Calpains are a family of calcium-dependent cysteine proteases that are ubiquitously expressed in mammals and play critical roles in neuronal death by catalyzing substrate proteolysis. Here, we ...developed two-dimensional gel electrophoresis-based protease proteomics to identify putative calpain substrates. To accomplish this, cellular lysates from neuronal cells were first separated by pI, and the immobilized sample on a gel strip was incubated with a recombinant calpain and separated by molecular weight. Among 25 altered protein spots that were differentially expressed by at least 2-fold, we confirmed that arsenical pump-driving ATPase, optineurin, and peripherin were cleaved by calpain using in vitro and in vivo cleavage assays. Furthermore, we found that all of these substrates were cleaved in MN9D cells treated with either ionomycin or 1-methyl-4-phenylpyridinium, both of which cause a calcium-mediated calpain activation. Their cleavage was blocked by calcium chelator or calpain inhibitors. In addition, calpain-mediated cleavage of these substrates and its inhibition by calpeptin were confirmed in a middle cerebral artery occlusion model of cerebral ischemia, as well as a stereotaxic brain injection model of Parkinson disease. Transient overexpression of each protein was shown to attenuate 1-methyl-4-phenylpyridinium-induced cell death, indicating that these substrates may confer protection of varying magnitudes against dopaminergic injury. Taken together, the data indicate that our protease proteomic method has the potential to be applicable for identifying proteolytic substrates affected by diverse proteases. Moreover, the results described here will help us decipher the molecular mechanisms underlying the progression of neurodegenerative disorders where protease activation is critically involved.
Background: It is important to assess contribution of calpain activation and identify substrates affected during neurodegeneration.
Results: Gel-based protease proteomics identified novel substrates that were cleaved in neurotoxin-treated culture and rat brain disease models.
Conclusion: These novel calpain substrates may confer protection against neurodegeneration.
Significance: Our findings contribute to better deciphering the molecular mechanism underlying the progression of protease-mediated neurodegeneration.
Objective: SB4, SB2, and SB5 are biosimilars of etanercept (ETN), infliximab (INF), and adalimumab (ADA), respectively. This pooled analysis evaluated the immunogenicity of these treatments across ...three phase III randomized controlled trials of patients with rheumatoid arthritis (RA).
Methods: Patients had to have at least one anti-drug antibody (ADAb) assessment up to the time of the primary endpoint from each study (week 24 in SB4 and SB5 studies; week 30 in SB2 study). The effect of ADAbs on American College of Rheumatology 20% (ACR20) response and the incidences of injection-site reactions (ISRs)/infusion-related reactions (IRRs) were evaluated.
Results: The study included 1709 patients. The cumulative incidences of ADAbs were 30.3% in the all-treatments-combined group, 29.1% in the biosimilars combined group, and 31.5% in the reference products combined group. ACR20 response rates were significantly lower in ADAb-positive patients in the all-treatments-combined odds ratio (95% confidence interval) 1.77 (1.37, 2.27), p < 0.0001, biosimilars combined 2.24 (1.53, 3.30), p < 0.0001, and reference products combined 1.49 (1.06, 2.09), p = 0.0225 groups. ADAb-positive patients also had a higher likelihood of developing ISRs/IRRs in the all-treatments-combined group 0.56 (0.31, 1.01), p = 0.0550, predominantly due to the results observed with SB2 + INF combined rather than with SB4 + ETN or SB5 + ADA combined.
Conclusion: In this pooled analysis, ADAbs were associated with reduced efficacy in patients with RA treated with biosimilars (SB4, SB2, and SB5) or their reference products (ETN, INF, and ADA). ADAbs were associated with an increased incidence of ISRs/IRRs in those treated with SB2 + INF.
Clinical trial registration numbers: NCT01936181 (SB2 study), NCT01895309 (SB4 study), and NCT02167139 (SB5 study).
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
The stopping power of liquid water was measured for the first time for carbon ions in the energy range between 1 and 6 MeV using the inverted Doppler shift attenuation method. The feasibility study ...carried out within the scope of the present work shows that this method is well suited for the quantification of the controversial condensed phased effect in the stopping power for heavy ions in the intermediate energy range. The preliminary results of this work indicate that the stopping power of water for carbon ions with energies prevailing in the Bragg-peak region is significantly lower than that of water vapor. In view of the relatively high uncertainty of the present results, a new experiment with uncertainties less than the predicted difference between the stopping powers of both water phases is planned.
Biporous silicates with both a MCM‐41 mesophase and a skeletal macrophase (see Figure) are synthesized using a new and simple method. Transmission electron microscopy confirms the existence of both ...mesopores and macropores, and a model of the formation of skeletal macroporosity is proposed. The synthetic method could lead to larger‐scale production of powdered biporous silicates.
Near-infrared spectroscopy assesses cerebral tissue oxygen saturation (Scto2) based on the absorption spectra of oxygenated and deoxygenated hemoglobin. It has been reported that IV-administered dyes ...including methylene blue, indigo carmine, and indocyanine green (ICG) may cause falsely low-pulse oximetry readings (Spo2). Although methylene blue and indigo carmine may also decrease Scto2, the effect of ICG has not been documented.
Simultaneous changes in the heart rate, arterial blood pressure, Scto2, and Spo2 were measured after IV administration of ICG (12.5 mg diluted in 5.0 mL 0.9% NaCl) in 15 patients undergoing carotid endarterectomy under sevoflurane-remifentanil anesthesia.
After the dye administration, no change in heart rate or arterial blood pressure was observed in any patient. Scto2 increased by 13.3±4.0 percentage points, reaching the peak at 42.0±28.4 seconds after the administration, whereas Spo2 decreased by 1.9±1.2 percentage points, reaching the peak at 64.0±42.5 seconds (P<0.0001 both).
