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V565 is an engineered TNFα-neutralising single domain antibody formulated into enteric coated mini-tablets to enable release in the intestine after oral administration as a possible oral treatment for inflammatory bowel disease (IBD). Following oral administration, ileal recovery of V565 was investigated in four patients with terminal ileostomy. Intestinal and systemic pharmacokinetics were measured in six patients with Crohn’s disease and evidence of target engagement assessed in five patients with ulcerative colitis. Following oral administration, V565 was detected at micromolar concentrations in ileal fluid from the ileostomy patients and in stools of the Crohn’s patients. In four of the five ulcerative colitis patients, biopsies taken after 7d dosing demonstrated V565 in the lamina propria with co-immunostaining on CD3+ T-lymphocytes and CD14+ macrophages. Phosphorylation of signalling proteins in biopsies taken after 7d oral dosing was decreased by approximately 50%. In conclusion, enteric coating of V565 mini-tablets provided protection in the stomach with gradual release in intestinal regions affected by IBD. Immunostaining revealed V565 tissue penetration and association with inflammatory cells, while decreased phosphoproteins after 7d oral dosing was consistent with V565-TNFα engagement and neutralising activity. Overall these results are encouraging for the clinical utility of V565 in the treatment of IBD.

Anti-TNFα and anti-IL-23 antibodies are highly effective therapies for Crohn’s disease or ulcerative colitis in a proportion of patients. V56B2 is a novel bispecific domain antibody in which a llama-derived IL-23p19-specific domain antibody, humanised and engineered for intestinal protease resistance, V900, was combined with a previously-described TNFα-specific domain antibody, V565. V56B2 contains a central protease-labile linker to create a single molecule for oral administration. Incubation of V56B2 with trypsin or human faecal supernatant resulted in a complete separation of the V565 and V900 monomers without loss of neutralising potency. Following oral administration of V900 and V565 in mice, high levels of each domain antibody were detected in the faeces, demonstrating stability in the intestinal milieu. In ex vivo cultures of colonic biopsies from IBD patients, treatment with V565 or V900 inhibited tissue phosphoprotein levels and with a combination of the two, inhibition was even greater. These results support further development of V56B2 as an oral therapy for IBD with improved safety and efficacy in a greater proportion of patients as well as greater convenience for patients compared with traditional monoclonal antibody therapies.

Background Cortical visual impairment (CVI) is the most common cause of pediatric visual impairment in developed countries, and cerebral palsy (CP) is diagnosed in approximately half of children with CVI. It is unknown whether children with CVI who also have CP (CVI+CP) have different characteristics and outcomes (with regard to visual acuity, strabismus, and response to strabismus surgery) than children with CVI without CP (CVI−CP). Methods The medical records of all children with CVI, with and without CP, evaluated at our institution between 2013 and 2019 were retrospectively reviewed. Presentation and outcomes of children with CVI+CP were compared to those with CVI−CP. Results A total of 151 children with CVI+CP and 153 children with CVI−CP were included. Children with CVI+CP were more likely to be diagnosed with significant refractive error (53.6% vs 41.2%; P = 0.03), optic atrophy (46.4% vs 32.7%; P = 0.01), and strabismus (82.8% vs 72.5%; P = 0.03) at presentation. Good ocular alignment after strabismus surgery was achieved in 30% of children with CVI+CP and 63.6% of children with CVI−CP (P = 0.48). Of 9 children with long-term (≥8 years) postoperative follow-up, 100% of CVI−CP patients achieved good outcomes compared with 0% of CVI+CP patients (P = 0.0079). Visual acuity at presentation and the percentage of patients who experienced improvement in visual acuity did not differ between groups. Conclusions In our study cohort, children with CVI+CP had a higher likelihood of ophthalmic comorbidities and may have worse long-term strabismus surgery outcomes than children with CVI−CP.

