Biomedical subjects

W T Liu

Publications and source records attributed to W T Liu.

At least 19 recordsLinked to original sources

PubMed2006

Microbial succession of glycogen accumulating organisms in an anaerobic-aerobic membrane bioreactor with no phosphorus removal.

The succession of glycogen accumulating organisms (GAOs) has been observed in an acetate-fed, anaerobic-aerobic sequencing membrane bioreactor (MBR) operated for 260 days without enhanced biological phosphorus removal (EBPR) activity. Semi-quantitative fluorescence in situ hybridization results showed that a gammaproteobacterial lineage GB frequently observed in EBPR processes was initially the numerically dominant species (50-66% of total cells) of the GAO in the MBR from day 1 to day 38. During this period, succession of two different subgroups of group GB was also observed. On day 85 onward, a population shift from GB group to 'Defluvicoccus'-related tetrad-forming organisms (TFO) occurred. This microbial succession was suspected to be related to the applied operating conditions (long hydraulic retention time and long solid residence time) which favored the proliferation of 'Defluvicoccus'-related TFO rather than the GB group. Application of terminal restriction fragment length polymorphism on selected samples further revealed that the microbial diversity of the seeding sludge as determined by the number of terminal restriction fragments was higher than that of sludge samples taken after day 85.

Aerobiosis↗

PubMed2006

Proliferation of glycogen accumulating organisms induced by Fe(III) dosing in a domestic wastewater treatment plant.

To meet the effluent requirements given for the sensitive receiving body, the Southpest Wastewater Treatment Plant of Budapest, Hungary uses a combined activated sludge-biofilter system with chemical precipitation for P removal. Causes of the proliferation of glycogen accumulating organisms (GAOs) observed in the unaerated/oxic activated sludge unit of this system were investigated both in full-scale and in lab-scale experiments combined with a detailed analysis of the microbial communities. Concentration profile measurements throughout the 8-stage activated sludge unit indicated anaerobic conditions in the first two unaerated reactors and low orthophosphate level (< 1 mg l(-1)) in all of the stages that could not be attributed to the influent quality, but to Fe (III)-dosing to the returned activated sludge. Microbiological analysis revealed the presence of GAOs from the GB group in the Gammaproteobacteria and occasionally tetrad-forming organisms from Actinobacteria, and the absence of Rhodocyclus-related polyphosphate accumulating organisms (PAOs) in the activated sludge samples. Comparative lab-scale studies carried out in two identically arranged UCT-systems with staged anoxic reactors also confirmed that Fe (III)-dosing may result in phosphorus deficiency of the microbial niche, leading to the suppression of growth and EBPR activity of PAOs and to the proliferation of GAOs.

Base Sequence↗

PubMed2006

A novel conformation-dependent monoclonal antibody specific to the native structure of beta-lactoglobulin and its application.

Molten globules are thought to be general intermediates in protein folding and unfolding. beta-lactoglobulin (beta-LG) is one of the major bovine whey proteins, constituting approximately 10 to 15% of total milk proteins. We have recently identified beta-LG as a superior marker for evaluating thermally processed milk. Strand D of beta-LG participates in irreversible thermal unfolding as probed by a monoclonal antibody (mAb) specific to thermally denatured beta-LG. In the present study, we used native beta-LG as an immunogen to test the hypothesis that a specific mAb against the native beta-LG could be established. As result, a mAb (4H11E8) directed against the native structure of beta-LG was made. The antibody did not recognize the heat-denatured form of beta-LG, such as its dimer and aggregates. Immunoassay using this "native" mAb showed that the stability of beta-LG was at temperatures < or =70 degrees C. beta-Lactoglobulin began to deteriorate between 70 and 80 degrees C over time. The denaturation was correlated with the transition temperature of beta-LG. Further chemical modification of Cys (carboxymethylation) or positively charged residues (acetylation) of beta-LG totally abolished its immunoreactivity, confirming the conformation-dependent nature of this mAb. Using competitive ELISA, the 4H11E8 mAb could determine the native beta-LG content in commercially processed milks. Concentrations of native beta-LG varied significantly among the local brands tested. From a technological standpoint, the mAb prepared in this study is relevant to the design and operation of appropriate processes for thermal sanitation of milk and of other dairy products.

