Both 5q-linked spinal muscular atrophy (SMA) and Duchenne muscular dystrophy (DMD) are fatal monogenic neuromuscular disorders caused by loss-of-function mutations. SMA is an autosomal recessive disorder affecting motor neurons that is typically caused by homozygous whole-gene deletions of SMN1. DMD is an X-linked recessive muscle disease most often due to exon deletions, but also duplications and smaller sized variants within the DMD gene. Gene replacement therapy offers the opportunity to correct the underlying genetic defect by the introduction of a functional gene. We review the transformative work from clinical trials to United States Food and Drug Administration approval of onasemnogene abeparvovec-xioi in SMA and its application in clinical practice and the early results of microdystrophin delivery in DMD. We also review the introduction of antisense oligonucleotides to alter pre-mRNA splicing to promote exon inclusion (as in nusinersen in SMA) or exclusion (as in eteplirsen in DMD) into neuromuscular therapeutics. There are multiple promising novel genetically mediated therapies on the horizon, which in aggregate point towards a hopeful future for individuals with SMA and DMD.

Team-Based Learning (TBL) is a pedagogical tool that has great potential to develop student engagement, accountability, and equity in the online classroom. TBL is rooted in evidence-based educational theories and practices that underlie many active learning approaches such as self-testing, team discussion, and application of knowledge. The use of these approaches is associated with better student performance, retention, and sense of belonging in the classroom, aspects that are often reported to be especially lacking in online courses. Here, we describe how we implemented TBL in a face-to-face and an online introductory level evolution and biodiversity course. Our experiences using TBL approaches in the online course have been rewarding, students are engaged and accountable for their learning, and performed well in the course. Our goal is to provide an example of how we designed a life science course using TBL approaches and transitioned the course to an online environment. With the current switch to remote instruction and online learning, we recommend the use of TBL as a course design approach that can improve the students online learning experience.

Abstract Background Differentiating viral from bacterial acute respiratory infections (ARIs) remains challenging, due to the non-specific clinical manifestations. The COVID-19 pandemic is putting extraordinary strain on healthcare resources. To date, molecular testing is available but has a long turnaround time and therefore cannot provide results at the point-of-care (POC) thereby exposing COVID-19/Non-COVID-19 patients to each other while awaiting diagnosis. Methods This observational study prospectively evaluated the utility of a triage strategy including FebriDx, a POC fingerstick blood test that differentiates viral from bacterial ARIs through simultaneous detection of Myxovirus-resistance protein A (MxA) and C-reactive protein (CRP), in rapidly determining viral cases requiring immediate isolation and confirmatory molecular testing, from non-infectious patients or bacterial infections requiring antibiotics. Results 75 consecutive patients were screened, 48 eligible cases were tested with FebriDx, 36 were confirmed viral infection and 35/36 had COVID-19. 31/35 COVID-19 cases tested positive for SARS-CoV-2 via rRT-PCR and (4/35) had a clinical diagnosis of probable COVID-19 based on symptoms, epidemiological history, and chest imaging (PPV 100% (35/35)). 13 cases were FebriDx viral negative and rRT-PCR was also negative. In one case, it was not possible to determine the exact cause of infection, although a viral infection could not be excluded. Including this patient, FebriDx NPV was 92.3% (12/13), exceeding the NPV of rRT-PCR a 68.3% (13/19), and diagnostic sensitivity was conservatively calculated at 97% (35/36) compared to 82.9% (29/35) for initial rRT-PCR. The diagnostic specificity of both FebriDx and rRT-PCR was 100%. Conclusions: FebriDx could be deployed as part of a reliable triage strategy for identifying possible COVID-19 patients with symptomatic ARI in the COVID-19 pandemic. Key words: Pandemic; COVID-19; SARS-CoV-2; pneumonia; viral; point of care; infection