ICG falsely increases the spectroscopy-determined cerebral oxygen saturation for up to 12 minutes but dampens pulse oximetry readings.
After 7-valent pneumococcal conjugate vaccine (PCV7) introduction, non-vaccine serotypes such as 19A are increasing among Streptococcus pneumoniae. However, only limited data on 19A S. pneumoniae are ...available in Asian countries.
Out of 1637 S. pneumoniae clinical pneumonia isolates collected during 2008 and 2009 from 10 Asian countries (Korea, Malaysia, Taiwan, Thailand, Saudi Arabia, Hong Kong, India, Japan, the Philippines and Vietnam), 91 serotype 19A S. pneumoniae isolates were identified. Capsular swelling reaction identified serotype 19A isolates. Antimicrobial susceptibility testing was performed on the serotype 19A isolates using the broth microdilution method, and the genotypes of the isolates were assessed using multilocus sequence typing.
Thirty different sequence types (STs) were identified. The most prevalent clone was ST320 (46 isolates, 51.1%). ST320 was found in Hong Kong, India, Korea, Malaysia, Saudi Arabia and Taiwan. ST320 isolates were mostly multidrug resistant (MDR) and showed significantly higher resistance rates than other STs for cefuroxime, clindamycin, and trimethoprim/sulfamethoxazole.
Although diverse clones were identified among 19A S. pneumoniae isolates, MDR ST320 was the predominant clone in Asian countries. Its predominance, even in countries with no or low coverage of PCV7, may indicate that its emergence and dissemination was due to more than just vaccine selection pressure in Asian countries. A longitudinal investigation of the change of serotypes and genotypes since the introduction of PCV7 is required to understand the emergence and dissemination mechanisms of a certain clone of 19A S. pneumoniae isolates.
Background
The pandemic strain of the influenza A virus (pH1N1) in 2009 caused many complications in patients. In this study, we introduce asthmatic symptoms as a complication of pH1N1 infection in ...children, not having a relationship with asthma history. The aim of this study was to quantify asthmatic symptoms in pH1N1‐infected children and elucidate the underlying mechanisms of airway hyper‐responsiveness (AHR) induced in a murine model of pH1N1 infection.
Methods
As a retrospective study, pH1N1‐infected children who were hospitalized with moderate to severe acute asthmatic symptoms were enrolled and administered a methacholine challenge test (MCT) at 3 months post‐discharge. Additionally, the induction of AHR by pH1N1 infection was measured by MCT in wild‐type and Rag1−/− mice. The effect of the innate immune response on the development of AHR following pH1N1 infection was investigated.
Results
More than 70% of the pH1N1‐infected children without a pre‐infection diagnosis of asthma had a negative response on the MCT. None of these children had recurrent wheezing or asthma during the 3 years following pH1N1 infection. The development of AHR in pH1N1‐infected mice was associated with an elevation in IL‐33 and innate lymphoid cells 2 (ILC2).
Conclusions
This study demonstrates that pH1N1 infection directly induces transient asthmatic symptoms in patients regardless of their medical history. pH1N1 infection was shown to stimulate the rapid development of AHR and Th2‐type cytokine secretion in mice via the activation of ILC2; it may be activated independently of adaptive immunity.
Purpose: The commercialization of sleep activity tracking devices has made it possible to manage sleep quality at home. However, it is necessary to verify the reliability and accuracy of wearable ...devices through comparison with polysomnography (PSG), which is the standard for tracking sleep activity. This study aimed to monitor overall sleep activity using Fitbit Inspire 2™ (FBI2) and to evaluate its performance and effectiveness through PSG under the same conditions. Patients and Methods: We compared the FBI2 and PSG data of nine participants (four male and five female participants; average age, 39 years) without severe sleeping problems. The participants wore FBI2 continuously for 14 days, considering the period of adaptation to the device. FBI2 and PSG sleep data were compared using paired t-tests, Bland–Altman plots, and epoch-by-epoch analysis for 18 samples by pooling data from two replicates. Results: The average values for each sleep stage obtained from FBI2 and PSG showed significant differences in the total sleep time (TST), deep sleep, and rapid eye motion (REM). In the Bland–Altman analysis, TST (P = 0.02), deep sleep (P = 0.05), and REM (P = 0.03) were significantly overstated in FBI2 compared to PSG. In addition, time in bed, sleep efficiency, and wake after sleep onset were overestimated, while light sleep was underestimated. However, these differences were not statistically significant. FBI2 showed a high sensitivity (93.9%) and low specificity (13.1%), with an accuracy of 76%. The sensitivity and specificity of each sleep stage was 54.3% and 62.3%, respectively, for light sleep, 84.8% and 50.1%, respectively, for deep sleep, and 86.4% and 59.1%, respectively for REM sleep. Conclusion: The use of FBI2 as an objective tool for measuring sleep in daily life can be considered appropriate. However, further research is warranted on its application in participants with sleep-wake problems.
The effect of the desorption-recombination temperature on the microstructure and magnetic properties of hydrogenation-disproportionation-desorption-recombination (HDDR) processed Nd-Fe-B powders was ...studied. The Nd
Ga
Nb
Fe
(x=12.5-13.5, at.%) casting alloys were pulverized after homogenizing annealing, and then subjected to HDDR treatment. During the HDDR process, desorption-recombination (DR) reaction was induced at two different temperature, 810°C and 820°C. The higher Nd content resulted in enhanced coercivity of the HDDR powder, and which was attributed to the thicker and more uniform Nd-rich phase along grain boundaries. But this uniform Nd-rich phase induced faster grain growth. The remanence of the powder DR-treated at 820°C is higher than that DR-treated at 810°C. In addition, it was also confirmed that higher DR temperature is much more effective to improve squareness.
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