Nasopharyngeal Research Articles

A study to evaluate the role of antioxidants and iron as diagnostic markers in oral cancer

Oral cavity (OC) and oropharyngeal (OP) cancers afflict about 500,000 people globally. Dental health care practitioners need to understand the causes, diagnosis, and treatment of oral cavities and oropharyngeal cancer. The constant and direct exposure of oral mucosal cells to tobacco product chemical carcinogens such as polynuclear aromatic hydrocarbons and nitrosamines causes the generation of free radicals/reactive oxygen species. Changes in salivary antioxidant and micronutrient levels may prove to be an important factor in the pathogenesis of oral premalignant and malignant disorders. The present study aimed to evaluate the levels of Vitamin C, MDA and iron levels in oral cancer, pre-malignant group and the oral cancer group. 15 oral cancer patients, 30 premalignant condition patients, and 15 normal controls were analyzed in the study. Among the 30 patients of the premalignant conditions- group (Group S1) 12 were diagnosed with oral submucous fibrosis, and 18 of them were diagnosed with oral leukoplakia. The oral cancer patients (Group S2) were segregated based on the TNM system from stages I to IV. The oral submucous fibrosis patients were subdivided into four grades. The oral leukoplakia patients were categorized into homogenous and speckled types. A detailed history was obtained from the patients. After obtaining informed consent, saliva samples will be collected from patients who have not consumed any food or any form of tobacco 2 hours before collection. Following a thorough mouth rinse using distilled water, saliva is allowed to accumulate in his or her mouth for 5 minutes. Accumulated saliva is collected by the spit method. 5ml of blood sample will also be collected from these patients, Serum was separated by Centrifugation. Collected serum and saliva were stored at -80℃ until further analysis. The serum and salivary samples were then analyzed for Vitamin C, iron, and malonaldehyde (lipid peroxidation) according to the standardized protocol. In the present study, a total of 60 samples were screened for Vitamin C, Iron, and MDA in both serum and saliva of normal control, premalignant, and oral cancer patients. We found that there was a decreased level of Vitamin C and iron in both serum and saliva of premalignant and oral cancer patients. MDA level was higher in premalignant, and oral cancer patients and there was a significant increase in the level of MDA in oral cancer patients. Understanding antioxidants can help reduce the incidence of oral cancer in its early stages. More study is needed before this supplement may be recommended as an adjuvant treatment.

Unlocking the Therapeutic Potential of LncRNA BLACAT1 in Hypopharynx Squamous Cell Carcinoma.

Identifying the key molecular targets in hypopharynx squamous cell carcinoma (HSCC) is crucial for understanding this prevalent and highly fatal type of head and neck tumor. The study aims to enhance comprehension of the HSCC process by accurately identifying these key molecular targets. In this study, we examined 47 clinical tissue samples from individuals diagnosed with HSCC using RNA-seq high-throughput assay. Quantitative real-time PCR (RT-PCR) was used to compare long non-coding RNA (lncRNA) bladder cancer-associated transcript 1 (BLACAT1) expression in HSCC tissues versus adjacent non-tumor tissues. The influence of highly expressed lncRNA BLACAT1 on prognostic survival was assessed. Subsequently, we cultured human pharynx squamous cell carcinoma FaDu cells. After reducing lncRNA BLACAT1 expression, we assessed FaDu cell proliferation, invasion, and migration using Cell Counting kit-8 (CCK-8) assay, colony formation assay, EUD assay, Transwell assay, and scratch assay. Additionally, liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS) and western blotting analysis were used to analyze proteins that bind to lncRNA BLACAT1. During in vivo experiments, mice received subcutaneous injections of FaDu cells transfected with lncRNA BLACAT1 shRNA or Scr plasmid (Control) in the dorsal region to observe and compare tumor growth. Lastly, tumor tissues underwent hematoxylin-eosin (HE) and immunohistochemical (IHC) staining. lncRNA BLACAT1 was screened as one of the most significant genes among the group of differentially expressed lncRNAs. RT-PCR exhibited elevated lncRNA BLACAT1 expression in HSCC tissues when compared to non-tumor tissues (p < 0.001). Furthermore, increased lncRNA BLACAT1 expression correlated with advanced clinical stages, heightened lymphatic invasion, and a poor prognosis. Subsequent in vitro experiments solidified our observations, demonstrating lncRNA BLACAT1's promotion of HSCC cell proliferation (p < 0.05), migration (p < 0.01), and invasion (p < 0.01) compared with the control group. Moreover, LC-MS/MS identified signal transducer and activator of transcription 3 (STAT3) and Prohibitin 2 (PHB2) as lncRNA BLACAT1-binding proteins and sh-lncRNA BLACAT1 inhibits STAT3/AKT phosphorylation (p < 0.01) and alters the subcellular distribution of PHB2 and P21 compared with the control group (p < 0.01). Moreover, in vivo experiments showed that lncRNA BLACAT1 inhibition suppresses tumorigenicity in an HSCC xenograft model compared to the control group (p < 0.01). lncRNA BLACAT1 is highly expressed in HSCC tumor tissues and plays a crucial role in the development of HSCC in vitro and in vivo. This increased expression may be caused by STAT3/AKT pathway activation, consequently inhibiting P21 expression through PHB2.

