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DOI: 10.14457/cu.the.2011.155
title: Oxidative enzyme activities and protein profile of transgenic rice Oryza sativa L. overxpressing calmodulin gene OsCam1-1

description: Rice (Oryza sativa L.) in which a large family of CaM and CaM-related proteins has been found, is one of the most important crops in the world and is the staple food for about three billion people. Thailand is one of the most important rice-exporter and rice-producer countries in the rice-world market. However, rice production is confronting the saline soil problem and recently coping with the low yielding. Of five OsCam genes, OsCam1-1 was shown to be rapidly and strongly increased in leaves under osmotic stress and is shown to play important roles in salt-stress responses. A proteomic study was performed to characterize the responses of transgenic rice Oryza sativa L. cv. KDML105, harboring OsCam1-1 gene to simulated salt stress. The emphasis of the present study was to investigate the overall protein expression changes when exposed to salt. Three sets of rice seeds (wild-type, transgenic line with vector alone and transgenic line with OsCam1-1 gene) were germinated in MS media for 3 weeks and stressed by 150 mM NaCl. Proteins in the shoot and the root of the stressed rice plants along with their controls were extracted and in-gel digested. Peptides of these samples were analysed by mass spectrometry. Each identified protein was functionally classified according to their known and putative functions by using STRAP software program. These proteins were categorized in to 7 groups based on their biological functions including cellular process (42%), metabolic process (14%), regulation (12%), developmental process (2%), localization (2%), response to stimulus (2%) while the rest are proteins in other processes or of unknown function. Many of the proteins in shoot and root responded to the induced salt stress were localized in nucleus (24%) and also in chromosome (5%). Significant fractions were also located in mitochondria (5%), ER (3%), and other intracellular organelles (5%) while only a small fraction was located in cytoplasm. Some are part of macromolecular complex (8%). Comparison of protein expression profiles among the three rice lines in both shoots and roots by using Venn diagram revealed the unique salt-responsive proteins in transgenic rice lines that constitutively over-express the OsCam1-1 gene when compared to the wild type and the control transgenic lines without OsCam1-1 over-expression. In addition, the complementary data on enzyme activity analysis (Peroxidase, Ascorbate peroxidase, Glutathione-S-transferase, and Adenosine Triphosphatase) in the the wild-type and the transgenic rice lines were also investigated. These results suggested that these proteins could play important roles in mediating plant response to salt stress and indicated that OsCam1-1 overexpression probably contributes to salt resistance in rice through these proteins.
DOI: 10.14457/cu.the.2011.156
title: Method development of automated flow analysis coupled with anodic stripping voltammetry for determination of heavy metals

description: In this research, automated flow analysis procedures for trace heavy metal determination by a differential pulse anodic stripping voltammetry (DPASV) and square wave anodic stripping voltammetry (SWASV) were developed. A micro-flow sensor consisted of preconcentration membrane and three-electrode electrochemical part with a carbon working electrode was used as a detector. Detection limits (DL) of Cd2+ and Pb2+ were 2.37 and 0.15 µg L–1 by DPASV and 0.02 and 0.01 µg L–1 by SWASV. The results indicated that DPASV and SWASV at the carbon electrode in micro-flow sensor coupled with on-line automated flow system have a good efficiency for the heavy metal ion determination. Furthermore, a low-cost electrochemical flow-through cell based on a carbon paste electrode (CPE) was designed and constructed for the highly sensitive determination of heavy metals by automated flow analysis coupled with SWASV. The sensitivity of the proposed flow-through cell for Cd2+ and Pb2+ ion detection was improved by using the small channel height. Under the optimum conditions, the DL of Cd2+ and Pb2+ ions (0.08 and 0.07 µg L–1, respectively) were 13.8- and 11.4-fold lower than that of a commercial flow cell (1.1 and 0.8 µg L–1, respectively). Moreover, the percentage recoveries of Cd2+ and Pb2+ for the in-house designed flow-through cell were higher than those for the commercially available flow cell in all tested water samples, and within the acceptable range. Finally, an environmentally friendly graphene-modified carbon paste electrode (GCPE) for determination of Cd2+ and Pb2+ levels in an automated flow system was successfully developed. The electrochemical behavior of the developed electrode was studied by cyclic voltammetry and SWASV. The in situ bismuth-modified graphene carbon paste electrode (Bi-GCPE) exhibited excellent electrooxidation of Cd2+ and Pb2+ in the automated flow system with a significantly higher peak current for both metal ions compared with the unmodified CPE. The DLs were 0.07 and 0.04 µg L-1 for Cd2+ and Pb2+, respectively. The Bi-GCPE was also applied for the determination of Cd2+ and Pb2+ in low- (tap water) and high- (sea bass fish and undulated surf clam tissues) matrix complexity samples by SWASV in the automated flow system. The recoveries were acceptable and ranged from 70.4 to 120% for Cd2+ and 65.8 to 113.5% for Pb2+.
DOI: 10.14457/cu.the.2011.157
title: The role of house flies (Musca Domestica) in transmission of avian influenza virus Subtype H5N1

