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2.
Zhonghua Yi Xue Za Zhi ; 100(32): 2481-2487, 2020 Aug 25.
Article in Chinese | MEDLINE | ID: mdl-32829592

ABSTRACT

Objective: To explore the relationship between family with sequence similarity 13 member A (FAM13A) gene and small airway remodeling in chronic obstructive pulmonary disease (COPD), and the effect of interference with FAM13A gene expression on the apoptosis and proliferation phenotype of human airway epithelial cells (16HBE). Methods: From January 2018 to January 2020, 74 patients in the Department of Thoracic Surgery of General Hospital of Ningxia Medical University were treated by surgery for lung tumors or pulmonary bullae. According to the lung function and smoking history, the 74 patients were divided into four groups: non-smoking group with normal lung function (normal group, 23 patients), smoking group with normal lung function (smoking group, 24 patients), non-smoking group with COPD (11 patients) and smoking group with COPD (16 patients). The expression of FAM13A in small airway of each group was detected by immunohistochemistry, and the correlation between FAM13A and the airflow restriction indexes by pulmonary function was analyzed. The shRNA fragment of FAM13A gene was designed, and the shRNA lentivirus vector of FAM13A gene was constructed and packaged. The expression level of FAM13A gene was detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot, and the best shRNA sequence was screened. Flow cytometry was used to detect apoptosis rate and the fluorescence intensity of proliferation marker Ki-67 in 16HBE cells. Results: FAM13A was mainly expressed in the cytoplasm of small airway epithelial cells. The levels of FAM13A absorbance (A) of small airway epithelial cells in non-smoking group and smoking group with COPD were higher than those in normal group and smoking group (0.365±0.026, 0.412±0.053 to 0.113±0.018, 0.105±0.009, all P<0.05), and they were negatively correlated with forced expiratory volume in 1s/forced vital capacity (FEV(1)/FVC) and FEV(1)% pre (r=-0.48 and r=-0.40, all P<0.05). The FAM13A shRNA lentiviral vector was successfully constructed, and FAM13A interference was successfully achieved in the 16HBE cell line. After infection of 16HBE cells, the results of qRT-PCR and Western blot showed that the expression of FAM13A in shRNA-target-2 group decreased (all P<0.01). Compared with the negative control group (shRNA-NC), the apoptosis rate of FAM13A shRNA group decreased (P=0.023), and the fluorescence intensity of Ki-67 also decreased (P=0.042). Conclusions: FAM13A gene expression is increased in COPD small airway epithelial cells, and it is related to COPD airflow limitation. FAM13A gene may participate in the process of COPD remodeling by affecting the apoptosis and proliferation of human airway epithelial cells.


Subject(s)
Airway Remodeling , Pulmonary Disease, Chronic Obstructive , Apoptosis , Cell Proliferation , Epithelial Cells , Humans , Lung
3.
Eur Rev Med Pharmacol Sci ; 22(7): 2005-2014, 2018 04.
Article in English | MEDLINE | ID: mdl-29687856

ABSTRACT

OBJECTIVE: Granulocyte colony-stimulating factor (G-CSF) plays a role in regulating phosphatidylinositol-3-kinase/serine/threonine kinase (PI3K/AKT) pathway, affecting cell proliferation and apoptosis, and inducing vascular endothelial growth factor (VEGF) expression. This study investigated the mechanism of G-CSF on angiogenesis and neural protection after intracerebral hemorrhage (ICH). MATERIALS AND METHODS: The rats were divided into four groups, including sham, ICH, ICH+G-CSF, and ICH+G-CSF+LY294002 (PI3K/AKT signaling pathway specific inhibitor). Cerebral neurological dysfunction was tested by Garcia scoring. Cell apoptosis was detected by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay. Angiogenesis marker CD34 expression, PI3K/AKT signaling pathway, B-cell lymphoma-2 (Bcl-2), and VEGF expressions were compared by IHC. Rat cerebral nerve RN-c cells were divided into four groups, including control, oxygen-glucose deprivation (OGD), OGD+G-CSF, and OGD+G-CSF+LY294002. RESULTS: Neurological dysfunction was more evident; CD34+ cell number, VEGF expression, and cell apoptosis significantly increased; phosphorylated AKT (p-AKT) and Bcl-2 levels markedly reduced in ICH group compared with sham group. G-CSF apparently up-regulated p-AKT and Bcl-2 expressions, attenuated cell apoptosis, and elevated CD34+ cell number. LY294002 significantly decreased p-AKT, Bcl-2, and VEGF expressions, and alleviated the cell apoptosis protective and angiogenesis effect induced by G-CSF. OGD treatment induced RN-c cell apoptosis, down-regulated p-AKT and Bcl-2 expressions, and enhanced the tube capacity of vascular endothelial cells (VEC). G-CSF markedly elevated p-AKT and Bcl-2 contents in RN-c cells, declined cell apoptosis, increased p-AKT and VEGF levels in VEC, and enhanced tube capacity. CONCLUSIONS: G-CSF enhanced PI3K/AKT signaling pathway activity, promoted Bcl-2 and VEGF expression, reduced nerve cell apoptosis, and enhanced tube capacity of VECs, which may be the mechanism of G-CSF in improving neurological function and angiogenesis after ICH.