Animals↗

PubMed2004

In-situ characterization of microbial community in an A/O submerged membrane bioreactor with nitrogen removal.

The bacterial community involved in removing nitrogen from sewage and their preferred DO environment within an anoxic/oxic membrane bioreactor (A/O MBR) was investigated. A continuously operated laboratory-scale A/O MBR was maintained for 360 d. At a sludge age of 150 d and a C/N ratio of 3.5, the system was capable of removing 88% of the influent nitrogen from raw wastewater through typical nitrogen removal transformations (i.e. aerobic ammonia oxidation and anoxic nitrate reduction). Characterization of the A/O MBR bacterial community was carried out using fluorescence in situ hybridization (FISH) techniques. FISH results further showed that Nitrosospira spp. and Nitrospira spp. were the predominant groups of ammonia and nitrite oxidizing group, respectively. They constituted up to 11% and 6% of eubacteria at DO below 0.05 mg/l (low DO), respectively, and about 14% and 9% of eubacteria at DO between 2-5 mg/l (sufficient DO), respectively, indicating preference of nitrifiers for a higher DO environment. Generally low counts of the genus Paracoccus were detected while negative results were observed for Paracoccus denitrificans, Alcaligenes spp, and Pseudomonas stutzeri under the low and sufficient DO environments. The overall results indicate that Nitrosospira spp., Nitrospira spp. and members of Paracoccus spp. can be metabolically functional in nitrogen removal in the laboratory-scale A/O MBR system.

Alcaligenes↗

PubMed2003

Removal of MS2 bacteriophage using membrane technologies.

Removals of MS2 bacteriophage virus using different membrane materials under different operating pressures were investigated. The results obtained in this study suggested that a better log removal in terms of MS2 bacteriophage virus could be achieved using Polyamide RO membrane under the optimum operating pressure of 100 psi. It is further noted that variable MS2 influent concentration levels resulted in corresponding variable log removals of the bacteriophages by the Polyamide RO membrane. The presence of MS2 bacteriophage virus in the effluent could possibly be due to leakage of bacteriophages through the membranes structure. Investigations using SEM and AFM showed that there were gaps or pores present in the membrane structure which were sufficiently large for the MS2 viruses to pass through.

Bacteria, Anaerobic↗

PubMed2002

Phylogenetic and physiological diversity of tetrad-forming organisms in deteriorated biological phosphorus removal systems.

Polyhydroxyalkanoate (PHA)- and polyphosphate-accumulating traits of different taxonomic tetrad-forming organisms (TFOs) in two anaerobic-aerobic sequential batch reactors (SBRs) were characterized by the simultaneous use of fluorescence in-situ hybridization, PHB stain and DAPI stain. The two SBRs with glucose as the main carbon source were operated under different P:total organic carbon feeding ratios for more than 300 days, but both exhibited no enhanced biological phosphorus removal (EBPR) activity. Microscopic observations on sludge samples taken at various times from those two SBRs revealed that TFOs consistently accounted for more than 50% of total cells, and were mostly affiliated with the beta- and gamma-subclasses of Proteobacteria and the high G+C phylum of gram-positive bacteria (HGC). Those TFOs from the beta-Proteobcateria exhibited PHB stain positive and DAPI stain negative, indicating that they could utilize compounds other than polyphosphate (i.e. glycogen) as reducing power for PHA synthesis from glucose. In contrast, two types of TFOs within the HGC group showed negative PHB stain and positive DAPI stain, indicating their capacity to accumulate polyphosphate without the synthesis and degradation of PHA. This metabolic trait was different from the widely accepted biochemical model of EBPR and non-EBPR metabolisms. Other TFOs within the HGC group and gamma-Proteobacteria showed negative responses to both PHA and DAPI stains, and their function in the deteriorated EBPR system need to be further clarified. Overall findings suggested that the phylogenic and physiological heterogeneity of TFOs in anaerobic-aerobic activated sludge systems were diverse and greatly exceeded the current understanding.