V565 is a novel oral anti-tumor necrosis factor (TNF)-α domain antibody being developed for topical treatment of inflammatory bowel disease (IBD) patients. Protein engineering rendered the molecule resistant to intestinal proteases. Here we investigate the formulation of V565 required to provide gastro-protection and enable optimal delivery to the lower intestinal tract in monkeys.Enteric-coated V565 mini-tablets were prepared and dissolution characteristics tested in vitro. Oral dosing of monkeys with enteric-coated mini-tablets containing V565 and methylene blue dye enabled in vivo localization of mini-tablet dissolution. V565 distribution in luminal contents and feces was measured by enzyme-linked immunosorbent assay (ELISA). To mimic transit across the damaged intestinal epithelium seen in IBD patients an intravenous (i.v.) bolus of V565 was given to monkeys and pharmacokinetic parameters of V565 measured in serum and urine by ELISA.Enteric-coated mini-tablets resisted dissolution in 0.1 M HCl, before dissolving in a sustained release fashion at neutral pH. In orally dosed monkeys methylene blue intestinal staining indicated the jejunum and ileum as sites for mini-tablet dissolution. Measurements of V565 in monkey feces confirmed V565 survival through the intestinal tract. Systemic exposure after oral dosing was very low consistent with limited V565 mucosal penetration in healthy monkeys. The rapid clearance of V565 after i.v. dosing was consistent with renal excretion as the primary route for elimination of any V565 reaching the circulation.These results suggest that mini-tablets with a 24% Eudragit enteric coating are suitable for targeted release of orally delivered V565 in the intestine for topical treatment of IBD.

Introduction Monoclonal antibodies to TNF transformed treatment options for patients with Inflammatory Bowel Disease (IBD). V565 is a novel oral anti-TNF domain antibody (Vorabody) engineered to be resistant to intestinal proteases. It is in development as a potential oral treatment for IBD. In vitro it suppressed phosphorylation of tyrosine kinases and signalling proteins and inhibited the release of inflammatory cytokines following culture with biopsies taken from patients with CD (Crowe et al. 18th International Congress of Mucosal Immunology, July 19–22 2017, Washington DC, USA). It was safe and well tolerated after high single and multiple doses in healthy volunteers and patient volunteers with CD and resulted in high concentrations of active drug in ileal fluid and faeces. Aims & Methods This open label study was designed to demonstrate that V565 enters GI mucosa and exerts a beneficial effect on inflammatory processes following oral dosing for 7 days to patients with Ulcerative Colitis. Patients with a Mayo score of 3–10 including an endoscopy score of ≥1 had up to 7 days of oral dosing with 555 mg tid V565. Sigmoidoscopy with biopsies was performed before and after the dosing period. The primary outcomes of interest were presence of V565 in the mucosa and reduction from baseline in phosphorylation of tyrosine kinases and signalling proteins. Detection of V565 was determined by immunohistochemistry. Phosphorylation was determined using PathScan RTK signalling arrays (Vossenkaemper et al 2014. Gastroenterology 147:172–83). Results Five patient volunteers were treated Due to visit scheduling, most received 6 days treatment. Presence of V565 was confirmed in the inflamed lamina propria and co-localised with CD14 +macrophages in post-treatment biopsies. Overall phosphorylation of the panel of kinases and signalling proteins was reduced by approximately 50% in four of the five patients. There were no treatment induced ADAs. Conclusion V565, an oral anti-TNF domain antibody engineered to be resistant to intestinal proteases, was demonstrated bound to CD14 +macrophages in the lamina propria of UC patients and resulted in inhibition of mucosal inflammatory processes after 6–7 days oral dosing. The reduction of 50% in overall phosphorylation is similar to that seen in an earlier study of UC biopsy cultures with infliximab at a concentration of 67 nM (10 µg/ml), a serum concentration associated with mucosal healing (Ungar et al, Clin Gastroenterol Hepatol. 2016 Apr;14(4):550–557). These results provide encouragement that oral dosing with V565 will be a beneficial oral treatment option for patients with IBD.

