A Pilot Study Comparing HPV-Positive and HPV-Negative Head and Neck Squamous Cell Carcinomas by Whole Exome Sequencing.

Background. Next-generation sequencing of cancers has identified important therapeutic targets and biomarkers. The goal of this pilot study was to compare the genetic changes in a human papillomavirus- (HPV-)positive and an HPV-negative head and neck tumor. Methods. DNA was extracted from the blood and primary tumor of a patient with an HPV-positive tonsillar cancer and those of a patient with an HPV-negative oral tongue tumor. Exome enrichment was performed using the Agilent SureSelect All Exon Kit, followed by sequencing on the ABI SOLiD platform. Results. Exome sequencing revealed slightly more mutations in the HPV-negative tumor (73) in contrast to the HPV-positive tumor (58). Multiple mutations were noted in zinc finger genes (ZNF3, 10, 229, 470, 543, 616, 664, 638, 716, and 799) and mucin genes (MUC4, 6, 12, and 16). Mutations were noted in MUC12 in both tumors. Conclusions. HPV-positive HNSCC is distinct from HPV-negative disease in terms of evidence of viral infection, p16 status, and frequency of mutations. Next-generation sequencing has the potential to identify novel therapeutic targets and biomarkers in HNSCC

Endobronchial Carcinoid Tumour with Extensive Ossification: An Unusual Case Presentation

Carcinoid tumour is a well-known primary endobronchial lung neoplasm. Although calcifications may be seen in up to 30% of pulmonary carcinoid tumours, near complete ossification of these tumours is an unusual finding. Such lesions can prove diagnostically challenging at the time of intraoperative frozen section as the latter technique requires thin sectioning of the lesion for microscopic assessment. We present an unusual case of endobronchial carcinoid tumour with extensive ossification in a 45-year-old male. Preliminary intraoperative diagnosis was achieved through the alternative use of cytology scrape smears. The final diagnosis was confirmed after decalcification of the tumour. The prognostic implications of heavily ossified carcinoid tumours remain elusive. Long-term clinical follow-up of these patients is recommended

Variable recombination dynamics during the emergence, transmission and ‘disarming’ of a multidrug-resistant pneumococcal clone

Background: Pneumococcal β-lactam resistance was first detected in Iceland in the late 1980s, and subsequently peaked at almost 25% of clinical isolates in the mid-1990s largely due to the spread of the internationally-disseminated multidrug-resistant PMEN2 (or Spain6B-2) clone of Streptococcus pneumoniae. Results: Whole genome sequencing of an international collection of 189 isolates estimated that PMEN2 emerged around the late 1960s, developing resistance through multiple homologous recombinations and the acquisition of a Tn5253-type integrative and conjugative element (ICE). Two distinct clades entered Iceland in the 1980s, one of which had acquired a macrolide resistance cassette and was estimated to have risen sharply in its prevalence by coalescent analysis. Transmission within the island appeared to mainly emanate from Reykjavík and the Southern Peninsular, with evolution of the bacteria effectively clonal, mainly due to a prophage disrupting a gene necessary for genetic transformation in many isolates. A subsequent decline in PMEN2’s prevalence in Iceland coincided with a nationwide campaign that reduced dispensing of antibiotics to children in an attempt to limit its spread. Specific mutations causing inactivation or loss of ICE-borne resistance genes were identified from the genome sequences of isolates that reverted to drug susceptible phenotypes around this time. Phylogenetic analysis revealed some of these occurred on multiple occasions in parallel, suggesting they may have been at least temporarily advantageous. However, alteration of ‘core’ sequences associated with resistance was precluded by the absence of any substantial homologous recombination events. Conclusions: PMEN2’s clonal evolution was successful over the short-term in a limited geographical region, but its inability to alter major antigens or ‘core’ gene sequences associated with resistance may have prevented persistence over longer timespans

Evidence for Soft Selective Sweeps in the Evolution of Pneumococcal Multidrug Resistance and Vaccine Escape

The multidrug-resistant Streptococcus pneumoniae Taiwan19F-14, or PMEN14, clone was first observed with a 19F serotype, which is targeted by the heptavalent polysaccharide conjugate vaccine (PCV7). However, “vaccine escape” PMEN14 isolates with a 19A serotype became an increasingly important cause of disease post-PCV7. Whole genome sequencing was used to characterize the recent evolution of 173 pneumococci of, or related to, PMEN14. This suggested that PMEN14 is a single lineage that originated in the late 1980s in parallel with the acquisition of multiple resistances by close relatives. One of the four detected serotype switches to 19A generated representatives of the sequence type (ST) 320 isolates that have been highly successful post-PCV7. A second produced an ST236 19A genotype with reduced resistance to β-lactams owing to alteration of pbp1a and pbp2x sequences through the same recombination that caused the change in serotype. A third, which generated a mosaic capsule biosynthesis locus, resulted in serotype 19A ST271 isolates. The rapid diversification through homologous recombination seen in the global collection was similarly observed in the absence of vaccination in a set of isolates from the Maela refugee camp in Thailand, a collection that also allowed variation to be observed within carriage through longitudinal sampling. This suggests that some pneumococcal genotypes generate a pool of standing variation that is sufficiently extensive to result in “soft” selective sweeps: The emergence of multiple mutants in parallel upon a change in selection pressure, such as vaccine introduction. The subsequent competition between these mutants makes this phenomenon difficult to detect without deep sampling of individual lineages

