<i>Trypanosoma brucei</i> DHRF-TS revisited:characterisation of a bifunctional and highly unstable recombinant dihydrofolate reductase-thymidylate synthase

<div><p>Bifunctional dihydrofolate reductase–thymidylate synthase (DHFR-TS) is a chemically and genetically validated target in African trypanosomes, causative agents of sleeping sickness in humans and nagana in cattle. Here we report the kinetic properties and sensitivity of recombinant enzyme to a range of lipophilic and classical antifolate drugs. The purified recombinant enzyme, expressed as a fusion protein with elongation factor Ts (Tsf) in ThyA^- <i>Escherichia coli</i>, retains DHFR activity, but lacks any TS activity. TS activity was found to be extremely unstable (half-life of 28 s) following desalting of clarified bacterial lysates to remove small molecules. Stability could be improved 700-fold by inclusion of dUMP, but not by other pyrimidine or purine (deoxy)-nucleosides or nucleotides. Inclusion of dUMP during purification proved insufficient to prevent inactivation during the purification procedure. Methotrexate and trimetrexate were the most potent inhibitors of DHFR (<i>K</i>_i 0.1 and 0.6 nM, respectively) and FdUMP and nolatrexed of TS (<i>K</i>_i 14 and 39 nM, respectively). All inhibitors showed a marked drop-off in potency of 100- to 1,000-fold against trypanosomes grown in low folate medium lacking thymidine. The most potent inhibitors possessed a terminal glutamate moiety suggesting that transport or subsequent retention by polyglutamylation was important for biological activity. Supplementation of culture medium with folate markedly antagonised the potency of these folate-like inhibitors, as did thymidine in the case of the TS inhibitors raltitrexed and pemetrexed.</p></div

Validated stability-indicating spectrofluorimetric methods for the determination of ebastine in pharmaceutical preparations

Two sensitive, selective, economic, and validated spectrofluorimetric methods were developed for the determination of ebastine (EBS) in pharmaceutical preparations depending on reaction with its tertiary amino group. Method I involves condensation of the drug with mixed anhydrides (citric and acetic anhydrides) producing a product with intense fluorescence, which was measured at 496 nm after excitation at 388 nm

Female Institutional Directors on Boards and Firm Value

The aim of this research is to examine what impact female institutional directors on boards have on corporate performance. Previous research shows that institutional female directors cannot be considered as a homogeneous group since they represent investors who may or may not maintain business relations with the companies on whose corporate boards they sit. Thus, it is not only the effect of female institutional directors as a whole on firm value that has been analysed, but also the impact of pressure-resistant female directors, who represent institutional investors (investment, pension and mutual funds) that only invest in the company, and do not maintain a business relation with the firm. We hypothesize that there is a non-linear association, specifically quadratic, between institutional and pressure-resistant female directors on boards and corporate performance. Our results report that female institutional directors on boards enhance corporate performance, but when they reach a certain threshold on boards (11.72 %), firm value decreases. In line with female institutional directors, pressure-resistant female directors on boards also increase firm value, but only up to a certain figure (12.71 % on boards), above which they have a negative impact on firm performance. These findings are consistent with an inverted U-shaped relationship between female institutional directors and pressure-resistant female directors and firm performance

NEOSCOPE: A randomised Phase II study of induction chemotherapy followed by either oxaliplatin/capecitabine or paclitaxel/carboplatin based chemoradiation as pre-operative regimen for resectable oesophageal adenocarcinoma

Background: Both oxaliplatin/capecitabine-based chemoradiation (OXCAP-RT) and carboplatin-paclitaxel based radiation (CarPac-RT) are active regimens in oesophageal adenocarcinoma, but no randomised study has compared their efficacy and toxicity. This randomised phase II "pick a winner" trial will identify the optimum regimen to take forward to a future phase III trial against neo-adjuvant chemotherapy, the current standard in the UK. Methods/Design: Patients with resectable adenocarcinoma of the oesophagus or Siewert Type 1-2 gastro-oesophageal junction (GOJ), ≥T3 and/or ≥ N1 are eligible for the study. Following two cycles of induction OXCAP chemotherapy (oxaliplatin 130 mg/m2 D1, Cape 625 mg/m2 D1-21, q 3 wk), patients are randomised 1:1 to OXCAP-RT (oxaliplatin 85 mg/m2 Day 1,15,29; capecitabine 625 mg/m2 twice daily on days of RT; RT-45 Gy/25 fractions/5 weeks) or CarPac-RT (Carboplatin AUC2 and paclitaxel 50 mg/m2 Day 1,8,15,22,29; RT-45 Gy/25 fractions/5 weeks). Restaging CT/PET-CT is performed 4-6 weeks after CRT, and a two-phase oesophagectomy with two-field lymphadenectomy is performed six to eight weeks after CRT. The primary end-point is pathological complete response rate (pCR) at resection and will include central review. Secondary endpoints include: recruitment rate, toxicity, 30-day surgical morbidity/mortality, resection margin positivity rate and overall survival (median, 3- and 5-yr OS. 76 patients (38/arm) gives 90% power and one-sided type 1 error of 10% if patients on one novel treatment have a response rate of 35% while the second treatment has a response rate of 15%. A detailed RT Quality Assurance (RTQA) programme includes a detailed RT protocol and guidance document, pre-accrual RT workshop, outlining exercise, and central evaluation of contouring and planning. This trial has been funded by Cancer Research UK (C44694/A14614), sponsored by Velindre NHS Trust and conducted through the Wales Cancer Trials Unit at Cardiff University on behalf of the NCRI Upper GI CSG. Discussion: Following encouraging results from previous trials, there is an interest in neo-adjuvant chemotherapy and CRT containing regimens for treatment of oesophageal adenocarcinoma. NEOSCOPE will first establish the efficacy, safety and feasibility of two different neo-adjuvant CRT regimens prior to a potential phase III trial

