced form in vivo, indicating a special mechanism may be involved in its regeneration. The presence of an NADH-dependent reductase system, semidehydroascorbate reductase (SDR), for regeneration of ascorbate from its partially oxidized form, semidehydroascorbate (SDA), was demonstrated in bovine ocular tissues after extraction in Triton X-100. Highest SDR activity was detected in retinal extracts in the order of retina greater than pigment epithelium-choroid = ciliary body greater than iris. Minimal or no activity was observed in lens extracts or in aqueous fluid. Freezing and thawing, or boiling, destroyed the NADH-dependent SDR activity. NADH oxidation was significantly reduced (22% of total activity) when assays with retinal extracts were performed at 5 degrees C. Treatment with 4 mM, N-ethylmaleimide reduced the rate of NADH oxidation to 73 or 42% compared with control values with retinal or ciliary body extracts, respectively.

Animals↗
PubMed1997

Prognostic value of nuclear morphometry in feline mammary carcinomas.

This study of 30 cats with mammary carcinoma was designed to investigate the relationship between (1) six nuclear morphometric parameters (mean nuclear profile area [MNA], standard deviation of MNA [SDa], coefficient of variation of MNA [CVa], nuclear form factor (p2/4 pi area) [FF], standard deviation of FF [SDf], and coefficient of variation of FF [CVf]) assessed by image analysis, and (2) survival for > 1 year or < 1 year after surgical removal of the tumour. Only the SDf and the CVf appeared to be related to survival. Cats that died within 1 year had an SDf or CVf (or both) higher than the corresponding mean values (SDft and CVft) for all 30 cats; but only four of 16 cases (25%) with a SDf lower than SDft and five of 17 cases (29.4%) with a CVf lower than CVft died within 1 year. The authors conclude that SDf and CVf represent reliable prognostic parameters in feline mammary carcinomas.

Adenocarcinoma↗
PubMed2002

Enzymatic degradation of luteinizing hormone releasing hormone (LHRH) by mucosal homogenates from the intestine of the common brushtail possum (Trichosurus vulpecula).

The peptidolytic activity of fresh and frozen mucosal homogenates from five regions (duodenum, jejunum, ileum, caecum and colon) of possum intestine from Trichosurus vulpecula towards human Luteinizing Hormone Releasing Hormone (LHRH) was investigated. The rank of order of specific peptidolytic activity of the mucosal homogenates was jejunum > ileum > caecum> duodenum = colon, with a 3 to 4 fold difference between the least and the most active segment in both frozen and fresh samples. The formation of peptides LHRH (1-3), LHRH (1-4) and LHRH (1-5) suggest endopepetidase-24.18, endopeptidase-24.15 and angiotensin converting enzyme (ACE) might be responsible for the peptide degradation in mucosal homogenates. The inhibition of LHRH degradation by mucosal homogenates was evaluated in four regions (jejunum, ileum, caecum and colon) of possum intestine. Ethylenediaminetetraacetic acid (EDTA, 5 mM), sodium deoxycholate (SDA, 10 mM) and bacitracin (3.5 or 9 mM) inhibited the degradation of LHRH in mucosal homogenates from small intestine and hindgut. However, the serine protease inhibitor, soybean trypsin-chymotrypsin inhibitor (SBTI), did not prevent degradation of LHRH. It is concluded that combining peptides with inhibitors may enhance oral delivery of bioactive peptides or proteins to possums.

Animals↗
PubMedDate not supplied

Removal efficiencies of PCDDs/PCDFs by air pollution control devices in municipal solid waste incinerators.

Removal efficiencies of polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans (PCDDs/PCDFs) by air pollution control devices (APCDs) in the commercial-scale municipal solid waste (MSW) incinerators with a capacity of above 200 ton/day were evaluated. The removal efficiencies of PCDDs/PCDFs were up to 95% when the activated carbon (AC) was injected in front of electrostatic precipitator (EP). Spray dryer absorber/bag filter (SDA/BF) had high removal efficiency (99%)) of PCDDs/PCDFs when a mixture of lime and AC was sprayed into the SDA. When the AC was not added in scrubbing solution, the whole congeners of PCDDs/PCDFs were enriched in the wet scrubber (WS) with negative removal efficiencies of -25% to -5731%. Discharge of PCDDs/PCDFs was decreased with increasing the proportions of AC added in scrubbing solution. Selective catalytic reduction (SCR) system had the removal efficiencies of up to 93% during the test operation.