Oral flora in acute stroke patients: a prospective exploratory observational study

Objective: To describe the bacterial profile of the oral flora during the first two weeks following a stroke, examining changes in the condition of the oral cavity and infections. Background: Dysphagia is common after a stroke and can lead to aspiration pneumonia. Oral flora changes associated with stroke have been implicated as a possible source of bacteria that can cause systemic infections. Materials and methods: Seventy-seven participants were recruited over a period of nine months. Fifty participants had a complete set of swabs taken from four different oral sites and saliva at three time points over a 14 day period. Molecular identification of bacteria was performed on pooled DNA extracted from swabs. Results: A total of 103 bacterial phylotypes were identified, 29 of which were not in the Human Oral Microbiome Database (HOMD). Fourteen of the twenty most common bacterial phylotypes found in the oral cavity were Streptococcal species with Streptococcus salivarius being the most common. The condition of the oral cavity worsened during the study period. Fifteen (30%) patients had at least one infection. Conclusions: It is unknown whether the 29 phylotypes identified that were not in the HOMD indicate a particular change in the oral flora associated with stroke, or the incomplete nature of the HOMD. The Holistic and Reliable Oral Assessment Tool detailed how the condition of the oral cavity following a stroke worsened over the fourteen days in hospital. Further research is needed to explore oral care methods to improve patient safety and comfort following a stroke

The reciprocal relationship between body mass index categories and physical fitness: a 4‐year prospective cohort study of 20 000 Chinese children

Introduction: Body mass index (BMI) categories and physical fitness are associated but the reciprocal relationship between BMI categories and physical fitness has not been investigated. This study aims to investigate the longitudinal reciprocal relationship between BMI categories and physical fitness. Methods: This is a population-based 4-year cohort study in 48 elementary schools. Children aged 6 to 9 years at recruitment were included. BMI categories and physical fitness including handgrip strength, core muscle endurance, flexibility, and cardiorespiratory fitness were measured using standard equipment and protocol. Results: Among 26 392 eligible participants, 19 504 (73.9%) were successfully followed for 3 years. Baseline obesity prevalence was 5.9%. After 3 years, those who were unfit at baseline had an increased risk of obesity (risk ratio [RR] 1.41, 95% CI 1.16-1.71, P < .001) and those who were fit at baseline had a decreased risk of obesity (RR 0.69, 95% CI 0.60-0.80, P < .001) compared with moderately fit children. Furthermore, improvement of fitness predicted decreased risk of obesity. Similarly, normal body weight also predicted better physical fitness. The path analysis confirmed a strong reciprocal relationship between physical fitness and obesity. Conclusions: Better physical fitness was prospectively associated with normal weight and vice versa. Physically fit children were more likely to maintain a healthy weight and those with a healthy weight were more likely to be physically fit, which is important for healthy development

Age- and sex-specific physical fitness reference and association with body mass index in Hong Kong Chinese schoolchildren

There is lacking a population-based study on the fitness level of Hong Kong schoolchildren, and it seems that increasing childhood obesity prevalence has shifted the classification of healthy fitness, with ‘underfit’ as normal. This cross-sectional territory study aimed to develop an age- and sex-specific physical fitness reference using a representative sample of children aged 6–17 and to determine the associations with body mass index in schoolchildren. The study analyzed Hong Kong School Physical Fitness Award Scheme data covering grade 1 to grade 12 students’ physical fitness and anthropometric measurements from 2017 to 2018. This reference was established without the impact due to COVID-19. Four aspects of physical fitness tests were measured using a standardized protocol, including (i) upper limb muscle strength, (ii) one-minute sit-up, (iii) sit-and-reach, and (iv) endurance run tests. The generalized additive model for location, scale, and shape was used to construct the reference charts. A Mann–Whitney U test was used to compare the mean differences in age, weight, and height, and a Pearson’s chi-square test was used to examine the distributions of sex groups. A Kruskal–Wallis test was used to compare the group differences in BMI status, followed by the Dunn test for pairwise comparisons. A 5% level of significance was regarded as statistically significant. Data of 119,693 students before the COVID-19 pandemic were included in the analysis. The association between physical fitness level and BMI status varied depending on the test used, and there were significant differences in fitness test scores among BMI groups. The mean test scores of the obese group were lower in most of the tests for both boys and girls, except for handgrip strength. The underweight group outperformed the obese group in push-ups, one-minute sit-ups, and endurance run tests, but not in handgrip strength. In conclusion, a sex- and age-specific physical fitness reference value for Hong Kong Chinese children aged 6 to 17 years old is established, and this study demonstrated a nonlinear relationship between BMI status and physical fitness. The reference will help to identify children with poor physical fitness to offer support and guidance on exercise training. It also serves as a baseline for assessing the impact of the COVID-19 pandemic on Hong Kong students’ physical fitness

