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Biomedical subjects

C Chang

Publications and source records attributed to C Chang.

At least 19 recordsLinked to original sources

Antimicrobial-resistant Staphylococcus aureus isolated from Australian wildlife admitted to a veterinary hospital.

Although antimicrobial resistance (AMR) is a growing One Health concern, little is known about AMR in Staphylococcus aureus from Australian wildlife. This study investigated the occurrence, phenotypic AMR profiles, and genetic characteristics of S. aureus from six representative Australian wildlife species admitted to a wildlife hospital in Western Australia, including the western grey kangaroo (Macropus fuliginosus), quenda (Isoodon fusciventer), pelican (Pelecanus conspicillatus), galah (Eolophus roseicapilla), shingleback skink (Tiliqua rugosa) and long-necked turtle (Chelodina colliei). Staphylococcus aureus was isolated from 11.7% (21/180, 95% CI: 7.4%-17.3%) of the animals on admission. Whole genome sequencing identified 13 multi-locus sequence types (STs) and various virulence factors, including the human-specific immune evasion cluster (IEC). Resistance to at least one antimicrobial class was observed in 63.6% of the isolates. The blaZ, erm(T), aac(6')-aph(2″), and tet(L) AMR genes were detected in 63.6%, 13.6%, 4.5%, and 4.5% of S. aureus, respectively. After 7 days of hospitalisation, S. aureus was isolated from 16.5% (16/97, 95% CI: 9.7%-25.4%) of the animals, including two methicillin-resistant S. aureus (MRSA) isolated from two pelicans. The two MRSA were identified as community-associated MRSA clones (mecA-positive ST1-IV and ST93-IV), suggesting direct or indirect transmission between humans and wildlife during hospitalisation may have occurred. This study highlighted Australian wildlife may be a potential reservoir for genetically diverse antimicrobial-resistant S. aureus. AMR surveillance including wildlife using a One Health approach may be required.

Animals↗

Trapped conformational states of semiquinone (D+*QB-*) formed by B-branch electron transfer at low temperature in Rhodobacter sphaeroides reaction centers.

The reaction center (RC) from Rhodobacter sphaeroides captures light energy by electron transfer between quinones QA and QB, involving a conformational gating step. In this work, conformational states of D+*QB-* were trapped (80 K) and studied using EPR spectroscopy in native and mutant RCs that lack QA in which QB was reduced by the bacteriopheophytin along the B-branch. In mutant RCs frozen in the dark, a light induced EPR signal due to D+*QB-* formed in 30% of the sample with low quantum yield (0.2%-20%) and decayed in 6 s. A small signal with similar characteristics was also observed in native RCs. In contrast, the EPR signal due to D+*QB-* in mutant RCs illuminated while freezing formed in approximately 95% of the sample did not decay (tau >107 s) at 80 K (also observed in the native RC). In all samples, the observed g-values were the same (g = 2.0026), indicating that all active QB-*'s were located in a proximal conformation coupled with the nonheme Fe2+. We propose that before electron transfer at 80 K, the majority (approximately 70%) of QB, structurally located in the distal site, was not stably reducible, whereas the minority (approximately 30%) of active configurations was in the proximal site. The large difference in the lifetimes of the unrelaxed and relaxed D+*QB-* states is attributed to the relaxation of protein residues and internal water molecules that stabilize D+*QB-*. These results demonstrate energetically significant conformational changes involved in stabilizing the D+*QB-* state. The unrelaxed and relaxed states can be considered to be the initial and final states along the reaction coordinate for conformationally gated electron transfer.

Cold Temperature↗

The study of state-selected ion-molecule reactions using the vacuum ultraviolet pulsed field ionization-photoion technique.

