Structural basis underlying the synergism of NADase and SLO during group A Streptococcus infection.

Group A Streptococcus (GAS) is a strict human pathogen possessing a unique pathogenic trait that utilizes the cooperative activity of NAD+-glycohydrolase (NADase) and Streptolysin O (SLO) to enhance its virulence. How NADase interacts with SLO to synergistically promote GAS cytotoxicity and intracellular survival is a long-standing question. Here, the structure and dynamic nature of the NADase/SLO complex are elucidated by X-ray crystallography and small-angle scattering, illustrating atomic details of the complex interface and functionally relevant conformations. Structure-guided studies reveal a salt-bridge interaction between NADase and SLO is important to cytotoxicity and resistance to phagocytic killing during GAS infection. Furthermore, the biological significance of the NADase/SLO complex in GAS virulence is demonstrated in a murine infection model. Overall, this work delivers the structure-functional relationship of the NADase/SLO complex and pinpoints the key interacting residues that are central to the coordinated actions of NADase and SLO in the pathogenesis of GAS infection.

Intelligent Healthcare System Using Mathematical Model and Simulated Annealing to Hide Patients Data in the Low-Frequency Amplitude of ECG Signals.

Healthcare is an important medical topic in recent years. In this study, the novelty we propose is the intelligent healthcare system using an inequality-type optimization mathematical model with signal-to-noise ratio (SNR) and wavelet-domain low-frequency amplitude adjustment techniques to hide patients' confidential data in their electrocardiogram (ECG) signals. The extraction of the hidden patient information also utilizes the low-frequency amplitude adjustment. The detailed steps of establishing the system are as follows. To integrate confidential patient data into ECG signals, we first propose a nonlinear model to optimize the quality of ECG signals with the embedded patients' confidential data including patient name, patient birthdate, date of medical treatment, and medical history. Then, we apply Simulated Annealing (SA) to solve the nonlinear model such that the ECG signals with embedded patients' confidential data have good SNR, good root mean square error (RMSE), and high similarity. In other words, the distortion of the PQRST complexes and the ECG shape caused by the embedded patients' confidential data is very small, and thus the quality of the embedded ECG signals meets the requirements of physiological diagnostics. In the terminals, one can receive the ECG signals with the embedded patients' confidential data. In addition, the embedded patients' confidential data can be received and extracted without the original ECG signals. The experimental results confirm the efficiency that our method maintains a high quality of each ECG signal with the embedded patient confidential data. Moreover, the embedded confidential data shows a good robustness against common attacks.

Effectiveness and response differences of a multidisciplinary workplace health promotion program for healthcare workers.

Background: Workplace health promotion (WHP) in the healthcare industry is an important yet challenging issue to address, given the high workload, heterogeneity of work activities, and long work hours of healthcare workers (HCWs). This study aimed to investigate the effectiveness and response differences of a multidisciplinary WHP program conducted in HCWs. Methods: This retrospective cohort study included HCWs participating in a multidisciplinary WHP program in five healthcare facilities. The 20-week intervention included multiple easy-to-access 90-min exercise classes, one 15-min nutrition consultation, and behavioral education. Pre- and post-interventional anthropometrics, body composition, and physical fitness (PF) were compared with paired sample t-tests. Response differences across sex, age, weight status, and shiftwork status were analyzed with a generalized estimating equation. Results: A total of 302 HCWs were analyzed. The intervention effectively improved all anthropometric (body mass index, waist circumference, waist-hip ratio, and waist-to-height ratio), body composition (body fat percentage, muscle weight, visceral fat area), and PF (grip strength, high jump, sit-up, sit-and-reach, step test) parameters in all participants (all p < 0.05). Subgroup analyses revealed shift workers had a more significant mean reduction in body mass index than non-shift workers (adjusted p = 0.045). However, there was no significant response difference across sex, age, and weight subgroups. Conclusion: This study suggested that a multidisciplinary WHP program can improve anthropometric and PF profiles regardless of sex, age, and weight status for HCWs, and shifter workers might benefit more from the intervention.

Application of deep machine learning for the radiographic diagnosis of periodontitis.