description: The objective of this study was to determine the potential role of the house flies in harboring and transmission avian influenza virus H5N1 (AIVs). To achieve this goal, this study was divided into three parts. The purpose of the first part was to determine the potential of the house flies as a vector of AIVs. In this part, the laboratory reared flies were experimentally fed with mixture containing the AIVs and then the external body surface of exposed flies was washed and sterile prior to preparation of homogenate whole fly between whole fly and brain heart infusion broth (BHI). The homogenate was inoculated into 10-day-old embryonated chicken eggs. Allantoic fluids were collected for determining virus concentration by HA test, RT-PCR assay. Moreover, the minimum of the house flies carrying AIVs was examined by inoculating one house fly homogenate into 10-day-old embryonated chicken eggs. In addition, the duration time of AIVs survival in house flies was determined by collecting exposed flies at different times of post exposure. The results in this part revealed that the house flies could carry the AIVs, one house fly was sufficient to carry and able to kill embryonated chicken eggs and the AIVs could survive within house flies for up to 96 hr. However, the virus titers were decreased over time. The second part that was the competence of the house flies as a transmission vector of AIVs in chickens under laboratory condition was determined. Three groups of 32-day-old chickens were individually inoculated AIVs infected house fly homogenate via oral route except the chickens in group 1 were inoculated with BHI, serving as a negative control group. Oropharyngeal and cloacal swabs were collected. Clinical signs and mortality were observed for 14 days post-inoculation (DPI). The experiment was terminated on 14 DPI and the tissues samples were collected for histopathological and immunohistochemistry evaluations. The results of this part indicated that AIVs infected house fly homogenate could induce the clinical signs and mortalities in inoculated chickens. Finally, the objective of last part was to examine the presence of the AIVs naturally infected house flies in epidemic area in Thailand during 2008. The result of this part revealed that none of AIVs naturally infected house flies was found. Nevertheless, all of the results in this study were conceivable illustrated to suggest that the house flies served as a mechanical vector of the AIVs and could transfer AIVs to chickens under experimental condition.
DOI: 10.14457/cu.the.2011.158
title: Effects of lead on paraoxonase ensymes in HEPG2 cells an THP-1 differentiated macrophage cells

description: Lead, an ubiquitous contaminant in the environment, is recognized as one of the important health problems. Its toxicities manifests in various pathological symptoms and diseases including cardiovascular diseases. The aim of this study was to explore effects of lead on paraoxonase (PON) enzymes. Effects of lead on PON1, PON2 and PON3 were assessed in HepG2 cells while effect of lead on PON2 was also assessed in THP-1 differentiated macrophage cells. Cell viability was determined by exposing cells to various concentrations (0, 0.05, 0.1, 0.5, 1, 10, 100 and 1000 µg/ml) of lead acetate for 24, 48 and 72 hours. Reactive oxygen species (ROS) generation and lipid peroxidation were assessed by exposing cells to various concentrations (0, 0.05, 0.1, 0.5, 1, 10 and 100 µg/ml) of lead acetate for 1 hour. Effects of lead acetate on PON1, PON2, and PON3 activities were determined using specific substrates after HepG2 cells or THP-1 differentiated macrophage cells were exposed to various concentrations (0, 0.05, 0.1, 0.5, 1, 10 and 100 µg/ml) of lead acetate for 24, 48 and 72 hours. PON protein and mRNA expressions were assessed by Western blot analysis and real time RT-PCR, respectively. The results showed that lead did not significantly decrease cell viability of both cells types at concentrations of lead acetate up to 100 µg/ml. Significant increase of ROS was shown in both cell types. Significant decrease of PON2 activity after lead exposure was observed in a concentration- and time-dependent manner in HepG2 cells but not in THP-1 differentiated macrophage cells. No effect of lead was observed on PON1 activity while decreases of PON3 activity were observed only at 24 hours of lead exposure in HepG2 cells. Modulation of PON2 activity by lead exposure was not associated to the modulation of both PON2 protein and PON2 mRNA expression. Calcium could restore the inhibitory effect of lead on PON2 activity suggesting that lead decreased PON2 activity via replacement of Ca2+ in the structure of PON2 enzyme. In contrast to PON2, modulation of PON3 activity by lead exposure was partly associated to the decrease of PON3 protein and mRNA expression.
DOI: 10.14457/cu.the.2011.159
title: Antioxidant and cytoprptective effects of Tamarindus indica seed coat extract