An effective vaccine against Toxoplasma gondii is an ideal strategy for controlling acute or chronic toxoplasmosis. In order to boost immune reactions to various antigens, liposomes may be utilized as immunoadjuvants. We encapsulated soluble Toxoplasma antigen (SA) and imiquimod adjuvant in 1, 2-Dioleoyl-3-trimethylammonium Propane (DOTAP) liposomes to evaluate the immune response induced by this vaccine. Three times with 2-week intervals, BALB/C mice were immunized subcutaneously with different formulations. The type of generated immune reaction, as well as the protection extent, was assessed through the percent survival survey of BALB/c mice after challenge with Toxoplasma gondii, the evaluation immune reaction with the generation of cytokine (IFN-γ, IL-4), and titration of IgG isotypes. Less mortality was observed in the immunized mice by liposome DOTAP + imiquimod + SA that was meaningfully different (P<0.01) in comparison to other groups. The IgG2a and IFN-γ secretion highest levels were seen with liposome DOTAP + imiquimod + SA more than the control group (P<0.001) and (P<0.0001), respectively. The results of this research reveal that a cellular immune reaction is produced by the formulation of liposome DOTAP + imiquimod + SA, which is protective facing T. gondii challenge

Chronic Granulomatous Disease (CGD) is a rare inherited disorder in which phagocytes lack NADPH oxidase activity. The most common form is the X-linked CGD (X-CGD), caused by mutations in the CYBB gene. Clinical, functional and genetic characterizations of 16 CGD cases of male patients and their relatives were done. We classified them as suffering from different variants of CGD (X910, X91− or X91+) according to NOX2 expression and NADPH oxidase activity in neutrophils. Twelve mutations were novel (10 X910-CGD and 2 X91− -CGD). One X910-CGD was due to a new and extremely rare double missense mutation Thr208Arg-Thr503Ile. We investigated the pathological impact of each single using stable transfection of each mutated cDNA in the NOX2 knock-out PLB-985 cell line. Both mutations leading to X91−-CGD were also novel; one deletion -67delT was localized in the promoter region of CYBB, the second one c.253-1879A>G mutation activates a splicing donor site, which unveils a cryptic acceptor site, leading to the inclusion of a 124-nucleotide pseudo-exon between exons 3 and 4 and responsible for the partial loss of NOX2 expression. Both X91−-CGD mutations were characterized by a low cytochrome b558 expression and a faint NADPH oxidase activity. The functional impact of new missense mutations is discussed in the context of a new 3D-model of the dehydrogenase domain of NOX2. Our study demonstrates that low NADPH oxidase activity found in both X91−-CGD patients correlates with mild clinical forms of CGD whereas X910-CGD and X91+-CGD cases remain the most clinically severe forms.

Aim. The multi-part Unified Parkinson’s Disease Rating Scale is the standard instrument in clinical trials. A sum of scores for all items in one or more parts of the instrument is usually analyzed. Without accounting for relative importance of individual items, this sum of scores conceivably does not optimize the power of the instrument. The aim was to compare the ability to detect drug effect in slowing down motor function deterioration, as measured by Part III of the Scale - motor examinations - between the item scores and the sum of scores. Methods. We used data from 423 patients in a Parkinson’s disease progression trial to estimate the symptom severity by item response modelling; modelled symptom progression using the severity and the sum of scores; and conducted simulations to compare the sensitivity of detecting a broad range of hypothetical drug effects on progression using the severity and the sum of scores. Results. The severity endpoint was far more sensitive than the sum of scores for detecting treatment effects, e.g., requiring 280 versus 570 patients per arm to achieve 60% Probability of Success for detecting a range of potential effects in a 2-year trial. Items related to the left side of the body were most informative; and the domain relevance of tremor items was questionable. Conclusion. This analysis generated clear evidence that longitudinal modelling of item scores can enhance trial efficiency and success. It also prompted the needs for a consensus on the placement of the tremor items in the instrument.

The use of passive infrared thermography comprises great opportunities to improve understanding the fatigue damage process of crack-patched structures. Quasi-static and cyclic coupon tests are performed using metallic specimens with single-sided bonded patches and monitored with passive infrared thermography. Different test setups help to differentiate between metallic crack growth and adhesive damage on thermal images. Results show that metallic crack growth can be monitored from the patched side, also in combination with local delamination at the patch/metal interface. Thus, it is possible to analyse the overall degradation progress of the crack patched component under loading conditions and thereby to identify the driving damage mechanism of the particular repair configuration. Being able to understand the overall damage behaviour of crack patched components is essential to improve the ability of predicting its long-term behaviour.

