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  1. Fulltext Comparative genomics of the nonlegume Parasponia reveals insights into evolution of nitrogen-fixing rhizobium symbioses
    van Velzen R, Holmer R, Bu F, Rutten L, van Zeijl A, Liu W, et al.
    Proc Natl Acad Sci U S A, 2018 May 15;115(20):E4700-E4709.
    PMID: 29717040 DOI: 10.1073/pnas.1721395115
    Nodules harboring nitrogen-fixing rhizobia are a well-known trait of legumes, but nodules also occur in other plant lineages, with rhizobia or the actinomycete Frankia as microsymbiont. It is generally assumed that nodulation evolved independently multiple times. However, molecular-genetic support for this hypothesis is lacking, as the genetic changes underlying nodule evolution remain elusive. We conducted genetic and comparative genomics studies by using Parasponia species (Cannabaceae), the only nonlegumes that can establish nitrogen-fixing nodules with rhizobium. Intergeneric crosses between Parasponia andersonii and its nonnodulating relative Trema tomentosa demonstrated that nodule organogenesis, but not intracellular infection, is a dominant genetic trait. Comparative transcriptomics of P. andersonii and the legume Medicago truncatula revealed utilization of at least 290 orthologous symbiosis genes in nodules. Among these are key genes that, in legumes, are essential for nodulation, including NODULE INCEPTION (NIN) and RHIZOBIUM-DIRECTED POLAR GROWTH (RPG). Comparative analysis of genomes from three Parasponia species and related nonnodulating plant species show evidence of parallel loss in nonnodulating species of putative orthologs of NIN, RPG, and NOD FACTOR PERCEPTION Parallel loss of these symbiosis genes indicates that these nonnodulating lineages lost the potential to nodulate. Taken together, our results challenge the view that nodulation evolved in parallel and raises the possibility that nodulation originated ∼100 Mya in a common ancestor of all nodulating plant species, but was subsequently lost in many descendant lineages. This will have profound implications for translational approaches aimed at engineering nitrogen-fixing nodules in crop plants.
    Matched MeSH terms: Plant Proteins/genetics*
  2. Evaluation of dietary taste patterns as assessed by FFQ against 24-h recalls and biomarkers of exposure
    van Langeveld AWB, Teo PS, Mars M, Feskens EJM, de Graaf C, de Vries JHM
    Eur J Clin Nutr, 2019 01;73(1):132-140.
    PMID: 30254242 DOI: 10.1038/s41430-018-0300-1
    BACKGROUND/OBJECTIVE: Taste is of key importance in food choice and dietary patterns, but studies on taste profiles are limited. We previously assessed dietary taste patterns by 24 h recalls (24hR), but for epidemiological studies food frequency questionnaires (FFQ) may also be suitable. This study compared dietary taste patterns based on FFQ against 24hR and biomarkers of exposure.

    SUBJECTS/METHODS: A taste database including 467 foods' sweet, sour, bitter, salt, umami and fat sensation values was combined with food intake data to assess dietary taste patterns: the contribution to energy intake of 6 taste clusters. The FFQ's reliability was assessed against 3-d 24hR and urinary biomarkers for sodium (Na) and protein intake (N) in Dutch men (n = 449) and women (n = 397) from the NQplus validation study (mean age 53 ± 11 y, BMI 26 ± 4 kg/m2).

    RESULTS: Correlations of dietary taste patterns ranged from 0.39-0.68 between FFQ and 24hR (p 

    Matched MeSH terms: Dietary Proteins/urine*
  3. Fulltext Mutation-specific pathophysiological mechanisms define different neurodevelopmental disorders associated with SATB1 dysfunction
    den Hoed J, de Boer E, Voisin N, Dingemans AJM, Guex N, Wiel L, et al.
    Am J Hum Genet, 2021 02 04;108(2):346-356.