Physiological Studies of the Human Pharynx, with a Focus on Upward Pharyngeal Movement

Physiological Studies of the Human Pharynx, with a Focus on Upward Pharyngeal Movement

Efficacy of SARS-CoV-2 detection from used surgical masks compared with standard detection method

The real-time reverse transcription-polymerase chain reaction (RT-PCR) test is the gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection. Proper specimen collection and obtaining a sufficient specimen are the most essential steps for laboratory diagnosis. The nasopharyngeal (NP) swab is recommended as the reference collection method. However, NP swab collection is invasive and uncomfortable for patients and poses some risk to healthcare workers. This study aimed to compare the efficacy of SARS-CoV-2 RNA detection from surgical masks with the NP swab method using RT-PCR testing. Of 269 patients, RT-PCR RNA from NP swabs was detected among 82 patients (30.5%) and was undetected among 187 patients (69.5%). All patients were tested for SARS-CoV-2 RNA from surgical masks. SARS-CoV-2 RNA was detected in 25/82 (30.5%) surgical mask filters, while undetected among 57 (69.5%). For the surgical mask with an average use time of 7.05 h, the sensitivity was 30.5%, the specificity was 100.0%, with positive predictive value of 100.0% and negative predictive value of 76.2%. Therefore, surgical masks could be an alternative non-invasive specimen source for SARS-CoV-2 RT-PCR testing. The results of our study suggest that the test could be employed after wearing surgical masks for at least 8-12 h, with increased sensitivity when used for more than 12 h.

Pharyngeal Cavity Electrical Stimulation-Assisted Swallowing for Post-stroke Dysphagia: A Systematic Review and Meta-analysis of Randomized Controlled Studies.

Stroke is the leading cause of death and disability among adults. The incidence of stroke per 100, 000 patient-years was 2875. As many as 37% to 78% of patients with acute strokes suffer dysphagia. Dysphagia can easily lead to inhalation pneumonia, dehydration, malnutrition, and other serious complications, affecting the quality of life of stroke patients and increasing their mortality. Effective prevention and treatment of post-stroke dysphagia are of great significance to improving the prognosis and quality of life of patients. Some studies have shown that Pharyngeal cavity electrical stimulation-assisted swallowing (PCES-assisted swallowing) has a positive effect on patients with post-stroke dysphagia. This study will evaluate the effects of PCES-assisted swallowing on post-stroke dysphagia, including swallowing function, withdrawal rate of nasal feeding tubes, duration of hospitalization, and so on. Randomized controlled trials (RCTs) of PCES-assisted swallowing in the treatment of post-stroke dysphagia were searched in eight databases, including Cochrane Library, Embase, PubMed, Web of Science, Chinese Biomedical Literature Database, VIP Information Resource System, CNKI, and Wanfang Medical Science. The retrieval time was from the database establishment to June 2022. Rayyan was used to screen the retrieved literature risk of bias for included studies and was calculated using ROB2.0. The RevMan 5.3 software was used for the meta-analysis with the standard mean difference (SMD) and 95% confidence interval (CI). The model type was a random effect model, The risk ratio (RR) was used as the effect size for the two categorical variables. The swallowing function scores, withdrawal rate of nasal feeding tubes, and Length of stay (LOS) of the intervention and control groups were extracted, and the results of the meta-analysis were presented using a forest plot. Six studies from 2010 to 2018 with a total of 341 people were included in the meta-analysis. All studies reported quantitative outcome measures for the severity of dysphagia, and some reported the withdrawal rate of nasal feeding tubes, LOS, and penetration-aspiration-scale (PAS). The overall swallowing function of the PCES group was better than that of the control group (SMD = -0.20, 95%CI -0.38 to -0.03, P = 0.02). In terms of the severity of dysphagia, there was a statistically significant difference in the Dysphagia Severity Rating scale (DSRS) between the Pharyngeal cavity electrical stimulation (PCES) group and the control group (SMD = -0.24, 95%CI -0.48 to 0, P = 0.05). The PCES group nasal feeding withdrawal rate of nasal feeding tubes was higher than the control group (RR = 2.88, 95% CI 1.15 to 7.26, P = 0.02). There was no significant difference in the LOS between the PCES group and the control group (SMD = -0.19, 95%CI -0.44 to 0.07, P = 0.15). This systematic review and meta-analysis provide reasonably reliable evidence that PCES-assisted swallowing can improve nasogastric feeding swallowing function and the withdrawal rate of nasal feeding tubes in patients with post-stroke dysphagia. However, the evidence for reducing oral feeding, aspiration, and length of hospitalization stay is lacking, and further studies are needed.

