This study was conducted to explore the anti-inflammatory effect of Jungia sellowii (Asteraceae) using a murine model of pleurisy induced by carrageenan (Cg). This plant is used in southern Brazil to treat inflammatory diseases. J. sellowii leaves were extracted with ethanol/water to obtain the crude extract (CE), which was fractionated with different solvents, yielding n-hexane (Hex), dichloromethane (DCM), ethyl acetate (EtOAc) and n-butanol (BuOH) fractions, and aqueous fraction (Aq). The major compounds succinic acid (SA) and lactic acid (LA) were isolated from Aq fraction, and their structures were determined by (1)H and (13)C NMR. Pleurisy was induced by Cg (Saleh et al. 1996). The leukocytes, exudation, myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities, metabolites of nitric oxide (NO x ) levels, protein levels and mRNA expression for interleukin 1 beta (IL-1β), tumour necrosis factor alpha (TNF-α), interleukin 17A (IL17A) and inducible of nitric oxide synthase (iNOs), and p65 protein phosphorylation (NF-κB) were analysed 4 h after pleurisy induction. Animals pre-treated with CE, BuOH, Aq, SA, or LA inhibited leukocytes, exudation, MPO and ADA activities, NO x , IL-1β, TNF-α, and IL-17A levels, and the mRNA expression for IL-1β, TNF-α, IL-17A, iNOS, and p65 protein phosphorylation (NF-κB) (p < 0.05). Our study demonstrated that J. sellowii can protect against inflammation induced by Cg by decreasing the leukocytes and exudation. Its effects are related to the decrease of either proinflammatory cytokines and/or NO x . The isolated compounds SA and LA may play an important role in this anti-inflammatory action by inhibiting all the studied parameters. The anti-inflammatory properties of these compounds are due to the downregulation of NF-κB.

Chronic alcohol consumption is associated with pathological effects on bone, and it is correlated with the increasing risk of osteoporosis and fractures. The negative effects of alcohol intake also influence bone repair processes and the osseointegration of implants. The aim of the present in vitro study was to investigate the proliferation and synthetic activity of osteoblasts isolated from the trabecular bone of rats previously exposed to 7-week intermittent exposure to ethanol vapour (EE-OB), and sham-aged rats (SA-OB), when cultured on standard commercially pure Ti (cpTi). Osteoblast proliferation (WST-1), alkaline phosphatase (ALP), osteocalcin (OC), collagen type I (CICP), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and transforming growth factor-beta1 (TGF-beta1) were measured at 1, 7, and 14 days of culture. Our results showed a decrease in the cell viability and synthetic activity of osteoblasts exposed to ethanol when cultured on cpTi. Moreover, the release of local regulatory factors from osteoblasts was imbalanced: TGF-beta1 production was reduced and TNF-alpha and IL-6 were up-regulated. These in vitro data suggest that alcohol abuse affects bone repair and decreases the ability to form bone around standard cpTi. Innovative surfaces and adjuvant therapies could be useful when implants are required in alcoholics.

Imaging techniques including computed tomography (CT), magnetic resonance imaging (MRI), single photon emission computed tomography (SPECT), and positron emission tomography (PET) have been widely applied to the investigation of patients with acute or chronic ataxias. Fundamentally, CT has a role in the emergency evaluation of the patient with acute ataxia to ascertain brainstem or cerebellar hemorrhage and to exclude a mass lesion in the posterior cranial fossa. Conventional MRI is the most frequently performed imaging investigation in patients with ataxia. It can support the diagnosis of acute cerebellitis and Wernicke encephalopathy by revealing T2 signal changes with a typical distribution. In patients with inherited or sporadic chronic ataxia it reveals three fundamental patterns of atrophy of the brainstem, cerebellum, and spinal cord which match the gross neuropathological descriptions. These are represented by olivopontocerebellar atrophy (OPCA), cortical cerebellar atrophy (CCA), and spinal atrophy (SA). A substantial correspondence exists among these patterns of atrophy shown by MRI and the etiological classification of inherited or acquired chronic ataxias. This, along with demonstration of T2 signal changes characteristic of some diseases, makes conventional MRI potentially useful for the diagnostic work-up of the single patient, especially in the case of a sporadic disease. Non-conventional MR techniques including diffusion MR, spectroscopy, and functional MR have been used in patients with acute or chronic ataxia, but their exact role in the evaluation of the single patient is not established yet. They are currently investigated as potential tools to monitor progression of neurodegeneration in chronic ataxia and to serve as "surrogate markers" in clinical trials. Several radiotracers have been utilized in combination with SPECT and PET in patients with ataxia. Perfusion SPECT can reveal cerebellar blood flow abnormalities early in the course of cerebellitis. It has also been utilized to investigate perfusion of the brain in several inherited or sporadic chronic ataxic diseases, contributing to improved understanding of the pathophysiology of these conditions. Recently, perfusion SPECT has been tested as a "surrogate marker" to verify the effects of newly developed therapies in patients with a variety of chronic ataxias. Whole-body FDG-PET is recommended in patients with suspected paraneoplastic cerebellar degeneration to detect the primary malignancy. Brain FDG-PET has provided important information on the pathophysiology of several acquired and inherited conditions. PET and SPECT with radiotracers able to assess the nigrostriatal system or the density of D2 dopamine receptors in the striatum are increasingly used in patients with adult-onset sporadic ataxia for the differential diagnosis between multiple system atrophy in which overt striatal abnormalities are found and idiopathic late-onset cerebellar ataxia in which no abnormality is detected.