Air Pollution↗
PubMed2002

Enhanced antimicrobial effects of combination of lactate and diacetate on Listeria monocytogenes and Salmonella spp. in beef bologna.

The antimicrobial activities of salts of organic acids such as lactate and acetate are well documented, but there is limited information on their effect when used in combination. We previously reported enhanced inhibition of Listeria monocvtogenes and Salmonella enterica serovar Enteritidis in sterile comminuted beef at 5 and 10 degrees C by combinations of sodium lactate (SL) (2.5%) and sodium diacetate (SDA) (0.2%). The present study was undertaken to evaluate the inhibitory effect of these salts, alone and in combination, in ready-to-eat (RTE) meat. Single strains and six-strain mixtures of each of the pathogens ( approximately 3 log CFU/g) were tested in beef bologna during aerobic storage at 5 and 10 degrees C for up to 60 days. The growth rate of the six-strain mixture of Listeria was faster than that of the single strain (Scott A) in the lactate/diacetate-free product. While each of the salts delayed growth of the listeriae at 5 degrees C, the effect of their combination was listericidal for the single strain and listeriostatic for the six-strain mixture. Enhanced inhibition by the salt combination was also observed at 10 degrees C. Salmonella numbers declined to undetectable levels in the untreated meat product and in each of the treatments after 20-30 days. However, the decline was more rapid in meat with the combination of the salts during storage at both 5 and 10 degrees C. Each of the salts further delayed the growth of the background microflora during storage at 5 degrees C, with their combinations showing the most effect.

Animals↗
PubMed2002

Platelet serotonin transporter in schizophrenic patients with and without neuroleptic treatment.

Among various hypotheses put forth to account for the etiology of schizophrenia, the abnormal function of serotonergic system has recently gained marked interest. Our previous study showed that drug-free schizophrenic patients had a significant increase in maximum numbers (B(max)) of platelet 5-HT(2A) receptors that declined to normal level after treatment with different neuroleptic drugs. To elucidate the role of the serotonin system in schizophrenia, the serotonin transporters on human platelets were examined in this study. Platelet serotonin transporters obtained from normal control subjects and schizophrenic patients were identified by using [(3)H]imipramine as the radioligand and fluoxetine to define the non-specific binding. The data showed that the mean B(max) of serotonin transporter sites for schizophrenic patients without neuroleptic therapy was significantly higher than in normal controls. The B(max) values for schizophrenic patients on phenothiazine, butyrophenone, thioxanthene and serotonin-dopamine antagonist (SDA) therapies were significantly lower than the B(max) values obtained from schizophrenic patients without neuroleptic therapy, and were comparable to those found in normal control subjects. The dissociation equilibrium constant (K(d)) values in all subject groups remained unchanged. The effect of various medication periods on platelet serotonin transporters was also studied. We found that, B(max) values of 1-4 weeks, 1-4 months, 4-12 months and >1 year of neuroleptic therapies were significantly decreased when compared with the unmedicated group. Significant reduction of brief psychiatric rating scale (BPRS) occurred in all types of neuroleptics and every period of drug treatments compared with the unmedicated group. The present results indicate that alteration of platelet serotonin transporters is associated with schizophrenia. Treatment with various types of neuroleptics suppresses the hypersensitivity of platelet serotonin transporters. The mechanisms of how neuroleptics achieve their therapeutic effects, whether they act via or modulate serotonin system in certain brain area, still need to be further evaluated.

Adolescent↗
PubMed1997

Machines for automated evolution experiments in vitro based on the serial-transfer concept.