This paper presents the methodology to generate beams of ions in single quantum states for bimolecular ion-molecule reaction dynamics studies using pulsed field ionization (PFI) of atoms or molecules in high-n Rydberg states produced by vacuum ultraviolet (VUV) synchrotron or laser photoexcitation. Employing the pseudocontinuum high-resolution VUV synchrotron radiation at the Advanced Light Source as the photoionization source, PFI photoions (PFI-PIs) in selected rovibrational states have been generated for ion-molecule reaction studies using a fast-ion gate to pass the PFI-PIs at a fixed delay with respect to the detection of the PFI photoelectrons (PFI-PEs). The fast ion gate provided by a novel interleaved comb wire gate lens is the key for achieving the optimal signal-to-noise ratio in state-selected ion-molecule collision studies using the VUV synchrotron based PFI-PE secondary ion coincidence (PFI-PESICO) method. The most recent development of the VUV laser PFI-PI scheme for state-selected ion-molecule collision studies is also described. Absolute integral cross sections for state-selected H2+ ions ranging from v+ = 0 to 17 in collisions with Ar, Ne, and He at controlled translational energies have been obtained by employing the VUV synchrotron based PFI-PESICO scheme. The comparison between PFI-PESICO cross sections for the H2+(HD+)+Ne and H2+(HD+)+He proton-transfer reactions and theoretical cross sections based on quasiclassical trajectory (QCT) calculations and three-dimensional quantum scattering calculations performed on the most recently available ab initio potential energy surfaces is highlighted. In both reaction systems, quantum scattering resonances enhance the integral cross sections significantly above QCT predictions at low translational and vibrational energies. At higher energies, the agreement between experiment and quasiclassical theory is very good. The profile and magnitude of the kinetic energy dependence of the absolute integral cross sections for the H2+(v+ = 0-2,N+ = 1)+He proton-transfer reaction unambiguously show that the inclusion of Coriolis coupling is important in quantum dynamics scattering calculations of ion-molecule collisions.

Journal Article↗

Crystal structures of delta1-pyrroline-5-carboxylate reductase from human pathogens Neisseria meningitides and Streptococcus pyogenes.

L-proline is an amino acid that plays an important role in proteins uniquely contributing to protein folding, structure, and stability, and this amino acid serves as a sequence-recognition motif. Proline biosynthesis can occur via two pathways, one from glutamate and the other from arginine. In both pathways, the last step of biosynthesis, the conversion of delta1-pyrroline-5-carboxylate (P5C) to L-proline, is catalyzed by delta1-pyrroline-5-carboxylate reductase (P5CR) using NAD(P)H as a cofactor. We have determined the first crystal structure of P5CR from two human pathogens, Neisseria meningitides and Streptococcus pyogenes, at 2.0 angstroms and 2.15 angstroms resolution, respectively. The catalytic unit of P5CR is a dimer composed of two domains, but the biological unit seems to be species-specific. The N-terminal domain of P5CR is an alpha/beta/alpha sandwich, a Rossmann fold. The C-terminal dimerization domain is rich in alpha-helices and shows domain swapping. Comparison of the native structure of P5CR to structures complexed with L-proline and NADP+ in two quite different primary sequence backgrounds provides unique information about key functional features: the active site and the catalytic mechanism. The inhibitory L-proline has been observed in the crystal structure.

Amino Acid Sequence↗

Interactions between cytochrome c2 and the photosynthetic reaction center from Rhodobacter sphaeroides: the cation-pi interaction.

The cation-pi interaction between positively charged and aromatic groups is a common feature of many proteins and protein complexes. The structure of the complex between cytochrome c(2) (cyt c(2)) and the photosynthetic reaction center (RC) from Rhodobacter sphaeroides exhibits a cation-pi complex formed between Arg-C32 on cyt c(2) and Tyr-M295 on the RC [Axelrod, H. L., et al. (2002) J. Mol. Biol. 319, 501-515]. The importance of the cation-pi interaction for binding and electron transfer was studied by mutating Tyr-M295 and Arg-C32. The first- and second-order rates for electron transfer were not affected by mutating Tyr-M295 to Ala, indicating that the cation-pi complex does not greatly affect the association process or structure of the state active in electron transfer. The dissociation constant K(D) showed a greater increase when Try-M295 was replaced with nonaromatic Ala (3-fold) as opposed to aromatic Phe (1.2-fold), which is characteristic of a cation-pi interaction. Replacement of Arg-C32 with Ala increased K(D) (80-fold) largely due to removal of electrostatic interactions with negatively charged residues on the RC. Replacement with Lys increased K(D) (6-fold), indicating that Lys does not form a cation-pi complex. This specificity for Arg may be due to a solvation effect. Double mutant analysis indicates an interaction energy between Tyr-M295 and Arg-C32 of approximately -24 meV (-0.6 kcal/mol). This energy is surprisingly small considering the widespread occurrence of cation-pi complexes and may be due to the tradeoff between the favorable cation-pi binding energy and the unfavorable desolvation energy needed to bury Arg-C32 in the short-range contact region between the two proteins.