To investigate the involvement of cytochrome P450s in the metabolism of plants treated with xenobiotic agrochemicals, bean leaves were treated with 3,5-dichlorosalicylic acid (DC-SA), a priming agent of plant defense and 2,6-dichloroisonicotinic acid (DC-INA), a chemical inducer of systemic acquired resistance. Through the use of directed differential display reverse transcription polymerase chain reactions, a differentially expressed cDNA amplicon, found to be up-regulated by both DC-SA and DC-INA treatment, was identified as a cytochrome P450 cDNA, CYP98A5. The nucleotide sequence indicates extensive homology to 3'-hydroxylases of p-coumaroyl esters. Dot blot analysis of leaves treated with various SA and isonicotinic acid derivatives showed enhanced expression of CYP98A5 due to DC-SA and DC-INA. Northern blot analysis of a time-dependent induction study of CYP98A5 in treated bean leaves indicated that DC-SA induces CYP98A5 mRNA transcripts earlier than DC-INA. Both inducers resulted in high transcript levels 24-48 h after treatment. The up-regulation of CYP98A5 is supportive of the conditioning and sensitizing effects of DC-SA and DC-INA to elicit a more rapid and effective defense response.

OBJECTIVE

To investigate the effects of salvionolic acid-A (SA-A), one of main effective components of Salvia miltiorrhiza for its antifibrotic action, on the cell proliferation and collagen production in cultured hepatic stellate cells (HSC).

METHODS

HSC were isolated through in situ perfusion of liver with pronase E and collagenase, and gradient centrifugation with Nycodenz. The cultured HSC were incubated with SA-A 0.1-100 mumol/L for 24 h. MTT spectrometric assay and intercellular incorporation of methyl-[3H]thymidine ([3H]TdR) was used to assess the cell proliferation. The amount of collagen was semi-quantified by ponceau staining and image analysis, the amount of type I collagen secretion was measured with ELISA and normalized by the total protein of cell layer. The total RNA was prepared from the control cells and the drug treated cells respectively, and the expression of pro-collagen alpha 2 (I) mRNA was semi-quantitatively analyzed with RT-PCR.

RESULTS

SA-A 100 mumol/L showed a little cytotoxity, SA-A 0.1-10 mumol/L did not influence cell morphology, and SA-A 1-100 mumol/L decreased the cell proliferation significantly in a concentration-dependent manner (P < 0.05). SA-A 1, 10, 100 mumol/L decreased the cell collagen deposition by 78.6%, 71.8%, and 61.3% of the control respectively (P < 0.05), and decreased type I collagen secretion to 53.1%, 52.6%, and 49.5% (P < 0.01 or P < 0.05). Both SA-A 1 and 10 mumol/L downregulated procollagen alpha 2 (I) mRNA expression remarkably (P < 0.05).

CONCLUSIONS

SA-A inhibited HSC proliferation and collagen expression. The inhibitory effect on HSC activation is the main mechanism of SA-A action against liver fibrosis.

