Contemporary Approaches to Addressing HIV Prevention Needs Among Sexual and Gender Diverse Individuals in Kazakhstan

Renewed efforts are needed to address rapidly rising HIV incidence among sexual and gender diverse (SGD) individuals—particularly cisgender gay, bisexual, and other men (MSM) and transgender and nonbinary individuals (TSM) who have sex with men—in Kazakhstan. Intervention research is uniquely positioned to advance HIV prevention through surveying factors shaping the HIV epidemic among MSM and TSM in Kazakhstan, developing and testing the effects of an HIV prevention intervention, and assessing overall social impacts of conducting research. This research proceeded to describe strategies and lessons learned during implementation of a stepped wedge clinical trial of an intervention designed to increase the number of MSM and TSM in the HIV care continuum in Kazakhstan cities of Almaty, Shymkent, and Nur-Sultan. Thus, this three-paper dissertation aimed to: (1) identify psychosocial factors associated with lifetime, past-12-month, and past-6-month HIV testing among a sample of MSM and TSM enrolled in the clinical trial; (2) describe the process of implementing remote training of facilitators for remotely delivering the HIV preventive intervention; and (3) assess social impacts of participating in the clinical trial. MSM and TSM from the study cities were recruited into the clinical trial and administered a structured behavioral survey at their primary visit and at follow-up visits every six months thereafter. After a period of no intervention implementation (‘pre-implementation period’), the intervention was implemented sequentially every six months in the study cities. Among 304 MSM and TSM enrolled in the clinical trial during the pre-implementation period, lifetime and past-12-month HIV testing were positively associated with polydrug use and negatively with sexual transmission HIV risk, and past-6-month HIV testing was negatively associated with sexual risk. The process of developing and implementing remote training of facilitators was guided by a protocol outlining phases involving formative assessment and planning, fundamentals training, and feedback loop and technical assistance. Out of 627 MSM and TSM who completed their primary assessment during the clinical trial, 579 (92%) returned for at least one follow-up visit; of these individuals, 88% reported at least one positive social impact, while 2% reported at least one negative social impact. Findings underscore the value of expanding access to substance use treatment for HIV prevention among MSM and TSM in Kazakhstan, the viability of remote training of facilitators for remote intervention delivery, and the feasibility of conducting HIV prevention research involving MSM and TSM with many benefits and few risks

Dynamical mean-field theory of Hubbard-Holstein model at half-filling: Zero temperature metal-insulator and insulator-insulator transitions

We study the Hubbard-Holstein model, which includes both the electron-electron and electron-phonon interactions characterized by $U$ and $g$, respectively, employing the dynamical mean-field theory combined with Wilson's numerical renormalization group technique. A zero temperature phase diagram of metal-insulator and insulator-insulator transitions at half-filling is mapped out which exhibits the interplay between $U$ and $g$. As $U$ ($g$) is increased, a metal to Mott-Hubbard insulator (bipolaron insulator) transition occurs, and the two insulating states are distinct and can not be adiabatically connected. The nature of and transitions between the three states are discussed.Comment: 5 pages, 4 figures. Submitted to Physical Review Letter

CHANGE IN MUSCLE ACITIVITY DURING SKIING USING 2 SIMULATORS

This research analyzed the change in lower muscle activity during skiing using two simulators. Participants in this study were 8 male adults who held a certificate issued by the Korea Ski Instructors Association. Kinematic factors when skiing on two types of simulators were compared and analyzed. Most subjects maintained a higher value of %MVIC while repeating the positions on the Pro Ski Simulator than Skiers Edge. Efficient skiing on ski-simulators requires the maximum use of skiing time on each foot and the ability to quickly flex and extend the hip and the knee joints. Furthermore, differences between individuals exist depending on the level of ski technique and the type of simulator and it is believed that the development of an additional adjustment apparatus in the existing equipment is necessary to enable safe and effective motions

Crystal Structure of Dps-1, a Functionally Distinct Dps Protein from Deinococcus radiodurans