Sketches of chimpanzee (Pan troglodytes) hoo’s: vowels by any other name?

In human speech, the close back rounded vowel /u/ (the vowel in “boot”) is articulated with the tongue arched toward the dorsal boundary of the hard palate, with the pharyngeal cavity open. Acoustic and perceptual properties of chimpanzee (Pan troglodytes) hoo’s are similar to those of the human vowel /u/. However, the vocal tract morphology of chimpanzees likely limits their phonetic capabilities, so that it is unlikely, or even impossible, that their articulation is comparable to that of a human. To determine how qualities of the vowel /u/ may be achieved given the chimpanzee vocal tract, we calculated transfer functions of the vocal tract area for tube models of vocal tract configurations in which vocal tract length, length and area of a laryngeal air sac simulacrum, length of lip protrusion, and area of lip opening were systematically varied. The method described is principally acoustic; we make no claim as to the actual shape of the chimpanzee vocal tract during call production. Nonetheless, we demonstrate that it may be possible to achieve the acoustic and perceptual qualities of back vowels without a reconfigured human vocal tract. The results, while tentative, suggest that the production of hoo’s by chimpanzees, while achieving comparable vowel-like qualities to the human /u/, may involve articulatory gestures that are beyond the range of the human articulators. The purpose of this study was to (1) stimulate further simulation research on great ape articulation, and (2) show that apparently vowel-like phenomena in nature are not necessarily indicative of evolutionary continuity per se.

Microevolution and Its Impact on Hypervirulence, Antimicrobial Resistance, and Vaccine Escape in Neisseria meningitidis.

Neisseria meningitidis is commensal of the human pharynx and occasionally invades the host, causing the life-threatening illness invasive meningococcal disease. The meningococcus is a highly diverse and adaptable organism thanks to natural competence, a propensity for recombination, and a highly repetitive genome. These mechanisms together result in a high level of antigenic variation to invade diverse human hosts and evade their innate and adaptive immune responses. This review explores the ways in which this diversity contributes to the evolutionary history and population structure of the meningococcus, with a particular focus on microevolution. It examines studies on meningococcal microevolution in the context of within-host evolution and persistent carriage; microevolution in the context of meningococcal outbreaks and epidemics; and the potential of microevolution to contribute to antimicrobial resistance and vaccine escape. A persistent theme is the idea that the process of microevolution contributes to the development of new hyperinvasive meningococcal variants. As such, microevolution in this species has significant potential to drive future public health threats in the form of hypervirulent, antibiotic-resistant, vaccine-escape variants. The implications of this on current vaccination strategies are explored.

Proposed clinical model for predicting speech outcomes in patients undergoing Furlow palatoplasty for velopharyngeal insufficiency after primary palatoplasty

This study aimed to validate the predictors of speech outcomes following Furlow palatoplasty in patients with velopharyngeal insufficiency (VPI) after primary palatoplasty and to propose and validate a model to predict the risk of persistent VPI.The study included patients with VPI after primary palatoplasty who underwent Furlow palatoplasty as a secondary surgery. Eleven variables were included: velar length, pharyngeal cavity depth, velopharyngeal gap, velopharyngeal closure pattern, sex, presence of cleft lip, existence of palatal fistula, surgeon, age at primary palatoplasty, age at secondary surgery, and time interval between primary palatoplasty and secondary surgery. Postoperative speech outcomes were assessed at least 1 year after the secondary surgery and classified as velopharyngeal competence (VPC) or VPI. Variables were analyzed using multivariate logistic regression analysis, and the area under the curve (AUC) was used to validate model accuracy.The study sample comprised 101 patients. Of the patients, 62 had VPC and 39 had VPI after secondary surgery. The results showed a younger age at secondary surgery, a smaller velopharyngeal gap, being female, having a coronal velopharyngeal closure pattern and a velopharyngeal closure ratio of 90% or greater produced a greater probability of VPC.Given the constraints of this study, it appears that the Furlow palatoplasty should be prioritized when the clinical model predicts a substantial likelihood of VPC post-surgery.