Salvianolic acid B (SalB) is an active component isolated from Chinese herbal medicine Salvia miltiorrhiza. The aim of this study was to investigate the extent of absolute oral bioavailability (F) of SalB in beagle dogs and the effect on blood viscosity after intravenous and oral administration of Salvianolic acids (SAs). A gradient elution HPLC method was developed and validated to determine the concentration of SalB and its three possible metabolites in plasma. After SAs (180 mg/kg, p.o.; 9 mg/kg, i.v.) were given, the AUCs of SalB were 1680 +/- 670 and 7840 +/- 1140 ng/mL.h, respectively. The F of SalB in dogs was calculated to be only 1.07 +/- 0.43%. The blood viscosity was remarkably decreased after a single intravenous injection of SAs (9 mg/kg). However, no significant change of blood viscosity was observed after a single oral administration of SAs (180 mg/kg). The results suggested that the F of SalB was extremely low and single oral administrated SAs had no effect on ameliorating blood viscosity in beagle dogs.

The human ribosomal protein SA (RPSA) is a multilocus protein, present in most cellular compartments. It is a multifunctional protein, which belongs to the ribosome but is also a membrane receptor for laminin, growth factors, prion, pathogenic microorganisms, toxins, and the anticarcinogen epigallocatechin gallate. It contributes to the crossing of the blood-brain barrier by neurotropic viruses and bacteria and is used as a biomarker of metastasis. RPSA includes an N-terminal domain, which is homologous to the prokaryotic ribosomal proteins S2, and a C-terminal extension, which is conserved in vertebrates. The structure of its N-domain has been determined from crystals grown at 17 °C. The structure of its C-domain remains unknown. We produced in Escherichia coli and purified the full-length RPSA and its N- and C-domains. We characterized the folding states of these recombinant proteins mainly by methods of fluorescence and circular dichroism spectrometry, in association with quantitative analyses of their unfolding equilibria, induced with heat or urea. The necessary equations were derived from first principles. The results showed that the N-domain unfolded according to a three-state equilibrium. The monomeric intermediate was predominant at the body temperature of 37 °C. It also existed in the full-length RPSA and bound ANS, a small fluorescent molecule. The C-domain was in an intrinsically disordered state. The recombinant N- and C-domains weakly interacted together. These results indicated a high plasticity of RPSA, which could be important for its multiple cellular localizations and functional interactions.

Graft-versus-host disease across minor histocompatibility barriers was induced in two different models by transplanting allogeneic bone marrow and spleen cells into irradiated H-2-compatible recipient mice. In this report, we show that administration of peptides with high binding affinity for the respective class II major histocompatibility complex molecules after transplantation is capable of preventing the development of graft-versus-host disease in two different murine models. The peptides used were myelin basic protein residues 1 through 11 with alanine at position 4 (Ac 1-11[4A]) for I-Au (A alpha uA beta u), and the antigenic core sequence 323 through 339 of ovalbumin with lysine and methionine extension (KM core) for I-As (A alpha sA beta s). In both systems, the mechanism of prevention was found to be major histocompatibility complex-associated, because nonbinding control peptides did not have any effect. Engraftment of allogeneic bone marrow cells was shown by polymerase chain reaction analysis of DNA polymorphisms in a microsatellite region within the murine interleukin-5 gene.

To study enterotoxigenic Escherichia coli (ETEC) association to the gut of pigs, a simple and reproducible experimental model would be helpful. The aim of this experiment was to establish a model for studying the association of ETEC to the gut epithelium of pigs. Intestinal segments were prepared from 4 weaned pigs, which were tested susceptible to E. coli O149:F4 (homo- and heterozygotic; 2 pigs each) and O138:F18 (all homozygotic). Five segments were taken from 50% of the intestinal length measured from duodenum [mid small intestine (SI)], and 5 segments were taken from 90% distal to the duodenum (distal SI). The segments were immersed in Dulbecco's Modified Eagle Medium (DMEM) and kept on ice. Polyethylene tubing was inserted into either end of the segment and tied. The tissue was washed with 50 mL of PBS. The other end of segment was tied, 10 mL of DMEM alone or DMEM containing either E. coli F4 or F18 was inoculated, and the segment was sealed with Teflon plug. The segment was immersed in DMEM in a 300-mL infusion bottle in a shaking water bath at 37°C. After 1 h the segment was removed, tissue was washed with 50 mL of PBS, weighed, and homogenized in PBS. Final dilution of 10(-6) was prepared from the content and homogenate. The E. coli was enumerated on MacConkey agar. Data were analyzed according to a 2 × 3 × 2 parametric model including the effects of intestinal segment, E. coli strain, and site of SI with GLM procedure in SAS. A t-test was used to analyze the effect of genotype in F4-inoculated segment. The binding of E. coli on the tissue was 10 times higher (P < 0.001) for F4 than F18. The E. coli F18 was highest (P < 0.05) in mid SI whereas differences were not observed (P>> 0.05) between sites of SI for F4. Fewer (P < 0.001) bacteria bound in the control and they associated more (P = 0.10) at distal than mid SI. The E. coli did not differ (P>> 0.05) between genotypes in F4-inoculated segment. In conclusion, the ex vivo model may be feasible to investigate the ETEC association to the gut epithelium of pigs.

