Otopathogenic Staphylococcus aureus Invades Human Middle Ear Epithelial Cells Primarily through Cholesterol Dependent Pathway

Chronic suppurative otitis media (CSOM) is one of the most common infectious diseases of the middle ear especially affecting children, leading to delay in language development and communication. Although Staphylococcus aureus is the most common pathogen associated with CSOM, its interaction with middle ear epithelial cells is not well known. In the present study, we observed that otopathogenic S. aureus has the ability to invade human middle ear epithelial cells (HMEECs) in a dose and time dependent manner. Scanning electron microscopy demonstrated time dependent increase in the number of S. aureus on the surface of HMEECs. We observed that otopathogenic S. aureus primarily employs a cholesterol dependent pathway to colonize HMEECs. In agreement with these findings, confocal microscopy showed that S. aureus colocalized with lipid rafts in HMEECs. The results of the present study provide new insights into the pathogenesis of S. aureus induced CSOM. The availability of in vitro cell culture model will pave the way to develop novel effective treatment modalities for CSOM beyond antibiotic therapy

BHPR research: qualitative1. Complex reasoning determines patients' perception of outcome following foot surgery in rheumatoid arhtritis

Background: Foot surgery is common in patients with RA but research into surgical outcomes is limited and conceptually flawed as current outcome measures lack face validity: to date no one has asked patients what is important to them. This study aimed to determine which factors are important to patients when evaluating the success of foot surgery in RA Methods: Semi structured interviews of RA patients who had undergone foot surgery were conducted and transcribed verbatim. Thematic analysis of interviews was conducted to explore issues that were important to patients. Results: 11 RA patients (9 ♂, mean age 59, dis dur = 22yrs, mean of 3 yrs post op) with mixed experiences of foot surgery were interviewed. Patients interpreted outcome in respect to a multitude of factors, frequently positive change in one aspect contrasted with negative opinions about another. Overall, four major themes emerged. Function: Functional ability & participation in valued activities were very important to patients. Walking ability was a key concern but patients interpreted levels of activity in light of other aspects of their disease, reflecting on change in functional ability more than overall level. Positive feelings of improved mobility were often moderated by negative self perception ("I mean, I still walk like a waddling duck”). Appearance: Appearance was important to almost all patients but perhaps the most complex theme of all. Physical appearance, foot shape, and footwear were closely interlinked, yet patients saw these as distinct separate concepts. Patients need to legitimize these feelings was clear and they frequently entered into a defensive repertoire ("it's not cosmetic surgery; it's something that's more important than that, you know?”). Clinician opinion: Surgeons' post operative evaluation of the procedure was very influential. The impact of this appraisal continued to affect patients' lasting impression irrespective of how the outcome compared to their initial goals ("when he'd done it ... he said that hasn't worked as good as he'd wanted to ... but the pain has gone”). Pain: Whilst pain was important to almost all patients, it appeared to be less important than the other themes. Pain was predominately raised when it influenced other themes, such as function; many still felt the need to legitimize their foot pain in order for health professionals to take it seriously ("in the end I went to my GP because it had happened a few times and I went to an orthopaedic surgeon who was quite dismissive of it, it was like what are you complaining about”). Conclusions: Patients interpret the outcome of foot surgery using a multitude of interrelated factors, particularly functional ability, appearance and surgeons' appraisal of the procedure. While pain was often noted, this appeared less important than other factors in the overall outcome of the surgery. Future research into foot surgery should incorporate the complexity of how patients determine their outcome Disclosure statement: All authors have declared no conflicts of interes

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

Higher Order Calcium-Dependent Protein Kinases (CPKs) Mutant Lines Elucidate Roles of CPKs within Abscisic Acid Signal Transduction in Arabidopsis and In Vivo Interactions of Calcium-Dependent Protein Kinases, CPK6 and CPK23, with PP2C Protein Phosphatases within Abscisic Acid Signaling /