Subject(s)
Cerebral Hemorrhage/drug therapy , Granulocyte Colony-Stimulating Factor/pharmacology , Neovascularization, Physiologic/drug effects , Animals , Apoptosis/drug effects , Cerebral Hemorrhage/physiopathology , Endothelial Cells/drug effects , Endothelial Cells/physiology , Male , Phosphatidylinositol 3-Kinase/physiology , Proto-Oncogene Proteins c-akt/physiology , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Vascular Endothelial Growth Factor A/analysis
4.
Eur Rev Med Pharmacol Sci ; 21(17): 3924-3934, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28975969

ABSTRACT

OBJECTIVE: To study the structural and functional changes in mitochondria in astrocytes of the cerebral cortex of the rats in the simulated sepsis environment in vitro and the relationship between these changes and the biogenesis of mitochondria in astrocytes by establishing models of sepsis astrocytes. MATERIALS AND METHODS: The structural and functional changes in mitochondria in astrocytes of the cerebral cortex of the rats were evaluated. The ultra structural changes in the mitochondria, astrocytes, and ultrathin sections, were observed with a transmission electron microscope. The expression of the regulatory factors related to biogenesis of mitochondria in astrocytes of the cerebral cortex of the rats was evaluated in various experimental groups. RT-PCR and Western blot were used to evaluate the expression of the regulatory factors related to biogenesis of mitochondria in astrocytes of the cerebral cortex of the rats. The "point grid method" was used to evaluate the volume density of the mitochondria in the astrocytes of the cerebral cortex of the rats in various experimental groups. The Western blotting was used to evaluate the role of fusion and fission of mitochondria in the astrocytes of the cerebral cortex of the rats in various experimental groups in regulating the expression of the protein-OPAI and DRPI. RESULTS: In the sepsis astrocyte models established by co-incubation of LPS and IFN-γ and astrocytes of the cerebral cortex of the rats, the mitochondria with a minor injury in the 6 h group (2.97± 0.92) increased significantly when compared with those in the 0 h group (1.08±0.95), 12 h group (1.70±1.01), and 24 h group (1.59±0.55) (p<0.05); the concentration of adenosine triphosphate (ATP) in the astrocytes of the cerebral cortex of the rats in the 6 h, 12 h, and 24 h groups increased significantly when compared with that in the 0 h group (p<0.05). PGC-1α mRNA, NRF-1 mRNA, NRF-2α mRNA, NRF-2ß mRNA, and mitochondrial transcription factor A (TFAM) mRNA in the astrocytes of the cerebral cortex of the rats in the 6 h and 12 h groups increased significantly when compared with those in the 0 h group (p<0.05); the concentration of TFAM mRNA (1.20±0.19) increased significantly when compared with that in the 0 h group (p<0.05). The OPAI protein concentration (1.21±0.17:1.34±0.06) and DRPI protein concentration (1.04±0.05; 1.05±0.05) in the astrocytes of the cerebral cortex of the rats in the 12 h group (1.25±0.17), 24 h group (1.33±0.24), and 6 h group increased significantly when compared with that in the 0 h group (p <0.05). CONCLUSIONS: The experimental sepsis conditions can cause a minor injury of the ultrastructure of the mitochondria in the astrocytes of the cerebral cortex of the rats. The biogenesis of the mitochondria in the astrocytes of the cerebral cortex of the rats was strengthened to cater for the increased demand for energy of the astrocytes under the sepsis conditions and finally recover the ultrastructure of the mitochondria with a minor injury. In response to the increased mitochondrial biogenesis, the activities of the mitochondrial fusion and fission of the astrocytes of the cerebral cortex of the rats were increased.


Subject(s)
Mitochondria/metabolism , Organelle Biogenesis , Sepsis-Associated Encephalopathy/pathology , Adenosine Triphosphate/metabolism , Animals , Astrocytes/cytology , Astrocytes/metabolism , Biomarkers/analysis , Cerebral Cortex/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Interferon-gamma/pharmacology , Interleukin-6/analysis , Lipopolysaccharides/pharmacology , Mitochondria/drug effects , Mitochondria/ultrastructure , Mitochondrial Proteins/genetics , Mitochondrial Proteins/metabolism , NF-E2-Related Factor 1/genetics , NF-E2-Related Factor 1/metabolism , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Sepsis-Associated Encephalopathy/veterinary , Transcription Factors/genetics , Transcription Factors/metabolism , Tumor Necrosis Factor-alpha/analysis
5.
Oncogenesis ; 6(4): e316, 2017 Apr 10.
Article in English | MEDLINE | ID: mdl-28394355
6.
Eur J Trauma Emerg Surg ; 43(6): 875-882, 2017 Dec.
Article in English | MEDLINE | ID: mdl-28070608