Bioreactors↗

PubMed2001

The spatiotemporal relationship among Schwann cells, axons and postsynaptic acetylcholine receptor regions during muscle reinnervation in aged rats.

To morphologically define the aging-related features during muscle reinnervation the spatiotemporal relationships among the major components of the neuromuscular junctions (NMJs) were investigated. A total of 64 rats, 30 adults (4 months old) and 34 aged adults (24 months old), were used. Between 1 and 12 weeks after sciatic nerve-crushing injury, cryosections of skeletal muscle were single or double labeled for S100, a marker of Schwann cells (SCs), for protein gene product 9.5, a neuronal marker, and for alpha-bungarotoxin (alpha-BT), a marker of the acetylcholine receptor site (AChR site), and then observed by confocal laser microscopy. The most obvious age changes were noted: (1) the regenerating SCs and axons were delayed in their arrival at the NMJ, (2) the dimensions of terminal SCs and AChR sites displayed a drastic and long-lasting drop (for terminal SCs, during 1-8 weeks; for AChR sites, during 1-12 weeks); (3) the degree of spatial overlap between AChR sites and terminal SCs was markedly low until 8 weeks post-crush; (4) damage and poor formation in the SCs, terminal axons and AChR sites, together with poor process extension from the terminal SC or terminal axon, were pronounced; (5) persistent aberrant changes, such as multiple innervation and terminal axon sprouting, together with poorly formed collateral innervation, nerve bundles, and NMJs, more frequently occurred in the later reinnervation period. Thus, with aging, regeneration is impaired during the period in which regenerating SC strands and axons extend into NMJs and the subsequent establishment of nerve-muscle contact is in progress. A complex set of morphological abnormalities between or among the TSCs, terminal axons, and AChR sites may be important in slowing of regeneration and reinnervation in aged motor endplates.

Aging↗

PubMed2001

Up-regulation of Fas ligand expression by human cytomegalovirus immediate-early gene product 2: a novel mechanism in cytomegalovirus-induced apoptosis in human retina.

Human CMV (HCMV) is an important pathogen that causes widespread diseases in immunocompromised individuals. Among the opportunistic HCMV infections, HCMV retinitis is most common in transplant recipients and AIDS patients. It often leads to blindness if left untreated. The question as to how HCMV infection causes retinal pathogenesis remains unresolved. Here, we report that viral immediate-early gene product 2 (IE2), but not IE1, up-regulates the Fas ligand (FasL) expression in HCMV-infected human retinal pigment epithelium cells. Increased secretion of FasL from virally infected cells into cultured medium was observed upon HCMV infection. The capability of such cell-free medium to induce apoptosis of Fas (CD95)-expressing Jurkat cells further implies that Fas-FasL interaction might mediate cell death in the lesion of HCMV retinitis. To support this idea, we observed augmented soluble FasL levels in vitreous from AIDS patients with HCMV retinitis as compared with that from AIDS patients without HCMV infection. In addition, by in situ hybridization and immunohistochemistry, we detected enhanced signals of FasL, the existence of viral IE Ags and apoptotic cells at the same sites in the lesion of HCMV-infected retina. These results strongly suggest that IE2 induction of FasL expression in human retina might be an important event that takes place in the early stage of infection and finally leads to visual loss in individuals affiliated with HCMV retinitis.

AIDS-Related Opportunistic Infections↗

PubMed2001

Far-reaching effects of the Hawaiian Islands on the Pacific Ocean-atmosphere system.

Using satellite data, we detected a wind wake trailing westward behind the Hawaiian Islands for 3000 kilometers, a length many times greater than observed anywhere else on Earth. This wind wake drives an eastward ocean current that draws warm water from the Asian coast 8000 kilometers away, leaving marked changes in surface and subsurface ocean temperature. Standing in the path of the steady trade winds, Hawaii triggers an air-sea interaction that provides the feedback to sustain the influence of these small islands over a long stretch of the Pacific Ocean.