    PMID: 33513338 DOI: 10.1016/j.ajhg.2021.01.007
    Whereas large-scale statistical analyses can robustly identify disease-gene relationships, they do not accurately capture genotype-phenotype correlations or disease mechanisms. We use multiple lines of independent evidence to show that different variant types in a single gene, SATB1, cause clinically overlapping but distinct neurodevelopmental disorders. Clinical evaluation of 42 individuals carrying SATB1 variants identified overt genotype-phenotype relationships, associated with different pathophysiological mechanisms, established by functional assays. Missense variants in the CUT1 and CUT2 DNA-binding domains result in stronger chromatin binding, increased transcriptional repression, and a severe phenotype. In contrast, variants predicted to result in haploinsufficiency are associated with a milder clinical presentation. A similarly mild phenotype is observed for individuals with premature protein truncating variants that escape nonsense-mediated decay, which are transcriptionally active but mislocalized in the cell. Our results suggest that in-depth mutation-specific genotype-phenotype studies are essential to capture full disease complexity and to explain phenotypic variability.
    Matched MeSH terms: Matrix Attachment Region Binding Proteins/genetics*; Matrix Attachment Region Binding Proteins/metabolism; Matrix Attachment Region Binding Proteins/chemistry
  4. Fulltext FAR1 and FAR2 regulate the expression of genes associated with lipid metabolism in the rice blast fungus Magnaporthe oryzae
    bin Yusof MT, Kershaw MJ, Soanes DM, Talbot NJ
    PLoS One, 2014;9(6):e99760.
    PMID: 24949933 DOI: 10.1371/journal.pone.0099760
    The rice blast fungus Magnaporthe oryzae causes plant disease via specialised infection structures called appressoria. These dome-shaped cells are able to generate enormous internal pressure, which enables penetration of rice tissue by invasive hyphae. Previous studies have shown that mobilisation of lipid bodies and subsequent lipid metabolism are essential pre-requisites for successful appressorium-mediated plant infection, which requires autophagic recycling of the contents of germinated spores and germ tubes to the developing appressorium. Here, we set out to identify putative regulators of lipid metabolism in the rice blast fungus. We report the identification of FAR1 and FAR2, which encode highly conserved members of the Zn2-Cys6 family of transcriptional regulators. We generated Δfar1, Δfar2 and Δfar1Δfar2 double mutants in M. oryzae and show that these deletion mutants are deficient in growth on long chain fatty acids. In addition, Δfar2 mutants are also unable to grow on acetate and short chain fatty acids. FAR1 and FAR2 are necessary for differential expression of genes involved in fatty acid β-oxidation, acetyl-CoA translocation, peroxisomal biogenesis, and the glyoxylate cycle in response to the presence of lipids. Furthermore, FAR2 is necessary for expression of genes associated with acetyl-CoA synthesis. Interestingly, Δfar1, Δfar2 and Δfar1Δfar2 mutants show no observable delay or reduction in lipid body mobilisation during plant infection, suggesting that these transcriptional regulators control lipid substrate utilization by the fungus but not the mobilisation of intracellular lipid reserves during infection-related morphogenesis.
    Matched MeSH terms: Fungal Proteins/biosynthesis*; Fungal Proteins/genetics
  5. Comparative effects of indomethacin and nabumetone on urine and electrolyte output in conscious rats
    bin Long I, Singh HJ, Rao GJ
    J. Pharmacol. Sci., 2005 Nov;99(3):272-6.
    PMID: 16293937
    The effects of indomethacin and nabumetone on urine and electrolyte excretion in conscious rats were examined. Male Sprague-Dawley rats were housed individually for a five-week duration, consisting of acclimatization, control, experimental, and recovery phases. During the experimental phase, rats were given either indomethacin (1.5 mg . kg(-1) body weight . day(-1) in 0.5 ml saline, n = 10), nabumetone (15 mg . kg(-1) body weight . day(-1) 0.5 ml saline, n = 10), or 0.5 ml saline alone (n = 10) for a period of two weeks. Water and food intake, body weight, urine output, and electrolyte excretions were estimated. Data were analyzed using two-way ANOVA. Urine output in the indomethacin- and nabumetone-treated groups was not different from the controls, but was significantly different between the drug-treated groups (P<0.01). Sodium, potassium, calcium, and magnesium excretions were not different between nabumetone-treated and control rats. However, sodium and potassium excretion was significantly lower in rats receiving indomethacin when compared to the control rats. Calcium and magnesium outputs, although did not differ from the controls, nevertheless decreased significantly with indomethacin (P<0.01). It appears that indomethacin and nabumetone when given at maximum human therapeutic doses may affect urine and electrolyte output in conscious rats.