ABSTRACT

OBJECTIVE: High vasopermeability and excessive inflammation following severe burns may result in tissue edema, organ dysfunction and the loss of circulatory plasma volume, which can influence the doctor to do the prognosis to the patients. The study aims to examine whether Xuebijing injection (XBJ), an extracts of a traditional Chinese medicine used to treat sepsis in clinic, can reduces fluid requirements by inhibiting vasopermeability and tissue edema in a canine model after burn injury. METHODS: Twenty-four beagle dogs were subjected to 50% TBSA burns, and then were randomly allocated to the following three groups: lactated Ringer's resuscitation (LR) group (n = 8), immediate LR containing Xuebijing injection (LR/XBJ) group (n = 8), and operation control group (n = 8). Hemodynamic variables and net fluid accumulation were measured. Blood samples were collected for measurement of hematocrit and circulatory plasma volume (PV). At 24 h after burn injury, heart, lung, small intestine and kidney were harvested for evaluation of the activities of myeloperoxidase (MPO) and neutrophil elastase (NE), vasopermeability, tissue water content and the amount of neutrophil infiltration. RESULTS: XBJ treatment significantly reduced net fluid accumulation, and pulmonary vascular permeability index (PVPI), extravascular lung water index (ELWI), and water content of heart, small intestine, kidney and lung compared with LR group. Furthermore, XBJ infusion significantly reduced tissue activities of MPO and NE compared with LR group. The amount of neutrophil infiltration in LR/XBJ group was lower than that in LR group. CONCLUSIONS: These results indicate that XBJ injection can reduce fluid requirements by inhibition of neutrophil protease-induced high vasopermeability and tissue edema.


Subject(s)
Burns/drug therapy , Capillary Permeability/drug effects , Disease Models, Animal , Dogs/injuries , Drugs, Chinese Herbal/pharmacology , Fluid Therapy , Isotonic Solutions/pharmacology , Animals , Drugs, Chinese Herbal/administration & dosage , Drugs, Chinese Herbal/therapeutic use , Injections, Intravenous , Isotonic Solutions/administration & dosage , Isotonic Solutions/therapeutic use , Random Allocation , Resuscitation , Ringer's Lactate
7.
Oncogenesis ; 5: e217, 2016 04 11.
Article in English | MEDLINE | ID: mdl-27065453

ABSTRACT

Mogroside V is a triterpenoid isolated from the traditional Chinese medical plant Siraitia grosvenorii. Mogroside V has a high degree of sweetness and a low calorific content. Herein, we found that mogroside V possesses tumor growth inhibitory activity in in vitro and in vivo models of pancreatic cancer by promoting apoptosis and cell cycle arrest of pancreatic cancer cells (PANC-1 cells), which may in part be mediated through regulating the STAT3 signaling pathway. These results were confirmed in vivo in a mouse xenograft model of pancreatic cancer. In xenograft tumors, Ki-67 and PCNA, the most commonly used markers of tumor cell proliferation, were downregulated after intravenous administration of mogroside V. Terminal deoxynucleotidyl transferase dUTP nick end labeling assays showed that mogroside V treatment promoted apoptosis of pancreatic cancer cells in the xenograft tumors. Furthermore, we found that mogroside V treatment significantly reduced the expression of CD31-labeled blood vessels and of the pro-angiogenic factor vascular endothelial growth factor in the xenografts, indicating that mogroside V might limit the growth of pancreatic tumors by inhibiting angiogenesis and reducing vascular density. These results therefore demonstrate that the natural, sweet-tasting compound mogroside V can inhibit proliferation and survival of pancreatic cancer cells via targeting multiple biological targets.

8.
Sci Total Environ ; 551-552: 614-21, 2016 May 01.
Article in English | MEDLINE | ID: mdl-26897404

ABSTRACT

Animal enclosures are often constructed from wood treated with the pesticide chromated copper arsenate (CCA), which leaches arsenic (As) into adjacent soil during normal weathering. This study evaluated potential pathways of As exposure in 25 species of zoo animals living in CCA-wood enclosures. We analyzed As speciation in complete animal foods, dislodgeable As from CCA-wood, and As levels in enclosure soils, as well as As levels in biomarkers of 9 species of crocodilians (eggs), 4 species of birds (feathers), 1 primate species (hair), and 1 porcupine species (quills). Elevated soil As in samples from 17 enclosures was observed at 1.0-110mg/kg, and enclosures housing threatened and endangered species had As levels higher than USEPA's risk-based Eco-SSL for birds and mammals of 43 and 46mg/kg. Wipe samples of CCA-wood on which primates sit had dislodgeable As residues of 4.6-111µg/100cm(2), typical of unsealed CCA-wood. Inorganic As doses from animal foods were estimated at 0.22-7.8µg/kg bw/d. Some As levels in bird feathers and crocodilian eggs were higher than prior studies on wild species. However, hair from marmosets had 6.37mg/kg As, 30-fold greater than the reference value, possibly due to their inability to methylate inorganic As. Our data suggested that elevated As in soils and dislodgeable As from CCA-wood could be important sources of As exposure for zoo animals.