description: This study aimed to determine antioxidant activities of Tamarindus indica seed coat extracts and evaluate its protective activities against hydrogen peroxide (H2O2)-induced oxidative stress in human foreskin fibroblast CCD-1064Sk cells. The roasted seed coat of tamarind was extracted with various solvent systems: 70% ethanol (TSCE-E), ethyl acetate (TSCE-EA), 70% ethanol followed by partition with chloroform and then ethyl acetate (TSCE-EEA), and boiling-water followed by partition with ethyl acetate (TSCE-W). The highest phenolic content was observed in TSCE-W which revealed the free radical scavenging activities in cell-free system. TSCE-W possessed the highest DPPH● and ABTS●+ scavenging activities. TSCE-W was also exhibited superoxide anion radical, hydroxyl radical, H2O2, and nitric oxide scavenging activities, including reducing power and anti-lipid peroxidation activities comparable with other TSCEs. In cell-based assay, TSCE-W up to 1 mg/mL did not show cytoxicity against CCD-1064Sk cells as determined by various standard assays: MTT, neutral red, Hoechst 33342 staining and trypan blue dye exclusion assay. Cells treated with TSCE-W alone at the non-toxic concentration did not generate reactive oxygen species (ROS) and lipid peroxidation. Additionally, TSCE-W significantly increased intracellular glutathione (GSH) level and the activity of superoxide dismutase (SOD) and catalase (CAT). TSCE-W reduced the generation of ROS and lipid peroxidation as well as increased intracellular GSH level in H2O2-treated cells. The antioxidant mechanism of TSCE-W via protein expression was performed using Western blot analysis. The result showed that treatment of TSCE-W before H2O2 exposure for 3 h significantly up-regulated the protein expression of antioxidant enzymes including copper, zinc superoxide dismutase (Cu,ZnSOD), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) as compared with vehicle control. Treatment of TSCE-W prior to H2O2 exposure for 6 h maintained the protein expression of Cu,ZnSOD, GPx, and CAT comparable with vehicle control. The results suggest that an inexpensive and simple boiling-water extraction of TSCE-W may provide a good source of natural antioxidant. TSCE-W protected oxidative stress in cells by the regulation of antioxidant enzymes expression, TSCE-W potentially useful for health food additives and cosmeceuticals.
DOI: 10.14457/cu.the.2011.161
title: The role of reactive oxygen species in cisplatin-and doxorubicin-included apoptosis in keratinocytes and dermal papilla cells