OBJECTIVE: Successful application of deep machine learning could reduce time-consuming and labor-intensive clinical work of calculating the amount of radiographic bone loss (RBL) in diagnosing and treatment planning for periodontitis. This study aimed to test the accuracy of RBL classification by machine learning. MATERIALS AND METHODS: A total of 236 patients with standardized full mouth radiographs were included. Each tooth from the periapical films was evaluated by three calibrated periodontists for categorization of RBL and radiographic defect morphology. Each image was pre-processed and augmented to ensure proper data balancing without data pollution, then a novel multitasking InceptionV3 model was applied. RESULTS: The model demonstrated an average accuracy of 0.87 ± 0.01 in the categorization of mild (< 15%) or severe (≥ 15%) bone loss with fivefold cross-validation. Sensitivity, specificity, positive predictive, and negative predictive values of the model were 0.86 ± 0.03, 0.88 ± 0.03, 0.88 ± 0.03, and 0.86 ± 0.02, respectively. CONCLUSIONS: Application of deep machine learning for the detection of alveolar bone loss yielded promising results in this study. Additional data would be beneficial to enhance model construction and enable better machine learning performance for clinical implementation. CLINICAL RELEVANCE: Higher accuracy of radiographic bone loss classification by machine learning can be achieved with more clinical data and proper model construction for valuable clinical application.

Alterations in kinematics of temporomandibular joint associated with chronic neck pain.

BACKGROUND: Temporomandibular disorder (TMD) is an umbrella term for pain and dysfunction of the temporomandibular joint (TMJ) and its associated structures. Patients with TMD show changes in TMJ kinematics and masticatory muscle activation. TMD is commonly comorbid with non-specific chronic neck pain (NCNP), which may be one of the risk factors for TMD. OBJECTIVES: This study aimed to investigate whether patients with NCNP have altered TMJ kinematics and masticatory muscle activity. METHODS: This was a cross-sectional exploratory study including 19 healthy participants and 20 patients with NCNP but without TMD symptoms. TMJ kinematics was measured during mouth opening and closing, jaw protrusion and jaw lateral deviation. Surface electromyography was used to record the muscle activity of the anterior temporalis, masseter, sternocleidomastoid and upper trapezius while clenching. Furthermore, cervical posture, cervical range of motion (ROM) and pressure-pain threshold of the neck and masticatory muscles were measured. RESULTS: Compared with the healthy group, the NCNP group showed significantly reduced upper cervical rotation ROM (p = .041) and increased condylar path length (p = .02), condylar translation (opening p = .034, closing p = .011) and mechanical pain sensitivity of the upper trapezius (p = .018). Increased condylar translation was significantly correlated with reduced upper cervical mobility and poor cervical posture (r = -0.322 to -0.397; p = .012-.046). CONCLUSION: Increased condylar translation and path length in patients with NCNP may indicate poor control of TMJ articular movement, which may result from neck pain or may be a compensation for limited neck mobility. Evaluation of excessive TMJ translation may be considered in patients with NCNP.

Effect of Doxycycline in Decreasing the Severity of Clostridioides difficile Infection in Mice.

Background: Doxycycline possesses antibacterial activity against Clostridioides difficile and anti-inflammatory effects. Materials and Methods: The influence of doxycycline on the development of CDI was studied in an established animal model of CDI using C57BL/6 mice. Results: Mice intraperitoneally administered doxycycline had higher cecum weight (1.3 ± 0.1 vs. 0.5 ± 0.1 g; p < 0.001) and less body weight reduction (0.7 ± 0.5 g vs. -17.4 ± 0.2 g; p < 0.001) than untreated mice infected with C. difficile. Oral doxycycline, metronidazole, or vancomycin therapy resulted in less body weight reduction in mice with CDI than in untreated mice (1.1 ± 0.1 g, 1.3 ± 0.2 g, 1.2 ± 0.1 g, vs. 2.9 ± 0.3 g; p < 0.001). Doxycycline therapy led to lower expression levels of inflammatory cytokines, such as macrophage inflammatory protein-2 (0.4 ± 0.1 vs. 2.9 ± 1.3, p = 0.02), and higher levels of zonula occludens-1 (1.2 ± 0.1 vs. 0.8 ± 0.1, p = 0.02) in colonic tissues than in untreated mice. Conclusions: Concurrent intraperitoneal administration of doxycycline and oral C. difficile challenge does not aggravate the disease severity of CDI, and oral doxycycline may be a potential therapeutic option for CDI.

The emergence of Clostridioides difficile PCR ribotype 127 at a hospital in northeastern Taiwan.