Comparison of Standard and Lateral Methods of Laryngeal Mask Airway Insertion in the Management of Pediatrics Airway

Background: The Laryngeal Mask Airway (LMA) has traditionally become an alternative device for airway management. This clinical trial compared two standard and lateral techniques in inserting laryngeal masks in pediatric airway management.&#x0D; Methods: This single-blind clinical trial study was conducted on pediatrics aged 2-6 years who were candidates for elective inguinal herniorrhaphy with ASA I and II under general anesthesia. The pediatrics were randomly assigned to either the standard or lateral methods according to the random number table. The time required to insert LMA and airway pressure as well as the number of attempts to insert in both groups were measured. Data analysis was performed using SPSS version 25; the significance level was less than 0.05.&#x0D; Results: Chest movement and mean airway pressure had no significant difference in both groups (p&gt;0.05). The mean LMA insertion time in the lateral and standard methods was 22.94±7.89 and 65±15.27 seconds, respectively (p=0.001). There was no case of mucosal damage in the lateral method, but ten children had mucosal damage in the standard technique (p=0.001). In the lateral method, LMA was inserted for the first time in 32 pediatrics (94.1%) and two pediatrics (5.9%) in the second time but in the standard group, LMA was inserted in the second time in 11 pediatrics (32.4%) (p=0.006).&#x0D; Conclusion: The lateral method for inserting LMA is practically easy, requires less effort, and has the least complications. Another advantage of the lateral method compared to the standard technique is that inserting a finger into the pharyngeal cavity is unnecessary.

Lung Megakaryocytes Arise from a Flt3 - Cell Lineage and Contribute to Recovery of Platelet Counts in Thrombocytopenic States

Platelet-producing megakaryocytes (Mks) are present in the lung, and we have described an immune modulatory phenotype of lung Mks. It is assumed that the extravascular lung Mks are ‘seeded’ from the bone marrow (BM). To investigate lung Mk origin and how origin may impact function, we used complementary models to lineage trace lung Mks and their platelets. FlkSwitch mTmG mice label all progeny from Flt3 + progenitors with GFP while all other cells are labeled with Tomato. We leveraged the FlkSwitch mouse model to compare BM and lung Mk origins. In 18 week old mice, the vast majority of BM and spleen Mks were GFP +, while lung Mks were largely Tomato + (Figure 1A). These data implied that in the adult mouse lung Mks arise from a Flt3 - pathway that is distinct from BM megakaryopoiesis in homeostatic conditions. We next used a lineage tracing model (MDS1-Cre-ERT2 Tomato) where tamoxifen (TAM) administration induces a Tomato reporter in hematopoietic stem cell (HSC)-derived cells. Mice were dosed with TAM at 18 weeks and sacrificed at 32 weeks. Lung and BM Mks were largely Tomato + (~75%) and matched Tomato levels of BM Lineage - Sca1 + c-Kit + (LSK). Next, we dosed mice with TAM at 8 weeks, sacrificed the mice at 14 weeks and found similarly high levels of Tomato in BM LSK, BM Mk, and lung Mk. In MDS1-Cre-ERT2 Tomato mice, both BM and lung Mks were Tomato +/HSC-dependent, suggesting that lung Mks arise from HSCs in adult mice. Next, we used CFSE dye given oropharyngeal (OP) to label only lung cells. We detected CFSE + lung Mks for up to 120 days post administration without any CFSE + Mks in BM or spleen. These data suggested there is a long-lived Mk population in the lung. Taken together, these complementary models imply that lung Mks arise from a Flt3 - lineage that is HSC-dependent, which yields Mks that can live in the lung for extended periods of time. The CFSE OP method labeled only lung cells, which enabled us to track lung Mk-derived platelets. We found that about 10% of the steady-state circulating platelet pool is lung derived. In a model of thrombocytopenia using diphtheria toxin mediated platelet depletion in PF4-Cre-iDTR mice, we observed a doubling of CFSE + lung-derived circulating platelets (about 20%). We also used Plasmodium yoelli murine malaria as a model of infection associated with more chronic thrombocytopenia. Using CFSE OP labeling of lung Mk-derived platelets, we saw a marked increase in CFSE + lung-derived platelets early post- P. Yoelli infection (Figure 1B). Our studies suggest that lung Mks arise from a Flt3 - HSC-dependent pathway and contribute relatively more platelets during acute and chronic thrombocytopenia. Ongoing work aims to describe the role of lung megakaryopoiesis via a Flt3 - HSC-dependent pathway in early development and in models of thrombocytopenia.