DNA protection during starvation (Dps) proteins play an important role in protecting cellular macromolecules from damage by reactive oxygen species (ROS). Unlike most orthologs that protect DNA by a combination of DNA binding and prevention of hydroxyl radical formation by ferroxidation and sequestration of iron, Dps-1 from the radiation-resistant Deinococcus radiodurans fails to protect DNA from hydroxyl radical-mediated cleavage through a mechanism inferred to involve continuous release of iron from the protein core. To address the structural basis for this unusual release of Fe2+, the crystal structure of D. radiodurans Dps-1 was determined to 2.0 Å resolution. Two of four strong anomalous signals per protein subunit correspond to metal-binding sites within an iron-uptake channel and a ferroxidase site, common features related to the canonical functions of Dps homologs. Similar to Lactobacillus lactis Dps, a metal-binding site is found at the N-terminal region. Unlike other metal sites, this site is located at the base of an N-terminal coil on the outer surface of the dodecameric protein sphere and does not involve symmetric association of protein subunits. Intriguingly, a unique channel-like structure is seen featuring a fourth metal coordination site that results from 3-fold symmetrical association of protein subunits through α2 helices. The presence of this metal-binding site suggests that it may define an iron-exit channel responsible for the continuous release of iron from the protein core. This interpretation is supported by substitution of residues involved in this ion coordination and the observation that the resultant mutant protein exhibits significantly attenuated iron release. Therefore, we propose that D. radiodurans Dps-1 has a distinct iron-exit channel. © 2006 Elsevier Ltd. All rights reserved

Molecular basis of the fructose-2,6-bisphosphatase reaction of PFKFB3: Transition state and the C-terminal function

The molecular basis of fructose-2,6-bisphosphatase (F-2,6-P 2ase) of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) was investigated using the crystal structures of the human inducible form (PFKFB3) in a phospho-enzyme intermediate state (PFKFB3-P•F-6-P), in a transition state-analogous complex (PFKFB3•AlF 4), and in a complex with pyrophosphate (PFKFB3•PP i) at resolutions of 2.45, 2.2, and 2.3 Å, respectively. Trapping the PFKFB3-P•F-6-P intermediate was achieved by flash cooling the crystal during the reaction, and the PFKFB3•AlF 4 and PFKFB3•PP i complexes were obtained by soaking. The PFKFB3•AlF 4 and PFKFB3•PP i complexes resulted in removing F-6-P from the catalytic pocket. With these structures, the structures of the Michaelis complex and the transition state were extrapolated. For both the PFKFB3-P formation and break down, the phosphoryl donor and the acceptor are located within ∼5.1 Å, and the pivotal point 2-P is on the same line, suggesting an in-line transfer with a direct inversion of phosphate configuration. The geometry suggests that NE2 of His253 undergoes a nucleophilic attack to form a covalent N-P bond, breaking the 2O-P bond in the substrate. The resulting high reactivity of the leaving group, 2O of F-6-P, is neutralized by a proton donated by Glu322. Negative charges on the equatorial oxygen of the transient bipyramidal phosphorane formed during the transfer are stabilized by Arg252, His387, and Asn259. The C-terminal domain (residues 440-446) was rearranged in PFKFB3•PP i, implying that this domain plays a critical role in binding of substrate to and release of product from the F-2,6-P 2ase catalytic pocket. These findings provide a new insight into the understanding of the phosphoryl transfer reaction. © 2011 Wiley Periodicals, Inc

A Direct Substrate-Substrate Interaction Found in the Kinase Domain of the Bifunctional Enzyme, 6-Phosphofructo-2-kinase/Fructose-2,6-bisphosphatase