The development of photoreceptors and two putative neurotransmitter systems in the pineal organ and retina was studied during embryogenesis in the three-spined stickleback Gasterosteus aculeatus L. The investigation was performed by aid of immunocytochemistry using well characterized antisera to the retinal proteins alpha-transducin (TD alpha) and S-antigen (SA) (photoreceptor-markers), antisera against L-glutamic acid decarboxylase (GAD), gamma-aminobutyric acid (GABA), choline-O-acetyltransferase (ChAT) and with acetylcholinesterase (AChE) histochemistry (neurotransmitter-markers). It was possible to set up the following developmental time-table concerning the first appearance of positive immuno- and enzyme-reactive cells in the pineal organ and retina: I AChE-activity and TD alpha- and SA-immunoreactive cells in the pineal organ; II GAD- and GABA-immunoreactive cells in the pineal organ and retina; ChAT immunoreactivity and AChE activity in the retina; III hatching; IV SA-immunoreactive cells in the retina. The obtained results provide good evidence that while photoreceptor cells develop much earlier in the pineal organ than in the retina, neurons develop simultaneously in the pineal organ and retina.

Harzianin HA V and saturnisporin SA IV are alpha-amino isobutyric-containing peptides with 18- and 20-residue chain length, respectively. They were isolated from in vitro cultures of Trichoderma species and their sequences were determined by the combined use of positive ion FAB mass spectrometry and NMR. In organic solvent solution, both peptides exhibited the same predominant alpha-helical secondary structure including a hinge at the level of the central Pro residue, as deduced from NMR data. Their interaction with neutral phospholipid bilayers was shown to induce leakage of the material entrapped in small unilamellar vesicles composed of egg phosphatidylcholine/cholesterol (7/3). When incorporated into neutral planar lipid bilayers, they promoted voltage-gated channels. The concentration- and voltage-dependences of the ionic conductances induced by these peptides were studied in macroscopic current-voltage experiments. Single-channel measurements showed that whilst SA IV developed non-integral multi-open states similar to those induced by alamethicins, but with faster kinetics, the shorter analogue, HA V promoted much smaller-sized conducting aggregates in agreement with macroscopic conductance data.

A new analogue of coenzyme B12 (5'-deoxyadenosylcobalamin, AdoCbl), in which the configuration of the N-glycosidic bond in the Ado ligand is inverted [(alpha-ribo)AdoCbl], has been synthesized and its crystal structure determined by X-ray diffraction [MoKalpha, lambda = 0.71073 A, monoclinic P212121, a = 16.132(12) A, b = 21. 684(15) A, c = 27.30(3) A, 9611 independent reflections, R1 = 0. 0708]. As suggested by molecular mechanics modeling before the structure was known, the Ado ligand lies over the southern quadrant of the molecule, as is the case for AdoCbl. The most striking feature of the structure is disorder in the orientation of the adenine (Ade) moiety relative to the ribose of the Ado ligand. This was resolved with a two-state model in which in the major (0.57 occupancy) conformer the A16(O)-A11-A9(N)-A8 dihedral angle is 1.9 degrees and the Ade is virtually perpendicular to the corrin ring; in the minor conformer, the Ade is tilted down, and this dihedral is -48.7 degrees. The Co-C and axial Co-N bond lengths and the Co-C-C bond angle are quite similar to those in AdoCbl. The corrin ring is considerably flatter than that of AdoCbl, with a fold angle of 11.7 degrees. The molecule was successfully modeled by molecular mechanics (MM), and rotation of the Ado ligand relative to the corrin gave rise to four locally minimum structures with the Ado in the southern, eastern, northern, or western quadrant, with the southern conformation as the global minimum, as is the case with AdoCbl itself. Nuclear Overhauser effects (nOe's) observed by two-dimensional (2D) NMR were incorporated as restraints in molecular dynamics (MD) and simulated annealing (SA) calculations. A MD simulation at 300 K showed that only the southern conformation is populated with the Ado ligand confined to an arc from over C15 to over C12, while the Ade ring oscillates from perpendicular to parallel to the corrin ring. Twenty-seven structures were collected by MD-SA. Most of these annealed into the southern conformation, but examples of the other conformations were also found. The new analogue is a partially active coenzyme for the ribonucleotide reductase from Lactobacillus leichmanii with maximal activity that is 9.7% of that of AdoCbl itself, and a very high Km value (245 microM compared to 0.54 microM for AdoCbl). In addition, the rate constant for enzyme-induced carbon-cobalt bond cleavage of (alpha-ribo)AdoCbl is 160-fold smaller than that for AdoCbl, and only 1/3 as much cob(II)alamin is produced at the active site.