Subject(s)
Animals, Zoo/metabolism , Arsenic/metabolism , Environmental Exposure/analysis , Hazardous Substances/metabolism , Animals , Arsenates/metabolism , Biomarkers/metabolism , Environmental Exposure/statistics & numerical data , Housing , Wood/chemistry
9.
Sci Total Environ ; 532: 812-20, 2015 Nov 01.
Article in English | MEDLINE | ID: mdl-26116410

ABSTRACT

Previous studies have established in vivo-in vitro correlations (IVIVC) between arsenic (As) relative bioavailability (RBA) and bioaccessibility in contaminated soils. However, their ability to predict As-RBA in soils outside the models is unclear. In this study, As bioaccessibility and As-RBA in 12 As-contaminated soils (22.2-4172 mg kg(-1) As) were measured using five assays (SBRC, IVG, DIN, PBET, and UBM) and a mouse blood model. Arsenic RBA in the soils ranged from 6.38 ± 2.80% to 73.1 ± 17.7% with soils containing higher extractable Fe showing lower values. Arsenic bioaccessibility varied within and between assays. Arsenic bioaccessibility was used as input values into established IVIVC to predict As-RBA in soils. There were significant differences between predicted and measured As-RBA for the 12 soils, illustrating the inability of established IVIVC to predict As-RBA in those contaminated soils. Therefore, a new IVIVC was established by correlating measured As-RBA and As bioaccessibility for the 12 soils. The strength of the predictive models varied from r(2) = 0.50 for PBET to r(2) = 0.83 for IVG, with IVG assay providing the best prediction of As-RBA. When IVIVC were compared to those of Juhasz et al. (2014a), slopes of the relationships were significantly higher possibly due to different As-RBA measurements. Our research showed that IVG has potential to measure As bioavailability in contaminated soils from China though UBM and SBRC assays were also suitable. More research is needed to verify their suitability to predict As-RBA in soils for refining health risk assessment.


Subject(s)
Environmental Monitoring/methods , Soil Pollutants/analysis , Arsenic/analysis , China , Environmental Pollution/statistics & numerical data , Risk Assessment , Soil/chemistry
10.
Environ Int ; 82: 35-40, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26004992

ABSTRACT

Concern about children's exposure to arsenic (As) from wood treated with chromated-copper-arsenate (CCA) led to its withdrawal from residential use in 2004. However, due to its effectiveness, millions of American homes still have CCA-wood decks on which children play. This study evaluated the effects of three deck-cleaning methods on formation of dislodgeable As and hexavalent chromium (CrVI) on CCA-wood surfaces and in leachate. Initial wipes from CCA-wood wetted with water showed 3-4 times more dislodgeable As than on dry wood. After cleaning with a bleach solution, 9.8-40.3µg/100cm(2) of CrVI was found on the wood surface, with up to 170µg/L CrVI in the leachate. Depending on the cleaning method, 699-2473mg of As would be released into the environment from cleaning a 18.6-m(2)-deck. Estimated As doses in children aged 1-6 after 1h of playing on a wet CCA-wood deck were 0.25-0.41µg/kg. This is the first study to identify increased dislodgeable As on wet CCA-wood and to evaluate dislodgeable CrVI after bleach application. Our data suggest that As and CrVI in 25-year old CCA-wood still show exposure risks for children and potential for soil contamination.


Subject(s)
Arsenates/adverse effects , Arsenic/adverse effects , Child , Chromium , Chromium Compounds , Housing , Humans , Play and Playthings , Risk , Wood
11.
J Hazard Mater ; 295: 145-52, 2015 Sep 15.
Article in English | MEDLINE | ID: mdl-25897696