TNFα is an important cytokine in inflammatory bowel disease. V565 is a novel anti-TNFα domain antibody developed for oral administration in IBD patients, derived from a llama domain antibody and engineered to enhance intestinal protease resistance. V565 activity was evaluated in TNFα-TNFα receptor-binding ELISAs as well as TNFα responsive cellular assays and demonstrated neutralisation of both soluble and membrane TNFα with potencies similar to those of adalimumab. Although sensitive to pepsin, V565 retained activity after lengthy incubations with trypsin, chymotrypsin, and pancreatin, as well as mouse small intestinal and human ileal and faecal supernatants. In orally dosed naïve and DSS colitis mice, high V565 concentrations were observed in intestinal contents and faeces and immunostaining revealed V565 localisation in mouse colon tissue. V565 was detected by ELISA in post-dose serum of colitis mice, but not naïve mice, demonstrating penetration of disrupted epithelium. In an ex vivo human IBD tissue culture model, V565 inhibition of tissue phosphoprotein levels and production of inflammatory cytokine biomarkers was similar to infliximab, demonstrating efficacy when present at the disease site. Taken together, results of these studies provide confidence that oral V565 dosing will be therapeutic in IBD patients where the mucosal epithelial barrier is compromised.

This case report describes an immune-competent patient with acute upper extremity ischemia caused by thromboembolism from an Aspergillus-infected ascending aortic pseudoaneurysm. Efforts to identify the source of an acute arterial thromboembolic occlusion should be made, and a high index of suspicion for mycotic infection should be maintained in patients with an atypical presentation, such as fevers of unknown origin. Additional measures, such as pathologic examination of thromboembolic debris, blood cultures, and positron emission tomography, should be performed to identify the etiology in these unexplained situations.

Numerical simulations were conducted to evaluate the efficacy of in situ field scale chemical oxidation (ISCO) (with permanganate) for trichloroethylene (TCE) and tetrachloroethylene (PCE) DNAPL in eight idealized heterogeneous porous media sites. The goal of this work was to study the effects of DNAPL source zone remediation at the site scale utilizing a high-resolution three-dimensional numerical model. The model results demonstrated that both dissolution (no treatment) and treatment were unable to completely deplete DNAPL mass or boundary mass flux for any of the sites over a 10 year period. Performance was highly variable due to competition with oxidizable organic aquifer materials, manganese dioxide (rind) formation, DNAPL architecture, and geologic characteristics. When compared to dissolution only, DNAPL mass depletion at the end of treatment ranged from adverse impacts to a three-fold improvement, but diminished with post-treatment monitoring. Similarly, the relative reduction in boundary mass flux varied between unity and a factor of two at the termination of treatment, but at 10 years ranged from a 10-fold reduction to adverse impacts. During the post-treatment phase of the simulations, most of the benefits achieved during active treatment were negated due to dissolution tailing.

We hypothesized that full-time orthopedic traumatologists would increase interfacility transfer patients with pelvic ring injuries at a trauma center seeking American College of Surgeons Level I verification. A trauma registry query identified 790 patients with pelvic ring injuries admitted to a trauma service over 10 years. The study groups included 173 patients 3 years before and 405 patients 3 years after recruitment of two full-time orthopedic traumatologists. Interfacility transfers of patients with pelvic ring injuries requiring operative procedures were increased over 230 per cent ( P < 0.01) with full-time orthopedic traumatologists. Financial analysis showed enhanced hospital margin per patient and decreased direct costs ( P < 0.01). Complications, mortality rates, and length of stay were decreased ( P < 0.01). Full-time orthopedic traumatologists enhance pelvic ring injury patient interfacility transfers, operative procedures, outcomes, and financials at a Level I trauma center in a rural setting.

The influence of source zone concentration reduction on solute plume detachment and recession times in fractured rock was investigated using new semianalytical solutions to transient solute transport in the presence of advection, dispersion, sorption, matrix diffusion, and first-order decay. Novel aspects of these solutions are: (1) the source zone concentration behavior is simulated using a constant concentration with the option for either an instantaneous reduction to zero concentration or an exponentially decaying source zone concentration initiated at some time (t*) after the source is introduced, and (2) different biodegradation rates in the fracture and rock matrix. These solutions were applied for sandstone bedrock and revealed that biodegradation in the matrix, not the fracture, may be the most significant attenuation mechanism and therefore may dictate remediation time scales. Also, instantaneous and complete source concentration reduction in aged plumes may not be beneficial with respect to plume response because back-diffusion can sustain plume migration for long periods of time. Moderate source zone concentration reduction has a similar impact on the rate of advance of the leading edge of the plume as aggressive concentration reduction. If the source zone concentration reduction half-life is less than the plume decay half-life, then volatile organic compound (VOC) mass sequestered in the rock matrix will ultimately dictate plume persistence and not the presence of the source zone.