Journal Article↗

PubMed2001

Detection of Kaposi sarcoma-associated herpesvirus in bone marrow biopsy samples from patients with multiple myeloma.

BACKGROUND: Kaposi sarcoma-associated herpesvirus (KSHV) recently has been identified in the bone marrow (BM) dendritic cell of multiple myeloma (MM) patients. However, whether or not KSHV is associated with MM remains controversial because many studies have failed to detect the presence of KSHV DNA sequences in the BM of their MM patients. METHODS: We have assayed for KSHV DNA sequences in the BM biopsy samples from 49 patients with MM and from 8 patients with normal BM, using nested polymerase chain reaction and dot blot analysis. The polymerase chain reaction product of KSHV was further determined by single-strand conformation polymorphism and sequence analyses. RESULTS: KSHV DNA was detectable in 22 of 49 patients (44.9%) with MM but was not detectable in normal BM cells. Single-strand conformation polymorphism and sequence analyses showed that there were interpatient specific mutations. Sixteen out of 22 KSHV DNA sequences belonged to a previously defined subgroup, and the other 6 remain unclassified and may represent distinct strains of KSHV in Taiwan. CONCLUSIONS: Data strongly supported that KSHV infection did exist in the BM of the current study patients with MM. However, the role of KSHV in the pathogenesis of multiple myeloma remains to be determined.

Adult↗

PubMed2001

Structure of microbial communities in activated sludge: potential implications for assessing the biodegradability of chemicals.

Various methods used to assess the biodegradability of chemicals often employ activated sludge as an inoculum since chemicals that ultimately enter the environment are often discharged through wastewater. Differences in the structure and function of activated sludge microbial communities that may complicate interpretation of biodegradation tests could arise from differences in wastewater composition, wastewater treatment plant (WWTP) operation, or manipulations done after collection of the activated sludge. In this study, various methods were used to characterize the structure of microbial communities found in freshly collected activated sludge from WWTPs in Japan, Europe, and the United States, as well as sludge that had been continuously fed either sewage or a glucose-peptone mixture for several weeks after collection. Comparisons of biomass levels, whole-community substrate utilization (determined using Biolog GN and GP plates), and phospholipid fatty acid (PLFA) profiles indicated there were both geographical and temporal differences among freshly collected activated sludge samples. Moreover, marked shifts in the structure of activated sludge microbial communities occurred upon continuous cultivation in the laboratory for 5 weeks using a glucose-peptone feed. These shifts were evident from whole-community substrate utilization and PLFA profiles as well as differences in the profiles of 16S rDNA genes from numerically dominant populations obtained by denaturing gradient gel electrophoresis and terminal restriction fragment analyses. Further studies are needed to better define the variability within and between activated sludge from wastewater treatment plants and laboratory reactors and to assess the impact of such differences on the outcome of biodegradability tests.

Bacteria↗

PubMed2001

A quantitative assay for measuring human foamy virus using an established indicator cell line.

In order to improve the accuracy for detecting human foamy virus (HFV), an indicator cell line was established by co-transfecting baby hamster kidney-21 cells with two plasmids: one containing a G418 antibiotic resistance marker and the other including the luc gene which was placed downstream of the inducible HFV long terminal repeat promoter (from -533 to +20). Among 11 independent subclones, IdB14 was found to be stable with a low basal level of luciferase activity. Although the changes in luciferase activity in infected clones showed time-dependency and peaked at day 8, it is possible to differentiate infected and uninfected cells on day 2. The sensitivity of the foamy virus activated luciferase (FAL) assay was 400 times higher than the end-point syncytium formation by TCID(50). The HFV LTR promoter in the IdB14 cell line was specific for this virus. Moreover, a linear relationship was found between the MOI and the activated intensity of luciferase expression. These findings suggest that the FAL assay using the IdB14 indicator cell line is a simple and useful technique for rapid diagnosis and quantitation of active HFV infection.

Animals↗

PubMed2001

An indicator cell assay for detection of human cytomegalovirus based on enhanced green fluorescent protein.

An indicator cell line (ML-UL54-EGFP) for the detection of human cytomegalovirus (HCMV) by a simple and direct method was developed. The stable line was constructed by introducing into mink lung cells an expression cassette that contains the enhanced green fluorescent protein (EGFP) reporter gene under the control of an HCMV-inducible promoter. The promoter was from the upstream region of the HCMV UL54 (pol) gene, an early gene promoter that is activated in the early phase of HCMV infection. Following infection with HCMV for 48 h, the stable line expressed well detectable level of the EGFP as observed under a fluorescence microscope. The sensitivity of the indicator cell assay is at least comparable with that of a plaque assay as assessed with a panel of HCMV strains. There were no detectable fluorescent cells after inoculations with several viruses other than HCMV, indicating high specificity. Analysis with flow cytometry revealed that the induced fluorescence from the infected cells was proportional to the titer of HCMV inoculated, making it possible to quantify HCMV infectious particles. In summary, the EGFP-based indicator cell line is of potential use for rapid detection and quantification of HCMV in clinical specimens.

Animals↗

PubMed2001

In situ identification of polyphosphate- and polyhydroxyalkanoate-accumulating traits for microbial populations in a biological phosphorus removal process.

Polyphosphate- and polyhydroxyalkanoate (PHA)-accumulating traits of predominant microorganisms in an efficient enhanced biological phosphorus removal (EBPR) process were investigated systematically using a suite of non-culture-dependent methods. Results of 16S rDNA clone library and fluorescence in situ hybridization (FISH) with rRNA-targeted, group-specific oligonucleotide probes indicated that the microbial community consisted mostly of the alpha- (9.5% of total cells), beta- (41.3%) and gamma- (6.8%) subclasses of the class Proteobacteria, Flexibacter-Cytophaga (4.5%) and the Gram-positive high G+C (HGC) group (17.9%). With individual phylogenetic groups or subgroups, members of Candidatus Accumulibacter phosphatis in the beta-2 subclass, a novel HGC group closely related to Tetrasphaera spp., and a novel gamma-proteobacterial group were the predominant populations. Furthermore, electron microscopy with energy-dispersive X-ray analysis was used to validate the staining specificity of 4,6-diamino-2-phenylindole (DAPI) for intracellular polyphosphate and revealed the composition of polyphosphate granules accumulated in predominant bacteria as mostly P, Ca and Na. As a result, DAPI and PHA staining procedures could be combined with FISH to identify directly the polyphosphate- and PHA-accumulating traits of different phylogenetic groups. Members of Accumulibacter phosphatis and the novel gamma-proteobacterial group were observed to accumulate both polyphosphate and PHA. In addition, one novel rod-shaped group, closely related to coccus-shaped Tetrasphaera, and one filamentous group resembling Candidatus Nostocoidia limicola in the HGC group were found to accumulate polyphosphate but not PHA. No cellular inclusions were detected in most members of the alpha-Proteobacteria and the Cytophaga-Flavobacterium group. The diversified functional traits observed suggested that different substrate metabolisms were used by predominant phylogenetic groups in EBPR processes.

Acetates↗

PubMed2001

Optimization of an oligonucleotide microchip for microbial identification studies: a non-equilibrium dissociation approach.