1678. Non-Invasive and invasive Pediatric Group A Streptococcus Infections (iGAS) Admissions During 2021 and 2022 at a Tertiary Health Care Center: Resurgence and Concomitant Viral Infections

Abstract Background A resurgence of invasive Group A Streptococcus (iGAS) infection in children was observed in Colorado and Minnesota by the Centers For Disease Control and Prevention (CDC) during the fall of 2022. Cases were accompanied by hospitalizations for respiratory syncytial virus (RSV) and influenza. We describe hospital admissions with iGAS and non iGAS infections at a tertiary health care center in Augusta, GA during 2021 and 2022; the only children’s Hospital with 150 licensed beds and a population of 201.196 as of 2021. Methods We used the iGAS definition by the CDC as a Group A Streptococcus (GAS) isolated by culture in specimens obtained from a sterile body site, or GAS isolated from wound culture and necrotizing fasciitis or streptococcal toxic shock syndrome. We conducted a retrospective chart review of iGAS and non iGAS cases admitted to the Children’s Hospital of Georgia during January to December 2021 and January to December 2022. We collected demographics, site of infection, concomitant viral infections diagnosed by nasopharyngeal (NP) PCR, intensive care unit (ICU) admission and mortality. Results During 2021, only 1 patient was admitted with non iGAS, a 7-year-old with a wound infection. In 2022, 15 patients were diagnosed with GAS infection, 7 of them had iGAS and 8 had noninvasive disease. We included positive peritoneal and pleural fluid, joint/bone aspirates, bronchoalveolar lavage samples, and blood cultures for iGAS. The age range for iGAS was 6-17 years and for noninvasive, 16 mo to 11 years. In 2022, 25% of noninvasive cases had concomitant influenza compared to 43% of iGAS cases. Only one patient with non iGAS tested positive for RSV. 2 patients had rhinovirus, one had adenovirus. One patient with iGAS was positive for metapneumovirus (HMPV). 57% of iGAS cases were admitted to the ICU (n=4). 50% of noninvasive cases (n=4) were admitted to the ICU. One iGAs case died. All noninvasive cases survived. Conclusion There was increase in iGAS and noninvasive GAS infections during 2022 compared to 2021 in our institution. iGAS was more common in older children. Of iGAS cases, the only coexisting viruses were influenza and HMPV. Most cases were seen during Fall of 2022. The cause of reappearance of iGAS and non iGAS is still unknown but believed to be related to end of mask mandates for COVID 19 infections. Disclosures All Authors: No reported disclosures

854. Performance of a Nasopharyngeal-Based Host Gene Expression Classifier at Distinguishing Bacterial versus Viral Respiratory Infection

Abstract Background Acute respiratory infections (ARI) present a diagnostic challenge due to overlapping clinical features of bacterial and viral infections, leading to unnecessary use of antibiotics. We have previously shown that a host-based blood mRNA expression test based on a 24-gene classifier has excellent performance differentiating bacterial and viral (B vs V) ARI. Since obtaining venous blood can be invasive and impractical in some settings, we examined the performance of this previously derived host mRNA test on nasopharyngeal (NP) samples. Methods We enrolled patients presenting with ARI to a tertiary care hospital in Sri Lanka from November 2019 to October 2020 and collected blood samples in PAXgene RNA tubes paired with NP samples in RNAlater. We selected a convenient sample of 21 subjects with a confirmed bacterial or viral ARI, extracted RNA using commercially available kits, and measured 24 host mRNA transcripts using a multiplex RT-PCR-based assay optimized for venous blood (Biomeme Franklin M1, Philadelphia, PA). Due to missing values, one target was removed from the 24-gene classifier. Two models, one for each sample type, were built using elastic net logistic regression as implemented in the glmnet R package to classify B vs V infections. Nested leave-one-out cross-validation was used to conduct both parameter regularization (lambda) and predictions of bacterial probability, with alpha set to 0.01. Results With 21 subjects [median age (IQR) 64 (56-66) years, 13 female, 8 male], the model trained on the PAXgene samples was able to correctly classify 8 of 9 bacterial and 8 of 12 viral ARIs using a 23 gene classifier, with an area under the curve (AUC) of 0.824 [confidence interval (CI) of 0.64-1]. Due to significantly different cycle thresholds in the normalizers, the NP sample set necessitated a second model that used 16 subjects [median age 61 (49-64) years, 10F, 6M]. This model correctly classified all 11 viral and 3 of 5 bacterial ARIs with an AUC of 0.818 (CI 0.57-1). Conclusion These results suggest that a host mRNA expression test based on non-invasive sampling from the nasopharynx shows promise in classifying B vs V ARI. These preliminary findings need to be confirmed in a larger cohort of subjects to develop tests that can help reduce antibiotic overuse for ARI. Disclosures Thomas W. Burke, PhD, Biomeme Inc: Advisor/Consultant|Biomeme Inc: Methods to diagnose and treat acute respiratory infections|Biomeme Inc: Ownership Interest Micah T. McClain, MD, PhD, Biomeme Inc: Methods to diagnose and treat acute respiratory infections Christopher W. Woods, MD, MPH, Biomeme Inc: Methods to diagnose and treat acute respiratory infections|Biomeme Inc: Chief Medical Consultant|Biomeme Inc: Ownership Interest