Canagliflozin and Renal Outcomes in Type 2 Diabetes and Nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to 300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m 2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Dellen formation as a complication of subconjunctival silicone oil following microincision vitrectomy

Mohamed M Mahgoub,1,2 Maged M Roshdy,1,2 Sherine S Wahba1,2 1Ophthalmology Department, Ain Shams University, 2Ophthalmology Department, Al Watany Eye Hospital, Cairo, Egypt Purpose: To study the subconjunctival silicone oil (SCSO) parameters associated with dellen formation following microincision vitrectomy (MIV).Patients and methods: This was a case&ndash;control study of 20 eyes with SCSO following MIV. Ten of them suffered postoperative dellen formation. Dellen occurrence, their sizes, number of loci, and distance between SCSO and the limbus were recorded. The outcome after silicone-oil removal was evaluated.Results: The SCSO was at 3.1&plusmn;1.2 mm from the limbus. All cases with dellen had SCSO within 2 mm of the limbus (P&lt;0.001). No other factors were found to be associated with dellen formation (age, P=0.414; sex, P=0.656; laterality, P=1; indication for pars plana vitrectomy, P=0.655; instrument gauge, P=0.211; circumference involved by SCSO, P=0.252). All the dellen healed after surgical evacuation of SCSO (P&lt;0.001), leaving scars.Conclusion: Dellen can be associated with MIV secondary to SCSO near the limbus. Resolution with scarring occurred following SCSO evacuation. Therefore, SCSO should be evacuated as early as possible to avoid long-term peripheral corneal morbidity. Keywords: silicone oil-removal, subconjunctival silicone oil, dellen, vitrectom

Visual performance of the Tecnis one-piece lens ZCB00

Sherein S Wahba, Riad F Riad, Fathy F Morkos, Abdallah K Hassouna, Maged M RoshdyNational Eye Hospital and Ain Shams University, Cairo, EgyptSynopsis: The aspheric, one-piece design of the Tecnis&amp;reg; ZCB00 intraocular lens provided higher mesopic contrast sensitivity and relatively constant refraction at different pupil sizes compared to the Sensar&amp;reg; AR40e.Purpose: Comparing the impact on functional vision of the aspheric Tecnis&amp;reg; ZCB00 one-piece lens compared to the spherical Sensar&amp;reg; AR40e three-piece lens.Setting: National Eye Hospital, Cairo, Egypt.Methods: In this prospective study, we implanted 32 Tecnis ZCB00 (Abbott Medical Optics, Santa Ana, CA) and 30 Sensar AR40e (Pharamacia and Upjohn LLC, North Peapack, NJ) lenses. Twenty-three patients in the ZCB00 group and 13 patients in the AR40e group provided reliable, reproducible data, and were therefore included in the statistical analysis. After 8 weeks, we measured photopic contrast sensitivity (PCS) at 85 cd/m&amp;sup2; and mesopic contrast sensitivity (MCS) at 5 cd/m&amp;sup2;. Uncorrected and best-corrected visual acuity were measured. Myopic shift was analyzed by comparing the manifest refraction spherical equivalent at 3 days and 8 weeks after surgery in the normal and dilated pupil.Results: The Tecnis ZCB00 group showed significantly better MCS than the Sensar AR40e group at a spatial frequency of six cycles per degree (cpd) (P = 0.037), but not at 3, 12, or 18 cpd (P = 0.299, P = 0.226, and P = 0.396, respectively). There were no significant differences between groups in corrected distance visual acuity (P = 0.175) or PCS at 3, 6, 12, or 18 cpd spatial frequencies (P = 0.440, P = 0.176, P = 0.365, and P = 0.251, respectively). The ZCB00 group showed less myopic shift in normal and dilated pupils between 3 days and 8 weeks after surgery compared to the AR40e group, but this difference was not significant.Conclusion: The Tecnis one-piece ZCB00 provided higher MCS at moderate spatial frequency than the three-piece Sensar AR40e, and was associated with relatively constant refraction at different pupil sizes.Keywords: Tecnis ZCB00, Sensar AR40e, contrast sensitivity, myopic shif