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To understand how the risk of different assisted reproductive technology (ART) failure types varies by ethnic group and explore the role of mediation by maternal age and suspected etiology.An observational study of 48,750 women who undertook treatment with ART in the United Kingdom between January 2017 and December 2018.The Human Fertilisation and Embryology Authority national ART registry of the United Kingdom.Women who commenced a first cycle of ART for the purpose of primary fresh embryo transfer using their own oocytes were included.Maternal ethnic group.The ART failure types were modeled on the maternal ethnic group using the Poisson regression to produce relative risks (RRs) with 95% confidence intervals. The potential indirect effects of maternal age and etiology of subfertility were estimated, and the RRs with 95% confidence intervals were produced.Black women were at greater risk of treatment failure with respect to live birth than women who were white: cycle cancellation, RR of 2.15 (1.78-2.62); failed fertilization, RR of 2.36 (1.90-2.93); unintended freeze-all, RR of 1.71 (1.43-2.05); failed implantation, RR of 1.23 (1.12-1.34); and pregnancy loss, RR of 1.38 (1.15-1.64). Women who were Asian were at moderately increased risk: RRs of 1.31 (1.17-1.47), 1.60 (1.42-1.80), 1.25 (1.14-1.38), 1.11 (1.07-1.16), and 1.13 (1.03-1.23), across the same outcomes, respectively. Inequality may have been reduced had women of all ethnicities initiated treatment at the same age.Black women were at greatest risk of all failure types, and women who were Asian were at intermediate risk compared with women who were white. Some of the risks among women who were black may be mediated by maternal age.

Objective\ud Predicting the outcomes of assisted reproductive technology (ART) treatments is desirable, but adopting prediction models into clinical practice remains limited. We aimed to review available prediction models for ART treatments by conducting a systematic review of the literature to identify the best performing models for their accuracy, generalisability and applicability.\ud \ud Evidence review\ud We searched electronic databases (MEDLINE, EMBASE, and CENTRAL) until June 2020. We included studies reporting on the development or evaluation of models predicting the reproductive outcomes before (pre-ART) or after starting (Intra-ART) treatment in couples undergoing any ART treatment. We evaluated the models’ discrimination, calibration, type of validation, and any implementation tools for clinical practice.\ud \ud Results\ud We included 69 cohort studies reporting on 120 unique prediction models. Half the studies reported on pre-ART (48%) and half on intra-ART (56%) prediction models. The commonest predictors used were maternal age (90%), tubal factor subfertility (50%), and embryo quality (60%).\ud \ud Only fourteen models were externally-validated (14/120, 12%) including eight pre-ART models (Templeton, Nelson, LaMarca, McLernon, Arvis, and the Stolwijk A/I,C,II models), and five intra-ART models (Cai, Hunault, van Loendersloot, Meijerink, Stolwijk B, and the McLernon post-treatment model) with a reported c-statistics ranging from 0.50 to 0.78. Ten of these models provided implementation tools for clinical practice with only two reported online calculators.\ud \ud Conclusion\ud We identified externally validated prediction models that could be used to advise couples undergoing ART treatments on their reproductive outcomes. The quality of available models remains limited and more research is needed to improve their generalizability and applicability into clinical practice.

The principle of proportionality protects civilians and civilian objects against expected incidental harm from an attack that is excessive to the military advantage anticipated from the attack. However, despite its status as a fundamental norm of IHL, key terms are not defined in relevant treaties nor benefit from critical judicial explanation. This has caused challenges for both academics and military commanders alike in explaining and applying the test for proportionality. The article expands upon two points that were raised and generated interesting discussion at The Second Israel Defense Forces International Conference on the Law of Armed Conflict during a panel that dealt with contemporary issues in proportionality. Those two issues are: a. What does the “reasonable military commander” standard for assessing proportionality entail? b. Should “reverberating effects” (i.e., collateral effects that are only expected to materialize in the long term) be accounted for as part of the assessment of collateral damage?