Aim: The retinal nerve fiber layer (RNFL) thickness and the choroidal thickness is compared to healthy volunteers in vitiligo patients. Material and method: The study included the right eyes of 60 healthy individuals and 60 generalized vitiligo patients. Following routine ophthalmological examination, the RNFL thickness and the choroidal thickness were analyzed with spectral domain optical coherence tomography (SD-OCT). Results: It was determined that the mean macular choroidal thickness decreased significantly in vitiligo patients, while the RNFL thickness did not change (p<0.05). Conclusion: As the fact that the RNFL thickness does not change despite the significantly reduced choroidal thickness in vitiligo patients may indicate that the melanin cells that are dense in the choroid are affected, the non-invasive, simple and rapid measurement of the choroidal thickness with OCT can help in determining the damage in vitiligo patients. Keywords: Vitiligo, choroid, nerve fibers, optical coherence tomography. 1. What is already known about this topic? Melanocytes are found in uveal tissues such as the eyelash, retinal pigment epithelium, choroid, ciliary body, and iris. Melanin in these tissues can disappear when destruction occurs in cutaneous melanocytes in vitiligo patients. As a result, some studies reported hypopigmented spots on the iris and retina, atrophic changes in the peripapillary area, degeneration in the retinal pigment epithelium and chorioretinal areas. Melanin, produced in melanocytes in the choroidal layer and stored in melanosomes, has a crucial role in protecting and absorbing light from intraocular reflection. 2. What does this article add? Although the visual acuity of the patients with vitiligo is at the same level as the control group, we found that the average macular choroid thickness was thinner. We found no significant changes in the choroidal thickness in patients with periocular attitude. We found a negative correlation between the choroidal thickness and the duration of the disease.

Plant roots interact with rhizosphere microbes to accelerate soil organic matter (SOM) mineralization and promote nutrient acquisition. Root-mediated changes in SOM turnover largely depend on root-C input and soil nutrient availability. Hence, interspecific competition and nutrient uptake dynamics over plant development stages as well as spatiotemporal variability in C input may modify SOM turnover. To investigate the effect of intraspecific competition on SOM mineralization at three growth stages (heading, flowering and ripening), we grew maize (C4 plant) under three planting densities on a C3 soil. 13C-natural abundance and 15N-pool dilution were applied in situ to determine C- and N-mineralization rates. Soil C- and N-mineralization rates were tightly coupled and peaked at maize flowering. However, the C-to-N-mineralization ratio increased with N, indicating that microbes mineralize N-rich components to mine SOM for N. Furthermore, intraspecific competition did not affect root biomass; instead, plants shaped root morphology towards higher specific root length as an efficient strategy competing for nutrient. Hence, root morphologic traits rather than root biomass per se were positively related to C- and N-mineralization. Overall, plant competition for nutrients controlled the intensity and mechanisms of soil C- and N-turnover by the adaptation of root traits and nutrient depletion.

Light is important for plant life as a source of energy to drive photosynthesis but also as an environmental signal regulating development or cellular events such as resetting of the circadian clock. Light itself can cause stress such as excess light, fluctuating light, photoperiod and ultraviolet light stress. Light quality, quantity and light duration are important sources of information to prepare plants for future light stress events. Recurring light stress results in acclimation processes to the changing light environment. Furthermore, light regulates the responses of plants to diverse biotic and abiotic stresses. For example, short day conditions or shady environments prime thermotolerance and increase cold acclimation. Similarly, during drought stress, light signaling is important for the plant´s stress response. Additionally, the light environment affects the plant´s responses to biotic intruders such as pathogens or insect herbivores. Light influence many stress responses resulting in positive growth-defense trade-offs. Under shade, however, plants prioritize growth over defense and stress responses. In this review, we summarize the impact of light as a stressor and its influences on abiotic and biotic stress responses with special focus on the role of the different light receptors and the crosstalk between light signaling components and stress response pathways.