A three-dimensional two-phase flow model is coupled to a non-linear reactive transport model to study the efficacy of potassium permanganate treatment on dense, non-aqueous phase liquid (DNAPL) source removal in porous media. A linear relationship between the soil permeability ( k ) and concentration of manganese dioxide precipitate ([MnO 2(s) ]), k = k o + S rind [MnO 2(s) ], is utilized to simulate nodal permeability reductions due to precipitate formation. Using published experimental column studies, an S rind = −5.5 × 10 −16 m 2 L/mg was determined for trichloroethylene (TCE) DNAPL. This S rind was then applied to treatment simulations on three-dimensional TCE DNAPL source zones comprising either DNAPL at residual saturations, or DNAPL at pooled saturations. DNAPL dissolution without oxidation treatment, simulated using equilibrium and the Nambi and Powers [Nambi I, Powers S. Mass transfer correlations for non-aqueous phase liquid dissolution from regions with high initial saturations. Water Resour Res 2003;39(2):1–11, SBH 4] mass transfer expression, required 31 and 36 years, respectively, to eliminate the residual source zone. For equilibrium dissolution with continuous treatment and no precipitate influence ( S rind = 0 m 2 L/mg), the residual source zone was removed after 11 years. However, when considering the precipitate influence (i.e., S rind = −5.5 × 10 −16 m 2 L/mg), 21 years of treatment were necessary to remove the DNAPL. When considering pulse treatments of 1 and 2 years duration followed by only dissolution, approximately 36 and 38 years, respectively, were required before the source zone was depleted, suggesting that there is no benefit to pulse treatment. Similar trends were observed when allowing 10 years of dissolution prior to treatment initiation. The treatment behaviour of the pooled TCE source, while slightly more efficient than the residual saturation source, was similar. Based on simulation findings, the precipitate (rind) formation significantly influences DNAPL treatment with permanganate; the most significant reductions in efficacy were observed for single pulse treatments (of 1 and 2 years), which exhibited times to source depletion similar to the case of dissolution without treatment.

Objective To investigate the role of CXCL13 in the development and pathogenesis of collagen-induced arthritis (CIA), and to determine the mechanisms involved in the modulation of arthritogenic response by CXCL13 neutralization. Methods Mice were immunized with type II collagen (CII) and treated with anti-CXCL13 or control antibodies during boosting. Mice were monitored for the development and severity of arthritis. The effects of CXCL13 neutralization on immune response to CII were evaluated by cytokine production by CII-specific T cells and CII-specific antibody production. Follicular response in the spleen and in synovial tissue was determined by in situ immunohistology. Results Mice receiving neutralizing antibodies to CXCL13 developed significantly less severe arthritis compared with mice injected with phosphate buffered saline or control antibodies. Follicular response both in the spleen and in synovial tissue was inhibited by anti-CXCL13 treatment. Injection with anti-CXCL13 antibodies did not significantly affect antigen-specific recall lymphocyte proliferation or type 1 cytokine production in vitro. Antibody response specific to CII was not inhibited by anti-CXCL13 treatment. However, anti-CXCL13 treatment induced significantly higher levels of interleukin-10 production after in vitro CII stimulation. Conclusion Neutralization of CXCL13 inhibits the development of CIA and reduces follicular response in both lymphoid and nonlymphoid tissues. These findings may have important implications regarding the pathogenesis and treatment of autoimmune arthritis.