In civil engineering, composite construction has become increasingly widespread due to the improvement of mechanical properties it provides. Recently, research on the development of post installed or retrofitted shear connectors has been conducted. The benefits of this are the ease of installation and the ease of removal for decommissioning structures at the end of their life. Most of the research in this area is concerned with modified versions of welded shear studs or various threaded rod and nut configurations and refers to only one type of shear connector. Therefore, the suitability of the proposed models across differing shear connection types is unknown. In the first of two companion papers an experimental study has been undertaken to ascertain the dynamic behaviour of identical steel–concrete composite beams with differing shear connection systems. Two blind bolt connector types were used as shear connection systems in steel–concrete composite beams. Alongside these, a welded shear stud specimen, and a non-composite specimen were tested for comparison.

In the first paper of these two companion papers an experimental study was undertaken to ascertain the dynamic behaviour of identical steel–concrete composite beams with differing shear connection systems. Two blind bolt connector types were used as shear connection systems in steel–concrete composite beams. Alongside these, a welded shear stud specimen, and a non-composite specimen were tested for comparison. In this, the second paper a Timoshenko beam model for steel–concrete composite beams is developed and is compared with the experimental results. An uncertain boundary condition is investigated using the Timoshenko beam model and an empirical relation between the displacements at the beam supports and the rotation of the cross section face is proposed.

The use of blind bolts in steel–concrete composite beams is beneficial for promoting sustainable design and for retrofitting existing steel structures. This paper presents an experimental study as well as finite element (FE) modelling analyses for the time-dependent behaviour of composite beams with blind bolts subjected to sustained loads. Four full-scale simply supported beams utilising different types of bolts and studs were tested under long-term static loads. The mid-span deflections were monitored for a period of over 260 days. Short-term push-out tests were also carried out on the connectors used in the composite beams, and their slip deformation was recorded for determining the initial stiffness. The experimental results were modelled by using a three-dimensional FE model, in which the creep of the concrete was simulated through defining a viscoelastic response and the shrinkage of concrete was incorporated by means of notional thermal expansion. The FE model was validated against the experimental results reported herein and other independent results of composite beams using conventional welded studs reported elsewhere, and it was subsequently applied to carry out parametric studies. An extensive body of parameters was considered to clarify their effects on the time-dependent behaviour of composite beams with blind bolts, including the mechanical properties and configuration details of shear connectors, the concrete strength, the loading conditions, the span-to-depth ratio and the reinforcement ratio. It was demonstrated that the time-dependent behaviour was sensitive to the stiffness and the bolt-to-hole clearance of connectors, and some other parameters also possessed effects with different degrees. The research findings implied that using blind bolts in composite beams was beneficial to the time-dependent response due to their relatively lower deflections resulting from the creep and shrinkage of the concrete over time. The outcome may provide an important basis and guidance for designing such composite beams when considering their long-term response with time effects.

Introduction There is a paucity of evidence in diagnostic radiography evaluating a career path into industrial imaging despite several higher education institutes stating this route as a career option on graduation. The link between a career in industrial radiography and diagnostic routes is unknown although there are anecdotal examples of individuals transferring between the two. Successfully obtaining a first post job following graduation in diagnostic radiography can be challenging in the current financial climate. A partnership was formed with an energy sector company that offered non-destructive testing/non-destructive evaluation (NDT/NDE) employing industrial radiographic technicians. Method As an initial pilot, 5 ( n = 5) final year diagnostic radiography students visited an industrial radiography site and underwent theoretical and practical training. Following this placement they engaged in a focus group and the student perceptions/responses were explored and recorded. Results Common themes were identified and categorised via a thematic analysis. These were; radiation safety, physics and technology, widening access, graduate attributes/transferable skill sets and working conditions. Conclusion Student discussion focussed around the benefits of working conditions in healthcare, the value of technology, safety and physics education in alternative placements and the transferability of skills into other/industrial sectors (e.g. NDT/NDE). Contemporary practice placements are a useful pedagogical approach to develop complex conceptual theoretical constructs, such as radiation physics. An in depth evaluation between the two industries skill sets is postulated. Additionally, this could offer alternative/emerging roles to interested diagnostic radiographers potentially meeting the skill shortage in industrial radiography.