description: Massive apoptosis of hair follicle cells has been implicated in the pathogenesis of chemotherapy-induced alopecia (CIA), but the underlying mechanisms of regulation are not well understood. The present study investigated the apoptotic effect of cisplatin and doxorubicin on human hair follicle dermal papilla cells and HaCaT keratinocytes, and determined the identity and role of specific reactive oxygen species (ROS) involved in the process. Treatment of the cells with cisplatin and doxorubicin induced ROS generation and a parallel increase in caspase activation and apoptotic cell death. Inhibition of ROS generation by antioxidants inhibited the apoptotic effect of cisplatin and doxorubicin, indicating the role of ROS in the process. ROS inhibition studies showed the essential role of different ROS in apoptotic effects of cisplatin and doxorubicin. Hydroxyl radical is the primary oxidative species responsible for the apoptotic effect of cisplatin, while superoxide anion is responsible for apoptosis by doxorubicin. Electron spin resonance studies confirmed the formation of hydroxyl radical and superoxide induced by cisplatin and doxorubicin, respectively. The mechanisms by which these specific ROS mediate the apoptotic effect of cisplatin and doxorubicin were shown to involve down-regulation of Bcl-2 via ubiquitin-proteasomal degradation. Ubiquitination and degradation of Bcl-2 was further shown to be mediated by dephosphorylation of Bcl-2. Together, our results indicate the essential role of ROS in cisplatin- and doxorubicin-induced cell death of hair follicle cells through Bcl-2 regulation. Since CIA is a major side effect of cisplatin, doxorubicin and many other chemotherapeutic agents with no known effective treatments, the knowledge gained from this study could be useful in the design of preventive treatment strategies for CIA through localized therapy without compromising the chemotherapy efficacy
DOI: 10.5517/ccdc.csd.cc1nq7k5
title: CCDC 1540717: Experimental Crystal Structure Determination

description: Related Article: L. Singla, H.R. Yadav, A. Roy Choudhury|2020|Acta Crystallogr.,Sect.B:Struct.Sci.,Cryst.Eng. and Mat.|76|604|doi:10.1107/S2052520620006873
DOI: 10.5517/ccdc.csd.cc1nq7p9
title: CCDC 1540721: Experimental Crystal Structure Determination

description: Related Article: L. Singla, H.R. Yadav, A. Roy Choudhury|2020|Acta Crystallogr.,Sect.B:Struct.Sci.,Cryst.Eng. and Mat.|76|604|doi:10.1107/S2052520620006873
DOI: 10.14457/cu.the.2011.162
title: Effect of cortical spreading depression on synaptic transmission in rat hippocampus

description: There is a relationship between migraine aura and amnesic attack. Cortical spreading depression (CSD), a phenomenon underlying migraine attack, may be responsible for hippocampus-related symptoms. However, the precise role of CSD on hippocampal activity has not been examined. The present study aimed to investigate the alteration of hippocampal synaptic plasticity and basal synaptic transmission induced by CSD. Furthermore, this study also aimed to examine whether capsaicin could modulate hippocampal plasticity that was altered by CSD. Male Wistar rats were divided into CSD and control groups. Repetitive CSDs were induced in vivo by topical application of solid KCl. Forty-five minutes later, the ipsilateral hippocampus was removed, and hippocampal slices were prepared for a series of in vitro electrophysiological studies. After CSD induction, SD waves also appeared in the hippocampus. Repetitive CSDs led to a decrease in the magnitude of long-term potentiation (LTP) in the hippocampus. CSD also reduced hippocampal synaptic efficacy, as shown by a reduction of postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor responses. In contrast, postsynaptic N-methyl-D-aspartate (NMDA) receptor responses remained unchanged. Furthermore, there were no changes in paired-pulse profiles between the groups, indicating that CSD did not induce any presynaptic alterations. Despite unaltered NMDA receptor responses, CSD elevated the ratio of GluN2A to GluN2B subunit of NMDA receptor (Glu2A/B ratio). Moreover, capsaicin administration recovered LTP that was suppressed by CSD. These findings suggest that a reduction of postsynaptic AMPA receptor responses and an increase of Glu2A/B ratio are the mechanisms responsible for impaired hippocampal LTP induced by CSD. This impaired LTP could be improved by capsaicin.
DOI: 10.14457/cu.the.2011.163
title: Removal of haloacetonitriles by adsorption on modified inorganic porous materials