    Matched MeSH terms: Membrane Proteins/antagonists & inhibitors
  6. Natural Variation in Freezing Tolerance and Cold Acclimation Response in Arabidopsis thaliana and Related Species
    Zuther E, Lee YP, Erban A, Kopka J, Hincha DK
    Adv Exp Med Biol, 2018 10 6;1081:81-98.
    PMID: 30288705 DOI: 10.1007/978-981-13-1244-1_5
    During low-temperature exposure, temperate plant species increase their freezing tolerance in a process termed cold acclimation. The molecular mechanisms involved in cold acclimation have been mostly investigated in Arabidopsis thaliana. In addition, other Brassicaceae species related to A. thaliana have been employed in recent years to study plant stress responses on a phylogenetically broader basis and in some cases with extremophile species with a much higher stress tolerance. In this paper, we briefly summarize cold acclimation responses in A. thaliana and current knowledge about cold acclimation in A. thaliana relatives with special emphasis on Eutrema salsugineum and two closely related Thellungiella species. We then present a transcriptomic and metabolomic analysis of cold acclimation in five A. thaliana and two E. salsugineum accessions that differ widely in their freezing tolerance. Differences in the cold responses of the two species are discussed.
    Matched MeSH terms: Arabidopsis Proteins
  7. Fulltext Continuous genotyping for human rotavirus group a
    Zuridah H
    Med J Malaysia, 2012 Oct;67(5):548.
    PMID: 23770884
    Matched MeSH terms: Capsid Proteins
  8. Sequence and phylogenetic analysis of S1, S2, M, and N genes of infectious bronchitis virus isolates from Malaysia
    Zulperi ZM, Omar AR, Arshad SS
    Virus Genes, 2009 Jun;38(3):383-91.
    PMID: 19242786 DOI: 10.1007/s11262-009-0337-2
    Two Malaysian infectious bronchitis virus isolates, MH5365/95 and V9/04 were characterized based on sequence and phylogenetic analyses of S1, S2, M, and N genes. Nucleotide sequence alignments revealed many point mutations, short deletions, and insertions in S1 region of both IBV isolates. Phylogenetic analysis of S1 gene and sequences analysis of M gene indicated that MH5365/95 and V9/04 belong to non-Massachusetts strain. However, both isolates share only 77% identity. Analysis based on S1 gene showed that MH5365/95 shared more than 87% identity to several Chinese strains. Meanwhile, V9/04 showed only 67-77% identity to all the previously studied IBV strains included in this study suggesting it is a variant of IBV isolate that is unique to Malaysia. Phylogenetic analysis suggests, although both isolates were isolated 10 years apart from different states in Malaysia, they shared a common origin. Analysis based on S2 and N genes indicated that both strains are highly related to each other, and there are fewer mutations which occurred in the respective genes.
    Matched MeSH terms: Viral Proteins/genetics*
  9. Fulltext Calculated parameters for the diagnosis of Wilson disease
    Zulkufli NS, Sthaneshwar P, Chan WK
    Singapore Med J, 2023 Mar;64(3):188-195.
    PMID: 35139628 DOI: 10.11622/smedj.2022019
    INTRODUCTION: The diagnosis of Wilson disease (WD) is plagued by biochemical and clinical uncertainties. Thus, calculated parameters have been proposed. This study aimed to: (a) compare the diagnostic values of non-caeruloplasmin copper (NCC), NCC percentage (NCC%), copper-caeruloplasmin ratio (CCR) and adjusted copper in WD; and (b) derive and evaluate a discriminant function in diagnosing WD.

    METHODS: A total of 213 subjects across all ages who were investigated for WD were recruited. WD was confirmed in 55 patients, and the rest were WD free. Based on serum copper and caeruloplasmin values, NCC, NCC%, CCR and adjusted copper were calculated for each subject. A function was derived using discriminant analysis, and the cut-off value was determined through receiver operating characteristic analysis. Classification accuracy was found by cross-tabulation.

