Analysis of IgA1 N-glycosylation and its contribution to FcalphaRI binding

The IgA isotype of human antibodies triggers inflammatory responses via the IgA-specific receptor FcalphaRI (CD89). Structural studies have suggested that IgA1 N-glycans could modulate the interaction with FcalphaRI. We have carried out detailed biophysical analyses of three IgA1 samples purified from human serum and recombinant IgA1-Fc and compared their binding to FcalphaRI. Analytical ultracentrifugation revealed wide variation in the distribution of polymeric species between IgA1 samples, and Fourier transform ion cyclotron resonance mass spectrometry showed overlapping but distinct populations of N-glycan species between IgA1 samples. Kinetic and equilibrium data from surface plasmon resonance experiments revealed that variation in the IgA1 C H2 N-glycans had no effect on the kinetics or affinity constants for binding to FcalphaRI. Indeed, complete enzymatic removal of the IgA1 N-glycans yielded superimposable binding curves. These findings have implications for renal diseases such as IgA nephropathy.     

Use of doubled haploids in maize breeding: implications for intellectual property protection and genetic diversity in hybrid crops

Through the concerted use of doubled haploidy (DH), molecular markers and off-season nurseries, maize (Zea mays L.) breeders have unprecedented capabilities to quickly and precisely create progeny with desired levels of similarity to either parents of a commercial hybrid. Genotypic data from both simulated and from actual populations created either by single seed descent or through doubled haploidy were examined for the initial and subsequent generations. Simulation data showed that DH progeny inherited larger blocks of parental chromosomes; approximately seven out of 10 chromosomes had intact segments of 50% or greater. By the third DH generation progeny can be selected that are more than 90% similar to either parent of the initial commercial hybrid. Actual marker data from the initial DH generation showed a maximum parental contribution of 88.4% compared to 78.7% for progeny developed by single seed descent (SSD). The number of intact chromosomes was higher among DH progeny than among progeny bred by SSD. Use of DH facilitates access to germplasm that is already present in commercial maize hybrids. Available technologies coupled with the intellectual property protection regime will influence decisions made by plant breeders in the balance of exotic compared to well-adapted germplasm they choose to access for further cycles.     

Dissemination of Cronobacter spp. (Enterobacter sakazakii) in a powdered milk protein manufacturing facility

The microbial contamination of air filters and possible links to contaminated product in a powdered milk protein-processing facility were investigated. Over a 10-month period, seven air filters, the environment, and powdered product were analyzed for the presence of Cronobacter spp. The effects of air filter installation, maintenance, and subsequent dissemination of Cronobacter were investigated. A total of 30 isolates were characterized by pulsed-field gel electrophoresis (PFGE). PFGE revealed the presence of three clonal populations distributed throughout the manufacturing site. This study highlights the need for proper installation of air filters to limit the dissemination of microorganisms into processing sites.     

Improved ECG detection of presence and severity of right ventricular pressure load validated with cardiac magnetic resonance imaging

The study aimed to assess whether the 12-lead ECG-derived ventricular gradient, a vectorial representation of ventricular action potential duration heterogeneity directed toward the area of shortest action potential duration, can improve ECG diagnosis of chronic right ventricular (RV) pressure load. ECGs from 72 pulmonary arterial hypertension patients recorded <30 days before onset of therapy were compared with ECGs from matched healthy control subjects (n = 144). Conventional ECG criteria for increased RV pressure load were compared with the ventricular gradient. In 38 patients a cardiac magnetic resonance (CMR) study had been performed within 24 h of the ECG. By multivariable analysis, combined use of conventional ECG parameters (rsr' or rsR' in V1, R/S > 1 with R > 0.5 mV in V1, and QRS axis >90 degrees ) had a sensitivity of 89% and a specificity of 93% for presence of chronic RV pressure load. However, the ventricular gradient not only had a higher diagnostic accuracy for chronic RV pressure load by receiver operating characteristic analysis [areas under the curve (AUC) = 0.993, SE 0.004 vs. AUC = 0.945, SE 0.021, P < 0.05], but also discriminated between mild-to-moderate and severe RV pressure load. CMR identified an inverse relation between the ventricular gradient and RV mass, and a trend toward a similar relation with RV volume. In conclusion, chronically increased RV pressure load is electrocardiographically reflected by an altered ventricular gradient associated with RV remodeling-related changes in ventricular action potential duration heterogeneity. The use of the ventricular gradient allows ECG detection of even mildly increased RV pressure load.     

Tilt determination in MEMS inertial vestibular prosthesis

BACKGROUND: There is a clear need for a prosthesis that improves postural stability in the balance impaired. Such a device would be used as a temporary aid during recovery from ablative inner-ear surgery, a postural monitor during rehabilitation (for example, hip surgery), and as a permanent prosthesis for those elderly prone to falls. METHOD OF APPROACH: Recently developed, small instruments have enabled wearable prostheses to augment or replace vestibular functions. The current prosthesis communicates by vibrators mounted on the subject's trunk. In this paper we emphasize the unique algorithms that enable tilt indication with modestly performing micromachined gyroscopes and accelerometers. RESULTS: For large angles and multiple axes, gyro drift and unwanted lateral accelerations are successfully rejected. In single-axis tests, the most dramatic results were obtained in standard operating tests where balance-impaired subjects were deprived of vision and proprioceptive inputs. Balance-impaired subjects who fell (into safety restraints) when not aided were able to stand with the prosthesis. Initial multiaxis tests with healthy subjects have shown that sway is reduced in both forward-back and sideward directions. CONCLUSIONS: Positive results in initial testing and a sound theoretical basis for the hardware warrant continued development and testing, which is being conducted at three sites.     

Bioprocess development for scalable production of cultivated meat

Traditional farm-based products based on livestock are one of the main contributors to greenhouse gas emissions. Cultivated meat is an alternative that mimics animal meat, being produced in a bioreactor under controlled conditions rather than through the slaughtering of animals. The first step in the production of cultivated meat is the generation of sufficient reserves of starting cells. In this study, bovine adipose-derived stem cells (bASCs) were used as starting cells due to their ability to differentiate towards both fat and muscle, two cell types found in meat. A bioprocess for the expansion of these cells on microcarriers in spinner flasks was developed. Different cell seeding densities (1,500, 3,000, and 6,000 cells/cm²) and feeding strategies (80%, 65%, 50%, and combined 80%/50% medium exchanges) were investigated. Cell characterization was assessed pre- and postbioprocessing to ensure that bioprocessing did not negatively affect bASC quality. The best growth was obtained with the lowest cell seeding density (1,500 cells/cm²) with an 80% medium exchange performed (p < .0001) which yielded a 28-fold expansion. The ability to differentiate towards adipogenic, osteogenic, and chondrogenic lineages was retained postbioprocessing and no significant difference (p > .5) was found in clonogenicity pre- or postbioprocessing in any of the feeding regimes tested.