Biomedical subjects
Publications and source records attributed to Z Hu.
A novel HLA-DRB1*09 allele, HLA-DRB1*090103, has been identified in a Guangdong Han person. Its sequence was confirmed by sequencing polymerase chain reaction products and clones. This allele differed by one nucleotide from HLA-DRB1*090102 at position 50(G-->C), which represents a silent mutation.
Several techniques have been proposed for biokinetic estimation of nitrification. Recently, an extant respirometric assay has been presented that yields kinetic parameters for both nitrification steps with minimal physiological change to the microorganisms during the assay. Herein, the ability of biokinetic parameter estimates from the extant respirometric assay to adequately describe concurrently obtained NH4+-N and NO(2-)-N substrate depletion profiles is evaluated. Based on our results, in general, the substrate depletion profiles resulted in a higher estimate of the maximum specific growth rate coefficient, micro(max) for both NH4+-N to NO(2-)-N oxidation and NO(2-)-N to NO(3-)-N oxidation compared to estimates from the extant respirograms. The trends in the kinetic parameter estimates from the different biokinetic estimation techniques are paralleled in the nature of substrate depletion profiles obtained from best-fit parameters. Based on a visual inspection, in general, best-fit parameters from optimally designed complete respirograms provided a better description of the substrate depletion profiles than estimates from isolated respirograms. Nevertheless, the sum of the squared errors for the best-fit respirometry based parameters was outside the 95% joint confidence interval computed for the best-fit substrate depletion based parameters. Notwithstanding the difference in kinetic parameter estimates determined in this study, the different biokinetic estimation techniques still are close to estimates reported in literature. Additional parameter identifiability and sensitivity analysis of parameters from substrate depletion assays revealed high precision of parameters and high parameter correlation. Although biokinetic estimation via automated extant respirometry is far more facile than via manual substrate depletion measurements, additional sensitivity analyses are needed to test the impact of differences in the resulting parameter values on continuous reactor performance.
The 9Be(32Ar, 31Ar)X reaction, leading to the 5/2+ ground state of a nucleus at the proton drip line, has a cross section of 10.4(13) mb at a beam energy of 65.1 MeV/nucleon. This translates into a spectroscopic factor that is only 24(3)% of that predicted by the many-body shell-model theory. We introduce refinements to the eikonal reaction theory used to extract the spectroscopic factor to clarify that this very strong reduction represents an effect of nuclear structure. We suggest that it reflects correlation effects linked to the high neutron separation energy (22.0 MeV) for this state.
Previous studies indicate that light information reaches the suprachiasmatic nucleus through a subpopulation of retinal ganglion cells that contain both glutamate and pituitary adenylyl cyclase-activating peptide (PACAP). Although the role of glutamate in this pathway has been well studied, the involvement of PACAP and its receptors is only beginning to be understood. To investigate the functions of PACAP in vivo, we developed a mouse model in which the gene coding for PACAP was disrupted by targeted homologous recombination. RIA was used to confirm a lack of detectable PACAP protein in these mice. PACAP-deficient mice exhibited significant impairment in the magnitude of the response to brief light exposures with both light-induced phase delays and advances of the circadian system impacted. This mutation equally impacted phase shifts induced by bright and dim light exposure. Despite these effects on phase shifting, the loss of PACAP had only limited effects on the generation of circadian oscillations, as measured by rhythms in wheel-running activity. Unlike melanopsin-deficient mice, the mice lacking PACAP exhibited no loss of function in the direct light-induced inhibition of locomotor activity, i.e., masking. Finally, the PACAP-deficient mice exhibited normal phase shifts in response to exposure to discrete dark treatments. The results reported here show that the loss of PACAP produced selective deficits in the light response of the circadian system.
Using soft-x-ray absorption spectroscopy at the Co L(2,3) and O K edges, we demonstrate that the Co3+ ions with the CoO5 pyramidal coordination in the layered Sr2CoO3Cl compound are unambiguously in the high spin state. Our result questions the reliability of the spin state assignments made so far for the recently synthesized layered cobalt perovskites and calls for a reexamination of the modeling for the complex and fascinating properties of these new materials.