    RESULTS: Caeruloplasmin, total copper, NCC, NCC%, CCR, adjusted copper and discriminant function were significantly lower in WD compared to non-WD. Discriminant function showed the best diagnostic specificity (99.4%), sensitivity (98.2%) and classification accuracy (99.1%). Caeruloplasmin levels <0.14 g/L showed higher accuracy than the recommended 0.20 g/L cut-off value (97.7% vs. 87.8%). Similarly, molar NCC below the European cut-off of 1.6 umol/L showed higher accuracy than the American cut-off of 3.9 umol/L (80.3% vs. 59.6%) (P < 0.001). NCC%, mass NCC, CCR and adjusted copper showed poorer performances.

    CONCLUSION: Discriminant function differentiates WD from non-WD with excellent specificity, sensitivity and accuracy. Performance of serum caeruloplasmin <0.14 g/L was better than that of <0.20 g/L. NCC, NCC%, CCR and adjusted copper are not helpful in diagnosing WD.

    Matched MeSH terms: Repressor Proteins
  10. Fulltext Acute phase proteins, interleukin 6, and heat shock protein 70 in broiler chickens administered with corticosterone
    Zulkifli I, Najafi P, Nurfarahin AJ, Soleimani AF, Kumari S, Aryani AA, et al.
    Poult Sci, 2014 Dec;93(12):3112-8.
    PMID: 25306460 DOI: 10.3382/ps.2014-04099
    An experiment was conducted to determine the effect of corticosterone (CORT) administration on serum ovotransferrin (OVT), α1-acid glycoprotein (AGP), ceruloplasmin (CPN), and IL-6 concentrations, and brain heat shock protein (HSP) 70 expression in broiler chickens. From 14 to 20 d of age, equal numbers of birds were subjected to either (i) daily intramuscular injection with CORT in ethanol:saline (1:1, vol/vol) at 6 mg/kg of BW, or (ii) daily intramuscular injection with 0.5 mL ethanol:saline (1:1, vol/vol; control). Blood samples were collected before CORT treatment (14 d old), 3 and 7 d after CORT injections, and 4 d after cessation of CORT administration for determination of serum levels of CORT, OVT, AGP, CPN, and IL-6. Brain samples (whole cerebrum) were collected to measure HSP 70 density. Although CORT administration significantly increased feed intake, weight gain was significantly depressed. Administration of CORT also increased CORT, OVT, CPN, AGP, IL-6, and HSP 70 expression. Four days following cessation of CORT administration, OVT declined to the basal level but not CPN and AGP. In conclusion, an elevation in CORT can induce an acute-phase response and HSP 70 expression. Thus, APP and HSP 70 may be of value as indicators of stress in poultry.
    Matched MeSH terms: Acute-Phase Proteins/genetics; Acute-Phase Proteins/metabolism*; HSP70 Heat-Shock Proteins/genetics; HSP70 Heat-Shock Proteins/metabolism*
  11. Fulltext Crating and heat stress influence blood parameters and heat shock protein 70 expression in broiler chickens showing short or long tonic immobility reactions
    Zulkifli I, Al-Aqil A, Omar AR, Sazili AQ, Rajion MA
    Poult Sci, 2009 Mar;88(3):471-6.
    PMID: 19211514 DOI: 10.3382/ps.2008-00287
    Two hundred thirty-five 1-d-old broiler chickens showing short or long tonic immobility responses were classified as low fear (LF) or high fear (HF) responders, respectively. On d 41, they were subjected to either crating or heat challenge (34 +/- 1 degrees C) for 3 h and its effect on plasma corticosterone concentration, heterophil/lymphocyte ratios, and heat shock protein (HSP) 70 expression in brain tissue were determined. Crating and heat exposure elevated heterophil/lymphocyte ratios in both LF and HF birds. Circulating corticosterone, however, was greater in HF than LF birds after crating and heat challenge. Although differences between fear responder group for HSP 70 were negligible before heat challenge, after 3 h of heat exposure, the response was greater for the HF than the LF group. Both LF and HF showed similar increases in HSP 70 after crating.
    Matched MeSH terms: HSP70 Heat-Shock Proteins/genetics; HSP70 Heat-Shock Proteins/metabolism*
  12. The effect of early-age food restriction on heat shock protein 70 response in heat-stressed female broiler chickens
    Zulkifli I, Che Norma MT, Israf DA, Omar AR
    Br Poult Sci, 2002 Mar;43(1):141-5.