In this study the cucumber and wheat plants were used to clarify the mechanism of redroot pigweed’s allelopathic effects on photosynthesis. In order to reach that goal, plants were cultivated hydroponically, treated by redroot pigweed’s leachate and finally chlorophyll fluorescence and photosynthetic gas exchange parameters, photosynthetic pigments content and the expression of photosynthetic genes (PsbA and PsbS) and allelochemical interaction with proteins of studied genes were analyzed. After exposure to the allelopathic stress, significant differences in the majority of photosynthetic characteristic were observed in the studied species. Redroot pigweed allelopathy led to alteration in photosynthesis performance, photochemistry, and photosynthetic genes expression of cucumber and wheat plants, ultimately its results were observed in morphological traits of plants. Molecular docking study strongly confirmed the possibility of direct binding and action of allelopathic compounds with allelopathic action’s target proteins. Overall, this study showed that compared to cucumber, the wheat plant is able to withstand the damaging effects of amaranth allelopathy on photosynthesis in order to resist and survive.

2.4 The relationship between levels of IL-10, IL-1β, IL-6, MCP-1, TNF-α, IP-10 and IL-4 and antibody concentration in serum samples of COVID-19 patientsThe correlation analysis of levels of IL-10, IL-1β, IL-6, MCP-1, TNF-α, IP-10 and antibody concentration in serum samples of COVID-19 patients were shown in Table 3. It was found that the levels of IP-10 in COVID-19 patients were positively correlated with the level of IgM (r=0.255, P<0.05), while the levels of TNF-α were negatively correlated with the level of IgG (r=-0.217, P<0.05), but there were no significant correlation between the levels of IL-10, IL-1β, IL-6, MCP-1 and IL-4 and the concentrations of IgG and IgM.

Conventional wisdom suggests any effort to control pests and parasites is better than none. However, growing evidence demonstrates weak or moderate effort can backfire, fail, and potentially worsen outcomes versus doing nothing. A central challenge is anticipating these potential failures before inducing environmental damage. We built a resource-explicit individual-based model of human schistosome and snail host transmission to evaluate biocontrol effort by simulating 22 predator stocking densities of native prawns. We test two host mortality scenarios and show intense biocontrol effort can succeed. However, weak to moderate effort can backfire by allowing large, prolific hosts to escape predation and drive overcompensation due to three interacting ecological mechanisms—resource competition among hosts, resource-dependent infectiousness, and predator gape limits. Ultimately, integrating physiology, ecology, and epidemiology can identify the risks of weak or moderate control effort when evaluating potential ‘do or do not’ control designs for future management of wildlife pests and diseases.

As neuronal data accumulates worldwide, accessible - yet rigorous - resources to develop hands-on experience with modern data analysis techniques are required. We present here an online educational resource for neural data analysis (https://mark-kramer.github.io/Case-Studies-Python). To reach the biologists, psychologists, and clinicians collecting neuronal data, we assume only a basic mathematics background, common to those trained in biological sciences. Through an interdisciplinary case-study approach, we use real-world data to motivate the study of modern quantitative analysis methods in Python. A modular format provides multiple coherent learning paths through the material, and thereby allows personalized learning for individuals with varying quantitative backgrounds and research interests, and flexible curation of material for redeployment in other curricula. Developed using Jupyter notebooks, the material supports fully interactive environments in most web browsers, and hosted on GitHub, the material is freely available for reuse, modification, and further development by the community.

The H10-H12 subtypes are designated as the “waterfowl-associated” subtypes and are not frequently detected in nature, but these viruses can highly reassort with other subtypes of AIVs. It has shown that very few H10-H12 subtypes were isolated from wild birds in China, it is essential to conduct the extensive surveillance of these rare subtypes in wild birds to narrow this knowledge gap in this region. In this study, 12 AIVs of H10-H12 subtypes were identified with the routine surveillance in wild birds in Shanghai, China from 2016 to 2019. There were 2 H10 subtypes, 3 H11 subtypes and 7 H12 subtypes, and the HA-NA combinations were H10N4, H11N2, H11Nx, H11N9, H12N2, H12N5 and H12N8. Sequence and phylogenetic analysis showed these gene segments of the 12 strains had high levels of genetic diversity among them, and most of them were closely related to the Eurasian lineage, and shared high sequence identity with those isolated from wild birds and domestic ducks in Japan, Korea, Bangladesh, Vietnam and China located at the East Asian-Australasian Flyway route. However, part of the gene segments of the two H12N2 strains (NH112319-H12N2 and NH101807-H12N2) were belonged to the North American lineage, which indicated that the gene flow and reassortment had occured between the Eurasian and American lineages in H12 subtypes. To better understand the ecological and phylodynamic features of these H10-H12 subtypes in wild birds, it is necessary to continuously conduct large-scale surveillance of wild birds in future.