BACKGROUND: Several studies have highlighted the incidence of Clostridioides difficile infections (CDIs) in Taiwan and certain ribotypes have been related to severe clinical diseases. A study was conducted to investigate the polymerase chain reaction (PCR) ribotypes and genetic relatedness of clinical C. difficile strains collected from January 2009 to December 2015 at a hospital in northeastern Taiwan. MATERIAL AND METHODS: A modified two-step typing algorithm for C. difficile was used by combining a modified 8-plex and 3'-truncated tcdA screening PCR. In addition, MLVA typing was adopted for investigation of bacterial clonality and transmission. RESULTS: Among a total of 86 strains, 24 (28%) were nontoxigenic and 62 (72%) had both tcdA and tcdB (A + B+). No tcdA-negative and tcdB-positive (A-B+) strains were identified. Binary toxin (CDT)-producing (cdtA+/cdtB+) strains were started to be identified in 2013. The 21 (34%) A+B+ clinical strains with binary toxin and tcdC deletion were identified as RT127 strains, which contained both RT078-lineage markers and fluoroquinolone (FQ)-resistant mutations (Thr82Ile in gyrA). Multiple loci variable-number tandem repeat analysis (MLVA) for phylogenetic relatedness of RT127 strains indicated that 20 of 21 strains belonged to a clonal complex that was identical to a clinical strain collected from southern Taiwan in 2011, suggestive of a clonal expansion in Taiwan. CONCLUSION: A two-step typing method could rapidly confirm species identification and define the toxin gene profile of C. difficile isolates. The clonal expansion of RT127 strains in Taiwan indicates monitoring and surveillance of toxigenic C. difficile isolates from human, animal, and environment are critical to develop One Health prevention strategies.

Corrigendum: The Role of Toll-Like Receptor-2 in Clostridioides difficile Infection: Evidence From a Mouse Model and Clinical Patients.

[This corrects the article DOI: 10.3389/fimmu.2021.691039.].

Intrusion detection by machine learning for multimedia platform.

The multimedia service company, Netflix, increased the number of new subscribers during the Coronavirus pandemic age. Intrusion detection systems for multimedia platforms can prevent the platform from network attacks. An intelligent intrusion detection system is proposed for the security IP Multimedia Subsystem (IMS) based on machine learning technology. For increasing the accuracy of the classifiers, it is vital to select the critical features to construct the intrusion detection system. Two-class classifiers, including the Decision Tree, Support Vector Machine, and Naive Bayesian, are selected to evaluate intrusion detection accuracy. According to the three classifiers' accuracy values, the most critical features are selected based on the features' ranking orders. Six critical features are selected:Service, dst_host_same_srv_rate, Flag, Protocol Type, Dst_host_rerror_rate, and Count. Numerical comparison with state_of_the_art shows that critical features improve intrusion detection accuracy, which can be better than the deep learning method.

The Role of Toll-Like Receptor-2 in Clostridioides difficile Infection: Evidence From a Mouse Model and Clinical Patients.

Background: Clostridioides difficile is the leading cause of nosocomial infectious diarrhea. Toll-like receptors (TLRs) are the major components of innate immunity that sense pathogens. The relationship between TLRs and C. difficile infection (CDI) was analyzed in clinical patients and a mouse model. Materials and Methods: A prospective investigation was conducted in medical wards of Tainan Hospital, Ministry of Health and Welfare, Tainan, Taiwan, from January 2011 to January 2013. Adult patients were followed up for the development of CDI. Single nucleotide polymorphisms (SNPs) of TLR2 and TLR4 were analyzed to assess the relationship between genetic polymorphisms and the development of CDI. A mouse model of CDI was used to investigate the pathogenic role of TLRs in CDI, TLR2 and TLR4 knockout (Tlr2-/- and Tlr4-/-) mice. Results: In the prospective study, 556 patients were enrolled, and 6.5% (36) of patients, accounting for 3.59 episodes per 1000 patient-days, developed CDI. Of 539 patients with available blood samples, the TLR2 rs3804099 polymorphism was more often noted in those with CDI than in those without CDI (64.5% vs. 46.1%; P = 0.046) but was not significant in multivariate analysis. Because the TLR2 rs3804099 polymorphism was moderately associated with CDI, the role of TLR2 and TLR4 was further evaluated in a mouse model. Both Tlr2-/- and Tlr4-/- mice showed more severe CDI disease than wild-type mice in terms of body weight change and fecal content five days after oral challenge with C. difficile. Furthermore, Tlr2-/- mice suffered from more severe disease than Tlr4-/- mice, as evidenced by stool consistency, cecum weight, and survival rate. Conclusion: The TLR2 rs3804099 polymorphism is marginally associated with the development of CDI, and the pathogenic role of TLR2 is further supported by a mouse model.