    PMID: 12003331
    1. This study was conducted to determine the effect of early-age food restriction on heat shock protein (hsp) 70 synthesis in the brains of female broiler chickens exposed to high ambient temperatures. 2. Chicks were brooded for 3 weeks and then maintained at 24+/-1 degrees C. 3. On d 0, chicks were assigned to one of 4 feeding regimens; each regimen was applied to 4 cages of chicks. The regimens were: (1) ad libitum feeding (AL); (2) 80% food restriction at 4, 5 and 6 d of age (F80); (3) 60% food restriction at 4, 5, and 6 d of age (F60); and (4) 40% food restriction at 4, 5 and 6 d of age (F40). From d 35 to d 41, all chicks were subjected to 38+/-1 degrees C for 2 h/d. 4. One day following food restriction (d 7), hsp 70 expression in the brain samples of F60 and F40 chicks was augmented but not those fed AL and F80. 5. Prior to the heat challenge (d 35), all chicks had similar hsp 70 response. Irrespective of feeding regimen, there was a marked increase in hsp 70 expression after 4 d of heat treatment (d 38). Following 7 d of heat exposure (d 41), except for the F60 chicks, the augmented hsp 70 expression in the brains of AL, F80 and F40 birds was not maintained. 6. Enhancement of hsp 70 expression was noted in birds subjected to F60, but not AL, F80 or F40, throughout the period of heat exposure.
    Matched MeSH terms: HSP70 Heat-Shock Proteins/biosynthesis*
  13. Dietary supplementation of L-glutamine and L-glutamate in broiler chicks subjected to delayed placement
    Zulkifli I, Shakeri M, Soleimani AF
    Poult Sci, 2016 Sep 1.
    PMID: 27587729
    This study was conducted to investigate the effect of dietary glutamine (Gln) + glutamic acid (Glu) supplementation on growth performance and physiological stress response in broiler chickens subjected to 24 h delay in placement. Equal number of day-old broiler chicks were assigned to either immediate placement or with 24 h delay in placement with no access to feed and water. Chicks from each placement group were fed either standard starter diet (control) or standard starter diet +1% AminoGut (AG; mixture of 10% Gln and 10% Glu) from 1 to 21 d. Blood and duodenal samples were collected at 21 d for analysis of serum levels of ceruloplasmin (CER), ovotransferin (OVT) and α-1 acid glycoprotein (AGP), duodenal heat shock protein (HSP) 70 expression, and villi length and crypt depth. Results showed that delayed placement for 24 h was detrimental to weight gain during the starter phase (1 to 21 d) but not thereafter. AG supplementation was not able to eliminate that reduction in weight gain and feed intake during the starter stage. However, the observed enhancement in villi length and crypt depth at d 21 resulted in improvement of FCR and weight gain during the finisher stage (22 to 42 d) and consequently the overall period (1 to 42 d). Broiler chickens supplemented with AG also showed lower mortality rate, and higher AGP, OVT, CER, and HSP 70 expression compared to their control counterparts. Based on AGP, OVT, CER, and HSP 70 expression, there is no indication that delayed placement was physiologically stressful to the broiler chickens at 21 d of age.
    Matched MeSH terms: HSP70 Heat-Shock Proteins
  14. Effects of low-protein diets on acute phase proteins and heat shock protein 70 responses, and growth performance in broiler chickens under heat stress condition
    Zulkifli I, Akmal AF, Soleimani AF, Hossain MA, Awad EA
    Poult Sci, 2018 Apr 01;97(4):1306-1314.