Construction of single chromosomal DNA libraries by means of chromosome microdissection and microcloning will be useful for genomic research, especially for those species that have not been extensively studied genetically. Application of the technology of microdissection and microcloning to woody fruit plants has not been reported hitherto, largely due to the generally small sizes of metaphase chromosomes and the difficulty of chromosome preparation. The present study was performed to establish a method for single chromosome microdissection and microcloning in woody fruit species using pomelo as a model. The standard karyotype of a pomelo cultivar ( Citrus grandis cv. Guanxi) was established based on 20 prometaphase photomicrographs. According to the standard karyotype, chromosome 1 was identified and isolated with fine glass microneedles controlled by a micromanipulator. DNA fragments ranging from 0.3 kb to 2 kb were acquired from the isolated single chromosome 1 via two rounds of PCR mediated by Sau3A linker adaptors and then cloned into T-easy vectors to generate a DNA library of chromosome 1. Approximately 30,000 recombinant clones were obtained. Evaluation based on 108 randomly selected clones showed that the sizes of the cloned inserts varied from 0.5 kb to 1.5 kb with an average of 860 bp. Our research suggests that microdissection and microcloning of single small chromosomes in woody plants is feasible.
Interindividual variation in lung cancer susceptibility may be modulated in part through genetic polymorphisms in the DNA repair genes, especially the genes involved in the nucleotide excision repair (NER) pathway. The xeroderma pigmentosum complementary group D (XPD) is one of the NER genes, and two of the XPD polymorphisms 751A --> C and 312G --> A have been extensively studied in the association with lung cancer, although published studies have been inconclusive. To clarify the impact of XPD polymorphisms on lung cancer risk, we performed a meta-analysis of the published data from nine (10 comparisons) individual case-control studies of 3725 lung cancer cases and 4152 controls. The results showed that individuals with the XPD 751CC genotype had a 21% (odds ratio (OR)= 1.21, 95% confidence interval (CI) = 1.02-1.43) increased risk of lung cancer compared with individuals with the 751AA genotype without any between-study heterogeneity (P = 0.26). There was also a significant association in the recessive model of 751 C allele by comparing the CC with AC + AA genotypes (OR = 1.19, 95% CI = 1.02-1.40). The results also showed a significantly increased risk of lung cancer associated with the 312AA homozygous genotype compared with the GG genotype and the 312 A allele in the recessive model (compared with GA + AA genotypes) (OR = 1.27, 95% CI = 1.04-1.56 and OR = 1.32, 95% CI = 1.09-1.59, respectively). These results support the hypothesis that both the XPD 751 C and 312 A are risk alleles and individuals with the XPD 751 CC and 312 AA genotypes are at higher risk of developing lung cancer. Large multi-center studies with precise design, and stratified/adjusted analyses of the gene-gene (haplotypes) and gene-environment interactions are needed.
Peripheral nerve inflammation is a common clinical problem that accompanies nerve injury and several diseases including Guillain-Barre syndrome and acute and chronic inflammatory demyelinating polyneuropathy. To determine if neuropeptides are induced in motor neurons after inflammation and to study the mechanisms involved, a nerve cuff soaked in complete Freund's adjuvant (CFA) was applied locally to the facial nerve of Balb/C mice. This procedure resulted in an influx of lymphocytes and macrophages to the affected area and a blockade of retrograde axonal transport distal, but not proximal, to the site of application. The same treatment resulted in a strong ipsilateral induction of pituitary adenylyl cyclase activating peptide (PACAP) gene expression in motor neurons in the facial motor nucleus. Because the changes could have occurred due to the loss of target-derived factors or to the production of new factors by immune cells, we studied the effect of the inflammatory stimulus on PACAP mRNA in mice with severe combined immunodeficiency (SCID). As expected, SCID mice showed a severely reduced influx of T-lymphocytes but not macrophages to the peripheral nerve. Moreover, although retrograde transport distal to the inflammation site was blocked similarly in control and SCID mice, the number of motor neurons expressing PACAP mRNA after CFA application was significantly reduced in SCID mice. The data indicate that the induction of PACAP mRNA during nerve inflammation requires the involvement of lymphocytes. However, because the induction of PACAP gene expression was only partially blocked in SCID mice, macrophages, loss of target-derived factors, or other mechanisms may also contribute to the upregulation of PACAP gene expression in motor neurons after nerve inflammation.