ABSTRACT

Arsenic bioaccessibility varies with in vitro methods and soils. Four assays including unified BARGE method (UBM), Solubility Bioaccessibility Research Consortium method (SBRC), in vitro gastrointestinal method (IVG), and physiologically based extraction test (PBET), were used to determine As bioaccessibility in 11 contaminated soils (22-4,172 mg kg(-1)). The objective was to understand how bioaccessible As by different methods was related to different As pools based on sequential extraction and 0.43 M HNO3 extraction. Arsenic bioaccessibility was 7.6-25, 2.3-49, 7.3-44, and 1.3-38% in gastric phase (GP), and 5.7-53, 0.46-33, 2.3-42, and 0.86-43% in intestinal phase (IP) for UBM, SBRC, IVG, and PBET, respectively, with HNO3-extractable As being 0.90-60%. Based on sequential extraction, As was primarily associated with amorphous (AF3; 17-79%) and crystallized Fe/Al oxides (CF4; 6.4-73%) while non-specifically sorbed (NS1), specifically sorbed (SS2), and residual fractions (RS5) were 0-10%, 3.4-20% and 3.2-25%. Significant correlation was found between As bioaccessibility by PBET and NS1+SS2 (R(2) = 0.55-0.69), and UBM-GP and NS1 + SS2 + AF3 (R(2) = 0.58), indicating PBET mostly targeted As in NS1+SS2 whereas UBM in NS1 + SS2 + AF3. HNO3-extractable As was correlated to bioaccessible As by four methods (R(2) = 0.42-0.72) with SBRC-GP having the best correlation. The fact that different methods targeted different As fractions in soils suggested the importance of validation by animal test. Our data suggested that HNO3 may have potential to determine bioaccessible As in soils.


Subject(s)
Arsenic/analysis , Soil/chemistry , Nitric Acid
12.
Br Poult Sci ; 55(2): 207-14, 2014.
Article in English | MEDLINE | ID: mdl-24404906

ABSTRACT

1. Ipriflavone (IP) has been widely studied in humans and is effective for inhibiting osteoclastic bone resorption and enhancing osteoblastic bone formation. The aim of this study was to investigate the effects of IP on the structural histology and histomorphometric parameters of tibia in laying hens fed on a low-calcium diet. 2. A total of 200 Hy-Line Brown laying hens, 24 weeks of age, were divided randomly into 4 groups which were fed on one of 4 diets (CaN, standard diet; CaL, low-calcium diet; IP1, low-calcium diet + 8 mg/kg of IP; IP2, low-calcium diet + 20 mg/kg of IP) for 60 d after which 10 hens of each group, chosen at random, were killed and the tibiae were processed and assessed under a fluorescent microscope 3. The cortical bone showing resorption cavities had become porous, and medullary bone did not completely fill the marrow cavity in the CaL group compared with the CaN group. However, in the IP2 group, the cortical bone showed a structure comparable to the CaN group with an absence of resorption cavities, and more medullary bone filled the marrow cavity compared with the CaL group. 4. The CaL hens had significantly lower trabecular bone area, trabecular perimeter, per cent trabecular area, trabecular number and trabecular thickness and significantly wider trabecular separation compared with CaN. These results indicated that the low-calcium diet led to increased loss of tibial bone, eventually causing osteoporosis. However, compared with the CaL group, the IP2 group caused a significant increase in trabecular bone area and bone quality. 5. It is concluded that even in hens fed on a low-calcium diet, the addition of 20 mg/kg IP exerted beneficial effects on histological structure and structural histomorphometric parameters of tibia, indicating an alleviation of caged layer osteoporosis (CLO).


Subject(s)
Calcium/deficiency , Chickens/anatomy & histology , Isoflavones/pharmacology , Tibia/anatomy & histology , Animal Feed/analysis , Animals , Chickens/physiology , Diet/veterinary , Dietary Supplements/analysis , Female , Random Allocation , Reproduction/drug effects
13.
Neuroscience ; 156(1): 89-98, 2008 Sep 22.
Article in English | MEDLINE | ID: mdl-18588949

ABSTRACT

The fat-derived hormone leptin regulates food intake and body weight in part by modulating the activity of neuropeptide Y (NPY) and proopiomelanocortin (POMC) neurons in the hypothalamic arcuate nucleus (ARC). To investigate the electrophysiological activity of these neurons and their responses to leptin, we recorded whole-cell calcium currents on NPY and POMC neurons in the ARC of rats, which we identified by morphologic features and immunocytochemical identification at the end of recording. Leptin decreased the peak amplitude of high voltage-activated calcium currents (I(HVA)) in the isolated neurons from ARC, which were subsequently shown to be immunoreactive for NPY. The inhibition was prevented by pretreatment with inhibitors of Janus kinase 2 (JAK2) and mitogen-activated protein kinases (MAPK). In contrast, leptin increased the amplitude of I(HVA) in POMC-containing neurons. The stimulations of I(HVA) were inhibited by blockers of JAK2 and phosphatidylino 3-kinase (PI3-k). Both of these effects were counteracted by the L-type calcium channel antagonist nifedipine, suggesting that L-type calcium channels were involved in the regulation induced by leptin. These data indicated that leptin exerted opposite effects on these two classes of neurons. Leptin directly inhibited I(HVA) in NPY neurons via leptin receptor (LEPR) -JAK2-MAPK pathways, whereas evoked I(HVA) in POMC neurons by LEPR-JAK2-PI3-k pathways. These neural pathways and intracellular signaling mechanisms may play key roles in regulating NPY and POMC neuron activity, anorectic action of leptin and, thereby, feeding.