Relationships were determined between the labeling of leucine and phenylalanine at the intracellular site of protein synthesis in the pancreas and labeling of plasma leucine, its keto acid, alpha-ketoisocaproate (KIC), and phenylalanine. Six healthy subjects were studied for 480 minutes during a primed constant infusion of [1-14C]leucine, and six healthy subjects were studied for 240 minutes with [1-14C]leucine, [4,5-3H]phenylalanine, and [1-13C]KIC. An oro-duodenal tube was placed and pancreatic exocrine fluid was sampled by duodenal aspiration during cholecystokinin stimulation. During the 480-minute study, in the final 120 minutes the specific activity (SA) of enzyme leucine (3.14 +/- 0.27 dpm/nmol) was lower than that of plasma leucine (4.18 +/- 0.30 dpm/nmol, P < .001), but was not different from that of plasma KIC (3.02 +/- 0.18 dpm/nmol). During the 240-minute study, protein synthesis rates of secreted pancreatic enzymes when calculated with [3H]phenylalanine were lower (P = .006) by 28% +/- 2% than rates based on [14C]KIC SA, and lower (P = .004) by 16% +/- 3% than those calculated using [14C]leucine SA. Incorporation of [13C]leucine into pancreatic enzymes was not different from that of [14C]leucine when [13C]leucine and [14C]KIC, respectively, were used to denote precursor labeling. The results indicate that plasma KIC SA reflects the precursor pool for pancreatic protein synthesis during leucine tracer infusion, and plasma leucine enrichment also reflects the precursor pool when [1-13C]KIC is infused in man. The precursor pool is erroneously overestimated when using plasma SA of [4,5-3H]phenylalanine or [1-14C]leucine.

BACKGROUND

Nurses' clinical competence is vital to ensure safe and high quality care, and the continuous assessment of nurses' clinical competence is of major concern. A validated instrument for the self-assessment of nurses' clinical competence at different educational levels across specialties and countries is lacking. The aim of this study was to test the reliability and construct validity of the new Professional Nurse Self-Assessment Scale (ProffNurse SAS) questionnaire in long term and home care contexts in Norway. The questionnaire is based on the Nordic Advanced Practice Nursing model, in which the nurse-patient relationship is central.

METHODS

The study has a cross-sectional survey design. A purposive sample of 357 registered nurses who worked in long term and home care contexts in two geographical regions encompassing eight municipalities and three counties was included. The respondents completed the 74-item ProffNurse SAS questionnaire and demographic background data was collected. Data collection was conducted in two phases: first region autumn 2011 and second region spring 2012. Exploratory factor analyses (EFA) were used to test the psychometric properties of the questionnaire and included the following steps: assessment of the factorality of the data, factor extraction by Principal Component Analysis (PCA), oblimin (oblique) factor rotation, and interpretation. Cronbach's alpha was used to estimate the internal consistency.

RESULTS

The PCA revealed a six-component structure, reducing the number of items in the questionnaire from 74 to 51. Based on the content of the highest-loading items, the six components were named: Direct Clinical Practice, Professional Development, Ethical Decision-Making, Clinical Leadership, Cooperation and Consultation, and Critical Thinking. The Cronbach's alpha values ranged from 0.940 (highest; Direct Clinical Practice) to 0.737 (lowest; Critical Thinking), leading to the estimation that the ProffNurse SAS is reliable.

CONCLUSIONS

The six components support the study's theoretical framework. The ProffNurse SAS showed acceptable reliability and construct validity and may therefore be a promising instrument for the assessment of practicing nurses' clinical competence. However, we recommend further psychometric testing in other countries and contexts and the inclusion of larger samples of nurses at various levels of education, particularly master's level APNs.

OBJECTIVE

This study aimed to investigate the actions of salviainolic acid A (SA-A), an antiperoxidative component of Salvia miltiorrhiza (Sm), on rat liver injury and fibrosis.