A fully integrated, standalone stretchable device platform with in-sensor adaptive machine learning for rehabilitation

Post-surgical treatments of the human throat often require continuous monitoring of diverse vital and muscle activities. However, wireless, continuous monitoring and analysis of these activities directly from the throat skin have not been developed. Here, we report the design and validation of a fully integrated standalone stretchable device platform that provides wireless measurements and machine learning-based analysis of diverse vibrations and muscle electrical activities from the throat. We demonstrate that the modified composite hydrogel with low contact impedance and reduced adhesion provides high-quality long-term monitoring of local muscle electrical signals. We show that the integrated triaxial broad-band accelerometer also measures large body movements and subtle physiological activities/vibrations. We find that the combined data processed by a 2D-like sequential feature extractor with fully connected neurons facilitates the classification of various motion/speech features at a high accuracy of over 90%, which adapts to the data with noise from motion artifacts or the data from new human subjects. The resulting standalone stretchable device with wireless monitoring and machine learning-based processing capabilities paves the way to design and apply wearable skin-interfaced systems for the remote monitoring and treatment evaluation of various diseases.

Molecular epidemiology and characteristics of respiratory syncytial virus among hospitalized children in Guangzhou, China

BackgroundHuman respiratory syncytial virus (RSV) is a leading cause of acute lower respiratory tract infection and hospitalization, especially in children. Highly mutagenic nature and antigenic diversity enable the RSV to successfully survive in human population. We conducted a molecular epidemiological study during 2017–2021 to investigate the prevalence and genetic characteristics of RSV.MethodsA total of 6499 nasopharyngeal (NP) swabs were collected from hospitalized children at Department of Pediatrics, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou, Guangdong, China. All NP swab specimens were preliminary screened for common respiratory viruses and then tested for RSV using specific PCR assays. Partial G genes of RSV were amplified for phylogenetic analysis and genetic characterization.ResultsThe overall detection rate for common respiratory viruses was 16.12% (1048/6499). Among those, 405 specimens (6.20%, 405/6499) were found positive for RSV. The monthly distribution of RSV and other respiratory viruses was variable, and the highest incidence was recorded in Autumn and Winter. Based on the sequencing of hypervariable region of G gene, 93 RSV sequences were sub-grouped into RSV-A (56, 60.2%) and RSV-B (37, 39.8%). There was no coinfection of RSV-A and RSV-B in the tested samples. Phylogenetic analysis revealed that RSV-A and RSV-B strains belonged to ON1 and BA9 genotypes respectively, indicating predominance of these genotypes in Guangzhou. Several substitutions were observed which may likely change the antigenicity and pathogenicity of RSV. Multiple glycosylation sites were noticed, demonstrating high selection pressure on these genotypes.ConclusionThis study illustrated useful information about epidemiology, genetic characteristics, and circulating genotypes of RSV in Guangzhou China. Regular monitoring of the circulating strains of RSV in different parts of China could assist in the development of more effective vaccines and preventive measures.

Robotic system for nasopharyngeal swab sampling based on remote center of motion mechanism.

In this study, a robotic system is proposed for nasopharyngeal (NP) swab sampling with high safety and efficiency. Most existing swab-sampling robots have more than six degrees of freedom (DOFs). However, not all six DOFs are necessarily required for NP swab sampling. A high number of DOFs can cause safety problems, such as collisions between the robot and patient. We developed a new type of robot with four DOFs for NP swab sampling that consists of a two DOFs remote center of motion (RCM) mechanism, a two DOFs insertion mechanism, and a nostril support unit. With the nostril support unit, the robot no longer needs to adjust the insertion position of the swab. The proposed robot enables the insertion orientation and depth to be adjusted according to different postures or facial shapes of the subject. For intuitive and precise remote control of the robot, a dedicated master device for the RCM and a visual feedback system were developed. The effectiveness of the robotic system was demonstrated by repeatability, RCM accuracy, tracking accuracy, and in vitro phantom experiments. The average tracking error between the master device and the robot was less than 2mm. The contact force exerted on the swab prior to reaching the nasopharynx was less than 0.04 N, irrespective of the phantom's pose. This study confirmed that the RCM-based robotic system is effective and safe for NP swab sampling while using minimal DOFs.