Swine wastewater (agricultural wastewater) is normally stored in a storage holding tank for a certain time before released to the other treatment units, such as anaerobic lagoon and aerobic wetland. One of the characteristics for this treatment approach is that all the processes are open systems that are generally more passive in design and operation. As a consequence, seasonal variability including temperature and precipitation can have substantial impact on treatment efficacy and effluent water quality. This paper examines seasonal impacts of temperature on swine wastewater quality and treatment efficacy at a farm in East Leicester, Nova Scotia, Canada. During warm temperatures denitrification was noticeable in the anaerobic conditions, which would reduce the TSS removal rate from 76.6% in moderate temperatures to 42.1% in the warmest period recorded. Rainfall improved final effluent water quality, although this was shown to be through dilution rather than improvement of treatment efficacy. Following precipitation events the contaminant removals were negatively impacted in the aerobic lagoon, as BOD5 removal decreased from 61.6% before rainfall to 41.5% after rainfall, TSS from 71.4% to 59.3%, VSS from 73.4% to 59.3%, TKN from 59.9% to 42.1%, and NH4+ -N from 51.3% to 41.6%. In comparison to the aerobic conditions, the removal rates were increased for anaerobic condition with the rainfall dilution (e.g., TSS from 18.2% to 34.3%), which lead to an overall treatment improvement for the entire system. Thus the case study data presented in this paper provides an assessment of the operational and design issues that are particularly relevant for passive treatment systems that are used in the agriculture industry.
Sedimentation is a widely used separation method for treating agricultural waste. There are several chemical and biological characteristics, which can affect the settling behavior and liquid waste. The optimization of cation balances and potential for nitrification are among these processes. In addition to sedimentation, it can also affect the dewaterability of the samples. Liquid hog manure was used during the laboratory based experiments to investigate the effects of Ca2+ and Mg2+ ions and nitrification inhibition on the overall settling and dewatering characteristics. The results indicated that settling and dewatering characteristics improved during the course of the experiments. However, the improvement in settling and dewatering characteristics was inconsistent and not statistically significant. Cation addition in aerated reactor increased the highest settling velocity (94%). The improvement in dewaterability, as quantified by capillary suction time, was also not consistent. The greatest filterability observed in the supernatant was a capillary suction time of 40 s for a M:D ratio of 2:1. Initial NH 4 + concentration was more important than the nitrification inhibitor, as the presence of nitrification inhibitor increased the nitrification rate by over 300% because of the high initial NH 4 + concentration and low volatile suspended solid. The results from these experiments provide the basis for further field evaluation of cation optimization.
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The system Li-V-N was studied by means of X-ray and neutron powder diffraction, thermal and chemical analyses, and XAS spectroscopy at the vanadium K-edge. Three polymorphs of Li(7)[VN(4)] have been established from X-ray and neutron powder diffraction (gamma-Li(7)[VN(4)], space group Pfourmacr;3n, No. 218, a = 960.90(4) pm, V = 887.23(6) x 10(6) pm(3), Z = 8; beta-Li(7)[VN(4)], space group Pathremacr;, No. 205, a = 959.48(3) pm, V = 883.31(5) x 10(6) pm(3), Z = 8; alpha-Li(7)[VN(4)], P4(2)/nmc, No. 137, a = 675.90(2) pm, c = 488.34(2) pm, V = 223.09(1) x 10(6) pm(3), Z = 2). Crystallographic and phase relations are discussed. All three modifications are diamagnetic, indicating vanadium in the oxidation state +5. The V-K XAS spectra support the oxidation state assignment, the non-centrosymmetric coordination (tetrahedral), and the nearly identical second coordination sphere of vanadium, made up from Li in all three phases. The 3d-related features of the spectra display strongly localized properties. The phase transitions appear to be reconstructive; no direct group-subgroup symmetry relations of the crystal structures exist. The formation of solid solutions between Li(2)O and beta-Li(7)[VN(4)] with the general formula Li(1.75)((V(0.25(1)(-)(x))Li(0.25)(x))(N(1)(-)(x)O(x)())) with 0 <or= x <or= 1 leads to increasing substitution of V by Li. At the approximate composition with x = 0.125 the Li-V-disorder seems to be complete: X-ray diffraction lines give rise to only one-half of the original unit cell dimensions, a' = 476.47(3) pm approximately (1/2)a(beta-Li(7)[VN(4)]).
Regions of extremely high sequence identity are recurrent in modular polyketide synthase (PKS) genes. Such sequences are potentially detrimental to the stability of PKS expression plasmids used in the combinatorial biosynthesis of polyketide metabolites. We present two different solutions for circumventing intra-plasmid recombination within the megalomicin PKS genes in Streptomyces coelicolor. In one example, a synthetic gene was used in which the codon usage was reengineered without affecting the primary amino acid sequence. The other approach utilized a heterologous subunit complementation strategy to replace one of the problematic regions. Both methods resulted in PKS complexes capable of 6-deoxyerythronolide B analogue biosynthesis in S. coelicolor CH999, permitting reproducible scale-up to at least 5-l stirred-tank fermentation and a comparison of diketide precursor incorporation efficiencies between the erythromycin and megalomicin PKSs.
