Biomedical subjects

M Fang

Publications and source records attributed to M Fang.

At least 37 records · Page 2Linked to original sources

PubMed2001

Toxicity evaluation of sewage sludges in Hong Kong.

Anaerobically digested sewage sludges collected from four wastewater treatment plants located in Sha Tin, Tai Po, Yuen Long, and Shek Wu Hui in Hong Kong were subjected to chemical characterization and toxicity testing to provide preliminary assessment of their suitability for land application. All sewage sludges were slightly alkaline with pH range of 8.3-8.7. Electrical conductivity (EC; 0.69 dS m(-1)) and soluble NH4-N content (996 mg kg(-1)) of sewage sludge from Yuen Long were lower than that of other plants. Concentrations of heavy metals were determined as total, extractable, and water-soluble fraction using mixed acid digestion, DTPA (pH 7.3), and distilled water, respectively. Yuen Long sludge was most polluted with Zn and Cr higher than the pollutant concentration limits listed in Part 503 of USEPA, owing to the effluent coming from the tannery industry. High concentration of Ni was found in sludge from Sha Tin that originated mainly from the electroplating industry. DTPA-extractable Zn contents were high in sludges from Yuen Long (1247 mg kg(-1)) and Shek Wu Hui (892 mg kg(-1)), while 3.7 mg kg(-1) of DTPA-extractable Cr was found in Yuen Long sludge. Metal speciation of sludges showed that Pb was major in residual phase while Cu, Cr, and Ni in organic and residual phases, and Zn did not show any dominant chemical phase. The sludge extracts did not exert significant adverse effect on seed germination of Brassica chinensis (Chinese cabbage), but Yuen Long sludge caused a reduction in root growth. In view of its lower EC and soluble ammonia contents, the high concentration of Zn and Cr in Yuen Long sludge would likely be responsible for this adverse effect on root growth. Therefore, Yuen Long sludge would likely have a more serious impact on soil quality and plant growth as compared to other sludges. This would require further verification from greenhouse and field experiments.

Ammonia↗

PubMed2001

Co-composting of soybean residues and leaves in Hong Kong.

The goal of this project was to evaluate the feasibility of co-composting of soybean residues and leaves and the effects of turning frequency on compost quality. Soybean residues were mixed with leaves and sawdust in 1:1:3 (w/w wet weight) for achieving a C/N ratio of about 30. Three heaps of about 4 m3 of compost mixtures were prepared receiving a turning frequency of daily (pile A), 3-day (pile B) and weekly (pile C) turning. Different turning frequencies did not significantly affect the changes in pH and volatile solids throughout the composting period. High turning frequency caused a lower electrical conductivity and NH4-N contents as well as a shorter duration of thermophilic phase, because of a high heat loss by evaporation and volatilization of ammonia in the pile. The highest C decomposition of 4% occurred in the pile with a 3-day turning period, which coincided with the higher-nitrogen content in this treatment. All treatments with different turning frequencies reached maturation at 63 days as indicated by the soluble organic carbon, soluble NH4-N, C/N ratio and cress seed germination index. However, increasing the aeration during composting period was beneficial in accelerating the maturation process. Taking into consideration less labour and lower operation costs as compared to daily turning, it can be suggested that a 3-day turning frequency would be more appropriate for reaching acceptable quality of compost and ease in operation.

Biodegradation, Environmental↗

PubMed2001

Effect of applying Hong Kong biosolids and lime on nutrient availability and plant growth in an acidic loamy soil.

A greenhouse experiment was conducted to evaluate the growth of Brassica chinensis L. in an acidic loamy soil amended with municipal biosolids with or without lime treatment (750 mg kg(-1)). The soil was amended with one of two municipal biosolids from Tai Po (TP) or Yuen Long (YL) wastewater treatment plants at application rates of 0, 5, 10, 25 and 50% (v/v). Initial NH(+)4-N and PO(3-)4-P concentrations in the amended soil increased with an increase in municipal biosolids loading rates and those with biosolids from TP had a greater increase in nutrients than those from YL. However, initial NO(-)3-N contents in both municipal biosolids amended soils decreased with an increase in biosolids loading rates, particularly for the soil amended with YL. Soluble K, Mg, Na and Ca concentrations in the soil increased with application rates. Municipal biosolids amended soil without lime treatment had higher NH(+)4-N, NO(-)3-N and major cation contents than those with lime. Addition of Tai Po municipal biosolids and lime significantly increased the dry weight yields of B. chinensis L., while YL municipal biosolids increased yields only at the 5% rate with lime amendment. The maximum growth was obtained at municipal biosolids application rates of 5 and 10% for TP and 5% for YL for limed soil. Addition of municipal biosolids increased the total N, P, Zn, Cr and major cation concentrations in the shoot tissue, while lime treatment reduced Zn and Cr contents. The high salt and metal concentrations would likely be factors inhibiting plant growth at high application rates, especially for YL biosolids. Further studies to determine the long-term effects of biosolid application on soil and crop quality are warranted.