Subject(s)
Arcuate Nucleus of Hypothalamus/metabolism , Calcium Channels, L-Type/metabolism , Leptin/metabolism , Neurons/metabolism , Neuropeptide Y/metabolism , Pro-Opiomelanocortin/metabolism , Animals , Animals, Newborn , Appetite Regulation/drug effects , Appetite Regulation/physiology , Arcuate Nucleus of Hypothalamus/cytology , Arcuate Nucleus of Hypothalamus/drug effects , Body Weight/drug effects , Body Weight/physiology , Calcium Channel Blockers/pharmacology , Calcium Channels, L-Type/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Feeding Behavior/drug effects , Feeding Behavior/physiology , Janus Kinase 2/antagonists & inhibitors , Janus Kinase 2/metabolism , Leptin/pharmacology , MAP Kinase Signaling System/drug effects , MAP Kinase Signaling System/physiology , Membrane Potentials/drug effects , Membrane Potentials/physiology , Neurons/drug effects , Patch-Clamp Techniques , Phosphatidylinositol 3-Kinases/drug effects , Phosphatidylinositol 3-Kinases/metabolism , Rats , Rats, Sprague-Dawley , Receptors, Leptin/drug effects , Receptors, Leptin/metabolism , Signal Transduction/drug effects , Signal Transduction/physiology
14.
Plant Biol (Stuttg) ; 10(3): 323-33, 2008 May.
Article in English | MEDLINE | ID: mdl-18426479

ABSTRACT

Salidroside, a novel effective adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor, can be derived from phenylalanine or tyrosine. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing production of salidroside by the plant. In this study, a cDNA clone encoding phenylalanine ammonia-lyase (PAL) was isolated from R. sachalinensis using rapid amplification of cDNA ends. The resulting cDNA was designated PALrs1. It is 2407-bp long and encodes 710 deduced amino acid residues. Southern blot analysis of genomic DNA indicated that the PAL gene family is composed of three to five genes in the R. sachalinensis genome. Northern blot analysis revealed that transcripts of PALrs1 were present in calli, leaves and stems, but expression in roots was very low. The PALrs1 under the 35S promoter with double-enhancer sequences from CaMV-Omega and TMV-Omega fragments was transferred into R. sachalinensis via Agrobacterium tumefaciens. PCR and PCR-Southern blot confirmed that the PALrs1 gene had been integrated into the genome of transgenic plants. Northern blot analysis revealed that the PALrs1 gene had been expressed at the transcriptional level. High-performance liquid chromatography indicated that overexpression of the PALrs1 gene resulted in a 3.3-fold increase in p-coumaric acid content, as expected. In contrast, levels of tyrosol and salidroside were 4.7-fold and 7.7-fold, respectively, lower in PALrs1 transgenic plants than in controls. Furthermore, overexpression of the PALrs1 gene resulted in a 2.6-fold decrease in tyrosine content. These data suggest that overexpression of the PALrs1 gene and accumulation of p-coumaric acid did not facilitate tyrosol biosynthesis; tyrosol, as a phenylethanoid derivative, is not derived from phenylalanine; and reduced availability of tyrosine most likely resulted in a large reduction in tyrosol biosynthesis and accumulation of salidroside.


Subject(s)
Glucosides/biosynthesis , Phenylalanine Ammonia-Lyase/metabolism , Phenylethyl Alcohol/analogs & derivatives , Rhodiola/metabolism , Amino Acid Sequence , Coumaric Acids/metabolism , Gene Expression , Molecular Sequence Data , Multigene Family , Phenols , Phenylalanine Ammonia-Lyase/genetics , Phenylethyl Alcohol/metabolism , Plants, Genetically Modified/metabolism , Propionates , Rhodiola/enzymology , Rhodiola/genetics , Sequence Analysis, DNA , Tyrosine/metabolism
15.
J Hazard Mater ; 136(3): 654-62, 2006 Aug 25.
Article in English | MEDLINE | ID: mdl-16487656

ABSTRACT

Immobilization of heavy metals from contaminated environments is an emerging field of interest from both resource conservation and environmental remediation points of view. This study investigated the feasibility of using phosphatic clay, a waste by-product of the phosphate mining industry, as an effective sorbent for Pb from aqueous effluents. The major parameters controlling aqueous Pb removal, viz. initial metal ion concentrations, solution pH, sorbent amounts, ionic strength and presence of both inorganic and organic ligands were evaluated using batch experiments. Results demonstrated that aqueous Pb removal efficiency of phosphatic clay is controlled mainly by dissolution of phosphatic clay associated fluoroapatite [Ca(10)(PO(4))(5)CaCO(3)(F,Cl,OH)(2)], followed by subsequent precipitation of geochemically stable pyromorphite [Pb(10)(PO(4))(6)(F,Cl,OH)(2)], which was confirmed by both X-ray diffraction (XRD) and scanning electron microscopic (SEM) analysis. Lead removal efficiency of phosphatic clay increased with increasing pH, sorbent amount and decreasing ionic strength. It also depends on the nature of complexing ligands. Formation of insoluble calcium oxalate and lead oxalate in the presence of oxalic acid explained high uptake of Pb by phosphatic clay from aqueous solution. However, Pb sorption kinetics onto phosphatic clay were biphasic, with initially fast reactions followed by slow and continuous Pb removal reactions. The slow reactions may include surface sorption, co-precipitation and diffusion. The exceptional capability of phosphatic clay to remove aqueous Pb demonstrated its potential as a cost effective way to remediate Pb-contaminated water, soils and sediments.


