101 research outputs found
Defensin-rich granules of human neutrophils: characterization of secretory properties
AbstractThe various granule subtypes of the human neutrophil differ in propensity for exocytosis. As a rule, granules formed at late stages of myelopoiesis have a higher secretory potential than granules formed in more immature myeloid cells. Neutrophils contain four closely related α-defensins, which are stored in a subset of azurophil granules. These defensin-rich azurophil granules (DRG) are formed later than defensin-poor azurophil granules, near the promyelocyte/myelocyte transition. In order to characterize the secretory properties of DRG, we developed a sensitive and accurate ELISA for detection of the neutrophil α-defensins HNP 1–3. This allowed us to quantify the exocytosis of α-defensins and markers of azurophil (myeloperoxidase), specific (lactoferrin) and gelatinase (gelatinase) granules from neutrophils stimulated with different secretagogues. The release pattern of α-defensins correlated perfectly with the release of myeloperoxidase and showed no resemblance to the exocytosis of lactoferrin or gelatinase. This finding was substantiated through subcellular fractionation experiments. In conclusion, despite a distinct profile of biosynthesis, DRG are indistinguishable from defensin-poor azurophil granules with respect to exocytosis. Thus, in contrast to peroxidase-negative granules, azurophil granules display homogeneity in their availability for extracellular release
Validity and reliability of seismocardiography for the estimation of cardiorespiratory fitness
BACKGROUND: Low cardiorespiratory fitness (ie, peak oxygen consumption [V.O2peak]) is associated with cardiovascular disease and all-cause mortality and is recognized as an important clinical tool in the assessment of patients. Cardiopulmonary exercise test (CPET) is the gold standard procedure for determination of V.O2peak but has methodological challenges as it is time-consuming and requires specialized equipment and trained professionals. Seismofit is a chest-mounted medical device for estimating V.O2peak at rest using seismocardiography.OBJECTIVE: The purpose of this study was to investigate the validity and reliability of Seismofit V.O2peak estimation in a healthy population.METHODS: On 3 separate days, 20 participants (10 women) underwent estimations of V.O2peak with Seismofit (×2) and Polar Fitness Test (PFT) in randomized order and performed a graded CPET on a cycle ergometer with continuous pulmonary gas exchange measurements.RESULTS: Seismofit V.O2peak showed a significant bias of -3.1 ± 2.4 mL·min-1·kg-1 (mean ± 95% confidence interval) and 95% limits of agreement (LoA) of ±10.8 mL·min-1·kg-1 compared to CPET. The mean absolute percentage error (MAPE) was 12.0%. Seismofit V.O2peak had a coefficient of variation of 4.5% ± 1.3% and an intraclass correlation coefficient of 0.95 between test days and a bias of 0.0 ± 0.4 mL·min-1·kg-1 with 95% LoA of ±1.6 mL·min-1·kg-1 in test-retest. In addition, Seismofit showed a 2.4 mL·min-1·kg-1 smaller difference in 95% LoA than PFT compared to CPET.CONCLUSION: The Seismofit is highly reliable in its estimation of V.O2peak. However, based on the measurement error and MAPE >10%, the Seismofit V.O2peak estimation model needs further improvement to be considered for use in clinical settings.</p
A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a disposable polymer microfluidic device.
A Disposable Polymer Lab-On-A-Slide For Point-Of-Care Diagnostics Of Methicillin-Resistant Staphylococcus Aureus (Mrsa)
Selective N-terminal acylation of peptides and proteins with a Gly-His tag sequence
His-tagged proteins can undergo N-terminal acylation as an undesired side-reaction. Here, the authors utilize this to develop a method for highly selective acylation and further modification of peptides and proteins using an optimized His sequence and 4-methoxyphenyl esters as acyl donors
Design and Synthesis of a Quintessential Self-Transmissible IncX1 Plasmid, pX1.0
DNA exchange in bacteria via conjugative plasmids is believed to be among the most important contributing factors to the rapid evolution- and diversification rates observed in bacterial species. The IncX1 plasmids are particularly interesting in relation to enteric bacteria, and typically carry genetic loads like antibiotic resistance genes and virulence factors. So far, however, a “pure” version of these molecular parasites, without genetic loads, has yet to be isolated from the environment. Here we report the construction of pX1.0, a fully synthesized IncX1 plasmid capable of horizontal transfer between different enteric bacteria. The designed pX1.0 sequence was derived from the consensus gene content of five IncX1 plasmids and three other, more divergent, members of the same phylogenetic group. The pX1.0 plasmid was shown to replicate stably in E. coli with a plasmid DNA per total DNA ratio corresponding to approximately 3–9 plasmids per chromosome depending on the growth phase of the host. Through conjugation, pX1.0 was able to self-transfer horizontally into an isogenic strain of E. coli as well as into two additional species belonging to the family Enterobacteriaceae. Our results demonstrate the immediate applicability of recent advances made within the field of synthetic biology for designing and constructing DNA systems, previously existing only in silica
Performance Comparison of Controllers with Fault-Dependent Control Allocation for UAVs
This paper combines fault-dependent control allocation with three different control schemes to obtain fault tolerance in the longitudinal control of unmanned aerial vehicles. The paper shows that fault-dependent control allocation is able to accommodate actuator faults that would otherwise be critical and it makes a performance assessment for the different control algorithms: an L1 adaptive backstepping controller; a robust sliding mode controller; and a standard PID controller. The actuator faults considered are the partial to total loss of the elevator, which is a critical component for the safe operation of unmanned aerial vehicles. During nominal operation, only the main actuator, namely the elevator, is active for pitch control. In the event of a partial or total loss of the elevator, fault-dependent control allocation is used to redistribute control to available healthy actuators. Using simulations of a Cessna 182 aircraft model, controller performance and robustness are evaluated by metrics that assess control accuracy and energy use. System uncertainties are investigated over an envelope of pertinent variation, showing that sliding mode and L1 adaptive backstepping provide robustness, where PID control falls short. Additionally, a key finding is that the fault-dependent control allocation is instrumental when handling actuator faults
- …