    PMID: 29381776 DOI: 10.3382/ps/pex436
    A study with a 4 × 2 factorial arrangement was conducted to investigate the effects of 4 dietary protein levels and 2 environmental conditions on acute phase proteins (APP), brain heat shock protein (HSP) 70 density, and growth performance of broiler chickens. Day-old broiler chicks (Cobb 500) were fed isocaloric diets but with various levels of crude protein (CP), namely, (1) 21.0 and 19.0% CP in starter and finisher diets, respectively (control), (2) 19.5 and 17.5% CP in starter and finisher diets, respectively (Diet A), (3) 18.0 and 16.0% CP in starter and finisher diets, respectively (Diet B), and (4) 16.5 and 14.5% CP in starter and finisher diets, respectively (Diet C). Equal numbers of birds from each diet were subjected to either 23±1°C throughout or 33±1°C for 6 h per d from 22 to 35 d of age. From d 1 to 21, feed intake (FI) and weight gain (WG) decreased linearly (P = 0.021 and P = 0.009, respectively), as CP level was reduced. During the heat treatment period (d 22 to 35), there were significant (P = 0.04) diet × heat treatment interactions for FCR. Diet had no effect on FCR among the unheated birds, but the ratio increased linearly (P = 0.007) as dietary CP level decreased. Irrespective of ambient temperature, there was a significant linear decrease in FI (P = 0.032) and WG (P < 0.001) as dietary CP level decreased. Low-CP diets improved the survivability of heat-stressed broilers when compared to those fed control diets. Low-CP diets linearly decreased (P < 0.01) APP (ovotransferrin and alpha-acid glycoprotein) responses. Both APP and HSP 70 reactions were elevated following heat treatment. In conclusion, feeding broilers with low-CP diets adversely affect the growth performance of broilers under heat stress condition. However, low-CP diets were beneficial in improving the survivability. Because APP are involved in the restoration of homeostasis, the adverse effect of low-CP diet on the synthesis of these proteins could be of concern.
    Matched MeSH terms: Acute-Phase Proteins/genetics*; Acute-Phase Proteins/metabolism; HSP70 Heat-Shock Proteins/genetics*; HSP70 Heat-Shock Proteins/metabolism; Avian Proteins/genetics*; Avian Proteins/metabolism
  15. Fulltext Food Consumption, Developmental Time, and Protein Profile of the Digestive System of the Red Palm Weevil, Rhynchophorus ferrugineus (Coleptera: Dryophthoridae) Larvae Reared on Three Different Diets
    Zulkifli AN, Zakeri HA, Azmi WA
    J Insect Sci, 2018 Sep 01;18(5).
    PMID: 30285257 DOI: 10.1093/jisesa/iey093
    The red palm weevil (RPW), Rhynchophorus ferrugineus Olivier (Coleoptera: Dryophthoridae) is one of the most dangerous pests of major cultivated palms including coconut, oil palm, and sago. The larval stage of the weevil causes the most destruction of the palms as it completely destroys the palm cabbage. In this study, the larvae were given three different diets-coconut cabbage, oil palm cabbage, and sago stem, under laboratory conditions for food consumption and developmental time experiment. The protein profiles of the digestive systems of the larvae fed on these three diets were also determined. Although the coconut diet was the most consumed by RPW larvae compared to oil palm and sago diets, the growth rate of RPW larvae on oil palm diet was however significantly shorter than those on the coconut and sago diets: the RPW only need 1 mo and 9 d to complete the larval duration. Proteins profiling of eight 2-DE gel protein spots that range 50-20 kDa were identified by mass spectrometry sequence analysis. Based on the Matrix Science Software, the most dominant protein was cationic trypsin. However, based on the NCBI BLAST tool, aminopeptidase N was the most dominant enzyme. This finding can lead to the development of pest control strategies based on the antinutritional protease inhibitors as potential biocontrol agents. Urgent action to find effective control methods should be taken seriously as this weevil is presumed to be one of the serious pests of oil palm industry in Malaysia.
    Matched MeSH terms: Insect Proteins/metabolism
  16. In Silico Docking of Vitamin E Isomers on Transport Proteins
    Zulkiflee NS, Awang SA, Ming WX, Kamilan MFW, Mariappan MY, Kit TJ
    Curr Comput Aided Drug Des, 2020;16(4):467-472.
    PMID: 31203808 DOI: 10.2174/1573409915666190614113733
    BACKGROUND: Vitamin E is comprised of α, β, γ and δ-tocopherols (Ts) and α, β, γ and δ- tocotrienols (T3s). Vitamin E has neuroprotective antioxidant, anti-cancer, and cholesterol-lowering effects. Intracellular trafficking of these isomers remains largely unknown, except for αT which is selectively transported by αT transfer protein (αTTP).

    OBJECTIVE: This study aimed to determine the binding of vitamin E isomers on transport proteins using in silico docking.