description: The main objective of this study was to investigate the removal of haloacetonitriles (HANs) in aqueous solution by adsorption on various surface-modified inorganic porous materials. Hexagonal mesoporous silicate (HMS) was synthesized and modified surface with various organic functional groups (amino-, mercapto- and octyl groups) to investigate the effect of surface functional group on adsorption efficiency and mechanism compared with powder activated carbon (PAC). In addition, HMS then modified by substitution of titanium in silicate structure to investigate the relationship between the porous structure and the resultant adsorption capacities, compared to the similar mesopore SBA-15. Zeolite NaY was used so as to investigate the effect of crystalline structure. The results indicated that the adsorption rate and capacity of HANs both significantly influenced by the different porous and crystalline structures as well as by the surface functional group of adsorbent. M-HMS provided higher adsorption capacity of HANs than the other organic-functionalized adsorbents and had a comparable the adsorption capacity to PAC. The different molecular structure of HANs obviously affected the adsorption capacity and selectivity over M-HMS whereas selective adsorption over PAC was not observed. Moreover, the effect of electrolyte in tap water and in the presence of co-existing DBPs (trihalomethanes and haloacetic acids) did not affect the selective adsorption of both adsorbents significantly. The adsorption mechanism is believed to occur via a more complex interplay between an ion-dipole electrostatic interaction and chemisorption. Adsorbent regeneration study by extraction method revealed an irreversible adsorption on adsorbent surface which confirmed the chemisorption mechanism. Furthermore, mesoporous silicate SBA-CHX was modified surface by polymerizable gemini surfactant (PG-SBA-CHX) adsorption to investigate the influence of adsolubilization on HAN adsorption efficiency. PG-SBA-CHX could efficiently adsorb the HANs especially hydrophobic HANs compared with unmodified adsorbent, however, hydrophilic HANs were the lower adsorption capacity. This indicated that the partition process (adsolubilization) plays crucial role on the adsorption related to hydrophilic / hydrophobicity of adsorbate.
DOI: 10.14457/cu.the.2011.164
title: Catalytic pyrolysis of corncob using metal supported SBA-15

description: In this study, prepared catalysts were divided into three groups consisting of microporous material (beta zeolite), mesoporous materials (SBA-15 and Al-SBA-15) and metal (Ni and Pd) loading on supported SBA-15 and beta zeolite. The catalysts were characterized by X-ray powder diffraction, nitrogen sorption analysis, solid state 27Al-nuclear magnetic resonance, scanning electron microscopy and transmission electron microscopy. All catalysts were applied for thermal and catalytic pyrolysis processes of agricultural waste as corncob in a stainless steel tube reactor under nitrogen atmosphere. Factors affected to reaction such as reaction temperature, catalytic type and amount of catalyst were studied. The pyrolysis temperature was the important parameter affected to the decomposition of biomass at low temperature as 300°C and short time were applied to be cracked corncob over various catalysts. The gas products were methane, the hydrocarbon having boiling point higher than n-pentane (C5+) and CO2 whereas the light hydrocarbon gases having 2-4 carbon atoms (C2-C4) were minor products. The distillated liquid products were divided into six functional groups including acid and alcohol groups, furan derivative group, phenolic compound group, light ketone group, light ester group and others. The Al-SBA-15(20) catalyst gave high light ester group such as 2-methyl-butyl propanoic acid ester and 2-methoxy phenyl acetate which most important used as intermediate products in sweet flavor and apple, banana aroma in the food industry.
DOI: 10.14457/cu.the.2011.165
title: Surface modification of fabrics by RF plasma for ink absorption

description: The physical and chemical properties of cotton and polyester fabrics were modified by three types of gas plasma pretreatment to improve ink absorption of pigmented water-based inks. Oxygen (O2), nitrogen (N2), and sulfur hexafluoride (SF6) gas plasmas were used in this research. Effects of each gas plasma parameter, namely, power, exposure time, and gas pressure during exposure were investigated on physical and chemical properties of the treated fabrics. X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy were used to identify functional groups on the fabrics. Attenuated total reflection Fourier transform infrared spectroscopy was used to study the functional groups of the pigments. Color spectroscopic technique was used to investigate color ink strength absorbed on the printed fabrics. The results indicated that O2 plasma and N2 plasma increased the fabric hydrophilicity and surface energy with the increases of C-O, C=O, and O-C=O groups on the fabrics after the treatment by O2 plasma. C-N and O=C-NH2 groups were given by the N2 plasma. The increased hydrophilicity of the fabric by O2 plasma induced greater water-based ink absorption than by N2 plasma. The K/S values were increased as well when comparing with those of the untreated fabrics. The SF6 plasma-treated fabrics were hydrophobic which caused less ink absorption. The fabric surface roughness, caused by plasma etching increased fabric surface areas to capture more ink and thus increased more ink adhesion. The cotton fabrics exhibited higher ink adhesion and wider color gamut after the O2 plasma treatment comparing with those from the N2 plasma. More ink adhesion and wider color gamut were found on printed polyester fabrics with N2 plasma treatment than the O2 plasma treatment did.