Two machine setups for automated evolution experiments in vitro are described. Both machines enable the monitoring of growing populations of RNA or DNA molecules in real time using high-sensitivity glass fiber laser fluorimeters and an automated sample handling facility for volumes in the microliter range. Growth conditions are kept constant by means of the serial-transfer technique, that is, the successive transfer of a small fraction of a growing population into a fresh solution containing no individuals prior to the transfer. The serial transfer technique was modified to work with large populations and constant growth conditions. In the single-channel evolution machine isothermal amplification reactions (Qbeta-system, 3SR, NASBA, SDA) are monitored successively in single test tubes. This machine is particularly well suited for the investigation of optimal adaptation to altered environmental conditions, as is experimentally demonstrated in the evolution of an RNA quasi-species using ribonuclease A as the selection pressure. The new variant of RNA appeared very rapidly (within approximately 80 generations) without stable intermediates, and it was selected by steadily increasing the RNaseA concentration during the serial-transfer experiment. The other machine, which is described in the second part of this article, is a consequent extension of the single-channel machine, and was designed to allow the multichannel detection of up to 960 samples simultaneously. Thus, high-throughput screening can be applied to evolution experiments. In addition to monitoring isothermal amplification reactions, it is also possible to follow PCR amplifications through thin plastic foils. Initial experiments have demonstrated the suitability of the apparatus for uniformly processing samples and for performing thermocycling.

Base Sequence↗
PubMed1997

The differentiation of Brucella species by substrate specific tetrazolium reduction.

In the development of a tetrazolium reduction assay to replace substrate stimulated oxygen uptake for the identification of Brucella species, nine tetrazolium salts were evaluated. Only the more readily reduced compounds (MTT and INT) detected increased metabolic activity with the more fastidious, slow growing strains and with B. suis strains on L-arginine and DL-ornithine. The assay was optimised with MTT. MTT reduction profiles offered with the medium on which the cells were grown. Cells grown on TSA gave profiles more similar to the published respirometric results than those grown on SDA. The optimal substrate concentration was 0.84 g l-1 and prolonged (> 3 h) exposure to substrate was necessary before adding MTT. MTT concentration was not critical and the OD was proportional to the MTT concentration between 0.03 and > 0.5 g l-1. MTT reduction was linear for 60 min after its addition. The reaction was, therefore, stopped after 60 min by adding formaldehyde solution. The optimised assay was evaluated with 71 strains of Brucella, representing all the species and biovars of the genus. Each strain was assigned to its previously identified species and sub-groups were defined.

Amino Acids↗
PubMed2002

The PDZ domain of TIP-2/GIPC interacts with the C-terminus of the integrin alpha5 and alpha6 subunits.

Different cDNA libraries were screened by the yeast two-hybrid system using as a bait the cytoplasmic sequence of integrin alpha6A or alpha6B subunits. Surprisingly, the same PDZ domain-containing protein, TIP-2/GIPC, was isolated with either of the variants, although their sequences are different. Direct interaction assays with the cytoplasmic domain of the integrin alpha1--7 subunits revealed that in addition to alpha6A and alpha6B, TIP-2/GIPC reacted also with alpha5, but not other alpha integrin subunits. The specificity of the interaction was confirmed by in vitro protein binding assays with purified peptides corresponding to integrin cytoplasmic domains. Further analysis with either truncation fragments of TIP-2/GIPC or mutated integrin cytoplasmic domains indicated that the interaction occurs between the PDZ domain of TIP-2/GIPC and a consensus PDZ domain-binding sequence, SDA, present at the C-terminus of the integrin alpha5 and alpha6A subunits. The integrin alpha6B subunit terminates with a different sequence, SYS, which may represent a new PDZ domain-binding motif.

Adaptor Proteins, Signal Transducing↗
PubMed2004

Characteristics of ashes from different locations at the MSW incinerator equipped with various air pollution control devices.