To understand the molecular basis of a phosphoryl transfer reaction catalyzed by the 6-phosphofructo-2-kinase domain of the hypoxia-inducible bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB3), the crystal structures of PFKFB3{radical dot}AMPPCP{radical dot}fructose-6-phosphate and PFKFB3{radical dot}ADP{radical dot}phosphoenolpyruvate complexes were determined to 2.7 Å and 2.25 Å resolution, respectively. Kinetic studies on the wild-type and site-directed mutant proteins were carried out to confirm the structural observations. The experimentally varied liganding states in the active pocket cause no significant conformational changes. In the pseudo-substrate complex, a strong direct interaction between AMPPCP and fructose-6-phosphate (Fru-6-P) is found. By virtue of this direct substrate-substrate interaction, Fru-6-P is aligned with AMPPCP in an orientation and proximity most suitable for a direct transfer of the γ-phosphate moiety to 2-OH of Fru-6-P. The three key atoms involved in the phosphoryl transfer, the β,γ-phosphate bridge oxygen atom, the γ-phosphorus atom, and the 2-OH group are positioned in a single line, suggesting a direct phosphoryl transfer without formation of a phosphoenzyme intermediate. In addition, the distance between 2-OH and γ-phosphorus allows the γ-phosphate oxygen atoms to serve as a general base catalyst to induce an associative phosphoryl transfer mechanism. The site-directed mutant study and inhibition kinetics suggest that this reaction will be catalyzed most efficiently by the protein when the substrates bind to the active pocket in an ordered manner in which ATP binds first. © 2007 Elsevier Ltd. All rights reserved

The antifungal activity and membrane-disruptive action of dioscin extracted from Dioscorea nipponica

AbstractDioscin is a kind of steroidal saponin isolated from the root bark of wild yam Dioscorea nipponica. We investigated the antifungal effect of dioscin against different fungal strains and its antifungal mechanism(s) in Candida albicans cells. Using the propidium iodide assay and calcein-leakage measurement, we confirmed that dioscin caused fungal membrane damage. Furthermore, we evaluated the ability of dioscin to disrupt the plasma membrane potential, using 3,3′-dipropylthiadicarbocyanine iodide [DiSC3(5)] and bis-(1,3-dibarbituric acid)-trimethine oxanol [DiBAC4(3)]. Cells stained with the dyes had a significant increase in fluorescent intensity after exposure to dioscin, indicating that dioscin has an effect on the membrane potential. To visualize the effect of dioscin on the cell membrane, we synthesized rhodamine-labeled giant unilamellar vesicles (GUVs) mimicking the outer leaflet of the plasma membrane of C. albicans. As seen in the result, the membrane disruptive action of dioscin caused morphological change and rhodamine leakage of the GUVs. In three-dimensional contour-plot analysis using flow cytometry, we observed a decrease in cell size, which is in agreement with our result from the GUV assay. These results suggest that dioscin exerts a considerable antifungal activity by disrupting the structure in membrane after invading into the fungal membrane, resulting in fungal cell death

Crystal structure of the hypoxia-inducible form of 6-phosphofructo-2- kinase/fructose-2,6-bisphosphatase (PFKFB3): A possible new target for cancer therapy

The hypoxia-inducible form of 6-phosphofructo-2-kinase/fructose-2,6- bisphosphatase (PFKFB3) plays a crucial role in the progression of cancerous cells by enabling their glycolytic pathways even under severe hypoxic conditions. To understand its structural architecture and to provide a molecular scaffold for the design of new cancer therapeutics, the crystal structure of the human form was determined. The structure at 2.1 Å resolution shows that the overall folding and functional dimerization are very similar to those of the liver (PFKFB1) and testis (PFKFB4) forms, as expected from sequence homology. However, in this structure, the N-terminal regulatory domain is revealed for the first time among the PFKFB isoforms. With a β-hairpin structure, the N terminus interacts with the 2-Pase domain to secure binding of fructose-6-phosphate to the active pocket, slowing down the release of fructose-6-phosphate from the phosphoenzyme intermediate product complex. The C-terminal regulatory domain is mostly disordered, leaving the active pocket of the fructose-2,6-bisphosphatase domain wide open. The active pocket of the 6-phosphofructo-2-kinase domain has a more rigid conformation, allowing independent bindings of substrates, fructose-6-phosphate and ATP, with higher affinities than other isoforms. Intriguingly, the structure shows an EDTA molecule bound to the fructose-6-phosphate site of the 6-phosphofructo-2-kinase active pocket despite its unfavorable liganding concentration, suggesting a high affinity. EDTA is not removable from the site with fructose-6-P alone but is with both ATP and fructose-6-P or with fructose-2,6-bisphosphate. This finding suggests that a molecule in which EDTA is covalently linked to ADP is a good starting molecule for the development of new cancer-therapeutic molecules. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc