Expression of scFv-anti-CHIKV-E2 in Escherichia coli with chaperones Co-expression, and its functional assay by electrochemical immunosensor.

Single Chain Variable Fragment (scFv), a small fragment of antibody can be used to substitute the monoclonal antibody for diagnostic purposes. Production of scFv in Escherichia coli host has been a challenge due to the potential miss-folding and formation of inclusion bodies. This study aimed to express anti-CHIKV E2 scFv which previously designed specifically for Asian strains by co-expression of three chaperones that play a role in increasing protein solubility; GroEL, GroES, and Trigger Factor. The scFv and chaperones were expressed in Origami B E. coli host under the control of the T7 promoter, and purified using a Ni-NTA column. Functional assay of anti-CHIKV-E2 scFv was examined by electrochemical immunosensor using gold modified Screen Printed Carbon Electrode (SPCE), and characterized by differential pulses voltammetry (DPV) using K3[Fe(CN)6] redox system and scanning microscope electron (SEM). The experimental condition was optimized using the Box-Behnken design. The results showed that co-expression of chaperone increased the soluble scFv yield from 54.405 µg/mL to 220.097 µg/mL (~5×). Furthermore, scFv can be used to detect CHIKV-E2 in immunosensor electrochemistry with a detection limit of 0.74048 ng/mL and a quantification limit of 2,24388 ng/mL. Thus, the scFv-anti-CHIKV-E2 can be applied as a bioreceptor in another immunoassay method.

Nrf2 Signaling Pathway as a Key to Treatment for Diabetic Dyslipidemia and Atherosclerosis.

Diabetes mellitus (DM) is a chronic endocrine disorder that affects more than 20 million people in the United States. DM-related complications affect multiple organ systems and are a significant cause of morbidity and mortality among people with DM. Of the numerous acute and chronic complications, atherosclerosis due to diabetic dyslipidemia is a condition that can lead to many life-threatening diseases, such as stroke, coronary artery disease, and myocardial infarction. The nuclear erythroid 2-related factor 2 (Nrf2) signaling pathway is an emerging antioxidative pathway and a promising target for the treatment of DM and its complications. This review aims to explore the Nrf2 pathway's role in combating diabetic dyslipidemia. We will explore risk factors for diabetic dyslipidemia at a cellular level and aim to elucidate how the Nrf2 pathway becomes a potential therapeutic target for DM-related atherosclerosis.

The Effect of Ethanol on Lipid Nanoparticle Stabilization from a Molecular Dynamics Simulation Perspective.

Lipid nanoparticles (LNPs) have emerged as a promising delivery system, particularly for genetic therapies and vaccines. LNP formation requires a specific mixture of nucleic acid in a buffered solution and lipid components in ethanol. Ethanol acts as a lipid solvent, aiding the formation of the nanoparticle's core, but its presence can also affect LNP stability. In this study, we used molecular dynamics (MD) simulations to investigate the physicochemical effect of ethanol on LNPs and gain a dynamic understanding of its impact on the overall structure and stability of LNPs. Our results demonstrate that ethanol destabilizes LNP structure over time, indicated by increased root mean square deviation (RMSD) values. Changes in the solvent-accessible surface area (SASA), electron density, and radial distribution function (RDF) also suggest that ethanol affects LNP stability. Furthermore, our H-bond profile analysis shows that ethanol penetrates the LNP earlier than water. These findings emphasize the importance of immediate ethanol removal in lipid-based systems during LNP production to ensure stability.

New Triterpenoids from Lansium domesticum Corr. cv kokossan and Their Cytotoxic Activity.

Lansium domesticum Corr. is a member of the Meliaceae family that is widely spread in tropical and subtropical region of Asia and America. Traditionally, the fruit of this plant has been consumed because of its sweet taste. However, the fruit peels and the seeds of this plant have been rarely utilized. The previous chemical investigation of this plant showed the presence of secondary metabolites with many biological activities, including cytotoxic triterpenoid. Triterpenoids is a class of secondary metabolites which contain thirty carbon atoms in the main skeleton. The high modification of this type of compound, including the ring opening, highly oxygenated carbons, and the degradation of its carbon chain to give the nor-triterpenoid structure, is responsible for its cytotoxic activity. In this paper, we isolated and elucidated the chemical structure of two new onoceranoid triterpenes, kokosanolides E (1) and F (2), from the fruit peels of L. domesticum Corr., along with a new tetranortriterpenoid, kokosanolide G (3), from the seeds of L. domesticum Corr. The structural determination of compounds 1-3 was undertaken through FTIR spectroscopic analysis, 1D and 2D NMR, mass spectrometry, as well as through a comparison of the chemical shifts of the partial structures of compounds 1-3 with the literature data. The cytotoxic properties of compounds 1-3 were tested against MCF-7 breast cancer cells using the MTT assay. Moderate activity was shown by compounds 1 and 3, with IC50 values of 45.90 and 18.41 µg/mL, respectively, while compound 2 showed no activity (IC50 168.20 µg/mL). For the onoceranoid-type triterpene, the high symmetrical structure of compound 1 is presumably the reason for its better cytotoxic activity compared with that of compound 2. Compound 3 showed moderate activity, mainly because of the presence of the furan ring, which, based on the literature, gives better cytotoxic activity in a tetranortriterpenoid-type structure. The findings of three new triterpenoid compounds from L. domesticum indicate the significant value of this plant as a source of new compounds.

Theoretical and Experimental Studies on the Evidence of 1,3-ß-Glucan in Marennine of Haslea ostrearia.

Marennine, a blue pigment produced by the blue diatom Haslea ostrearia, is known to have some biological activities. This pigment is responsible for the greening of oysters on the West Coast of France. Other new species of blue diatom, H. karadagensis, H. silbo sp. inedit., H. provincialis sp. inedit, and H. nusantara, also produce marennine-like pigments with similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms present a commercial potential for the aquaculture, food, cosmetics, and health industries. Unfortunately, for a hundred years, the exact molecular structure of this bioactive compound has remained a mystery. A lot of hypotheses regarding the chemical structure of marennine have been proposed. The recent discovery of this structure revealed that it is a macromolecule, mainly carbohydrates, with a complex composition. In this study, some glycoside hydrolases were used to digest marennine, and the products were further analyzed using nuclear magnetic resonance (NMR) and mass spectroscopy (MS). The reducing sugar assay showed that marennine was hydrolyzed only by endo-1,3-ß-glucanase. Further insight into the structure of marennine was provided by the spectrum of 1H NMR, MS, a colorimetric assay, and a computational study, which suggest that the chemical structure of marennine contains 1,3-ß-glucan.

Quantifications and predictions of sectoral pollutants emissions in Nigeria from 1980 to 2050.

The fast-economic development and population growth in Nigeria have resulted in huge quantities of air pollutants emission which have implications on the environment. Detailed sectoral emission inventory to serve as the basis for policy formation to mitigate the condition is still lacking. This study builds detailed sectoral emission inventory using the emission factor approach to estimates various pollutant emissions from different sources. Five major sources of pollutant emissions were identified which include transportation, energy, municipal solid waste, wood fuel, and agricultural sectors. An increasing trend in emissions from 1980 to 2020 was observed for total emission of CO, NOx, PM2.5, PM10, SO2, NH3 and NMVOC in Nigeria that increased from 1 736-6 210; 143-338; 126-551; 171-717; 19-60; 4-28; and 471-1 587 Gg, respectively. Wood fuel, transportation, and municipal waste sectors are the major sources that contributed to 63%, 16%, and 15% of the total CO emission. Three mitigation scenarios for emission reduction for the future were analyzed. CO emission reductions of 38%, 24%, and 38% will be obtained from the liquefied petroleum gas (LPG) intervention, waste to energy (WTE) technology, and vehicle inspection and maintenance (VIM) policy scenarios, respectively, through to the year 2050.

Synthesis, Anticancer Activity, Structure-Activity Relationship, and Molecular Modeling Studies of α-Mangostin Derivatives as hERα Inhibitor.

α-Mangostin is one of the secondary metabolites in mangosteen pericarp, which has been reported to have anti-breast cancer activity. In our previous study, three α-mangostin derivatives were computationally designed as hERα antagonists. In this present study, the designed compounds were synthesized undergoing a benzoylation reaction between α-mangostin with three benzoyl chloride derivatives to produce three derivatives, namely, AMB-1, AMB-2, and AMB-10. The synthesized compounds were then evaluated for their antiproliferative activity against the MCF-7 breast cancer cell model with hERα as the protein target. The in vitro assay shows moderate activity (57-126 µM) for all derivatives. The dynamic behaviors of all ligands, including α-mangostin and 4-hydroxytamoxifen (4-OHT), were studied with 100 ns of MD simulation. The structure-activity relationship shows that although it does not entirely concord with the expected design, it can explain the trend of α-mangostin and its derivatives antiproliferative activities against MCF-7, which associates with hERα antagonism.

Haematological Features of White Rats (Rattus norvegicus) Infected with S. pyogenes and Administered with Probiotics (Yogurt).

This study aimed to study the inhibition activity of lactic acid bacteria probiotics deriving from Acehnese fermented Etawa goat's milk (yogurt) against Streptococcus pyogenes bacterial infection in rats (Rattus norvegicus). Haematological analysis of the rats' blood was performed on the following parameters: platelets, leukocytes, lymphocytes, neutrophils, and monocytes, where the data were further processed using ANOVA and Duncan's test with a confidence level of 95% (0.05). The results revealed that administering yogurt containing probiotics could reduce infections in the throats of rats caused by S. pyogenes. Based on the haematology examination, the probiotic yogurt could maintain the number of platelets, leukocytes, lymphocytes, neutrophils, and monocytes. Statistical significance was obtained when the infected rats were administered with a ±1.00 mL/day dose for seven days of treatment (p < 0.05).

Residual Interactions of LL-37 with POPC and POPE:POPG Bilayer Model Studied by All-Atom Molecular Dynamics Simulation.

LL-37 is a membrane-active antimicrobial peptide (AMP) that could disrupt the integrity of bacterial membranes due to its inherent cationic and amphipathic nature. Developing a shorter derivative of a long peptide such as LL-37 is of great interest, as it can reduce production costs and cytotoxicity. However, more detailed information about the residual interaction between LL-37 and the membrane is required for further optimization. Previously, molecular dynamics simulation using mixed all-atom and united-atom force fields showed that LL-37 could penetrate the bilayer membrane. This study aimed to perform all-atom molecular dynamics simulations, highlighting the residual interaction of LL-37 with the simplest model of the bacterial membrane, POPE:POPG (2:1), and compare its interaction with the POPC, which represents the eukaryotic membrane. The result showed leucine-leucine as the leading residues of LL-37 that first contact the membrane surface. Then, the cationic peptide of LL-37 started to penetrate the membrane by developing salt bridges between positively charged amino acids, Lys-Arg, and the exposed phosphate group of POPE:POPG, which is shielded in POPC. Residues 18 to 29 are suggested as the core region of LL-37, as they actively interact with the POPE:POPG membrane, not POPC. These results could provide a basis for modifying the amino acid sequence of LL-37 and developing a more efficient design for LL-37 derivatives.

Insights to antimicrobial resistance: heavy metals can inhibit antibiotic resistance in bacteria isolated from wastewater.

The alarming upsurge in the co-existence of heavy metal and antibiotic resistance may have a devastating impact on humans, animals, and the environment. Four metal-resistant bacteria were isolated from hospital effluents and industrial drain. Heavy metal resistance and antimicrobial resistance were examined in the isolates followed by identification through 16S rRNA gene sequencing. Delftia tsuruhatensis strain FK-01 and Carnobacterium inhibens strain FK-02 tolerated arsenic with maximal tolerated concentration (MTC) of 30 mM and 10 mM, respectively. Staphylococcus hominis strain FK-04 tolerated copper up to 4 mM and lead-resistant Raoultella ornithinolytica strain FK-05 exhibited tolerance to 1 mM lead. The growth kinetics of bacteria were monitored in the presence of metals and the following antibiotics, tetracycline, chloramphenicol, and kanamycin. The presence of arsenate significantly enhanced tetracycline resistance in C. inhibens. Heavy metal-induced antibiotic resistance was also observed in S. hominis and R. ornithinolytica, against chloramphenicol and tetracycline respectively. D. tsuruhatensis showed resistance to kanamycin but when grown in the presence of arsenic and kanamycin, bacteria lost resistance to the antibiotic. Therefore, it is suggested that the novel arsenate-resistant strain Delftia tsuruhatensis FK-01 has a unique ability to inhibit antimicrobial resistance that can be harnessed in bioremediation.

Molecular Dynamics of Cobalt Protoporphyrin Antagonism of the Cancer Suppressor REV-ERBß.

Nuclear receptor REV-ERBß is an overexpressed oncoprotein that has been used as a target for cancer treatment. The metal-complex nature of its ligand, iron protoporphyrin IX (Heme), enables the REV-ERBß to be used for multiple therapeutic modalities as a photonuclease, a photosensitizer, or a fluorescence imaging agent. The replacement of iron with cobalt as the metal center of protoporphyrin IX changes the ligand from an agonist to an antagonist of REV-ERBß. The mechanism behind that phenomenon is still unclear, despite the availability of crystal structures of REV-ERBß in complex with Heme and cobalt protoporphyrin IX (CoPP). This study used molecular dynamic simulations to compare the effects of REV-ERBß binding to Heme and CoPP, respectively. The initial poses of Heme and CoPP in complex with agonist and antagonist forms of REV-ERBß were predicted using molecular docking. The binding energies of each ligand were calculated using the MM/PBSA method. The computed binding affinity of Heme to REV-ERBß was stronger than that of CoPP, in agreement with experimental results. CoPP altered the conformation of the ligand-binding site of REV-ERBß, disrupting the binding site for nuclear receptor corepressor, which is required for REV-ERBß to regulate the transcription of downstream target genes. Those results suggest that a subtle change in the metal center of porphyrin can change the behavior of porphyrin in cancer cell signaling. Therefore, modification of porphyrin-based agents for cancer therapy should be conducted carefully to avoid triggering unfavorable effects.

Source identification and health risk assessments of polycyclic aromatic hydrocarbons in settled dusts from different population density areas of Ilorin, Nigeria.

Polycyclic aromatic hydrocarbons (PAHs) have attracted significant attention in recent times on account of their reasonably high environmental burden and extreme toxicity. Samples of indoor dusts were obtained daily over a period of 2 weeks from 10 residences located within low, medium, and high density residential areas of Ilorin City. The concentration levels, potential sources, and cancer health risks of sixteen polycyclic aromatic hydrocarbons (PAHs) were investigated using gas chromatography/mass spectrometry. PAHs total concentrations varied from 3.95 ± 0.19 to 8.70 ± 0.43 µg/g with arithmetic mean of 6.09 ± 0.46 µg/g. Fluoranthene was the most dominant PAHs congener. High molecular weight (HMW) PAHs (4-6 rings) were the most prevalent PAHs and were responsible for 79.29% of total PAHs in sampled residences. Chrysene (Chry) was the most abundant compound among the 7 carcinogenic PAHs (CPAHs). Moreover, diagnostic ratios and positive matrix factorization (PMF) employed to apportion PAHs suggested that indoor dusts originated from indoor activities and infiltrating outdoor air pollutants. Diagnostic ratios revealed that PAHs are from mixed sources which include coal/wood combustion, non-traffic and traffic emissions, petroleum, petrogenic (gasoline), and petroleum combustion. Similarly, positive matrix factorization (PMF) model suggested five sources (factors) were responsible for PAHs in indoor dusts comprised of petroleum combustion and traffic emissions (60.05%), wood and biomass combustion emissions (20.84%), smoke from cooking, incense burning and tobacco (4.17%), gasoline combustion from non-traffic sources (13.89%), and emissions from coal burning and electronic devices (1.05%). The incremental lifetime carcinogenic risks (ILCR) of PAHs in adults and children estimated by applying benzo(a)pyrene (BaP) equivalent were within the satisfactory risk limits in Ilorin. Indoor PAHs emissions in Ilorin residences could be monitored and controlled by using data provided in this study.

Arylamide as Potential Selective Inhibitor for Matrix Metalloproteinase 9 (MMP9): Design, Synthesis, Biological Evaluation, and Molecular Modeling.

Previous studies have reported that compounds bearing an arylamide linked to a heterocyclic planar ring have successfully inhibited the hemopexin-like domain (PEX9) of matrix metalloproteinase 9 (MMP9). PEX9 has been suggested to be more selectively targeted than MMP9's catalytic domain in a degrading extracellular matrix under some pathologic conditions, especially in cancer. In this study, we aim to synthesize and evaluate 10 arylamide compounds as MMP9 inhibitors through an enzymatic assay as well as a cellular assay. The mechanism of inhibition for the most active compounds was investigated via molecular dynamics simulation (MD). Molecular docking was performed using AutoDock4.0 with PEX9 as the protein model to predict the binding of the designed compounds. The synthesis was carried out by reacting aniline derivatives with 3-bromopropanoyl chloride using pyridine as the catalyst at room temperature. The MMP9 assay was conducted using the FRET-based MMP9 kits protocol and gelatin zymography assay. The cytotoxicity assay was done using the MTT method, and the MD simulation was performed using AMBER16. Assay on MMP9 demonstrated activities of three compounds (2, 7, and 9) with more than 50% inhibition. Further inhibition on MMP9 expressed by 4T1 showed that two compounds (7 and 9) inhibited its gelatinolytic activity more than 50%. The cytotoxicity assay against 4T1 cells results in the inhibition of the cell growth with an EC50 of 125 µM and 132 µM for 7 and 9, respectively. The MD simulation explained a stable interaction of 7 and 9 in PEX9 at 100 ns with a free energy of binding of -8.03 kcal/mol and -6.41 kcal/mol, respectively. Arylamides have potential effects as selective MMP9 inhibitors in inhibiting breast cancer cell progression.

Emission factors of some common grass species in West Africa.

Frequent burnings occurring in the grasslands of the West African region during the dry seasons largely contribute to emissions of trace gases and particulates being released into the ambient environment, which has significantly impacted both regional and global climate patterns. Burning potentials of forty different grassland biomes were examined by determining their Net Heating Value (NHV) and Total Organic Carbon (TOC). Simulations of the field operations which involve open burning were performed in the laboratory using a fabricated combustion chamber for the determination of emission factors. Particulates were collected using Whatman quartz fibre filters and analyzed gravimetrically. Emissions of gaseous pollutants from open burning of these common grass species were measured with portable devices. The values of the NHV and TOC of the grass species ranged from 15,022.19 to 18,181.84 kJ/kg and 21.14 to 55.62%, respectively. The average Emission Factors (EFs) obtained for carbon dioxide (CO2), carbon monoxide (CO), sulphur dioxide (SO2), nitrogen dioxide (NO2), volatile organic compounds (VOC), and PM2.5 are 1465.55 g/kg, 40.99 g/kg, 0.39 g/kg, 0.02 g/kg, 7.78 g/kg, and 6.00 g/kg, respectively. The study has shown that Digitaria nuda, Digitaria eriantha, Panicum subalbidum, Paspalum polystratchyum, and Perotis indica have the highest emission factors for CO2, CO, SO2, NO2, VOC, and PM2.5, respectively. The result obtained would help in the quantification of the global warming forcing on the climate in the West African region from grassland burnings. The results will potentially serve as additional information for emission inventories and basis for the formulation of mitigation strategies.

Utilization of Leucaena leucocephala and Gliricidia sepium as supplements by goats fed Panicum maximum basal diet.