Can Anthropometry and Body Composition Explain Physical Fitness Levels in School-Aged Children?

Physical fitness (PF) is closely related to various health outcomes and quality of life among children. However, the associations between anthropometry, body composition (BC), and PF are not fully elucidated. This cross-sectional study aimed to investigate the associations between demographic metrics (age, sex), anthropometric measures (body mass index z-score (BMI z-score) waist/height ratio (WHtR)), BC parameters (body-fat percentage (BF%), muscle weight), and PF levels (800-m run, sit-and-reach, 1-min sit-ups, standing long jump) in school-aged children. Continuous variables were dichotomized by median splits. The results of 180 girls and 180 boys (mean age: 10.0 ± 0.7 years; mean BMI z-score: 0.366 ± 1.216) were analyzed. Multivariable linear regressions revealed that BF% (regression coefficient (B) = 3.4, 95% confidence interval (CI) = 2.5-4.3) was independently correlated with the 800-m run. Sex (B = 4.6, 95% CI = 3.0-6.3), age (B = 3.1, 95% CI = 1.9-4.3), and BMI z-score (B = -0.7, 95% CI = -1.4--0.1) were independently related to sit-and-reach. Age (B = 3.3, 95% CI = 2.0-4.7), BF% (B = -0.3, 95% CI = -0.4--0.2), and muscle weight (B = 0.7, 95% CI = 0.2-1.2) were independently associated with 1-min sit-ups. In addition to demography, anthropometry and BC provided additional information concerning some PF levels in school-aged children. Weight management and PF promotion should be addressed simultaneously in terms of preventive medicine and health promotion for children.

Ganoderma tsugae suppresses the proliferation of endometrial carcinoma cells via Akt signaling pathway.

Ganoderma is one of the common medicinal mushrooms in traditional Chinese medicine. Previous researches have unveiled the multifaceted biological activity of Ganoderma extract. Ganoderma tsugae has been investigated the potential on curing prostate, colon, lung, epidermoid, breast and ovarian cancers, but not including endometrial cancer. Endometrial cancer is a gynecological malignant tumor with serious drug resistance problem in clinical cancer treatment. This study aimed to demonstrate the first study of Ganoderma on treating endometrial cancer. The Ganoderma tsugae ethanol extract (GTEE) could suppress the proliferation of endometrial cancer cells HEC-1-A, KLE, and AN3 CA. GTEE also induced G1/S phase arrest and mitochondria-mediated apoptosis in endometrial cancer cells. Furthermore, the Akt signaling pathway could be suppressed by GTEE. Therefore, our results suggest for the first time that GTEE has the potential to be an adjuvant therapeutic agent in the treatment of endometrial cancer.

Are Body Composition Parameters Better than Conventional Anthropometric Measures in Predicting Pediatric Hypertension?

Body composition (BC) parameters are associated with cardiometabolic diseases in children; however, the importance of BC parameters for predicting pediatric hypertension is inconclusive. This cross-sectional study aimed to compare the difference in predictive values of BC parameters and conventional anthropometric measures for pediatric hypertension in school-aged children. A total of 340 children (177 girls and 163 boys) with a mean age of 8.8 ± 1.7 years and mean body mass index (BMI) z-score of 0.50 ± 1.24 were enrolled (102 hypertensive children and 238 normotensive children). Significantly higher values of anthropometric measures (BMI, BMI z-score, BMI percentile, waist-to-height ratio) and BC parameters (body-fat percentage, muscle weight, fat mass, fat-free mass) were observed among the hypertensive subgroup compared to their normotensive counterparts. A prediction model combining fat mass ≥ 3.65 kg and fat-free mass ≥ 34.65 kg (area under the receiver operating characteristic curve = 0.688; sensitivity = 66.7%; specificity = 89.9%) performed better than BMI alone (area under the receiver operating characteristic curve = 0.649; sensitivity = 55.9%; specificity = 73.9%) in predicting hypertension. In conclusion, BC parameters are better than anthropometric measures in predicting pediatric hypertension. BC measuring is a reasonable approach for risk stratification in pediatric hypertension.

Membrane Cholesterol Is Crucial for Clostridium difficile Surface Layer Protein Binding and Triggering Inflammasome Activation.