Transient and steady-state analytical solutions are derived to investigate solute transport in a fractured porous medium consisting of evenly spaced, parallel discrete fractures. The solutions incorporate a finite width strip source, longitudinal and transverse dispersion in the fractures, source decay, aqueous phase decay, one-dimensional diffusion into the matrix, sorption to fracture walls, and sorption within the matrix. The solutions are derived using Laplace and Fourier transforms, and inverted by interchanging the order of integration and utilizing a numerical Laplace inversion algorithm. The solutions are verified for simplified cases by comparison to solutions derived by Batu [Batu V. A generalized two-dimensional analytical solution for hydrodynamic dispersion in bounded media with the first-type condition at the source. Wat Resour Res 1989;25(6):1125] and Sudicky and Frind [Sudicky EA, Frind EO. Contaminant transport in fractured porous media: analytical solutions for a system of parallel fractures. Wat Resour Res 1982;18(6):1634]. The application of the solutions to a fractured sandstone demonstrates that narrower source widths and larger values of transverse dispersivity both lead to lower downstream concentrations in the fractures and shorter steady-state plumes. The incorporation of aqueous phase decay and source concentration decay both lead to lower concentrations and shorter plumes, with even moderate amounts of decay significantly shortening the persistence of contamination.

Dynamic Line Rating (DLR) is a smart grid technology that allows the rating of electrical conductors to be increased based on local weather conditions. Overhead lines are conventionally given a conservative rating based on worst case scenarios. We demonstrate that observing the conditions in real time leads to additional capacity and safer operation. This paper provides a report of a pioneering scheme in the United States of America in which DLR has been applied. Thereby, we demonstrate that observing the local weather conditions in real time leads to additional capacity and safer operation. Secondly, we discuss limitations involved. In doing so, we arrive at novel insights which will inform and improve future DLR projects. Third, we provide a policy background and discussion to clarify the technology's potential and identifies barriers to the imminent adoption of dynamic line rating systems. We provide suggestions for regulatory bodies about possible improvements in policy to encourage adoption of this beneficial technology.

In situ bioremediation (ISB) was not initially considered a feasible technology for treating chlorinated solvent source zones, and it still faces some skepticism. However, experience over the last decade demonstrates that it is a viable technology for treating some source zones, although realistic objectives must be set. It is reasonable to expect 90-99% reductions in groundwater concentrations and mass discharge from a source in many situations, but it is not likely that ISB can achieve complete cleanup of a source zone within a few years. ISB is best applied in stages, optimizing the design and operations over time. Some residual contamination will remain, and modeling suggests that contaminant concentrations may rebound after treatment, although no rebound has been observed so far at field sites, and it is likely that the accumulation of bacterial biomass and reduced minerals during ISB will sustain treatment for several years after active treatment ends. Practitioners considering ISB for a source zone should be aware of several potential difficulties. The electron donor demand can be so great that it is not feasible to supply enough donor and/or pH buffer, or adding large amounts of donor may cause other problems such as methane production, biofouling of wells or clogging of the subsurface. Using ISB to treat source zones requires careful design, monitoring and continuous optimization throughout treatment, and likely will require extended passive management after treatment.

Epithelial intercellular adhesion is fundamental to the formation of the airway epithelial protective barrier. In this respect, cadherins are important because these adhesion molecules regulate formation and maintenance of epithelial intercellular junctions. To study the importance of airway epithelial integrity in determining susceptibility to virus infection, we used a replication-incompetent adenovirus, RAd35, and an E-cadherin specific function-blocking antibody, SHE78-7, to disrupt intercellular contacts in human bronchial epithelial cell line 16HBE14o- and primary bronchial epithelial cells. After exposure of 16HBE14o- cell cultures to SHE78-7, disruption of the transepithelial permeability barrier was indicated by a loss of transepithelial electrical resistance and an associated increase of mannitol, inulin, and dextran paracellular flux. Subsequent exposure of SHE78-7-treated cell cultures to RAd35 showed a remarkable increase in adenoviral infection as assessed by beta-galactosidase reporter gene expression. In cultures exposed to SHE78-7, disruption of E-cadherin function resulted in infection equivalent to that in control cultures using 16-fold lower viral titers. These studies show that manipulation of E-cadherin function provides a specific means of altering epithelial integrity that in turn determines resistance of airway epithelia to adenoviral infection.