Subject(s)
Aluminum Silicates/chemistry , Lead/chemistry , Lead/isolation & purification , Phosphites/chemistry , Adsorption , Algorithms , Chemical Phenomena , Chemistry, Physical , Clay , Elements , Hydrogen-Ion Concentration , Inorganic Chemicals , Kinetics , Ligands , Organic Chemicals , Thermodynamics , X-Ray Diffraction
16.
Int J Phytoremediation ; 8(4): 311-22, 2006.
Article in English | MEDLINE | ID: mdl-17305305

ABSTRACT

A field study was conducted to determine the efficiency of Chinese brake fern (Pteris vittata L.), an arsenic hyperaccumulator, on removal of arsenic from soil at an arsenic-contaminated site. Chinese brake ferns were planted on a site previously used to treat wood with chromated copper arsenate (CCA). Arsenic concentrations in surface and profile soil samples were determined for 2000, 2001, and 2002. In both 2001 and 2002, senesced and senescing fronds only, as well as all fronds, were harvested. Frond arsenic concentrations were not significantly different between the three harvests. Compared to senesced fronds, live fronds resulted in the greatest amount of arsenic removal. There were no significant differences in soil arsenic concentrations between 2000, 2001, and 2002, primarily due to the extreme variability in soil arsenic concentrations. However, the mean surface soil arsenic was reduced from 190 to 140 mg kg(-1). Approximately 19.3 g of arsenic were removed from the soil by Chinese brake fern. Therefore, this fern is capable of accumulating arsenic from the CCA -contaminated site and may be competitive, in terms of cost, to conventional remediation systems. However, better agronomic practices are needed to enhance plant growth and arsenic uptake to obtain maximum soil arsenic removal and to minimize remediation time.


Subject(s)
Arsenates/pharmacokinetics , Arsenic/pharmacokinetics , Pteris/metabolism , Soil Pollutants/pharmacokinetics , Biodegradation, Environmental , Humans , Industrial Waste/prevention & control , Plant Roots , Plant Shoots , Pteris/growth & development
17.
Int J Phytoremediation ; 7(3): 231-40, 2005.
Article in English | MEDLINE | ID: mdl-16285413

ABSTRACT

Two hydroponic experiments were conducted to evaluate factors affecting plant arsenic (As) hyperaccumulation. In the first experiment; two As hyperaccumulators (Pteris vittata and P. cretica mayii) were exposed to 1 and 10 mg L(-1) arsenite (AsIII) and monomethyl arsenic acid (MMA) for 4 wk. Total As concentrations in plants (fronds and roots) and solution were determined In the second experiment P. vittata and Nephrolepis exaltata (a non-As hyperaccumulator) were exposed to 5 mgL(-1) arsenate (AsV) and 20 mgL(-1) AsIIIfor 1 and 15 d. Total As and AsIII concentrations in plants were determined Compared to P. cretica mayii, P. vittata was more efficient in arsenic accumulation (1075-1666 vs. 249-627mg kg(-1) As in the fronds) partially because it is more efficient in As translocation. As translocation factor (As concentration ratio in fronds to roots) was 3.0-5.6 for P. vittata compared to 0.1 to 4.8 for P. cretica. Compared to N. exaltata, P. vittata was significantly more efficient in arsenic accumulation (38-542 vs. 4.8-71 mg kg(-1) As in thefronds) as well asAs translocation (1.3-5.6 vs. 0.2-0.5). In addition, P. vittata was much more efficient in As reduction from AsV to AsIII (83-84 vs. 13-24% AsIII in the fronds). Little As reduction occurred after 1-d exposure to AsV in both species indicates that As reduction was not instantaneous even in an As hyperaccumulator. Our data were consistent with the hypothesis that both As translocation and As reduction are important for plant As hyperaccumulation.


