A novel repeat sequence-based PCR (rep-PCR) using specific repeat sequences of Mycobacterium intracellulare as a DNA fingerprinting

Repetitive sequence-based PCR (rep-PCR) is a potential epidemiological technique that can provide high-throughput genotype fingerprints of heterogeneous Mycobacterium strains rapidly. Previously published rep-PCR primers, which are based on nucleotide sequences of Gram-negative bacteria may have low specificity for mycobacteria. Moreover, it was difficult to ensure the continuity of the study after the commercial rep-PCR kit was discontinued. Here, we designed a novel rep-PCR for Mycobacterium intracellulare, a major cause of nontuberculous mycobacterial pulmonary disease with frequent recurrence. We screened the 7,645 repeat sequences for 200 fragments from the genome of M. intracellulare ATCC 13950 in silico, finally generating five primers with more than 90% identity for a total of 226 loci in the genome. The five primers could make different band patterns depending on the genome of three different M. intracellulare strains using an in silico test. The novel rep-PCR with the five primers was conducted using 34 bacterial samples of 7 species containing 25 M. intracellulare clinical isolates, compared with previous published rep-PCRs. This shows distinguished patterns depending on species and blotting assay for 6 species implied the sequence specificity of the five primers. The Designed rep-PCR had a 95–98% of similarity value in the reproducibility test and showed 7 groups of fingerprints in M. intracellulare strains. Designed rep-PCR had a correlation value of 0.814 with VNTR, reference epidemiological method. This study provides a promising genotype fingerprinting method for tracing the recurrence of heterogeneous M. intracellulare

Evaluation of Cortical Bone Formation on Mandibular Condyle in Asymptomatic Adolescents and Young Adults Using Cone-Beam Computed Tomography

The aim of this study was to evaluate cortical bone formation on the mandibular condyle using cone-beam computed tomography (CBCT) in asymptomatic adolescents and young adults and to evaluate the relationship between age and sex. CBCT images that can evaluate the shape of the mandibular condyle were selected from asymptomatic patients aged 13–25. The degree of cortication on the mandibular condyle (CMC) was evaluated using CBCT images reconstructed in the axial, sagittal, and coronal planes. CBCT data of 829 patients (413 males, 416 females) were selected and then the left and right images of all patients were acquired; consequently, a total of 1658 temporomandibular joint-related images were evaluated in this study. The degree of CMC was correlated with age in men and women (p < 0.05). The frequency of CMC 0 disappeared in woman aged 20 years and in men aged 21 years. Cortical bone formation of the mandibular condyle was completed at age 22 years in women and 24 years in men. The degrees of cortical bone formation of the mandibular condyle between men and women showed significant differences between the ages of 15–19 and 22 years. This difference can be interpreted as a different mandible growth period between the sexes

Neuronal ApoE Regulates the Cell-to-Cell Transmission of α-Synuclein

The presence of protein inclusions, called Lewy bodies (LBs) and Lewy neurites (LNs), in the brain is the main feature of Parkinson’s disease (PD). Recent evidence that the prion-like propagation of α-synuclein (α-syn), as a major component of LBs and LNs, plays an important role in the progression of PD has gained much attention, although the molecular mechanism remains unclear. In this study, we evaluated whether neuronal ApoE regulates the cell-to-cell transmission of α-syn and explored its molecular mechanism using in vitro and in vivo model systems. We demonstrate that neuronal ApoE deficiency attenuates both α-syn uptake and release by downregulating LRP-1 and LDLR expression and enhancing chaperone-mediated autophagy activity, respectively, thereby contributing to α-syn propagation. In addition, we observed that α-syn propagation was attenuated in ApoE knockout mice injected with pre-formed mouse α-syn fibrils. This study will help our understanding of the molecular mechanisms underlying α-syn propagation

Annexin A1 in plasma from patients with bronchial asthma: its association with lung function

Abstract Background Annexin-A1 (ANXA1) is a glucocorticoid-induced protein with multiple actions in the regulation of inflammatory cell activation. The anti-inflammatory protein ANXA1 and its N-formyl peptide receptor 2 (FPR2) have protective effects on organ fibrosis. However, the exact role of ANXA1 in asthma remains to be determined. The aim of this study was to identify the role of ANXA1 in bronchial asthma. Methods In mice sensitized and challenged with ovalbumin (OVA-OVA mice) and mice sensitized with saline and challenged with air (control mice), we investigated the potential links between ANXA1 levels and bronchial asthma using ELISA, immunoblotting, and immunohistochemical staining. Moreover, we also determined ANXA1 levels in blood from 50 asthmatic patients (stable and exacerbated states). Results ANXA1 protein levels in lung tissue and bronchoalveolar lavage fluid were significantly higher in OVA-OVA mice compared with control mice. FPR2 protein levels in lung tissue were significantly higher in OVA-OVA mice compared with control mice. Plasma ANXA1 levels were increased in asthmatic patients compared with healthy controls. Plasma ANXA1 levels were significantly lower in exacerbated patients compared with stable patients with bronchial asthma (p < 0.05). The plasma ANXA1 levels in controlled asthmatic patients were correlated with forced expiratory volume in 1 s (FEV1) (r = − 0.191, p = 0.033) and FEV1/forced vital capacity (FVC) (r = −0.202, p = 0.024). Conclusion These results suggest that ANXA1 may be a potential marker and therapeutic target for asthma

Communication: Comparison of Respiratory Specimens for the Detection of SARS-CoV-2

We compared SARS-CoV-2 detection rate of different respiratory specimens (nasopharyngeal swab [NPS], n=92; oropharyngeal swab [OPS], n=18; sputum, n=11). We also compared cycle threshold (Ct) values of paired specimen types obtained from the same patient on the same day. Then we characterized viral load kinetics of NPS (n=142), OPS (n=126), and sputum (n=75), during disease course. Sputum samples showed higher detection rate than NPS, and OPS exhibited the lowest detection rate. The median Ct values in NPS were significantly lower than in paired OPS, and higher than in paired sputum, respectively (P&lt;0.05). During the disease course, viral load was the lowest in OPS and the highest in sputum samples.N

The Importance of Porins and β-Lactamase in Outer Membrane Vesicles on the Hydrolysis of β-Lactam Antibiotics

Gram-negative bacteria have an outer membrane inhibiting the entry of antibiotics. Porins, found within the outer membrane, are involved in regulating the permeability of &beta;-lactam antibiotics. &beta;-lactamases are enzymes that are able to inactivate the antibacterial properties of &beta;-lactam antibiotics. Interestingly, porins and &beta;-lactamase are found in outer membrane vesicles (OMVs) of &beta;-lactam-resistant Escherichia coli and may be involved in the survival of susceptible strains of E. coli in the presence of antibiotics, through the hydrolysis of the &beta;-lactam antibiotic. In this study, OMVs isolated from &beta;-lactam-resistant E. coli and from mutants, lacking porin or &beta;-lactamase, were evaluated to establish if the porins or &beta;-lactamase in OMVs were involved in the degradation of &beta;-lactam antibiotics. OMVs isolated from E. coli deficient in &beta;-lactamase did not show any degradation ability against &beta;-lactam antibiotics, while OMVs lacking OmpC or OmpF showed significantly lower levels of hydrolyzing activity than OMVs from parent E. coli. These data reveal an important role of OMVs in bacterial defense mechanisms demonstrating that the OmpC and OmpF proteins allow permeation of &beta;-lactam antibiotics into the lumen of OMVs, and antibiotics that enter the OMVs can be degraded by &beta;-lactamase