To date, more than thirty animals were tested positive for SARS-CoV-2, all of them infected by humans with COVID-19. Some animal experiments suggested the possibility of an animal to animal transmission of SARS-CoV-2 and were seen in some cases of infected animals. Animal to human transmission was considered unlikely until investigations revealed the possibility of mink to human transmission of SARS-CoV-2 in the Netherlands. Studying the susceptibility of SARS-CoV-2 to domestic animals concluded that pigs, chicken, and ducks were not vulnerable to Covid-19; dogs showed less susceptibility to SARS-CoV-2 and cats as well as ferrets were seen susceptible to Covid-19. SARS-CoV-2 is seen crossing the species barrier, infecting humans from the wild with the source yet unclear, spreading from humans to humans quickly, humans to animals, animals to animals, and is likely to spread from animals to humans even though minimally. Animals appear somewhat resistant to SARS-CoV-2 transmission compared to humans who globally crossed eight million infection cases, and the infected animals mostly do not show many complications and recover quickly. Precautions are advised to prevent human to animal transmission of the virus, and in some areas, avoid animal to human spread of the virus. Further monitoring is required to assess the SARS-CoV-2 infection in animals as COVID-19 is a rapidly evolving condition worldwide.

Animal health information systems or risk analysis tools are indispensable not only for animal health surveillance, but also to observe the evolution and risk of disease incursion into a disease-free area. Given their essential function in animal disease prevention, different international and national organizations have created their own aforementioned systems/tools. Moreover, with the increase of technology and data storage, they have become more accessible and widely used by professionals in animal and human health sciences. This study aimed to establish their preferences, needs and constraints in respect of these tools. An online survey was conducted and answered by 213 respondents from 132 countries. The respondents were animal or public health professionals in different employment sectors (mostly in government, research and university institutions) and various fields of competency (highest for animal and public health). The majority of respondents used the animal health information systems frequently and on a weekly basis, with prevention measures of diseases being regarded as the most useful information. Descriptive epidemiology is more used/needed than analytical epidemiology. Risk analysis was performed by the majority of the respondents (70%), using a qualitative approach more than a quantitative or semi-qualitative. The primary objectives was to produce risk assessment and preparedness in areas involving origin and spread of animal diseases. The features most sought after in risk assessment tools were pathways of introduction and spread assessment. The level of satisfaction was higher for the platform which is most used by the respondents. Thus, the platform choice is most likely influenced by its efficiency and functionality. Overall, these results could be taken into consideration when improving an already available platform, or when creating a new efficient tool.

Author Information: Amit Jain M.D., Anesthesiology Institute, Cleveland Clinic Abu Dhabi, United Arab EmiratesD. John Doyle, M.D., Ph.D., Anesthesiology Institute, Cleveland Clinic Abu Dhabi, United Arab Emirates

Minimally Invasive Aortic Valve Replacement is not just a metric of the incision, but rather a holistic approach to minimize the surgical trauma: our technique tries to reproduce the gold-standard conventional procedure in terms of safety, effectiveness and operative times through a small and different incision. Thanks to a thorough optimization, the surgical times can be at least equal to the classic approach. We started this technique about 10 years ago and after more than 1000 of aortic valve replacement via right anterior mini-thoracotomy, we came up with some tips&tricks to make this technique our approach of choice.

Permanent pacemaker implantation is life saving but sometimes the procedure may itself become life threatening. Right ventricular perforation is a rare complication during pacemaker insertion that could be life-threatening and need immediate intervention. Chest X-ray, echocardiogram and computer tomogram of chest are used to diagnose lead migration and its complications. We present a rarest of the rare case in which a seventy six year old patient undergoing permanent pacemaker implantation suffered both iatrogenic cardiac injury leading to hemopericardium and pacemaker malfunction as well as injury to left pleura leading to massive pneumothorax and hemodynamic instability which was managed successfully.