Development of allosteric inhibitors into efficient drugs is hampered by their indirect mode-of-action and complex structure-kinetic relationships. To enable the design of efficient allosteric drugs targeting the polymerase of hepatitis C virus (NS5B), the interaction characteristics of three non-nucleoside compounds (filibuvir, VX-222, and tegobuvir) inhibiting HCV replication via NS5B have been analyzed. Since there was no logical correlation between the anti-HCV replicative and enzyme inhibitory effects of the compounds, surface plasmon resonance biosensor technology was used to resolve the mechanistic, kinetic, thermodynamic and chemodynamic features of their interactions with their target and their effect on its interaction with RNA. Tegobuvir could not be seen to interact with NS5B at all while filibuvir interacted in a single reversible step (except at low temperatures) and VX-222 in two serial steps, interpreted as an induced fit mechanism. Both filibuvir and VX-222 interfered with the interaction between NS5B and RNA. They competed for binding to the enzyme, suggesting that they had a common inhibition mechanism and identical or overlapping binding sites. The greater anti-HCV replicative activity of VX-222 over filibuvir is hypothesized to be due to a greater allosteric conformational effect, resulting in the formation of a less catalytically competent complex. In addition, the induced fit mechanism of VX-222 gives it a kinetic advantage over filibuvir, exhibited as a longer residence time. These insights have important consequences for the selection and optimization of new allosteric NS5B inhibitors.

A purification process for the manufacture of a recombinant C-terminus heavy chain fragment from botulinum neurotoxin serotype C [rBoNTC(H(c))], a potential vaccine candidate, has been defined and successfully scaled-up. The rBoNTC(H(c)) was produced intracellularly in Pichia pastoris X-33 using a three step fermentation process, i.e., glycerol batch phase, a glycerol fed-batch phase to achieve high cell densities, followed by a methanol induction phase. The rBoNTC(H(c)) was captured from the soluble protein fraction of cell lysate using hydrophobic charge induction chromatography (HCIC; MEP HyperCel™), and then further purified using a CM 650M ion exchange chromatography step followed by a polishing step using HCIC once again. Method development at the bench scale was achieved using 5-100mL columns and the process was performed at the pilot scale using 0.6-1.6L columns in preparation for technology transfer to cGMP manufacturing. The process yielded approximately 2.5 g of rBoNTC(H(c))/kg wet cell weight (WCW) at the bench scale and 1.6 g rBoNTC(H(c))/kg WCW at the pilot scale. The purified rBoNTC(H(c)) was stable for at least 3 months at 5 and -80°C as determined by reverse phase-HPLC and SDS-PAGE and was stable for 24 months at -80 °C based on mouse potency bioassay. N-Terminal amino acid sequencing confirmed that the N-terminus of the purified rBoNTC(H(c)) was intact.

The basis for efficacy of the recombinant botulinum vaccine, serotypes A and B (rBV A/B) is that neutralizing antibodies induced by vaccination bind to botulinum neurotoxin complex serotype A, subtype A1 (BoNT/A1) and serotype B, subtype B1 (BoNT/B1) and prevent their actions at cholinergic neurons. The protective capacity of BoNT/A1 and BoNT/B1 neutralizing antibodies derived from the serum of clinical volunteers vaccinated with rBV A/B was evaluated in a guinea pig passive transfer model and a mouse bioassay. Guinea pigs passively immunized to achieve circulating neutralizing antibody concentration (NAC) levels representing the lowest measurable concentrations for BoNT/A1 and BoNT/B1 were protected against an intramuscular (IM) challenge more than 10 times the guinea pig IM median lethal dosage for BoNT/A1 and BoNT/B1. The passively immunized guinea pigs were asymptomatic during the 14-day post-challenge observation period. Control guinea pigs died within 48 h after challenge. Calculation of neutralizing efficiency of antibodies using results from a mouse bioassay indicated that a simple linearly proportional relationship does not exist between NAC level and the amount of BoNT neutralized. Based on this finding, estimates of level of protection must consider variability in BoNT neutralizing efficiency at different NAC levels. The protective capacity of human BoNT/A1 and BoNT/B1 neutralizing antibodies induced by rBV A/B vaccination was verified in a guinea pig passive immunization model. Additionally, estimates of the neutralizing efficiency have been established for BoNT/A1 and BoNT/B1 neutralizing antibodies obtained from clinical volunteers vaccinated with the rBV A/B.