Clostridium difficile is a major cause of chronic antibiotic-associated diarrhea and a significant health care-associated pathogen that forms highly resistant and infectious spores. Spo0A is a highly conserved transcriptional regulator that plays a key role in initiating sporulation in Bacillus and Clostridium species. Here, we use a murine model to study the role of the C. difficile spo0A gene during infection and transmission. We demonstrate that C. difficile spo0A mutant derivatives can cause intestinal disease but are unable to persist within and effectively transmit between mice. Thus, the C. difficile Spo0A protein plays a key role in persistent infection, including recurrence and host-to-host transmission in mice.

A two-dimensional, transient-flow, and transport numerical model was developed to simulate in situ chemical oxidation (ISCO) of trichloroethylene and tetrachloroethylene by potassium permanganate in fractured clay. This computer model incorporates dense, nonaqueous phase liquid dissolution, reactive aquifer material, multispecies matrix diffusion, and kinetic formulations for the oxidation reactions. A sensitivity analysis for two types of parameters, hydrogeological and engineering, including matrix porosity, matrix organic carbon, fracture aperture, potassium permanganate dosage, and hydraulic gradient, was conducted. Remediation metrics investigated were the relative rebound concentrations arising from back diffusion and percent mass destroyed. No well-defined correlation was found between the magnitude of rebound concentrations during postremedy monitoring and the amount of contaminant mass destroyed during the application. Results indicate that all investigated parameters affect ISCO remediation in some form. Results indicate that when advective transport through the fracture is dominant relative to diffusive transport into the clay matrix (large System Peclet Number), permanganate is more likely to be flushed out of the system and treatment is not optimal. If the System Peclet Number is too small, indicating that diffusion into the matrix is dominant relative to advection through the fracture, permanganate does not traverse the entire fracture, leading to postremediation concentration rebound. Optimal application of ISCO requires balancing advective transport through the fracture with diffusive transport into the clay matrix.

A mathematical solution for solute transport in a three-dimensional porous medium with a patch source under steady-state, uniform ground water flow conditions was developed by Domenico (1987). The solution derivation strategy used an approximate approach to solve the boundary value problem, resulting in a nonexact solution. Variations of the Domenico (1987) solution are incorporated into the software programs BIOSCREEN and BIOCHLOR, which are frequently used to evaluate subsurface contaminant transport problems. This article mathematically elucidates the error in the approximation and presents simulations that compare different versions of the Domenico (1987) solution to an exact analytical solution to demonstrate the potential error inherent in the approximate expressions. Results suggest that the accuracy of the approximate solutions is highly variable and dependent on the selection of input parameters. For solute transport in a medium-grained sand aquifer, the Domenico (1987) solution underpredicts solute concentrations along the centerline of the plume by as much as 80% depending on the case of interest. Increasing the dispersivity, time, or dimensionality of the system leads to increased error. Because more accurate exact analytical solutions exist, we suggest that the Domenico (1987) solution, and its predecessor and successor approximate solutions, need not be employed as the basis for screening tools at contaminated sites.

Distribution of airway junctional complex proteins after antigen or lipopolysaccharide challenge in sensitized or naive mice, respectively, was investigated. E-cadherin immunoreactivity was detected continuously along neighboring epithelial cell borders and between adjacent alveolar epithelial cells in naive and saline-challenged mice. Occludin and ZO-1 immunoreactivity were observed in the tight junction areas. Both challenges induced changes in epithelial morphology and phenotype, accompanied initially by focal loss of epithelial E-cadherin that increased in size with time and number of allergen challenges. Allergen challenge also led to focal loss of occludin and ZO-1. Western blot analysis revealed increased levels of sE-cadherin in lavage fluid after either challenge, and this increase correlated with lavage neutrophil numbers (P = 0.002). Immunocytochemistry of lavage cells 6 h after either challenge revealed E-cadherin epitopes within cytoplasmic vacuoles of neutrophils, the major cell type. In contrast, peripheral blood neutrophils or tissue neutrophils before epithelial transmigration were negative, suggesting that in airway inflammation, E-cadherin extracellular domain is cleaved by neutrophils during epithelial penetration, instigating the destabilization of adherens and tight junctions. This junctional deterioration could lead to a progressive decrease in epithelial integrity and induce alterations in epithelial morphology, with consequent enhanced paracellular transit of antigens and pathogens.