    METHODS: Transport proteins were selected using AmiGo Gene Ontology tool based on the same molecular function annotation as αTTP. Protein structures were obtained from the Protein Data Bank. Ligands structures were obtained from ZINC database. In silico docking was performed using SwissDock.

    RESULTS AND DISCUSSION: A total of 6 transport proteins were found: SEC14-like protein 2, glycolipid transfer protein (GLTP), pleckstrin homology domain-containing family A member 8, collagen type IV alpha-3-binding protein, ceramide-1-phosphate transfer protein and afamin. Compared with other transport proteins, αTTP had the highest affinities for all isomers except βT3. Binding order of vitamin E isomers toward αTTP was γT > βT > αT > δT > αT3 > γT3 > δT3 > βT3. GLTP had a higher affinity for tocotrienols than tocopherols. βT3 bound stronger to GLTP than αTTP.

    CONCLUSION: αTTP remained as the most preferred transport protein for most of the isomers. The binding affinity of αT toward αTTP was not the highest than other isomers suggested that other intracellular trafficking mechanisms of these isomers may exist. GLTP may mediate the intracellular transport of tocotrienols, especially βT3. Improving the bioavailability of these isomers may enhance their beneficial effects to human.

    Matched MeSH terms: Carrier Proteins/metabolism*; Carrier Proteins/chemistry
  17. Photodynamic therapy mediates innate immune responses via fibroblast-macrophage interactions
    Zulaziz N, Azhim A, Himeno N, Tanaka M, Satoh Y, Kinoshita M, et al.
    Hum. Cell, 2015 Oct;28(4):159-66.
    PMID: 25997703 DOI: 10.1007/s13577-015-0118-2
    Antibacterial photodynamic therapy (PDT) has come to attract attention as an alternative therapy for drug-resistant bacteria. Recent reports revealed that antibacterial PDT induces innate immune response and stimulates abundant cytokine secretion as a part of inflammatory responses. However, the underlying mechanism how antibacterial PDT interacts with immune cells responsible for cytokine secretion has not been well outlined. In this study, we aimed to clarify the difference in gene expression and cytokine secretion between combined culture of fibroblasts and macrophages and their independent cultures. SCRC-1008, mouse fibroblast cell line and J774, mouse macrophage-like cell line were co-cultured and PDT treatments with different parameters were carried out. After various incubation periods (1-24 h), cells and culture medium were collected, and mRNA and protein levels for cytokines were measured using real-time PCR and ELISA, respectively. Our results showed that fibroblasts and macrophages interact with each other to mediate the immune response. We propose that fibroblasts initially respond to PDT by expressing Hspa1b, which regulates the NF-κB pathway via Tlr2 and Tlr4. Activation of the NF-κB pathway then results in an enhanced secretion of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) and neutrophil chemoattractant MIP-2 and KC from macrophages.
    Matched MeSH terms: HSP70 Heat-Shock Proteins/metabolism
  18. Selective inhibition of genes in methicillin resistant Staphylococcus aureus (MRSA) treated with Melastoma malabathricum methanol extract
    Zulaikah Mohamed, Nazlina Ibrahim, Ismail Ahmad
    Sains Malaysiana, 2008;37(1):107-113.
    Methanol extract of Melastoma malabathricum leaves inhibited the growth of Staphylococcus aureus and six clinical isolates of Methicilin Resistant Stapyhlococcus aureus (MRSA 1-6). The minimum inhibitory concentration (MIC) of test substance was 1.565mg/ml and the minimum bacteriocidal concentration (MBC) was 3.125 mg/ml. The methanol extract suppressed RNA synthesis at 10 mg/ml as shown by RNA profile which was devoid of three bands compared to the control. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis using seven primer pairs was only successful in amplifying four cDNA amplicons. The failure to amplify three cDNA amplicons for three primer pairs corresponding to gyrA, femA and nuc genes, implied the possibility of suppression of the corresponding mRNA. Electrophoretic separation of endogenous and exogenuos bacterial proteins showed that three and five protein, respectively were not expressed. One endogenous and three exogenous proteins were over-expressed in treated MRSA compared with untreated control. The results of the molecular and proteomic analyses are in agreement, and based on primers being used, methanol extract of M. malabathricum leaves possibly inhibits MRSA growth through inhibition of DNA synthesis, peptidoglycan production, and nuclease production.