The characteristics of ashes from different locations at a municipal solid waste incinerator (MSWI) equipped with a water spray tower (WST) as a cooling system, and a spray dryer adsorber (SDA), a bag filter (BF) and a selective catalytic reactor (SCR) as air pollution control devices (APCD) was investigated to provide the basic data for further treatment of ashes. A commercial MSWI with a capacity of 100 tons per day was selected. Ash was sampled from different locations during the normal operation of the MSWI and was analyzed to obtain chemical composition, basicity, metal contents and leaching behavior of heavy metals. Basicity and pH of ash showed a broad range between 0.08-9.07 and 3.5-12.3, respectively. Some major inorganics in ash were identified and could affect the basicity. This could be one of the factors to determine further treatment means. Partitioning of hazardous heavy metals such as Pb, Cu, Cr, Hg and Cd was investigated. Large portions of Hg and Cd were emitted from the furnace while over 90% of Pb, Cu and Cr remained in bottom ash. However 54% of Hg was captured by WST and 41% by SDA/BF and 3.6% was emitted through the stack, while 81.5% of Cd was captured by SDA/BF. From the analysis data of various metal contents in ash and leach analysis, such capturing of metal was confirmed and some heavy metals found to be easily released from ash. Based on the overall characteristics of ash in different locations at the MSWI during the investigation, some considerations and suggestions for determining the appropriate treatment methods of ash were made as conclusions.

Air Pollution↗
PubMed2002

An integrated, stacked microlaboratory for biological agent detection with DNA and immunoassays.

An integrated, stacked microlaboratory for performing automated electric-field-driven immunoassays and DNA hybridization assays was developed. The stacked microlaboratory was fabricated by orderly laminating several different functional layers (all 76 x 76 mm(2)) including a patterned polyimide layer with a flip-chip bonded CMOS chip, a pressure sensitive acrylic adhesive (PSA) layer with a fluidic cutout, an optically transparent polymethyl methacrylate (PMMA) film, a PSA layer with a via, a patterned polyimide layer with a flip-chip bonded silicon chip, a PSA layer with a fluidic cutout, and a glass cover plate layer. Versatility of the stacked microlaboratory was demonstrated by various automated assays. Escherichia coli bacteria and Alexa-labeled protein toxin staphylococcal enterotoxin B (SEB) were detected by electric-field-driven immunoassays on a single chip with a specific-to-nonspecific signal ratios of 4.2:1 and 3.0:1, respectively. Furthermore, by integrating the microlaboratory with a module for strand displacement amplification (SDA), the identification of the Shiga-like toxin gene (SLT1) from E. coli was accomplished within 2.5 h starting from a dielectrophoretic concentration of intact E. coli bacteria and finishing with an electric-field-driven DNA hybridization assay, detected by fluorescently labeled DNA reporter probes. The integrated microlaboratory can be potentially used in a wide range of applications including detection of bacteria and biowarfare agents, and genetic identification.

Biological Warfare↗
PubMed2002

Effects of temperature on the metabolic response to feeding in Python molurus.

As ectothermic vertebrates, reptiles undergo diurnal and seasonal changes in body temperature, which affect many biological functions. In conjunction with a general review regarding the effects of temperature on digestion in reptiles, we describe the effects of various temperatures (20-35 degrees C) on the metabolic response to digestion in the Burmese python (Python molurus). The snakes were fed mice amounting to 20% of their body weight and gas exchange (oxygen uptake and CO(2) production) were measured until digestion had ended and gas exchange returned to fasting levels. Elevated temperature was associated with a faster and larger metabolic increase after ingestion, and the time required to return to fasting levels was markedly longer at low temperature. The factorial increase between fasting oxygen consumption (VO(2)) and maximal VO(2) during digestion was, however, similar at all temperatures studied. Furthermore, the integrated SDA response was not affected by temperature suggesting the costs associated with digestion are temperature-independent. Other studies on reptiles show that digestive efficiency is only marginally affected by temperature and we conclude that selection of higher body temperatures during digestion (postprandial thermophilic response) primarily reduces the time required for digestion.

Animals↗
PubMed2001

Estimation of autonomic response based on individually determined time axis.