Amino Acids, Aromatic↗

Quinone (QB) reduction by B-branch electron transfer in mutant bacterial reaction centers from Rhodobacter sphaeroides: quantum efficiency and X-ray structure.

The photosynthetic reaction center (RC) from purple bacteria converts light into chemical energy. Although the RC shows two nearly structurally symmetric branches, A and B, light-induced electron transfer in the native RC occurs almost exclusively along the A-branch to a primary quinone electron acceptor Q(A). Subsequent electron and proton transfer to a mobile quinone molecule Q(B) converts it to a quinol, Q(B)H(2). We report the construction and characterization of a series of mutants in Rhodobacter sphaeroides designed to reduce Q(B) via the B-branch. The quantum efficiency to Q(B) via the B-branch Phi(B) ranged from 0.4% in an RC containing the single mutation Ala-M260 --> Trp to 5% in a quintuple mutant which includes in addition three mutations to inhibit transfer along the A-branch (Gly-M203 --> Asp, Tyr-M210 --> Phe, Leu-M214 --> His) and one to promote transfer along the B-branch (Phe-L181 --> Tyr). Comparing the value of 0.4% for Phi(B) obtained in the AW(M260) mutant, which lacks Q(A), to the 100% quantum efficiency for Phi(A) along the A-branch in the native RC, we obtain a ratio for A-branch to B-branch electron transfer of 250:1. We determined the structure of the most effective (quintuple) mutant RC at 2.25 A (R-factor = 19.6%). The Q(A) site did not contain a quinone but was occupied by the side chain of Trp-M260 and a Cl(-). In this structure a nonfunctional quinone was found to occupy a new site near M258 and M268. The implications of this work to trap intermediate states are discussed.

Bacteriochlorophylls↗

A pulsed-field ionization photoelectron secondary ion coincidence study of the H2+ (X,upsilon+=0-15,N+=1)+He proton transfer reaction.

The endothermic proton transfer reaction, H2+(upsilon+)+He-->HeH+ + H(DeltaE=0.806 eV), is investigated over a broad range of reactant vibrational levels using high-resolution vacuum ultraviolet to prepare reactant ions either through excitation of autoionization resonances, or using the pulsed-field ionization-photoelectron-secondary ion coincidence (PFI-PESICO) approach. In the former case, the translational energy dependence of the integral reaction cross sections are measured for upsilon+=0-3 with high signal-to-noise using the guided-ion beam technique. PFI-PESICO cross sections are reported for upsilon+=1-15 and upsilon+=0-12 at center-of-mass collision energies of 0.6 and 3.1 eV, respectively. All ion reactant states selected by the PFI-PESICO scheme are in the N+=1 rotational level. The experimental cross sections are complemented with quasiclassical trajectory (QCT) calculations performed on the ab initio potential energy surface provided by Palmieri et al. [Mol. Phys. 98, 1839 (2000)]. The QCT cross sections are significantly lower than the experimental results near threshold, consistent with important contributions due to resonances observed in quantum scattering studies. At total energies above 2 eV, the QCT calculations are in excellent agreement with the present results. PFI-PESICO time-of-flight (TOF) measurements are also reported for upsilon+=3 and 4 at a collision energy of 0.6 eV. The velocity inverted TOF spectra are consistent with the prevalence of a spectator-stripping mechanism.

Journal Article↗

Vibration-assisted electron tunneling in C140 transistors.

We measure electron tunneling in transistors made from C(140), a molecule with a mass-spring-mass geometry chosen as a model system to study electron-vibration coupling. We observe vibration-assisted tunneling at an energy corresponding to the stretching mode of C(140). Molecular modeling provides explanations for why this mode couples more strongly to electron tunneling than to the other internal modes of the molecule. We make comparisons between the observed tunneling rates and those expected from the Franck-Condon model.

Carbon↗

Extraperitoneal laparoscopic pyeloplasty for primary and secondary ureteropelvic junction obstruction.