Clostridium difficile, an obligate anaerobic gram-positive bacillus, generates spores and is commonly found colonizing the human gut. Patients with C. difficile infection (CDI) often exhibit clinical manifestations of pseudomembranous colitis or antibiotic-associated diarrhea. Surface layer proteins (SLPs) are the most abundant proteins in the C. difficile cell wall, suggesting that they might involve in immune recognition. Our previous results demonstrated that C. difficile triggers inflammasome activation. Here, we found SLPs as well as C. difficile induced inflammasome activation, and in a dose-dependent manner. In addition, the cholesterol-rich microdomains on the cell membrane (also referred to as lipid rafts) are thought to be crucial for bacterial adhesion and signal transduction. We demonstrated that lipid rafts participated in C. difficile SLPs binding to the cell membrane. Fluorescence microscopy showed that membrane cholesterol depletion by methyl-ß-cyclodextrin (MßCD) reduced the association of SLPs with the cell surface. The coalescence of SLPs in the cholesterol-rich microdomains was confirmed in C. difficile-infected cells. Furthermore, the inflammasome activations induced by SLPs or C. difficile were abrogated by MßCD. Our results demonstrate that SLPs recruit the lipid rafts, which may be a key step for C. difficile colonization and inducing inflammasome activation.

Genomic Differences between Listeria monocytogenes EGDe Isolates Reveal Crucial Roles for SigB and Wall Rhamnosylation in Biofilm Formation.

Listeria monocytogenes is a Gram-positive firmicute that causes foodborne infections, in part due to its ability to use multiple strategies, including biofilm formation, to survive adverse growth conditions. As a potential way to screen for genes required for biofilm formation, we harnessed the ability of bacteria to accumulate mutations in the genome over time, diverging the properties of seemingly identical strains. By sequencing the genomes of four laboratory reference strains of the commonly used L. monocytogenes EGDe, we showed that each isolate contains single nucleotide polymorphisms (SNPs) compared with the reference genome. We discovered that two SNPs, contained in two independent genes within one of the isolates, impacted biofilm formation. Using bacterial genetics and phenotypic assays, we confirmed that rsbU and rmlA influence biofilm formation. RsbU is the upstream regulator of the alternative sigma factor SigB, and mutation of either rsbU or sigB increased biofilm formation. In contrast, deletion of rmlA, which encodes the first enzyme for TDP-l-rhamnose biosynthesis, resulted in a reduction in the amount of biofilm formed. Further analysis of biofilm formation in a strain that still produces TDP-l-rhamnose but which cannot decorate the wall teichoic acid with rhamnose (rmlT mutant) showed that it is the decorated wall teichoic acid that is required for adhesion of the cells to surfaces. Together, these data uncover novel routes by which biofilm formation by L. monocytogenes can be impacted.IMPORTANCE Biofilms are an important mode of growth in many settings. Here, we looked at small differences in the genomes of the bacterium Listeria monocytogenes isolate EGDe and used them to find out how biofilms form. This important fundamental information may help new treatments to be developed and also highlights the fact that isolates of the same identity often diverge.

Antioxidant property of Taraxacum formosanum Kitam and its antitumor activity in non-small-cell lung cancer cells.

BACKGROUND: Non-small-cell lung cancer (NSCLC) is known to exhibit resistance to various therapeutic agents and become progressively incurable. Taraxacum formosanum is a medicinal Chinese herb that has been clinically used in Taiwan. However, the investigations of the effects of whole plant on lung cancer are limited. PURPOSE: This study evaluated the in vitro antioxidant, antiproliferative, and antimigration effects of the ethanol extract of T. formosanum (ETF). The possible molecular mechanism underlying its antitumor effects on cultured human NSCLC cell lines was also elucidated. METHODS: The antioxidant effects of the ETF were determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Trolox equivalent antioxidant capacity (TEAC) assays, and its antiproliferative and antimigration effects were determined using trypan blue exclusion and wound healing assays, respectively. In addition, changes in the mitogen-activated protein kinase (MAPK) signaling pathway were investigated using Western blot analyses. Various inhibitors were used to determine the roles of the MAPK signaling pathway involved in the molecular mechanism of the ETF. RESULTS: Our results showed that the ETF exhibited strong reducing power, a high Trolox equivalent antioxidant capacity (TEAC) value, and potent 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging and Fe+2-chelating abilities. The ETF also exerted antiproliferative and antimigration effects on NSCLC cells in a dose-dependent manner. These effects may be mediated by the inhibitory effects of the ETF on the activation of extracellular signal-regulated kinase. CONCLUSIONS: This study performed the first pharmacological exploration of T. formosanum. Our results demonstrated the antioxidant and antitumor effects of the ETF on NSCLC cell lines, indicating their potential preventive and therapeutic values for lung cancer.