Brassica↗

PubMed2001

Co-composting of pig manure with leaves.

The increase in the market price of sawdust makes it a less attractive bulking agent for pig manure composting. Hence, it was the aim of this project to evaluate the feasibility of co-composting pig manure with leaves with special emphasis on its effects on compost maturity and quality. Two piles were prepared with one pile (Pile A) just constituted of pig manureand sawdust at a mixing ratio of 3:2 (w/w, fresh weight), while the other one (Pile B) with pig manure, sawdust and leaves at 3:1:1 ratio (w/w, fresh weight) to achieve a C/N ratio of 30. Each 8 m3 heap was turned and mixed every 3 days to provide sufficient aeration and the moisture content was kept at 60-70% (w/w) throughout the composting period. After 49 days of composting, dissolved organic carbon (DOC), soluble NH4-N(solid), C/N(aqueous) and C/N(aqueous) of pile A decreased to <5%, 400 mgkg(-1), 20 and 6, respectively, indicating a high degree of maturity, while pile B with leaves required only 35days. Seed germination index (GI) of pile A increased to 66.5% at day 49, while pile B to 52.4% at day 35, which was higher than the phytotoxin-free level of 50% recommended for agricultural use. This demonstrated that the addition of leaves enhanced the humification process and shortened the time required for maturation and stabilization of pig manure composting. Therefore, it is recommended to co-compost pig manure with leaves to provide a means to reutilize this waste and in the same time to reduce the dependence on sawdust as a buLking agent.

Animals↗

PubMed2001

Evidence for an intrinsic toxicity of phosphatidylcholine to Sec14p-dependent protein transport from the yeast Golgi complex.

Yeast phosphatidylinositol-transfer protein (Sec14p) is essential for Golgi secretory function and cell viability. This requirement of Sec14p is relieved by genetic inactivation of the cytidine diphosphate-choline pathway for phosphatidycholine (PtdCho) biosynthesis. Standard phenotypic analyses indicate that inactivation of the phosphatidylethanolamine (PtdEtn) pathway for PtdCho biosynthesis, however, does not rescue the growth and secretory defects associated with Sec14p deficiency. We now report inhibition of choline uptake from the media reveals an efficient "bypass Sec14p" phenotype associated with PtdEtn-methylation pathway defects. We further show that the bypass Sec14p phenotype associated with PtdEtn-methylation pathway defects resembles other bypass Sec14p mutations in its dependence on phospholipase D activity. Finally, we find that increased dosage of enzymes that catalyze phospholipase D-independent turnover of PtdCho, via mechanisms that do not result in a direct production of phosphatidic acid or diacylglycerol, effect a partial rescue of sec14-1(ts)-associated growth defects. Taken together, these data support the idea that PtdCho is intrinsically toxic to yeast Golgi secretory function.

1-Phosphatidylinositol 4-Kinase↗

PubMed2001

Alcohol and retinoids.

This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Hirokazu Yokoyama and David Crabb. The presentations were (1) Roles of vitamin A, retinoic acid, and retinoid receptors in the expression of liver ALDH2, by J. Pinaire, R. Hasanadka, M. Fang, and David W. Crabb; (2) Alcohol, vitamin A, and beta-carotene: Adverse interactions, by M. A. Leo and Charles S. Lieber; (3) Retinoic acid, hepatic stellate cells, and Kupffer cells, by Hidekazu Tsukamoto, K. Motomura, T. Miyahara, and M. Ohata; (4) Retinoid storage and metabolism in liver, by William Bosron, S. Sanghani, and N. Kedishvili; (5) Characterization of oxidation pathway from retinol to retinoic acid in esophageal mucosa, by Haruko Shiraishi, Hirokazu Yokoyama, Michiko Miyagi, and Hiromasa Ishii; and (6) Ethanol in an inhibitor of the cytosolic oxidation of retinol in the liver and the large intestine of rats as well as in the human colon mucosa, by Ina Bergheim, Ina Menzl, Alexandr Parlesak, and Christiane Bode.