The objective of this study was to determine the effect of supplementations of Leucaena leucocephala or Gliricidia sepium by goats fed old Panicum maximum basal diets on their dry matter and nutrient intake, digestibility, and N balance. The experiment used four goats weighing of around 12 kg fed four kinds of diet in Latin square design. The diets were 1 young guinea grass, 2 old guinea grass, 3 old guinea grass 60% + Leucaena 40%, 4 old guinea grass 60% + Gliricidia 40%. Results of the study revealed that dry matter and crude protein intake, dry matter, crude protein, and NDF digestibility were significantly higher when the goats fed young grass, Leucaena- or Gliricidia-supplemented old guinea grass than goats fed old guinea grass only. N retained as proportion of ingested N by the goats fed young guinea grass, and old guinea grass supplemented with Leucaena or Gliricidia were positive (63.95, 68.85, and 67.88%, respectively), while the goats fed old guinea grass was negative (- 55.49%). Intake of young grass resulted in greater N excreted via urine while supplementation with Leucaena or Gliricidia resulted in greater N excreted via feces. It can be concluded that supplementation of Leucaena or Gliricidia is very beneficial to increase dry matter and nutrients intake, digestibility, and N retention in goats fed Panicum maximum basal diets. An important finding of this work is that young forage consumed by goats allows to reach important performances and that supplementation is not essential.

Microbiology and clinical characteristics of acute cholangitis with their impact on mortality; a retrospective cross sectional study.

OBJECTIVE: To evaluate microbiological and clinical characteristics of acute cholangitis along with their impact on mortality, and to compare the role of early versus late biliary drainage in the management of cholangitis. METHODS: The retrospective study was conducted at the Shaukat Khanum Memorial Cancer Hospital Research Centre, Lahore, Pakistan, and comprised records of all patients presenting with acute cholangitis from June, 2012, to June, 2017. The risk factors, presence of bacteremia, resistance pattern of microbial pathogens and severity were assessed according to Tokyo guidelines in addition to associated mortality and recurrence at 3 months. Data was analysed using SPSS 20. RESULTS: Of the 230 patients, 137(59.6%) were male. The overall mean age was 56±13 years. The most common isolated organism was Escherichia coli 54(70.1%). Clinical severity (p=0.001), late biliary drainage (p=0.001) and use of multiple stents (p=0.03) were associated with increased mortality. However, in multivariable analysis, only high body mass index (p=0.01) and Tokyo severity grades II (p=0.04) and III (p=0.001) were significant factors associated with mortality. CONCLUSIONS: Early identification of risk factors, administration of appropriate antibiotics and establishing early biliary drainage were found to be the key management steps to reduce cholangitis-related mortality.

Exploration of charge states of balanol analogues acting as ATP-competitive inhibitors in kinases.

BACKGROUND: (-)-Balanol is an ATP mimic that inhibits protein kinase C (PKC) isozymes and cAMP-dependent protein kinase (PKA) with limited selectivity. While PKA is a tumour promoter, PKC isozymes act as tumour promoters or suppressors, depending on the cancer type. In particular, PKCε is frequently implicated in cancer promotion, making it a potential target for anticancer drugs. To improve isozyme selectivity of balanol, exhaustive structural and activity relationship (SAR) studies have been performed in the last two decades, but with limited success. More recently, fluorination on balanol has shown improved selectivity for PKCε, although the fluorine effect is not yet clearly understood. Understanding the origin to this fluorine-based selectivity will be valuable for designing better balanol-based ATP mimicking inhibitors. Computational approaches such as molecular dynamics (MD) simulations can decipher the fluorine effect, provided that correct charges have been assigned to a ligand. Balanol analogues have multiple ionisable functional groups and the effect of fluorine substitutions on the exact charge state of each analogue bound to PKA and to PKCε needs to be thoroughly investigated in order to design highly selective inhibitors for therapeutic applications. RESULTS: We explored the charge states of novel fluorinated balanol analogues using MD simulations. For different potential charge states of these analogues, Molecular Mechanics Generalized Born Surface Area (MMGBSA) binding energy values were computed. This study suggests that balanol and the most potent fluorinated analogue (5S fluorine substitution on the azepane ring), have charges on the azepane ring (N1), and the phenolic (C6''OH) and the carboxylate (C15''O2H) groups on the benzophenone moiety, when bound to PKCε as well as PKA. CONCLUSIONS: To the best our knowledge, this is the first study showing that the phenolate group is charged in balanol and its analogues binding to the ATP site of PKCε. Correct charge assignments of ligands are important to obtain predicted binding energy values from MD simulations that reflect experimental values. Both fluorination and the local enzymatic environment of the ATP site can influence the exact charge states of balanol analogues. Overall, this study is highly valuable for further rational design of potent balanol analogues selective to PKCε.