METHODS

Acute and chronic rat liver injury models were established using carbon tetrachloride (CCl4). After 48 h (acute) or during 6 weeks of CCl4 injection, rats were further divided and treated with biphenyl dimethyl-dicarboxylate (BDD) or colchicine, as a control antifibrotic treatment, with Sm, a herbal compound, or SA-A, a water-soluble extract of Sm. Liver function was investigated by assessing alanine transaminase (ALT) and aspartate transaminase (AST) activities, histological analysis, hydroxyproline (Hyp) and malondiadehyde (MDA) content. In vitro, isolated cultured hepatocytes were injured with CCl4 gas for 24 h, followed by treatment with either vitamin E or various concentrations of SA-A. The extent of hepatocyte injury was monitored by analyzing various lipid peroxidative parameters such as superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), lactase dehydrogenase (LDH), and glutathione peroxidase (GSH-PX) levels in hepatocyte supernatants.

RESULTS

SA-A significantly decreased abnormal serum ALT activity both in acutely and chronically injured rat livers, decreased abnormal serum AST activity, Hyp and MDA content and attenuated hepatic collagen deposition. After CCl4 incubation and injury, the activities of AST, ALT CAT, GSH-PX and LDH and MDA content in hepatocyte supernatants increased significantly, but GSH levels decreased significantly. SA-A markedly improved these pathological changes in a dose-dependent manner. 10(-4) mol/l SA-A had stronger inhibitory action than vitamin E.

CONCLUSIONS

Our studies suggest that SA-A has antiperoxidative effects on injured hepatocytes in liver injury and fibrosis induced by CCl4.

Hepatic stellate cell (HSC) activation is an essential event during liver fibrogenesis. Phosphatase and tension homolog deleted on chromosome 10 (PTEN) is a negative regulator of this process. DNA methyltransferase 1 (DNMT1), which catalyzes DNA methylation and subsequently leads to the transcriptional repression of PTEN, is selectively induced in myofibroblasts from diseased livers. Sennoside A (SA), a major purgative constituent of senna and the Chinese herb rhubarb, is widely used in China and other Asian countries as an irritant laxative. SA is reported to improve hepatic steatosis. However, the effect and mechanism of SA on liver fibrosis remain largely unknown. We recently identified a novel strategy for protecting liver fibrosis via epigenetic modification by targeting DNMT1. A Surface Plasmon Resonance (SPR) assay first reported that SA could directly bind DNMT1 and inhibit its activity. Administration of SA significantly prevented liver fibrosis, as evidenced by the dramatic downregulation of α-smooth muscle actin (α-SMA) and type I collagen alpha-1 (Col1α1) protein levels in a CCl₄ -induced mouse hepatic fibrosis model and in TGF-β1-activated HSC-T6 cells, in vivo and in vitro. SA decreased the expression of Cyclin D1, CDK, and C-myc, indicating that SA may inhibit the activation and proliferation of TGF-β1-induced HSC-T6. Moreover, SA significantly promoted the expression of PTEN and remarkably inhibited the expression of p-AKT and p-ERK in vitro. Blocking PTEN or overexpressing DNMT1 could reduce the effect of SA on liver fibrosis. These data suggest that SA directly binds and inhibits the activity and that attenuated DNMT1-mediated PTEN hypermethylation caused the loss of PTEN expression, followed by the inhibition of the AKT and ERK pathways and prevented the development of liver fibrosis. Hence, SA might be employed as a promising natural supplement for liver fibrosis drug therapy.

<strong cl<em>a</em>ss="sub-title"> Keywords: </strong> DNMT1; PTEN; Sennoside A; epigenetic; hep<em>a</em>tic stell<em>a</em>te cell; liver fibrosis.

Sophora alopecuroides L. is a highly medicinal plant. The aim of the current study was to determine the phytochemical screening, pharmacological potentials and application of scanning electron microscope (SEM) of S. alopecuroides (SA) seeds. To achieve this purpose, six different solvents were used to prepare SA seed extracts. Phytochemical and antioxidant activities were determined calorimetrically. To investigate the antidiabetic activity, α-amylase inhibition assay was determined. Brine shrimp assay was used to determine cytotoxicity potential. Anti-leishmanial potential was confirmed using MTT assay. Disc-diffusion method was used to detect protein kinase inhibitory, antibacterial and antifungal activities and showed significant results. SEM analysis was used as an identification tool. Considerable amount of phenolic and flavonoid contents were identified in methanol extract (SASM) (93.76 ± 2.71 GAE/mg) and (77 ± 3.60 QE/mg). Highest DPPH scavenging potential (82%) was reported for SASM. Significant total antioxidant capacity (90.60 ± 1.55 alpha amylase enzyme [AAE]/mg) and total reducing power (94.44 ± 1.38 AAE/mg) were determined for LOSM. Highest α-amylase inhibition was reported in SASM (78.20 ± 1.58%). Highest LD₅₀ of brine shrimp was found for n-hexane extract (SASH) 13.03 μg/ml. All extracts showed strong anti-leishmanial activity except SASH. The seeds of SA were seen to be oblong to obovate, projections, wavy slightly straight, anticlinal wall was raised with apex acuminate. In conclusion, our experimental findings highly support the ethnomedicinal and biological potentials of the SA seeds. Moreover, SA seeds need to be explored for identification and isolation of bioactive compounds. In future, we recommend further in vivo toxicity assays and clinical efficacies to further evaluate its different biomedical properties.

Keywords: Sophora alopecuroides L. antimicrobial; anti-leishmanial; antioxidant; enzyme inhibition assays; scanning electron microscopy.

Fusarium graminearum is the major causal agent of fusarium head blight in wheat, a serious disease worldwide. Linoleic acid isomerase (LAI) catalyses the transformation of linoleic acid (LA) to conjugated linoleic acid (CLA), which is beneficial for human health. We characterised a cis-12 LAI gene of F. graminearum (FGSG_02668; FgLAI12), which was downregulated by salicylic acid (SA), a plant defence hormone. Disruption of FgLAI12 in F. graminearum resulted in decreased accumulation of cis-9,trans-11 CLA, enhanced sensitivity to SA, and increased accumulation of LA and SA in wheat spikes during infection. In addition, mycelial growth, accumulation of deoxynivalenol, and pathogenicity in wheat spikes were reduced. Re-introduction of a functional FgLAI12 gene into ΔFgLAI12 recovered the wild-type phenotype. Fluorescent microscopic analysis showed that FgLAI12 protein was usually expressed in the septa zone of conidia and the vacuole of hyphae, but was expressed in the cell membrane of hyphae in response to exogenous LA, which may be an element of LA metabolism during infection by F. graminearum. The cis-12 LAI enzyme encoded by FgLAI12 is critical for fungal response to SA, mycelial growth and virulence in wheat. The gene FgLAI12 is potentially valuable for biotechnological synthesis of cis-9,trans-11 CLA.

OBJECTIVE

Vision is considered important for academic performance in children; however, the evidence in this area tends to be inconsistent and inconclusive. This study explored the association between vision function and visual information processing measures and standardised academic achievement scores in Grade 3 Australian children.

METHODS

Participants included 108 Grade 3 primary school children (M = 8.82 ± 0.32 years) from three state primary schools in South-East Queensland. All participants underwent a standard vision screening, including distance visual acuity (VA), binocular vision testing and stereoacuity (SA). A computer-based battery of visual information processing tests including the Development Eye Movement (DEM) test, Visual Sequential Memory (VSM) and Symbol Search (SS) was also administered. Australian National Assessment Program for Literacy and Numeracy (NAPLAN) scores across five subtests of academic performance were obtained for each child: Reading, Writing, Spelling, Grammar/Punctuation and Numeracy.

RESULTS

The DEM adjusted horizontal and vertical times were most strongly associated with all of the NAPLAN subtest scores (p < 0.01), adjusted for age and the socio-economic status of the school; the DEM ratio was not significantly associated with any of the NAPLAN subtests. VSM and SS scores were significantly associated with one or more NAPLAN subtests, as were worse and better eye VA; SA showed no significant association with any of the NAPLAN subtests.

CONCLUSIONS

Performance on the horizontal and vertical DEM subtests was most strongly associated with academic performance. These data, in conjunction with other clinical data, can provide useful information to clinicians regarding their prescribing and management philosophy for children with lower levels of uncorrected refractive error and binocular vision anomalies.

This study is aimed to evaluate the potential effects of sodium aescinate (SA, the sodium salt of aescin) on wound healing in streptozotocin-induced diabetic rats. An excision skin wound was created in diabetic rats, and the wounded rats were divided into three groups: I) control group, II) gel-treated group, and III) SA-treated group. The control group wounds received topically normal saline once daily for 19 days. The gel-treated and SA-treated wounds received topically 400 μl of pluronic F-127 gel (25%) and 400 μl of SA (0.3%) in pluronic gel, respectively, once daily for 19 days. SA application in diabetic rats increased the wound contraction and significantly decreased the level of the inflammatory cytokine tumor necrosis factor-alpha (TNF-α) in comparison to the gel-treated group and control group. SA application in diabetic rats also resulted in a marked increase in the level of anti-inflammatory cytokine interleukin-10 (IL-10) and activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) compared to the other groups. Histopathologically, SA-treated wounds showed better granulation tissue dominated by marked fibroblast proliferation, and wounds were covered by thick regenerated epithelial layer. Additionally, the application of only pluronic gel produced some beneficial effects in some parameters in comparison to control group, but most of them were not significantly different. These findings demonstrated that SA may effectively control and improve wound healing in diabetic rats via its anti-inflammatory and antioxidant activities.

This study compared the gait characteristics of individuals walking in heat while wearing firefighting equipment in fatigued and non-fatigued states. Nineteen subjects performed a 50-min treadmill protocol in a heated room while gait patterns were recorded using a digital video camcorder. Forty gait cycles were analyzed near the beginning (9 min) and at the end (39-49 min) of exercise. Spatio-temporal gait variables including step frequency, step length, swing time, stance time, cycle time and double-support time were determined. Gait variability was quantified by the standard deviation (SD) and coefficient of variation (CV) of each variable. Left-right symmetry was calculated using the symmetry index (SI) and symmetry angle (SA). Paired t-tests (alpha = 0.05) were performed to identify difference between the beginning and the end of the protocol for each measured variable. Spatio-temporal gait characteristics did not differ between the beginning and the end of exercise. Gait variability of the double-support time increased at the end as measured by both SD (P = 0.037) and CV (P = 0.030) but no change was observed for other variables. Left-right symmetry measured using either SI or SA did not differ between sessions. In summary, spatio-temporal gait characteristics and symmetry while wearing firefighting equipment are insensitive to physiological fatigue. Prolonged walking in heat while wearing firefighting equipment may increase gait variability and therefore the likelihood of a fall. Future studies are needed to confirm the potential relationship between fatigue and gait variability and to investigate the possible influence of individual variation.

The structural features of rehmannan SA, a polysaccharide with remarkable reticuloendothelial system-potentiating activity obtained from the root of Rehmannia glutinosa, were investigated by methylation analysis and periodate oxidation. Rehmannan SA is mainly made up of arabino-3,6-galactan type structural units. Both rehmannan SA and rehmannan SB showed pronounced anti-complementary activity.

Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 ->> 3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p < 0.05) induced cytokine production IL-6>> TNF alpha>> IL-1 beta>> GM-CSF>> IL-10>> IFN gamma. The induction of these cytokines was significantly (p < 0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P < 0.05) LPS and SA induced cytokines. PTx further augmented (p>> 0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti-inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent.

Inhibition of oxidative stress and inflammation in vascular endothelial cells (ECs) may represent a new therapeutic strategy against endothelial activation. Sinapic acid (SA), a phenylpropanoid compound, is found in natural herbs and high-bran cereals and has moderate antioxidant activity. We aimed to develop new SA agents with the properties of antioxidation and blocking EC activation for possible therapy of cardiovascular disease. We designed and synthesized 10 SA derivatives according to their chemical structures. Preliminary screening of the compounds involved scavenging hydroxyl radicals and 2,2-diphenyl-1-picrylhydrazyl (DPPH(⋅)), croton oil-induced ear edema in mice, and analysis of the mRNA expression of adhesion molecules in ECs. 1-Acetyl-sinapic acyl-4-(3'-chlorine-)benzylpiperazine (SAad the strongest antioxidant and anti-inflammatory activities both in vitro and in vivo. Thus, the effect of SAas further studied. SAactor α-induced upregulation of adhesion molecules in ECs at both mRNA and protein levels, as well as the consequent monocyte adhesion to ECs. In vivo, result of face-to-face immunostaining showed that SAaccharide-induced expression of intercellular adhesion molecule-1 in mouse aortic intima. To study the molecular mechanism, results from luciferase assay, nuclear translocation of NF-κB, and Western blot indicated that the mechanism of the anti-inflammatory effects of SAacellular generation of ROS and inhibition of NF-κB activation in ECs. SAa prototype of a novel class of antioxidant with anti-inflammatory effects in ECs. It may represent a new therapeutic approach for preventing endothelial activation in cardiovascular disorders.