Aldehyde Dehydrogenase↗

PubMed2001

Medical management of a large aortic thrombus in a young woman with essential thrombocythemia.

Aortic thrombus formation is rare in the patients with essential thrombocytosis (ET); therefore, no guidelines for its management have been established. Embolism from ET-associated large vessel thrombi is potentially lethal and has been managed surgically in a few reported cases. We describe herein a 45-year-old black woman with ET found to have a 3.5-cm, pedunculated intra-aortic thrombus at the thoracoabdominal junction. How to treat this potentially devastating aortic thrombus was a management dilemma. We believed, based on the patient's diagnosis of ET and the histology of similar thrombi in 1 reported series, that the aortic thrombus was a "white thrombus" consisting primarily of aggregated platelets with a minimal fibrin network and almost no entrapped erythrocytes. The patient was treated with aspirin, 325 mg daily, as a platelet antiaggregating agent and hydroxyurea, 1,500 mg daily, to reduce the platelet count to less than 450 x 10(9)/L. The thrombus resolved without severe thromboembolic events. To our knowledge, this is the first reported case of a large intra-aortic thrombosis associated with ET that has been successfully managed with medical therapy alone.

Aorta, Abdominal↗

PubMed2001

[Advances in in vitro refolding of inclusion body proteins].

Overexpression of recombinant proteins in E. coli often results in formation of insoluble, inactive inclusion bodies. These inclusion bodies, which contain the recombinant proteins in a highly enriched form, can be isolated by solid/liquid separation. After solubilization, active proteins can be generated through an appropriate refolding process. Within the last decade, specific strategies and methods have been developed for preparing active recombinant proteins from inclusion bodies. Recent developments in renaturation procedure include the inhibition of aggregation during refolding by the application of low molecular weight additives and matrix-bound renaturation techniques.

Animals↗

PubMed2001

[Studies on the expression condition of human trefoil factor 3 in Pichia pastoris].

In order to enhance the expression level of human trefoil factor 3 (hTFF3) in Pichia pastoris, we optimized the transformant growth conditions in shake flasks including carbon sources in growth medium, inoculation ratio, methanol concentration, pH rotation speed and inducing time. The transformant could grow on the glucose to OD600 5.0 after 14 hours inoculation. The best inoculation ration of 100 mL growth medium to the induction medium was 1:1. The expression level of dimeric human trefoil factor 3 induction with 1% methanol for 48 hours at pH 6.0, agitation speed 240 r/min could reach 20 mg/L with OD600 15. The protein was expressed in 5-liter fermentor with 2% methanol induction for 32 hours, finally the cell density reached OD600 120. 100 mg/L of recombinant hTFF3 was obtained in the supernatant.

Cell Count↗

PubMed2001

Expression of Human Trefoil Factor 3 in Pichia pastoris and Its Biological Activity Analysis.

The human TFF3 (trefoil factor 3) DNA fragment was amplified by polymerase chain reaction (PCR) from human fetal placenta cDNA. The gene was cloned into the Pichia pastoris expression vector pPIC9K containing AOX 1 promoter and alpha-factor leader sequence. Multi-copies insertion transformants were screened on G418 plates. After the induction by 2% methanol for 48 hours, the expression of dimeric hTFF3 came up to 45% of total proteins in medium, as identified by SDS-PAGE and Western blot assay. The recombinant protein was further purified by S-Sepharose, Q-Sepharose ion-exchange chromatography and Sephacryl-S-100 gel filtration chromatography to the 95% purity, as shown by densitometric scanning. The N-terminus and molecular weight of the recombinant hTFF3 was in good agreement with the native hTFF3. The recombinant protein was proved to have good biological activity of preventing rats from the gastric ulcer induced by hydrochloric acid.

Journal Article↗

PubMed2000

A 72-base pair AT-rich DNA sequence element functions as a bacterial gene silencer.