The analysis of the time-dependence of autonomic response requires: 1. A reliable procedure for the quantification of autonomic activity under nonsteady conditions, such as an algorithm for time-frequency decomposition (ex. SDA. Wigner-Ville, or others). 2. The choice of an adequate time scale for focusing on the data: (a) the regular, universal time scale, independent of the unsteady physiological conditions, or (b) a time axis defined by specific events related to an applied perturbation, as the indicators of specific experimental or physiological conditions, so that each individual is considered according to his own intrinsic time scale. The alignment of the various subjects according to their intrinsic time scale, reflecting their individual response mechanisms, may help to disclose a common pattern of autonomic function. Using an absolute time scale to align and average results for different subjects may obscure the underlying mechanisms. Several examples of autonomic challenges are presented, in which the use of an individual time scale contributes to unveil a typical response pattern: tilt test in vasovagal syncope, the autonomic effect of active standing on hypertension, and the autonomic response to acute hypoxia.

Autonomic Nervous System↗
PubMed1984

Finnish twins reared apart. III: Personality factors.

This study is based on data from 165 adult twin pairs separated at 10 years or less. Information on personality factors: extraversion (E) and neuroticism (N) (EPI scale short from), life satisfaction (LS) (Allardt) and stress of daily activities (SDA) was obtained as part of the questionnaire study carried out in the entire Finnish Twin Cohort in 1975. Later in 1979 a questionnaire sent to the twins reared apart yielded a scale (range 7-30 points) measuring the environmental dissimilarities after separation (reliability 0.83). The effect of separation on personality factors by analysis of variance of individual data was studied. Sex, zygosity and age-at-separation were included in the models. The overall explanatory rates were low (2.1-4.4%). The definitive study group was formed by selecting those pairs with a dissimilarity score greater than 15. The following intraclass correlations were obtained.

Adult↗
PubMed1990

Bloodmeal digestion by strains of Anopheles stephensi liston (Diptera: Culicidae) of differing susceptibility to Plasmodium falciparum.

Blood digestion was studied in strains of Anopheles stephensi which had been genetically selected for either refractoriness or susceptibility to infection by Plasmodium falciparum. Females of the refractory Pb3-9a strain ingested more blood than selected (Sda-500) and unselected (Punjab) susceptible females and began to degrade the haemoglobin soon after feeding. In susceptible females, haemoglobin degradation started only after a significant post-feeding lag period. Total protein content of the midgut after the bloodmeal was correspondingly higher for refractory than for susceptible females, but absolute and relative rates of protein degradation were not significantly different between the different mosquito strains. Bloodmeal induction of midgut trypsin activity and the maximal trypsin activity were the same for the different strains. The residual aminopeptidase activity and its relative post-feeding activity (enzyme units per midgut) were significantly higher in refractory females. However, when converting to specific aminopeptidase activity, no differences between strains were evident. The results indicate that both the early initiation of haemoglobin degradation and higher aminopeptidase activity in the Pb3-9a refractory strain are important in the limitation of parasite development within the mosquito midgut, whereas trypsin plays no role in this process.

Aminopeptidases↗
PubMed2005

Analysis of sulfur in deposited aerosols by thermal decomposition and sulfur dioxide analyzer.

A thermal decomposition method that measures aerosol sulfur at the nanogram level directly from the collection substrate is described. A thermal decomposition apparatus was designed. A stainless steel strip was used as the aerosol collection substrate. A 0.1 mol/L MnCl2 solution was added as the thermal decomposition catalyst. Currents were passed through the strip where aerosol particles had been deposited. In this way, the strip was heated at 780 +/- 10 degrees C, and particulate sulfur was evaporated. A sulfur dioxide analyzer (SDA) with flame photometric detector (FPD) was used to detect gaseous sulfur. High sulfur recoveries from (NH4)2SO4 and other inorganic sulfates, such as NH4HSO4, K2SO4, MgSO4, and CaSO4, were obtained. From the sulfur blank and the calibration, a lower limited detection of 0.2 ng of sulfur and the determination range of 3.3-167 ng of sulfur were estimated. The method is effective for measuring the sulfate size distributions of urban aerosols in a small sample air volume of 50-60 L. The method is applicable to measuring the sulfur in aqueous extracts of size-segregated urban aerosols collected by impactor and comparing the results with the sulfate data measured by ion chromatography.

Journal Article↗