Antimicrobial Resistance in the Upper Respiratory Tract of Children Compared with Adults

Abstract Background Antimicrobial resistance (AMR) is a growing global public health threat. In 2019, resistant bacterial infections and respiratory tract infections were a leading cause of death in children worldwide. While the gut is a known reservoir of AMR genes, there is little data on the AMR burden in the respiratory tract. We used metagenomic RNA next-generation sequencing (mNGS) to describe bacterial AMR genes detected in the upper respiratory tract of children and compare with adults. Methods We leveraged two established cohort studies of children and adults diagnosed with acute respiratory illnesses in the inpatient and outpatient setting in California and Colorado. Nasopharyngeal (NP) swabs were collected between March to September 2020. Specimens underwent RNA extraction and paired-end Illumina sequencing. AMR genes were detected using the ARG-ANNOT database and the CZID pipeline. Children ≤18 years of age and adults≥ 40 years of age were eligible for inclusion in the analysis. We described and compared AMR genes and AMR gene classes detected in children and adults. P-values were calculated using the Wilcoxon rank-sum test. A multiple logistic regression model was fitted to evaluate the association of age cohort (children vs adults) with presence of AMR genes, while accounting for possible confounding from sex and encounter setting. 95% confidence intervals (CI) were calculated using the Wald CI. Results NP swab mNGS data from 236 patients (82 children, 154 adults) were analyzed. Children were a median age of 4 years (range: 0-17 years), and adults were a median age of 62 years (range: 40-89 years). Both cohorts were 50% male. AMR genes were detected in 44 (54%) children and 109 (71%) adults. Genes conferring resistance to beta-lactams (n=112, 47%), macrolides (n=89, 38%), and tetracyclines (n=72, 31%) were the most frequently detected. Among children, children ages 0 to 2 years had a higher number of AMR genes than children of other age groups, particularly when compared with children ages 3 to 10 years (p-value: 0.031) (Figure 1A). When comparing children with adults, children had fewer AMR genes (p-value: 0.014) as well as fewer AMR gene classes (p-value: 0.031) (Figure 1B). In the multiple logistic regression model, children still had lower odds of any AMR genes detected compared with adults (odds ratio: 0.39, 95% CI: 0.21-0.71). Figure 1. Box plots of (A) the number of antimicrobial resistance (AMR) genes detected in children stratified by age, and (B) the number of AMR genes and AMR gene classes detected in children (n=82) and adults (n=154). The size of the individual data points within the box plots correlates with the number of patients at each data point. Conclusion Children had fewer AMR genes and gene classes than adults in their upper respiratory tract microbiome. Despite this, over half of the children had detectable resistance genes, and children 2 years and younger had more AMR genes than other children. Almost half of the children had resistance genes to beta-lactams, the most frequently prescribed antibiotics to children. These findings suggest that presence of AMR organisms in the airway is common even in children, and that continued efforts to reduce AMR burden in this population are essential.

Selection of effective therapy for inflammatory diseases of the pharynx

Inflammatory processes of the upper respiratory tract and pharynx occupy a leading position both in terms of the frequency of visits to an otolaryngologist and in the variety of pathogenic factors that cause processes accompanied by sore throat. With an inflammatory process occurring in the lymphoid ring of the pharynx, symptoms are usually caused by local factors, so local treatment, both anti-inflammatory and analgesic, is especially important. However, according to the literature, in most cases systemic antibacterial therapy is prescribed. According to modern concepts, it is known that systemic antimicrobial agents should be prescribed strictly according to indications, since the viral etiology of the inflammatory process is the most common. Based on this point of view, non-steroidal anti-inflammatory drugs are effective. In turn, according to many studies, preparations of the lysine salt of ketoprofen have shown their effectiveness in relieving symptoms without leading to the development of obvious side effects. Based on a review of literature data, it is shown that the lysine salt of ketoprofen has a wide therapeutic potential, good tolerability, and ease of use, which allows us to recommend this drug for use in patients with acute inflammatory processes in the pharynx caused by both microbial and viral agents. This drug is one of the most effective because there is no interaction with other drugs, and it can be used as part of combination therapy with antiseptics. If it is necessary to carry out antibacterial therapy for the treatment of an acute or exacerbation of a chronic process in the pharyngeal cavity, the drug of choice can be considered a third-generation semisynthetic cephalosporin antibiotic for oral administration with a broad spectrum of action, cefixime.