The central role of natural killer cells in mediating acute myocarditis after mRNA COVID-19 vaccination

BACKGROUND: Vaccine-related acute myocarditis is recognized as a rare and specific vaccine complication following mRNA-based COVID-19 vaccinations. The precise mechanisms remain unclear. We hypothesized that natural killer (NK) cells play a central role in its pathogenesis. METHODS: Samples from 60 adolescents with vaccine-related myocarditis were analyzed, including pro-inflammatory cytokines, cardiac troponin T, genotyping, and immunophenotyping of the corresponding activation subsets of NK cells, monocytes, and T cells. Results were compared with samples from 10 vaccinated individuals without myocarditis and 10 healthy controls. FINDINGS: Phenotypically, high levels of serum cytokines pivotal for NK cells, including interleukin-1β (IL-1β), interferon α2 (IFN-α2), IL-12, and IFN-γ, were observed in post-vaccination patients with myocarditis, who also had high percentage of CD57 NK cells in blood, which in turn correlated positively with elevated levels of cardiac troponin T. Abundance of the CD57 NK subset was particularly prominent in males and in those after the second dose of vaccination. Genotypically, killer cell immunoglobulin-like receptor (KIR) KIR2DL5B(-)/KIR2DS3(+)/KIR2DS5(-)/KIR2DS4del(+) was a risk haplotype, in addition to single-nucleotide polymorphisms related to the NK cell-specific expression quantitative trait loci DNAM-1 and FuT11, which also correlated with cardiac troponin T levels in post-vaccination patients with myocarditis. CONCLUSION: Collectively, these data suggest that NK cell activation by mRNA COVID-19 vaccine contributed to the pathogenesis of acute myocarditis in genetically and epidemiologically vulnerable subjects

Normal values of regional left ventricular endocardial motion: multicenter color kinesis study. Am J Physiol Heart Circ Physiol 279: H2464– H2476

. Normal values of regional left ventricular endocardial motion: multicenter color kinesis study. Am J Physiol Heart Circ Physiol 279: H2464-H2476, 2000.-Our goal was to establish normal values for quantitative color kinesis indexes of left ventricular (LV) wall motion over a wide range of ages, which are required for objective diagnosis of regional systolic and diastolic dysfunction. Color-encoded images were obtained in 194 normal subjects (95 males, 99 females, age 2 mo to 79 yr) in four standard views. Quantitative indexes of magnitude and timing of systolic and diastolic function were studied for age-and genderrelated differences. Normal limits of all ejection and filling indexes were in a narrow range (Յ25% of the mean), with no major gender-related differences. Despite invariable ejection fractions, both peak filling and ejection rates decreased with age (30 and 20%, correspondingly) with a concomitant increase in mean filling and ejection times, resulting in fiveand twofold increases in the late to early filling and ejection ratios, correspondingly. Diastolic asynchrony increased with age (from 4.7 Ϯ 2.0 to 6.4 Ϯ 3.2 from the 2nd to 7th decade). The normal values of color kinesis indexes should allow objective detection of regional LV systolic and diastolic dysfunction. echocardiography; ultrasound imaging; ventricular function; wall motion COLOR KINESIS IS AN EMERGING echocardiographic technique based on acoustic quantification, which uses color-encoding to depict left ventricular (LV) systolic and diastolic endocardial motion However, a new technique, which detects abnormalities by comparing individual patient's data with normal values, relies on having these normal values established in a large sample of the normal population. Accordingly, normal values of different indexes of magnitude and timing of global and regional LV function derived from color kinesis images need to be established to allow objective detection of abnormalities with a high level of confidence. We also hypothesized that these indexes may be age and gender dependent, in which case normal values would need to be established for different demographic groups. Accordingly, the purpose of this collaborative multicenter effort was to establish normal values for magnitude and timing indexes of wall motion by acquiring and analyzing systolic and diastolic color kinesis images in a large group of normal subjects of both sexes over a wide range of ages. METHODS Study population. The protocol for this study was approved by the Institutional Review Board of the University of Chicago (protocol #9171). This protocol was initially designed to include eight age groups (8 decades between 0 and 80 yr) with a minimum of 20 normal subjects each (50% males and 50% Address for reprint requests and othe