H274Y's Effect on Oseltamivir Resistance: What Happens Before the Drug Enters the Binding Site.

Increased reports of oseltamivir (OTV)-resistant strains of the influenza virus, such as the H274Y mutation on its neuraminidase (NA), have created some cause for concern. Many studies have been conducted in the attempt to uncover the mechanism of OTV resistance in H274Y NA. However, most of the reported studies on H274Y focused only on the drug-bound system, so the direct effects of the mutation on NA itself prior to drug binding still remain unclear. Therefore, molecular dynamics simulations of NA in apo form, followed by principal component analysis and interaction energy calculations, were performed to investigate the structural changes of the NA binding site as a result of the H274Y mutation. It was observed that the disruption of the NA binding site due to the H274Y mutation was initiated by the repulsive effect of Y274 on the 250-loop, which in turn altered the hydrogen-bonding network around residue 274. The rotated W295 side chain caused the upward movement of the 340-loop. Consequently, sliding box docking results suggested that the binding pathway of OTV was compromised because of the disruption of this binding site. This study also highlighted the importance of the functional group at C6 of the sialic acid mimicry. It is hoped that these results will improve the understanding of OTV resistance and shed some light on the design of a novel anti-influenza drug.

A methodology for on-board CBCT imaging dose using optically stimulated luminescence detectors.

Cone-beam computed tomography CBCT systems are used in radiation therapy for patient alignment and positioning. The CBCT imaging procedure for patient setup adds substantial radiation dose to patient's normal tissue. This study pre-sents a complete procedure for the CBCT dosimetry using the InLight optically-stimulated-luminescence (OSL) nanoDots. We report five dose parameters: the mean slice dose (DMSD); the cone beam dose index (CBDIW); the mean volume dose (DMVD); point-dose profile, D(FOV); and the off-field Dose. In addition, CBCT skin doses for seven pelvic tumor patients are reported. CBCT-dose mea-surement was performed on a custom-made cylindrical acrylic body phantom (50cm length, 32cm diameter). We machined 25 circular disks (2 cm thick) with grooves and holes to hold OSL-nanoDots. OSLs that showed similar sensitivities were selected and calibrated against a Farmer-type ionization-chamber (0.6 CT) before being inserted into the grooves and holes. For the phantom scan, a standard CBCT-imaging protocol (pelvic sites: 125 kVp, 80 mA and 25 ms) was used. Five dose parameters were quantified: DMSD, CBDIW, DMVD, D(FOV), and the off-field dose. The DMSD for the central slice was 31.1 ± 0.85 mGy, and CBDIW was 34.5± 0.6 mGy at 16cm FOV. The DMVD was 25.6 ± 1.1 mGy. The off-field dose was 10.5 mGy. For patients, the anterior and lateral skin doses attributable to CBCT imaging were 39.04 ± 4.4 and 27.1 ± 1.3 mGy, respectively.OSL nanoDots were convenient to use in measuring CBCT dose. The method of selecting the nanoDots greatly reduced uncertainty in the OSL measurements. Our detailed calibration procedure and CBCT dose measurements and calculations could prove useful in developing OSL routines for CBCT quality assessment, which in turn gives them the property of high spatial resolution, meaning that they have the potential for measurement of dose in regions of severe dose-gradients.