The ability of scanning tunneling microscopy to probe the pathways of thermally activated high-barrier surface processes is frequently limited by competing low-barrier processes that can confuse measurement of the true initial and final configuration. We introduce an approach to circumvent this difficulty by driving the surface process with nanosecond laser heating. The method is applied to determine the pathway of recombinative desorption in the H/Si(001) system. The observed configuration of dangling bonds after laser heating reveals that the desorbed hydrogen molecules are not formed on single dimers, but rather from neighboring silicon dimers via an interdimer reaction pathway.
We measured the strength of the 0(+)(gs)-->2(+)(1) excitations in the radioactive mirror nuclei 32Ar and 32Si using the techniques of intermediate-energy Coulomb excitation for 32Ar and inelastic proton scattering in inverse kinematics for 32Si. The 32Ar measurement, taken together with previously existing Coulomb excitation data for 32Si, yields the isoscalar and isovector multipole matrix elements for the 0(+)(1)-->2(+)(1) transition between T = 2 states in the A = 32 system. The proton scattering measurement for 32Si, when combined with the Coulomb excitation data for this nucleus, yields a ratio of neutron and proton matrix elements, M(n)/M(p), for 32Si.
Dissociative adsorption of molecular hydrogen on clean Si(001) surfaces has been investigated by means of scanning tunneling microscopy. The dissociated hydrogen atoms are found to occupy Si atoms of adjacent dimers. In addition to this interdimer configuration associated with the adsorption of isolated hydrogen molecules, pairs of adjacent doubly occupied dimers are readily formed. They arise from the enhanced reactivity of partially occupied dimers following the initial H2 adsorption step. The results are considered in light of recent adsorption and desorption measurements.
The incidence of atherosclerosis and cardiovascular disease (CVD) in women increases following menopause and has been associated with a reduction in circulating estrogen. Increased CVD risk is also perpetuated by sedentary lifestyle. Growing evidence indicates that oxidation of lipoproteins leads to a powerful immune response, disruption of normal lipoprotein function, and deposition of atherosclerotic plaques. For example, once high-density lipoproteins (HDL) are oxidized, they lose the ability to a) participate in reverse transport of cholesterol to the liver, and b) protect low-density lipoproteins (LDL) against oxidation. The purpose of this study was to determine the effects of combining estrogen replacement and exercise upon lipid peroxidation of the HDL fraction (HDL-ox). Blood samples were drawn from 34 post-menopausal women from four groups: women who were not receiving estrogen replacement and who were sedentary (NSD) (n = 9); women who were not receiving estrogen replacement and who were participating in regular exercise (NEX) (n = 8); women who were receiving estrogen replacement and who were sedentary (ESD) (n = 8); and women who were receiving estrogen replacement and who were participating in regular exercise (EEX) (n = 9). Total-HDL cholesterol was significantly higher (p<0.05) in EEX when compared with NEX, NSD, and ESD. HDL-ox was assessed via malondialdehyde (MDA). Mean (+/- SEM) values for HDL MDA expressed in nM are as follows: NSD = 903.3 +/- 118.4; NEX = 1226.7 +/- 247.7; ESD = 876.7 +/- 116.3; EEX = 537.4 +/- 74.8. EEX lipid peroxidation was significantly (p = 0.02) lower than NEX. Lipid peroxidation tended to be lower in EEX than in NSD and ESD (p = 0.07). These data indicate that the combination of estrogen replacement and regular exercise in post-menopausal women may be most effective in reducing oxidation of HDL in vivo.
Here we describe a convenient method to generate homologous recombinant baculoviral genomes in E. coli. The recombination takes place with the aid of recombination enzymes provided by the phage lambda Red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. Proof of concept was provided when the cathepsin gene (v-cath) of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) was replaced by the chloramphenicol resistance gene (CmR). First, CmR was inserted between the flanking sequences of the HaS-NPV v-cath. Each of the flanking regions was about 1 kb. The fragment was linearized and electroporated into bacteria containing both the HaSNPV bacmid and the lambda Red system. Recombinant bacmids resistant to chloramphenicol were selected. In comparison to the standard co-transfection/plaque assays, this method significantly reduces the time required to construct baculovirus knockout mutants. It may also be useful in the manipulation of other large viral genomes.