The utility of a high-density oligonucleotide microarray (microchip) for identifying strains of five closely related bacilli (Bacillus anthracis, Bacillus cereus, Bacillus mycoides, Bacillus medusa and Bacillus subtilis) was demonstrated using an approach that compares the non-equilibrium dissociation rates ('melting curves') of all probe-target duplexes simultaneously. For this study, a hierarchical set of 30 oligonucleotide probes targeting the 16S ribosomal RNA of these bacilli at multiple levels of specificity (approximate taxonomic ranks of domain, kingdom, order, genus and species) was designed and immobilized in a high-density matrix of gel pads on a glass slide. Reproducible melting curves for probes with different levels of specificity were obtained using an optimized salt concentration. Clear discrimination between perfect match (PM) and mismatch (MM) duplexes was achieved. By normalizing the signals to an internal standard (a universal probe), a more than twofold discrimination (> 2.4x) was achieved between PM and 1-MM duplexes at the dissociation temperature at which 50% of the probe-target duplexes remained intact. This provided excellent differentiation among representatives of different Bacillus species, both individually and in mixtures of two or three. The overall pattern of hybridization derived from this hierarchical probe set also provided a clear 'chip fingerprint' for each of these closely related Bacillus species.

Bacillus↗

PubMed2001

Monoclonal antibodies against human ribosomal P proteins penetrate into living cells and cause apoptosis of Jurkat T cells in culture.

OBJECTIVE: This study was designed to determine the role of autoantibodies to the ribosomal P protein (anti-P Abs) in the pathogenesis of systemic lupus erythematosus (SLE) using monoclonal anti-P antibodies (anti-P mAbs). METHODS: Anti-P mAbs were prepared by a standard hybridoma procedure using recombinant human P1 and P2 proteins as immunogens. We studied the reactivities of these mAbs to P proteins, their binding and penetration capabilities in different cell lines and their apoptotic effects on Jurkat T cells. RESULTS: In addition to recognizing human P0, P1 and P2 proteins, the anti-P mAb 9B6-4 bound to 20-40% and penetrated 50-90% of astrocytes, Jurkat T cells and lung cancer cells via the P0 surface protein. Treatment with the mAb 9B6-4 also caused increases in the percentages of Jurkat T cells in the sub-G1 phase of the cell cycle (14.8%) and undergoing apoptosis (21.3%). CONCLUSION: Anti-P autoantibodies may play a role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE.

Animals↗

PubMed2001

Study of microbial community of brewery-treating granular sludge by denaturing gradient gel electrophoresis of 16S rRNA gene.

The microbial community structure of granular sludge from an upflow anaerobic sludge blanket (UASB) reactor treating brewery effluent was studied by denaturing gradient gel electrophoresis (DGGE). Twelve major bands were observed in the DGGE fingerprint for the Bacteria domain and four bands for the Archaea domain. Of the bacterial bands observed, six were successfully purified and sequenced. Among them, three were related to the gram-positive low G + C group, one to the Delta subclass of the Proteobacteria, one to the Gamma subclass, and one to the Cytophaga group with no close related sequence. The 16S rRNA sequences of the four archaeal bands were closely associated with Methanosaeta concilii and Methanobacterium formicum.

Bacteria, Anaerobic↗

PubMed2001

Human papillomavirus type 18 in colorectal cancer.

Human papillomaviruses (HPVs) have been recognized as the etiological agent of warts, and they may also be associated with many cancers. HPV-18 is very common both in genital papillomas and in large bowel cancer. The relation between HPV-18 infection and natural course of colorectal cancer has not been fully defined. In this study, normal mucosa and colorectal cancer tissue were evaluated for the presence of HPV gene to determine whether or not HPV was involved in the development of colon neoplasm. The DNA extracted from colon tissue was screened for HPV by polymerase chain reaction (PCR) to amplify the viral gene fragment. These PCR products were digested with restriction enzyme, and Southern blotting was then performed to confirm the existence of HPV-18. The nucleotide sequence related to HPV-18 DNA was detected in 53% (10/19) of the normal mucosa specimens and in 84% (16/19) of the colorectal cancer specimens. The correlation between cancer samples and positive rate of HPV-PCR was statistically significant by chi-square test (p < 0.01). These data indicate that HPV-18 can infect the normal mucosa of the colon, and that this infection may be a risk factor for the development of colorectal cancer. The presence of HPV-18 DNA in patients with colorectal cancer suggests that the pathogenesis of colorectal cancer includes viral involvement.

Colorectal Neoplasms↗