We have previously demonstrated that sequential activation of the bacterial ilvIH-leuO-leuABCD gene cluster involves a promoter-relay mechanism. In the current study, we show that the final activation of the leuABCD operon is through a transcriptional derepression mechanism. The leuABCD operon is transcriptionally repressed by the presence of a 318-base pair AT-rich upstream element. LeuO is required for derepressing the repressed leuABCD operon. Deletion analysis of the repressive effect of the 318-bp element has led to the identification of a 72-bp AT-rich (78% A+T) DNA sequence element, AT4, which is capable of silencing a number of unrelated promoters in addition to the leuABCD promoter. AT4-mediated gene silencing is orientation-independent and occurs within a distance of 300 base pairs. Furthermore, an increased gene-silencing effect was observed with a tandemly repeated AT4 dimer. The possible mechanism of AT4-mediated gene silencing in bacteria is discussed.

Base Pairing↗

PubMed2000

Pike. A nuclear gtpase that enhances PI3kinase activity and is regulated by protein 4.1N.

While cytoplasmic PI3Kinase (PI3K) is well characterized, regulation of nuclear PI3K has been obscure. A novel protein, PIKE (PI3Kinase Enhancer), interacts with nuclear PI3K to stimulate its lipid kinase activity. PIKE encodes a 753 amino acid nuclear GTPase. Dominant-negative PIKE prevents the NGF enhancement of PI3K and upregulation of cyclin D1. NGF treatment also leads to PIKE interactions with 4.1N, which has translocated to the nucleus, fitting with the initial identification of PIKE based on its binding 4.1N in a yeast two-hybrid screen. Overexpression of 4.1N abolishes PIKE effects on PI3K. Activation of nuclear PI3K by PIKE is inhibited by the NGF-stimulated 4.1N translocation to the nucleus. Thus, PIKE physiologically modulates the activation by NGF of nuclear PI3K.

Adenosine Triphosphate↗

PubMed2000

Cytotoxicity of PM(2.5) and PM(2.5--10) ambient air pollutants assessed by the MTT and the Comet assays.

Ambient air particulate matters are classified into two distinct modes in size distribution, namely the coarse and fine particles. Correlation between high particulate concentration and adverse effects on human populations has long been recognized, however, the toxicology of these adverse effects has not been clarified. In the current report, the cytotoxic effects of the solvent-extractable organic compounds (SEOC) from fine particles smaller than 2.5 microm (PM(2.5)) and from coarse particles between 2.5-10 microm (PM(2.5-10)) were studied. Nine 24h consecutive monthly samples were tested to determine the correlation between cytotoxicity and total SEOC in two size fractions of particulate air pollution. Cytotoxicity of SEOC was measured by two micro-scale mammalian cells-based bioassays: the MTT cell proliferation assay, and the Comet assay for the detection of DNA damage. A well-defined mammalian cell line - Rat 6 rodent fibroblast was employed in the study. The SEOC extracts of air particulate matters were sub divided into two equal parts. One part was dissolved in DMSO, the other in KOH/hexane and then conjugated with bovine serum albumin to produce a lipid-soluble fraction for testing. The DMSO fraction would contain mainly the polycyclic aromatic hydrocarbons (PAH), alkanes and alkanols, while the lipid-soluble fraction would be enriched with fatty acids. The results from MTT assay showed that cytotoxicity of the PM(2.5) was much more severe than the PM(2.5-10), suggesting that toxic SEOC were confined to the fine particles. By and large, the DMSO solubles were much more toxic than the lipid solubles. The degree of cytotoxicity of the DMSO soluble samples is positively correlated to the amount of particulates present in the ambient air. For the PM(2.5), the winter samples were significantly more toxic than the summer samples in terms of cell killing, which seemed to be a direct reflection of the total loading of organic matter in the samples. Results from Comet assays showed that SEOC samples of PM(2.5) derived from winter months induced DNA damage at dosages resulting in no obvious cell killing in the MTT assay. Thus, long-term exposure to non-killing dosage of air pollutants may lead to the accumulation of DNA lesions, which may be one of the mechanisms responsible for the chronic adverse health effects of particulate air pollution.

Aerosols↗

PubMed2000

ppGpp-dependent leuO expression in bacteria under stress.

Despite the known potential transcription regulatory role of leuO gene product, LeuO, the condition when leuO expresses during bacterial growth cycle remains unclear. Mechanistically, leuO expression was shown to be part of promoter relay mechanism, however, the factor(s) responsible for the regulation of leuO expression is not known. Combining Northern and Western results, we demonstrate in the present communication that leuO expression is normally low and enhanced when bacteria are in transition from exponential growth to stationary phase. The stationary phase-associated leuO expression is ppGpp dependent and rpoS (sigma(s) factor) independent.