The adsorption of recombinant botulinum neurotoxin (BoNT) protein-derived vaccine antigens to aluminum salt adjuvants has been previously studied for the development of a trivalent vaccine against the neurotoxins (Vessely et al., in press, J Pharm Sci). The current paper describes an investigation of the stability of recombinant BoNT antigens adsorbed to aluminum salt adjuvants during storage in aqueous solution. Both chemical and physical changes occurred during storage. Phosphate groups in the buffer exchanged with hydroxyl groups on the adjuvant surface. The resulting changes in solution pH and adjuvant surface chemistry promoted more favorable electrostatic interaction between the BoNT proteins and the surface, possibly increasing the affinity of the proteins for the surface during storage. Fluorescence and UV spectroscopy suggested changes to protein structure during storage, whereas differential scanning calorimetry showed changes to thermal processes related to protein conformation and/or surface adsorption. The consequence of the chemical and physical changes to the proteins was a decrease in the ability to desorb protein from the adjuvant surface during storage. Overall, the results of this study emphasize the utility of a thorough characterization of the interactions between protein antigens and aluminum salt adjuvants.

We identified a series of structurally novel SCD (Delta9 desaturase) inhibitors via high-throughput screening and follow-up SAR studies. Modification of the central bicyclic scaffold has proven key to our potency optimization effort. The most potent analog (8g) had IC(50) value of 50 pM in a HEPG2 SCD assay and has been shown to be metabolically stable and selective against Delta5 and Delta6 desaturases.

Highly potent and selective 4-amidofuran-3-one inhibitors of cathepsin S are described. The synthesis and structure-activity relationship of a series of inhibitors with a sulfonamide moiety in the P3 position is presented. Several members of the series show sub-nanomolar inhibition of the target enzyme as well as an excellent selectivity profile and good cellular potency. Molecular modeling of the most interesting inhibitors describes interactions in the extended S3 pocket and explains the observed selectivity towards cathepsin K.

A general solid-phase synthesis of 1,2,5-trisubstituted imidazolidin-4-ones is described. The key synthetic transformation incorporates a microwave-assisted condensation of an α-amino amide on solid support with an aldehyde in solution to give the corresponding resin-bound imidazolidin-4-one in a simple one-pot procedure.

A process was developed for production of a candidate vaccine antigen, recombinant C-terminal heavy chain fragment of the botulinum neurotoxin serotype E, rBoNTE(H(c)) in Pichia pastoris. P. pastoris strain GS115 was transformed with the rBoNTE(H(c)) gene inserted into pHILD4 Escherichia coli-P. pastoris shuttle plasmid. The clone was characterized for genetic stability, copy number, and BoNTE(H(c)) sequence. Expression of rBoNTE(H(c)) from the Mut(+) HIS4 clone was confirmed in the shake-flask, prior to developing a fed-batch fermentation process at 5 and 19 L scale. The fermentation process consists of a glycerol growth phase in batch and fed-batch mode using a defined medium followed by a glycerol/methanol transition phase for adaptation to growth on methanol and a methanol induction phase resulting in the production of rBoNTE(H(c)). Specific growth rate, ratio of growth to induction phase, and time of induction were critical for optimal rBoNTE(H(c)) production and minimal proteolytic degradation. A computer-controlled exponential growth model was used for process automation and off-gas analysis was used for process monitoring. The optimized process had an induction time of 9 h on methanol and produced up to 3 mg of rBoNTE(H(c)) per gram wet cell mass as determined by HPLC and Western blot analysis.

Reported results for patellar ACI include a mixed population of patients having or not a concomitant correction of the extensor mechanism. No study has reported separate outcome for those two patellar ACI subgroups. Forty four patients were included in this study designed to compare the outcome of patellar ACI with extensor realignment (group A) to patellar ACI with normal patellofemoral tracking (group B). Group A (n=22) had a higher increase in average modified Cincinnati knee score (4.5 vs. 1.7 points), better function (1.7 vs. 2.5), better SF-36 physical component scores (70.9 vs. 55.4 points) and higher IKDC scores (85.2 vs. 60.6 points) when compared to group B (n=22) at an average follow-up of 2 years. Patellar ACI with concomitant extensor realignment has superior outcome than patellar ACI with normal patellofemoral tracking. An unloading osteotomy could be desirable in selected patients with normal patellofemoral biomechanics to improve the outcome in this population.