A patient presented with torsade de pointes after inhaling tetrafluoroethane, a volatile gas propellant used to clean keyboards. She had a prior hospitalization for cardiac arrest without rhythm documentation after inhaling a similar product. Urine toxicology revealed methamphetamine. We are unaware of previous descriptions of torsade de pointes associated with volatile hydrocarbon inhalation or “huffing.” Although sudden death is associated with “huffing,” an arrhythmic mechanism has not been elucidated. Tetrafluoroethane, however, is chemically similar to chlorophorm (trichloromethane), which blocks the delayed rectifier potassium ion current (Ikr). Thus, we postulate that delayed repolarization with methamphetamine triggering may account for this arrhythmia.

Background and Purpose: Patients with community acquired pneumonia (CAP) may suffer from myocardial injury, which is associated with increased risk of major adverse cardiovascular events (MACE). Corticosteroids are often prescribed to CAP patients, but the relationship between their use, myocardial injury and outcomes in these patients is unknown. Experimental Approach: 541 CAP patients were recruited (334 males; mean age: 71.9±16.2 years). High-sensitivity troponin T (hs-cTnT) was measured at admission, during the hospital stay and at discharge. MACE occurrence was registered during a long-term follow-up. Key Results: Overall, 318 patients (59%) showed hs-cTnT elevation > 99th percentile (>0.014 µg/L). Patients with hs-cTnT elevation were older, more likely to be former smokers, and with a higher prevalence of cardiovascular comorbidities. In a median follow-up of 22.7 months, a multivariable Cox proportional hazard regression analysis showed age, heart failure and the increasing quintiles of hs-cTnT (HR: 2.16; 95% CI: 1.82-2.58; p<0.001) predicted MACE. In-hospital corticosteroid use was found in 137 (25%) patients. Among patients with hs-cTnT >0.014 µg/L at admission, 102 patients (31%) were on corticosteroids and showed lower intra-hospital hs-cTnT increase compared to untreated ones (p=0.003). Among patients with hs-cTnT >0.014 µg/L, corticosteroid-treated patients showed a lower incidence of MACE than untreated ones [29% (27/99) vs. 43% (92/213); p value =0.042]; no effect of corticosteroids on MACE was observed in CAP patients with normal troponin. Conclusion and Implications: The study provides evidence that corticosteroid use is associated with lower increase of hs-cTnT and incidence of MACE in CAP patients.

Background: Nosocomial infections (NI) are common complications after cardiac surgery. Till now, there are rare manuscripts investigating NI in intensive care unit (ICU) after cardiac surgery. Our article was designed to investigate the characteristics of distribution of pathogenic bacteria and antibiotics resistance of NI. Methods: 1360 patients received standard postoperative care including antibiotic prophylaxis. Microbiological examinations of the sputum, blood, catheter tips and excretion were performed as clinically indicated to isolate pathogens. Results: 89 patients (6.54%) acquired microbiological documented NI. There was a statistical difference in mortality between two groups with or without postoperative NI (23.60% vs 2.28%, p<0.00). 98 (73.13%) were isolated from sputum and 32 (23.88%) from blood. 3 (2.24%) SSI were detected (2 superficial SSI, 1 mediastinitis) and only 1 (0.75%) from urine. The most common pathogens were Gram-negative bacteria (78.36%), followed by Gram-positive bacteria (14.93%) and fungi (6.71%). The major species of pathogens had different levels of drug resistance and most of them exhibited multidrug-resistance (MDR). Conclusions: We analyzed the characteristics of distribution of pathogens and antibiotics resistance of NI in our center and provided some suggestions for clinical practice. Except for antibiotic treatment, aggressive infection control measures may be crucial to stop or prevent outbreaks.

Excitation energy transfer is a ubiquitous process of fundamental importance for understanding natural phenomena, such as photosynthesis, as well as advancing technologies ranging from photovoltaics to development of photosensitizers and fluorescent labels. This work provides an overview of recent advancements in excitation energy transfer modeling with the PyFREC software package. Computational methods currently implemented in PyFREC include molecular fragmentation techniques, as well as methods for electronic coupling computations, analysis of coupled electronic excited states, and quantum dynamics simulations. Advanced functionality and possible vectors for future development of the package are also explored.