PURPOSE: Greater experience with urological laparoscopy has lead to increasing interest in its use for reconstructive surgery, such as pyeloplasty. MATERIALS AND METHODS: A total of 124 cases of laparoscopic pyeloplasty were performed, of which 11 followed failed primary treatment done elsewhere, namely balloon dilation (3), endopyelotomy (3); open pyeloplasty (3), endopyelotomy plus balloon dilation (1) and open pyeloplasty plus balloon dilation (1). Nine patients had renal calculi. A 4 port, balloon dissecting, extraperitoneal laparoscopic approach was used in all except 1 patient, who had a horseshoe kidney, necessitating a transperitoneal approach. RESULTS: Operative time was 29 minutes longer in the secondary pyeloplasty group compared to primary cases (173.3 vs 144.0 minutes) but the conversion rate (0% vs 1.6%) and duration of postoperative hospitalization (2.8 nights each) were no greater. The complication rate was 3.6% and 9.1%, respectively. The success rate was 98.2% and 90.9% (p = 0.63) at a mean followup of 20.2 and 19.7 months, respectively. In the 9 patients with renal calculi a total of 18 calculi (94.7%) were successfully removed and the ureter was transposed medial to a crossing vessel in 50.0%. Trainee operating did not significantly prolong the procedure vs no training (162.0 vs 143.9 minutes, p = 0.06). CONCLUSIONS: Extraperitoneal laparoscopic dismembered pyeloplasty is capable of addressing all causes of ureteropelvic junction obstruction with excellent functional results and low morbidity, and with an operative time similar to that of open pyeloplasty. Secondary laparoscopic pyeloplasty does not increase hospitalization, conversion or complication rates.

Adolescent↗

Molecular requirements of the human nucleoside transporters hCNT1, hCNT2, and hENT1.

Concentrative nucleoside transporters (CNTs) and equilibrative nucleoside transporters (ENTs) are important in physiological and pharmacological activity and disposition of nucleosides and nucleoside drugs. A better understanding of the structural requirements of inhibitors for these transporters will aid in designing therapeutic agents. To define the relative and unified structural requirements of nucleoside analogs for interaction with hCNT1, hCNT2, and hENT1, we applied an array of structure-activity techniques. Unique pharmacophore models for each respective nucleoside transporter were generated. These models reveal that hCNT2 affinity is dominated by hydrogen bonding features, whereas hCNT1 and hENT1 displayed mainly electrostatic and steric features. Hydrogen bond formation over 3'-OH is essential for all nucleoside transporters. Inhibition of nucleoside transporters by a series of uridine and adenosine analogs and a variety of drugs was analyzed by comparative molecular field analysis. Cross-validated r2 (q2) values were 0.65, 0.52, and 0.74 for hCNT1, hCNT2, and hENT1, respectively. The predictive quality of the models was further validated by successful prediction of the inhibition of a set of test compounds. Addition of a hydroxyl group around the 2-position of purine (or 3-position of pyrimidine) may increase inhibition to hCNT2 transporter; addition of hydroxyl group around the 2,7-position of purine (or the 3,5-position of pyrimidine) would increase the inhibition to hENT1 transporter. Utilization of these models should assist the design of high-affinity nucleoside transporter inhibitors and substrates for both anticancer and antiviral therapy.

Biological Transport↗

Androgens and ovarian cancers.

Ovarian cancer is one of the most lethal malignancies in women. Numerous studies indicate that the steroid hormones have been implicated in the etiology and/or progression of epithelial ovarian cancer and support a role for androgens, including: 1) androgen receptor (AR) is present in primate ovaries at almost all stages of the menstrual cycle and involve folliculogenesis and ovulation; 2) high androgen serum levels show high risk of ovarian cancer and ovarian cancer occurring after menopause when the balance of ovarian steroid production shifts from estrogens to androgens; 3) ovarian cancer tissue shows a 90% AR positive rate and is associated with favorable outcomes; 4) androgens promote or inhibit ovarian cancer cell growth; 5) chemotherapy decreases androgen production from cancer cells. This review seeks to summarize our current understanding about the roles of androgens, AR and AR coregulators in the initiation and/or progression of ovarian cancers.

Androgen Antagonists↗

Recent advances in androgen receptor action.