The identification and evaluation of aryl-[1,4]diazepane ureas as functional antagonists of the chemokine receptor CXCR3 are described. Specific examples exhibit IC(50) values of approximately 60 nM in a calcium mobilization functional assay, and dose-dependently inhibit CXCR3 functional response to CXCL11 (interferon-inducible T-cell alpha chemoattractant/I-TAC) as measured by T-cell chemotaxis, with a potency of approximately 100 nM.

Between October 2000 and December 2003, 252 autologous chondrocyte implants were performed in 183 patients. Eighty lesions showed overgrowth of the subchondral bone plate under the chondral lesion, this was termed a "bone boss." Thirty-seven were on the medial femoral condyle (MFC), 18 on the lateral femoral condyle (LFC), 21 in the trochlea and 4 on the patella. There was a statistically significant association between the LFC and "bone boss." The lesions showing this finding were of a larger area (3.4 cm2 and 2.8 cm2 respectively, p=0.006), and had more diffuse chondral changes than lesions without. The patients with a "bone boss" had a tendency to longer duration of symptoms (85.3 months and 64.3 months respectively, p=0.089). The "bone bosses" were resected back to the level of the surrounding subchondral bone prior to implantation. Radiological and clinical follow-up showed no statistical difference between the two groups. A discussion of the possible aetiology of the "bone boss" is made.

High tibial osteotomy has been generally accepted as a useful treatment for unicompartment osteoarthritis of the knee to improve a patient's quality of life. Few studies have examined the outcome from the patient's perspective. A survival analysis of one to twenty-one years was conducted on 67 knees including analysis of the patients' satisfaction level with this procedure. Cumulative survival probability of 89.5% at 5 years, 74.7% at 10 years and 66.9 % for 15 and 20 years was reported. Ninety-one percent of patients had improvement in pain score and would choose to have this surgery again. Average patient satisfaction level was 75.5%. Forty-eight percent of patients were able to perform at a higher level of activity than before surgery, although none was able to perform at the level prior to the onset of knee pathology. It was concluded that this procedure was able to improve a patient's quality of life and achieved high satisfaction among patients. There is an increasing role of high tibial osteotomy as an adjunct to an autologous chondrocyte implantation procedure.

Structural conversion of the serotype A recombinant botulinum neurotoxin heavy chain fragment (rBoNTA(Hc)) produced intracellularly in Pichia pastoris yeast was observed and characterized during purification development efforts. A pH screening study captured the transformation stages of the original recovered species into its derived counterpart and a number of analytical tools such as peptide mapping by LC/MS confirmed the formation of a disulfide bond, especially in samples of neutral to basic pH. A cation exchange chromatographic method proved useful in following the incidence of the reaction in various rBoNTA(Hc) samples. The disulfide formation kinetics were characterized using a one-quarter quadratic factorial design, following the investigation and development of controlled oxidation conditions using cysteine and cystamine as the redox pair. Temperature, pH and concentration of the redox pair had a significant effect on the yield and rate of the disulfide formation. This controlled reaction was eventually introduced as a functional unit operation in the purification process. The summation of preliminary scale-up and potency data showed scalability and robustness in the production of an active disulfide-bonded form of a recombinant botulism vaccine candidate. The presence of the disulfide bond did not effect the vaccine potency and it enhanced the molecule's thermal stability.

Initial purification of two serotypic variants of recombinant botulinum neurotoxin toxin heavy chain fragment [rBoNT(Hc)], produced intracellularly in the yeast Pichia pastoris, using hydrophobic charge induction chromatography (HCIC) is reported. HCIC employs a matrix containing a weakly ionizable ligand that binds proteins through hydrophobic interactions at neutral pH and elutes the proteins by charge repulsion at acidic pH. HCIC optimization led to different purification conditions for each of the proteins even though they have 58% sequence similarity. The HCIC resin has a higher affinity for the fragment of serotype A than that of serotype B. The 10% dynamic breakthrough capacity for the serotype A fragment is >12.5 mg per ml of resin and is approximately 3.5 mg or the serotype B fragment per ml of resin. Stable elution conditions are also different for the two serotypes. The serotype A fragment is unstable when citrate is used to elute the product. However the serotype B fragment is stable when eluted with citrate buffer, and it is further purified by a overnight precipitation caused by the citrate buffer. This paper reports the development strategy, dynamic capacity breakthrough curves, resin and separation reproducibility, and preliminary scale-up data. The summation of the data demonstrates that HCIC is a scaleable process step for biopharmaceutical production of rBoNT(Hc) proteins.