Subject(s)
Arsenates/pharmacokinetics , Arsenic/pharmacokinetics , Ferns/metabolism , Water Pollutants/pharmacokinetics , Biodegradation, Environmental , Ferns/classification , Humans , Hydroponics , Plant Structures
18.
J Environ Qual ; 34(5): 1490-6, 2005.
Article in English | MEDLINE | ID: mdl-16091601

ABSTRACT

Knowledge of water movement in the plant-xylem system and contaminant bioavailability in the soil environment is crucial to evaluate the success of phytoremediation practices. This study investigated the removal of 2,4,6-trinitrotoluene (TNT) from a contaminated sandy soil by a single poplar (Populus fastigiata) tree through the examinations of temporal variations of xylem water potential, root water uptake, and soil TNT bioavailability. A mathematical model, CTSPAC (Coupled Transport of water, heat, and solutes in the Soil-Plant-Atmosphere Continuum), was modified for the purpose of this study. The model was calibrated using laboratory measurements before its application. Our simulations show that the xylem water potential was high in the roots and low in the leaves with a potential head difference of 3.55 cm H2O, which created a driving force for water flow and chemical transport upward from the roots through the stem to the leaves. The daily average root water uptake rate was 25 cm3 h(-1) when an equilibrium condition was reached after 24 h. Our simulations further reveal that no TNT was found in the stem and leaves and only about 1% of total TNT mass was observed in the roots due to the rapid biodegradation and transformation of TNT into its daughter products. About 13% of the soil TNT was removed by the poplar tree, resulting mainly from root uptake since TNT is a recalcitrant compound. In general, the soil TNT bioavailability decreased with time due to the depletion of soil solution TNT by the poplar tree. A constant bioavailability (i.e., 3.1 x 10(-6)) was obtained in 14 d in which the soil TNT concentration was about 10 mg L(-1). Our study suggests that CTSPAC is a useful model to simulate phytoremediation of TNT-contaminated sites.


Subject(s)
Environmental Pollution/prevention & control , Models, Biological , Plant Transpiration/physiology , Soil Pollutants/metabolism , Trinitrotoluene/metabolism , Biodegradation, Environmental , Computer Simulation , Soil Pollutants/analysis , Trinitrotoluene/analysis
19.
Environ Pollut ; 124(3): 407-17, 2003.
Article in English | MEDLINE | ID: mdl-12758021

ABSTRACT

Chromated copper arsenate (CCA)-treated wood has been widely used in the Southeastern United States to protect wood products from microbial and fungal decay. The aims of this study were to (1). determine the distribution of arsenic (As), chromium (Cr), and copper (Cu), in soils surrounding CCA-treated wood structures such as decks, fences and poles; and (2). evaluate the impacts of these structures on As, Cr and Cu loading of the soils. Profile and lateral soil samples were collected under CCA-treated decks and adjacent to poles and fences. The results showed elevation of As, Cr and Cu concentrations close to and under the structures, with mean As concentrations as high as 23 mg x kg(-1) close to utility poles compared with less than 3 mg x kg (-1) at distances of about 1.5 m away. Concentrations of As, Cr, and Cu decreased with depth in areas close to CCA-treated poles. However, these results were only apparent in relatively new structures. A combination of weathering and leaching with time may have reduced the impact in older poles. Increased concentrations of As, Cu and Cr were also observed close to CCA-treated decks and fences, with age showing a similar impact. These results are helpful for CCA-treated wood product users to determine the safe use of these structures.


Subject(s)
Arsenic/analysis , Chromium/analysis , Copper/analysis , Soil Pollutants/analysis , Wood , Arsenates , Chromium Compounds , Environmental Monitoring/methods
20.
J Environ Qual ; 32(1): 109-19, 2003.
Article in English | MEDLINE | ID: mdl-12549549

ABSTRACT

Arsenic contamination is of concern due to its effect as a carcinogen. Understanding the distribution of arsenic in urban soils is important for establishing baseline concentrations from which anthropogenic effects can be measured. The soil cleanup target level (SCTL) for arsenic in Florida (0.8 and 3.7 mg kg(-1) in residential and commercial areas, respectively) is lower than in most states and is near the arsenic background concentrations in Florida soils. The objective of this study was to characterize the distribution of arsenic in the soils of two Florida cities, Gainesville and Miami. More than 200 soil samples were collected from three land-use classes in each city (residential, commercial, and public land), digested with USEPA Method 3051a, and analyzed with graphite furnace atomic absorption spectrophotometry. Arsenic concentrations varied greatly in Gainesville, ranging from 0.21 to approximately 660 mg kg(-1) with a geometric mean (GM) of 0.40 mg kg(-1) (after discarding outliers), which was significantly lower than the GM of 2.81 mg kg(-1) in Miami, although Miami samples ranged only from 0.32 to approximately 110 mg kg(-1). Arsenic concentrations in 29 and 4% of the Gainesville soil samples and 95 and 33% of the Miami samples exceeded the Florida residential and commercial SCTL, respectively. This study is the first to provide information on arsenic distribution in urban soils of Florida, and the data are useful for assessing arsenic contamination and determining the need for remediation.


Subject(s)
Arsenic/analysis , Soil Pollutants/analysis , Cities , Environmental Monitoring , Environmental Pollution/prevention & control , Florida , Reference Values
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