Androgens, principally testosterone and 5 alpha-dihydrotestosterone, play critical roles in the development and growth of the male reproductive and nonreproductive systems. Their biological actions are mediated by the androgen receptor (AR), a ligand-dependent transcription factor, belonging to the nuclear receptor superfamily. These androgen-AR complexes interact with various factors (e.g. coactivators or corepressors) to modulate transcription of androgen target genes via specific DNA sequences. Many lines of evidence have also correlated AR with several mammalian disorders. Finally, recent advances in molecular biology have significantly impacted our knowledge of the role of AR in mammals. The aim of this review is to present recent emerging aspects of AR action.

Androgen-Insensitivity Syndrome↗

Motile chondrocytes from newborn calf: migration properties and synthesis of collagen II.

OBJECTIVE: To determine whether differentiated chondrocytes are motile. DESIGN: Calf articular chondrocytes isolated from six animals were cultured in spinner flasks and removed on days 3 and 7. Boyden chamber assays and time-lapse videomicroscopy were performed to monitor and quantify cell migration. A novel method for selectively harvesting and metabolically labeling the migrated cells was developed, based on cell movement to the underside of the Boyden chamber membranes. The 3H-collagen synthesized by these cells was purified and analyzed by SDS-PAGE and autoradiography either before or after cyanogen bromide cleavage. RESULTS: In Boyden chambers, locomotion of day 3 chondrocytes on fibronectin-coated membranes was approximately 3-fold higher than on bovine serum albumin-coated controls (39+/-15 vs 12+/-8 cells/mm(2), respectively (P=0.005)). Insulin-like growth factor-I (IGF-I, 10 ng/ml) was chemotactic, increasing motility to 87+/-16 cells/mm(-) (difference from fibronectin alone: P=0.0003). A similar response was observed for day 7 cells, but IGF-I activation was not as pronounced (P=0.055). The collagen patterns produced by the migrated cells closely resembled those of standard collagen type II, without any evidence of collagen I production. In videotracking experiments, motile cells attached on fibronectin exhibited typical lamellipodia and filopodia, and approximately 30% of attached cells were motile (speed >1 micro m/h and directional persistence >1h). Typical cell path lengths were 30-50 micro m, substantially greater than a full cell length displacement. CONCLUSION: A population of well-differentiated chondrocytes capable of matrix (COL II) synthesis are motile in vitro. This original finding opens new avenues to study the potential of motile cells for cartilage repair.

Animals↗

Segmental allotetraploidy and allelic interactions in buffelgrass (Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.) as revealed by genome mapping.

Linkage analyses increasingly complement cytological and traditional plant breeding techniques by providing valuable information regarding genome organization and transmission genetics of complex polyploid species. This study reports a genome map of buffelgrass (Pennisetum ciliare (L.) Link syn. Cenchrus ciliaris L.). Maternal and paternal maps were constructed with restriction fragment length polymorphisms (RFLPs) segregating in 87 F1 progeny from an intraspecific cross between two heterozygous genotypes. A survey of 862 heterologous cDNAs and gDNAs from across the Poaceae, as well as 443 buffelgrass cDNAs, yielded 100 and 360 polymorphic probes, respectively. The maternal map included 322 RFLPs, 47 linkage groups, and 3464 cM, whereas the paternal map contained 245 RFLPs, 42 linkage groups, and 2757 cM. Approximately 70 to 80% of the buffelgrass genome was covered, and the average marker spacing was 10.8 and 11.3 cM on the respective maps. Preferential pairing was indicated between many linkage groups, which supports cytological reports that buffelgrass is a segmental allotetraploid. More preferential pairing (disomy) was found in the maternal than paternal parent across linkage groups (55 vs. 38%) and loci (48 vs. 15%). Comparison of interval lengths in 15 allelic bridges indicated significantly less meiotic recombination in paternal gametes. Allelic interactions were detected in four regions of the maternal map and were absent in the paternal map.

Alleles↗

The androgen receptor in spermatogenesis.

Androgens are steroid hormones that are necessary for normal male phenotype expression, including the outward development of secondary sex characteristics as well as the initiation and maintenance of spermatogenesis. Many physiological actions of androgens are mediated by the androgen receptor (AR), a member of the nuclear receptor superfamily. AR functions as a ligand-dependent transcription factor, regulating expression of an array of target genes that are important in male pubertal development and fertility. In this review, the expression and necessity of AR in specific testicular cell types that are important in spermatogenesis will be discussed, and recent information obtained through the study of complete and cell type-specific AR null mouse models will be presented.

Animals↗