The cysteinyl proteinase cathepsin S is implicated as a key enzyme in the processing of major histocompatability complex (MHC) class II molecules expressed on antigen presenting cells and thus is a potential therapeutic target for modulation in immune system-based disease. We have identified a form of rat cathepsin S, similar to a published mouse form with an eight-amino acid extended presequence relative to the human enzyme and the previously published rat enzyme. In addition, we have expressed these mouse and rat proteins in baculovirally infected Sf9 insect cells along with "humanized" forms truncated by eight residues at the amino-terminus. All forms of the rodent proteinases were overexpressed and milligram per litre amounts of functional enzyme could be isolated from the cells and/or the cell culture supernatant. Furthermore, addition of a carboxy-terminal hexahistidine purification tag had no effect on the kinetic characteristics of any of the enzyme forms against the Boc-Val-Leu-Lys-AMC peptide substrate (rat k(cat) s(-1) approximately 30; mouse k(cat) s(-1) approximately 65). Differences were seen in the potency of the generic cysteine proteinase inhibitor, E64, against the human, mouse, or rat form of the enzyme (13.3 x 10(4), 43.2 x 10(4), and 25 x 10(4) K(obe)/[I] M(-1) s(-1), respectively). Such data highlights the need for greater awareness of species variation in inhibitor potency. These reagents are vital for confirming inhibitor potency against the endogenous form of the enzyme prior to evaluation of drug candidates in rodent model systems.

Mobile bearing knee replacement has been designed to provide an effective prosthetic tool for improving long-term survivorship. Although, its potential long-term superiority has not been proven, an alarming possibility of its early mechanical failure has been previously reported. In the present study on 27 patients with 35 replaced knees due to osteoarthritis, 97.1% of a 5-year survivorship of LCS mobile bearing cementless total knee prosthesis is reported. This high survival rate, with a satisfactory functional outcome at middle term follow-up has been achieved by following an optimal technique of implantation. Therefore, in the middle term this mobile bearing design of total knee replacement is no less effective than the conventional fixed bearing designs.

A chemoenzymatic approach for the synthesis of homoazasugars is described, utilizing an aldolase to catalyze the key asymmetric aldol addition reaction. This approach is illustrated by the synthesis of β-D-homomannonojirimycin (11) using D-fructose diphosphate (FDP) aldolase.

4-Chlorophenylsulfinylacetates of defined stereochemistry at sulfur were prepared from monochiral alcohols: compound 2 from (+)-menthol; 3 and 4 from (−)-8-phenylmenthol; 5 and 6 from (+)-trans-2-phenylcyclohexanol; and, 7 and 9 from (-)-10-dicyclohexylsulfamoyl-D-isoborneol. Reagents for which the sulfoxide asymmetry is matched with the inducing effect of the auxiliary give good diastereoselection in SPAC reactions affording γ-hydroxy-α,β-unsaturatod alcohols of high optical purity. Asymmetry at the sulfoxide has a greater effect than the auxiliaries on the stereochemical outcome of these reactions.

(1S, 2R, 7R, 7aR)- 1,2,7-Trihydroxypyrrolizidine ( 1 ) is prepared by elaboration of an erythrose derivative ( 4 ) via an asymmetric allylation reaction; this approach promises to be widely applicable to syntheses of polyhydroxylated indolizidine and pyrrolizidine alkaloids.

γ-Hydroxy-α,β-unsaturated compounds ( 3 ) (SPAC creation products) are resolved via irreversible biocatalytic acylations; this process can be used in series with asymmetric SPAC reactions to improve optical purities.

Biocatalytic resolutions of methyl sulfinylacetates fford sulfoxides (R)-( 1 ) - ( 6 ) in very high optical yields; the products have been used in a systematic study of the “SPAC” reaction, an asymmetric synthesis of γ-hydroxy-α,β-unsaturated esters.

Double diastereoselection in the S ulfoxide P iperidine A nd C arbonyl ( SPAC ) reactions of sulfinyl acetate esters (2) with α-unsubstituted aldehydes (3) has been investigated to facilitate asymmetric syntheses of γ-hydroxy-α,β-unsaturated esters (1) .