Digestive system and feeding of deep-sea acorn worm Quatuoralisia malakhovi (Enteropneusta: Torquaratoridae) from the Bering sea

We studied the structure of digestive system of deep-sea torquaratorid Quatuoralisia malakhovi including the food-obtaining apparatus, microanatomy of all regions of digestive tract, and fine structure of various regions of the gut. The mouth of Q. malakhovi is surrounded by two symmetrical triangular lateral collar lips and one midventral collar lip, which may retract into the mouth. The ventral ciliary grooves run along the anterior-ventral edges of lateral lips and lead into the internal canals of the lips. The pharyngeal cavity is divided by left and right parabranchial ridges into the median pharyngeal zone with gill bars and two symmetrical peripheral canals, which fuse behind the site of genesis of new gill bars. The gill pharynx is followed by the hepatic region with metameric hepatic sacculations comprising cells with numerous large phagosomes indicating active intracellular digestion. The intestine is filled with undigested food residues including detritus flakes and remnants of various organisms. The intestine content was examined using SEM and histological studies. We analyzed the ratio of organic and mineral components of the bottom sediment from the habitat of Q. malakhovi, and the stable isotope composition of carbon and nitrogen in the intestine content, in the tissues, and in the sediment from the habitat of the studied acorn worm. Using the obtained data and the photographs and videotapes in situ, we have proposed a hypothesis explaining the feeding mechanism of Q. malakhovi. The width of the ventral ciliary groove of the lateral collar lips (100–200 μm) regulates the maximum size of particles that enter the pharynx of the animal. The minimum size of the sediment particles selected by Q. malakhovi is determined by the width of the gill pores and is 1–3 μm. The selected sediment particles move through the peripheral pharyngeal canals to the hepatic region, where the digestion of the food is carried out. In the posthepatic intestine, weight of the gut contents might serve as an anchor that holds the heavily watered jelly-like torquaratorid body at the bottom during the feeding.

Dynamics of the nasopharyngeal microbiome of apparently healthy calves and those with clinical symptoms of bovine respiratory disease from disease diagnosis to recovery.

Bovine respiratory disease (BRD) is a multifactorial disease complex in which bacteria in the upper respiratory tract play an important role in disease development. Previous studies have related the presence of four BRD-pathobionts (Mycoplasma bovis, Histophilus somni, Pasteurella multocida, and Mannheimia haemolytica) in the upper respiratory tract to BRD incidence and mortalities in the dairy and beef cattle industry, but these studies typically only use one time point to compare the abundance of BRD-pathobionts between apparently healthy and BRD-affected cattle. The objective of this study was to characterize the longitudinal development of the nasopharyngeal (NP) microbiome from apparently healthy calves, and in calves with clinical signs of BRD, the microbiota dynamics from disease diagnosis to recovery. Deep nasopharyngeal swabs were taken from all calves immediately after transport (day 0). If a calf was diagnosed with BRD (n = 10), it was sampled, treated with florfenicol or tulathromycin, and sampled again 1, 5, and 10 days after antibiotic administration. Otherwise, healthy calves (n = 20) were sampled again on days 7 and 14. Bacterial community analysis was performed through 16S rRNA gene amplicon sequencing. The NP microbiome of the healthy animals remained consistent throughout the study, regardless of time. The NP microbiota beta diversity and community composition was affected by tulathromycin or florfenicol administration. Even though BRD-pathobionts were identified by 16S rRNA gene sequencing in BRD-affected animals, no difference was observed in their relative abundance between the BRD-affected and apparently healthy animals. The abundance of BRD-pathobionts was not predictive of disease development while the relative abundance of BRD pathobionts was unique to each BRD-affected calf. Interestingly, at the end of the study period, the genera Mycoplasma was the most abundant genus in the healthy group, while Lactobacillus was the most abundant genus in the animals that recovered from BRD. This study highlights that injected antibiotics seem to improve the NP microbiome composition (higher abundance of Lactobacillus and lower abundance of Mycoplasma), and that the relative abundance of BRD-pathobionts differs between individual calves but is not strongly predictive of BRD clinical signs, indicating that additional factors are likely important in the clinical progression of BRD.

3D-printed wearable BaTiO3/PDMS piezoelectric nanogenerator for self-powered body movement sensing

Flexible piezoelectric nanogenerators (PENGs) used in body movement real-time monitoring are of great interest for their wide application potential such as in the field of smart healthcare. In this work, a self-powered BaTiO3/Polydimethylsiloxane (PDMS) PENG for body movement sensing was successfully fabricated by extrusion 3D printing. Matrix system composed of different ratios of PDMS was selected based on the rheological property of materials. Experimental investigations were conducted to examine the impact of printing pressure and speed on the linewidth. Subsequently, the extrusion parameters for nanogenerators were determined based on the printed linewidth. The composite showed good ferroelectric property. After polarization, the nanogenerators exhibited an improvement in output performance of up to 55.2%. Additionally, the device demonstrated a good linear relationship between voltage and tapped force test by an electromechanical vibrator. Successful detection of body or muscle movement signals was achieved when the nanogenerator was mounted on the human finger, throat, or foot using a wearable sock, highlighting its potential for applications in self-powered wearable devices for smart healthcare.