Stomata, present in the aerial epidermis of land plants, provide gateways for regulating carbon dioxide and water exchange between plants and the atmosphere. The stress- induced phytohormone abscisic acid (ABA) reduces transpirational water loss crucial for the fitness of plants by inducing stomatal closure. Cytosolic Ca²⁺ has been reported to play a major role in ABA-induced stomatal closure through Ca²⁺ sensors, such as Calcium Dependent Protein Kinases (CPKs). Plants lacking transcripts of CPK3 and CPK6 (cpk3cpk6) have shown impaired ABA- and Ca²⁺- induced stomatal closing. Double knockout mutants of CPK4 and CPK11 (cpk4cpk11) exhibit similar ABA-hyposensitive phenotypes in ABA-induced stomatal closing response as well as a reduced ABA response in seed germination and seedling growth. Single mutant cpk23, however, demonstrated increased sensitivity to ABA, indicating a possible negative regulatory role in ABA signaling for CPK23. To improve the understanding of these proteins and their functions, two quadruple mutant plant lines, cpk3cpk4cpk6cpk11 and cpk5cpk6cpk11cpk23, were established. In Chapter 1, the quadruple mutant lines are phenotypically characterized by examining a variety of ABA -dependent biological responses. Other proteins important in ABA-dependent stomatal responses are Protein Phosphatases 2C (PP2Cs). Group A PP2Cs are known negative regulators within the Ca²⁺-independent ABA signaling pathway; however, experimental evidence suggests other regulatory roles exist for PP2Cs within the Ca²⁺-dependent branch of this signaling network which includes CPKs. In Chapter 2, yeast two-hybrid system is used to evaluate whether PP2Cs interact with CPKs. Observed interactions can provide additional insight to these proteins' functions within the Ca²⁺-dependent ABA signalin

Urban heat island effect on cicada densities in metropolitan Seoul

Background Urban heat island (UHI) effect, the ubiquitous consequence of urbanization, is considered to play a major role in population expansion of numerous insects. Cryptotympana atrata and Hyalessa fuscata are the most abundant cicada species in the Korean Peninsula, where their population densities are higher in urban than in rural areas. We predicted a positive relationship between the UHI intensities and population densities of these two cicada species in metropolitan Seoul. Methods To test this prediction, enumeration surveys of cicada exuviae densities were conducted in 36 localities located within and in the vicinity of metropolitan Seoul. Samples were collected in two consecutive periods from July to August 2015. The abundance of each species was estimated by two resource-weighted densities, one based on the total geographic area, and the other on the total number of trees. Multiple linear regression analyses were performed to identify factors critical for the prevalence of cicada species in the urban habitat. Results C. atrata and H. fuscata were major constituents of cicada species composition collected across all localities. Minimum temperature and sampling period were significant factors contributing to the variation in densities of both species, whereas other environmental factors related to urbanization were not significant. More cicada exuviae were collected in the second rather than in the first samplings, which matched the phenological pattern of cicadas in metropolitan Seoul. Cicada population densities increased measurably with the increase in temperature. Age of residential complex also exhibited a significantly positive correlation to H. fuscata densities, but not to C. atrata densities. Discussion Effects of temperature on cicada densities have been discerned from other environmental factors, as cicada densities increased measurably in tandem with elevated temperature. Several mechanisms may contribute to the abundance of cicadas in urban environments, such as higher fecundity of females, lower mortality rate of instars, decline in host plant quality, and local adaptation of organisms, but none of them were tested in the current study. Conclusions In sum, results of the enumeration surveys of cicada exuviae support the hypothesis that the UHI effect underlies the population expansion of cicadas in metropolitan Seoul. Nevertheless, the underlying mechanisms for this remain untested

Tracking the Development of Cerebrovascular Risk Factors Following Pregnancy With Preeclampsia

To evaluate the development and management of cerebrovascular risk factors following a pregnancy with preeclampsia. This is a retrospective chart review including women diagnosed with preeclampsia between 2012 and 2013 with later encounters within 2014-2016. For each subject that met inclusion criteria, the development of cerebrovascular risk factors was determined using ICD codes within the 2014-2016 electronic medical record (EMR). For subjects who developed risk factors, current treatment was determined from the EMR. Demographic data was also documented. Differences in the development and treatment of risk factors were compared among racial groups and age. Descriptive statistics were calculated using SAS statistical software. Compared to prepregnancy health status, the incidence of hypertension increased by 1.7 times (P < .05), hyperlipidemia increased by 4.5 (P < .05), migraines increased by 2.2 (P < .05), and diabetes mellitus increased by 2 (P < .05) after a pregnancy with preeclampsia. Black non-Hispanics had highest rates of hypertension, obesity, and migraines (20.5%, 9.1%, and 6.8%, respectively.) Of hypertensives, 73.6% (42/57) were prescribed medication. Of diabetics, 88.9% (16/18) were prescribed medication. No patients with hyperlipidemia were prescribed a statin. Black non-Hispanics had higher rates of risk factor management (74.3% of hypertensives and 100% of diabetics treated) than white Hispanics (55% and 77.8%, respectively). This study shows a significant increased risk of the development of cerebrovascular risk factors such as hypertension, hyperlipidemia, migraines, and diabetes following a diagnosis of preeclampsia. Opportunities exist for the early treatment of these risk factors, which could reduce the long-term rate of stroke in these wome