    Keywords: Methicillin resistant Staphylococcus aureus; Melastoma malabathricum; gene expression; protein production
    Matched MeSH terms: Proteins
  19. Fulltext Stepwise evolution of pandrug-resistance in Klebsiella pneumoniae
    Zowawi HM, Forde BM, Alfaresi M, Alzarouni A, Farahat Y, Chong TM, et al.
    Sci Rep, 2015;5:15082.
    PMID: 26478520 DOI: 10.1038/srep15082
    Carbapenem resistant Enterobacteriaceae (CRE) pose an urgent risk to global human health. CRE that are non-susceptible to all commercially available antibiotics threaten to return us to the pre-antibiotic era. Using Single Molecule Real Time (SMRT) sequencing we determined the complete genome of a pandrug-resistant Klebsiella pneumoniae isolate, representing the first complete genome sequence of CRE resistant to all commercially available antibiotics. The precise location of acquired antibiotic resistance elements, including mobile elements carrying genes for the OXA-181 carbapenemase, were defined. Intriguingly, we identified three chromosomal copies of an ISEcp1-bla(OXA-181) mobile element, one of which has disrupted the mgrB regulatory gene, accounting for resistance to colistin. Our findings provide the first description of pandrug-resistant CRE at the genomic level, and reveal the critical role of mobile resistance elements in accelerating the emergence of resistance to other last resort antibiotics.
    Matched MeSH terms: Bacterial Proteins
  20. Annona muricata leaves induce G₁ cell cycle arrest and apoptosis through mitochondria-mediated pathway in human HCT-116 and HT-29 colon cancer cells
    Zorofchian Moghadamtousi S, Karimian H, Rouhollahi E, Paydar M, Fadaeinasab M, Abdul Kadir H
    J Ethnopharmacol, 2014 Oct 28;156:277-89.
    PMID: 25195082 DOI: 10.1016/j.jep.2014.08.011
    ETHNOPHARMACOLOGICAL RELEVANCE: Annona muricata known as "the cancer killer" has been widely used in the traditional medicine for the treatment of cancer and tumors. The purpose of this study is to investigate the anticancer properties of ethyl acetate extract of Annona muricata leaves (EEAM) on HT-29 and HCT-116 colon cancer cells and the underlying mechanisms.
    MATERIALS AND METHODS: The effect of EEAM on the cell proliferation of HT-29 and HCT-116 cells was analyzed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) assay. High content screening system (HCS) was applied to investigate the cell membrane permeability, mitochondrial membrane potential (MMP), nuclear condensation and cytochrome c translocation from mitochondria to cytosol. Reactive oxygen species (ROS) formation, lactate dehydrogenase (LDH) release and activation of caspase-3/7, -8 and -9 were measured while treatment. Flow cytometric analysis was used to determine the cell cycle distribution and phosphatidylserine externalization. The protein expression of Bax and Bcl-2 was determined using immunofluorescence analysis. In addition, the potential of EEAM to suppress the migration and invasion of colon cancer cells was also examined.
    RESULTS: EEAM exerted significant cytotoxic effects on HCT-116 and HT-29 cells as determined by MTT and LDH assays. After 24 h treatment, EEAM exhibited the IC₅₀ value of 11.43 ± 1.87 µg/ml and 8.98 ± 1.24 µg/ml against HT-29 and HCT-116 cells, respectively. Flow cytometric analysis demonstrated the cell cycle arrest at G1 phase and phosphatidylserine externalization confirming the induction of apoptosis. EEAM treatment caused excessive accumulation of ROS followed by disruption of MMP, cytochrome c leakage and activation of the initiator and executioner caspases in both colon cancer cells. Immunofluorescence analysis depicted the up-regulation of Bax and down-regulation of Bcl-2 proteins while treated with EEAM. Furthermore, EEAM conspicuously blocked the migration and invasion of HT-29 and HCT-116 cells.
    CONCLUSIONS: These findings provide a scientific basis for the use of A. muricata leaves in the treatment of cancer, although further in vivo studies are still required.
    Matched MeSH terms: Proto-Oncogene Proteins c-bcl-2/metabolism
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