Bacterial Proteins↗

PubMed2000

The retinoid X receptor response element in the human aldehyde dehydrogenase 2 promoter is antagonized by the chicken ovalbumin upstream promoter family of orphan receptors.

Two tandem sites in the aldehyde dehydrogenase 2 promoter (designated FP330-5' and FP330-3') that bind members of the nuclear receptor superfamily were recently identified. Antibodies against apolipoprotein regulatory protein (ARP-1) altered DNA-protein interactions in electrophoretic mobility shift assays using oligonucleotides representing either promoter site and rat liver or cultured cell nuclear extracts. In vitro-translated chicken ovalbumin upstream promoter transcription factor (COUP-TFI), ARP-1, or ErbA-related protein 2 (Ear2) bound both sites. In addition, ARP-1/RXR, COUP-TFI/RXR, and ARP-1/COUP-TFI heterodimers bound the FP330-3' site. Mutagenesis of the FP330-3' site indicated that a DR-1 element was the preferred binding site for these factors. Transfected expression plasmids for these factors suppressed basal expression of reporter constructs containing the FP330-3' sites and the induction of the reporter by RXRalpha plus retinoic acid. Mutation of the two sites increased activity of a construct driven by 600 bp of the ALDH2 promoter in cell lines expressing COUP-TFs. The ALDH2 FP330-3' site appears to represent a complex nuclear receptor response element that is activated by RXRs and HNF-4 but repressed by members of the COUP-TF family.

Aldehyde Dehydrogenase↗

PubMed2000

Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition.

We report here the identification of a novel protein, Smac, which promotes caspase activation in the cytochrome c/Apaf-1/caspase-9 pathway. Smac promotes caspase-9 activation by binding to inhibitor of apoptosis proteins, IAPs, and removing their inhibitory activity. Smac is normally a mitochondrial protein but is released into the cytosol when cells undergo apoptosis. Mitochondrial import and cleavage of its signal peptide are required for Smac to gain its apoptotic activity. Overexpression of Smac increases cells' sensitivity to apoptotic stimuli. Smac is the second mitochondrial protein, along with cytochrome c, that promotes apoptosis by activating caspases.

Adult↗

PubMed2000

Differential modulation of paclitaxel-mediated apoptosis by p21Waf1 and p27Kip1.

The impact of the cyclin dependent kinase (CDK) inhibitors p21Waf1 and p27Kip1 on paclitaxel-mediated cytotoxicity was investigated in RKO human colon adenocarcinoma cells with the ecdysone-inducible expression of p21Waf1 or p27Kip1. Ectopic expression of p27Kip1 arrested cells at G1 phase, whereas p21Waf1 expression arrested cells at G1 and G2. Expression of p21Waf1 after paclitaxel treatment produced much greater resistance to paclitaxel than did expression of p27Kip1. We attributed this difference to the additional block at G2 induced by p21Waf1, which prevented cells from entering M phase and becoming paclitaxel susceptible. Expression of p21Waf1 inhibited p34cdc2 activity and markedly reduced paclitaxel-mediated mitotic arrest, from 87.5 to 23%. In contrast, p27Kip1 expression also inhibited p34cdc2 but reduced mitotic arrest only slightly, from 87. 4 to 74.5%. We concluded that the G2 block produced by p21Waf1, but not by p27Kip1, contributed to their unequal modulation of sensitivity to paclitaxel-mediated apoptosis in RKO cells, and there is no causal relationship between paclitaxel-mediated cytotoxicity and elevation of p34cdc2 activity.

Antineoplastic Agents, Phytogenic↗

PubMed2000

Decreased GDNF mRNA expression in dorsal spinal cord of unilateral arthritic rat.

It is now well established that nerve growth factor (NGF) plays a key role in inflammation-induced hyperalgesia. It was also reported that brain derived neurotrophic factor (BDNF), another member of neurotrophins, contributed to the pain pathway as a neurotransmitter in the CNS. The present work demonstrated a down-regulation of glial cell line-derived neurotrophic factor (GDNF) mRNA expression in dorsal spinal cord in complete Freund's adjuvant-induced unilateral arthritic rats serving as a chronic pain model. The fast occurring and long lasting down-regulations suggest that GDNF might contribute to pain pathway in a way different from neurotrophins and might play a role in the maintenance of chronic pain status.

Animals↗