Biological Breathalyzer

The aim of this research is the construction of a biological breathalyzer through synthetic biology, specifically through use of the metabolic pathways of a species of the Pichia taxa. The yeast utilized is able to metabolize both ethanol and methanol. However, when both ethanol and methanol are present, the yeast prefers to metabolize ethanol such that an AOX gene is not expressed because the first known by-product of methanol metabolism is the AO enzyme from the AOX gene. The AOX gene promoter is fused with a fluorescence protein gene so expression of the AOX gene can be visually detected. When the cell is supplied with both ethanol and methanol, the amount of time before fluorescence will correspond to the amount of ethanol given to the cell. In this way, the concentration of ethanol can be determined

Construction and Testing of a Biological Breathalyzer

The ultimate goal of this research was the construction of a biological breathalyzer using methods of synthetic biology. The metabolic pathways of Pichia taxa were utilized for this research. This yeast is able to metabolize both ethanol and methanol present in the environment. The first known by-product of methanol metabolism is the AO enzyme from the AOX gene. When both ethanol and methanol are present, the yeast preferentially metabolizes ethanol and the AOX gene is not expressed. By fusing the AOX gene promoter with a fluorescence protein gene, the expression of the AOX gene may be visually detected. When the organism is supplied with both ethanol and methanol, the amount of time before fluorescence will correspond to the amount of ethanol fed to the cell. In this way, the concentration of ethanol can be determined

SARS-CoV-2 specific antibody and neutralization assays reveal the wide range of the humoral immune response to virus.

Development of antibody protection during SARS-CoV-2 infection is a pressing question for public health and for vaccine development. We developed highly sensitive SARS-CoV-2-specific antibody and neutralization assays. SARS-CoV-2 Spike protein or Nucleocapsid protein specific IgG antibodies at titers more than 1:100,000 were detectable in all PCR+ subjects (n = 115) and were absent in the negative controls. Other isotype antibodies (IgA, IgG1-4) were also detected. SARS-CoV-2 neutralization was determined in COVID-19 and convalescent plasma at up to 10,000-fold dilution, using Spike protein pseudotyped lentiviruses, which were also blocked by neutralizing antibodies (NAbs). Hospitalized patients had up to 3000-fold higher antibody and neutralization titers compared to outpatients or convalescent plasma donors. Interestingly, some COVID-19 patients also possessed NAbs against SARS-CoV Spike protein pseudovirus. Together these results demonstrate the high specificity and sensitivity of our assays, which may impact understanding the quality or duration of the antibody response during COVID-19 and in determining the effectiveness of potential vaccines

Mainstreaming climate change education in UK higher education institutions

Key messages: • Mainstreaming Climate Change Education (CCE) across all learning and operational activities enables Higher Education Institutions (HEIs) to better serve their core purpose of preparing learners for their roles in work and wider society, now and in the future. • Student and employer demand for climate change education is growing, not just in specialist subjects but across all degree pathways. • The attitudes, mindsets, values and behaviours that graduates need to engage with climate change include the ability to deal with complexity, work collaboratively across sectors and disciplines and address challenging ethical questions. • The complexity of the climate crisis means all disciplines have a role to play in delivering education for the net-zero transition. Embedding interdisciplinarity is crucial to ensuring that our response to climate change makes use of all of the expertise HEIs have to offer and promotes knowledge exchange and integration for students and staff. • Student-centered CCE, including peer-to-peer learning, is a powerful tool for facilitating an inclusive and empowering learning experience, and developing graduates as change agents for the climate and ecological crisis. • HEIs should develop learning outcomes for CCE that include understanding the scale, urgency, causes, consequences and solutions of climate change; how social norms and practices are driving the climate crisis; and the ability to identify routes to direct involvement in solutions via every discipline. • Pedagogical approaches to teaching CCE should enable learners to engage with, and respond to, climate change as a “real-world” problem, such as through experiential learning. • Further recommendations for the HEI sector include developing a strategy for aligning CCE teaching provision with governance structures; partnering with industry, government and third sector organisations to enable context-specific CCE; and working with trade unions and accreditation bodies to enable curriculum reform

Mainstreaming Climate Change Education in UK Higher Education Institutions

Key messages• Mainstreaming Climate Change Education (CCE) across all learning and operational activities enables Higher Education Institutions (HEIs) to better serve their core purpose of preparing learners for their roles in work and wider society, now and in the future.• Student and employer demand for climate change education is growing, not just in specialist subjects but across all degree pathways.• The attitudes, mindsets, values and behaviours that graduates need to engage with climate change include the ability to deal with complexity, work collaboratively across sectors and disciplines and address challenging ethical questions.• The complexity of the climate crisis means all disciplines have a role to play in delivering education for the net-zero transition. Embedding interdisciplinarity is crucial to ensuring that our response to climate change makes use of all of the expertise HEIs have to offer and promotes knowledge exchange and integration for students and staff.• Student-centered CCE, including peer-to-peer learning, is a powerful tool for facilitating an inclusive and empowering learning experience, and developing graduates as change agents for the climate and ecological crisis.• HEIs should develop learning outcomes for CCE that include understanding the scale, urgency, causes, consequences and solutions of climate change; how social norms and practices are driving the climate crisis; and the ability to identify routes to direct involvement in solutions via every discipline.• Pedagogical approaches to teaching CCE should enable learners to engage with, and respond to, climate change as a “real-world” problem, such as through experiential learning.• Further recommendations for the HEI sector include developing a strategy for aligning CCE teaching provision with governance structures; partnering with industry, government and third sector organisations to enable context-specific CCE; and working with trade unions and accreditation bodies to enable curriculum reform

Constructing an Ethanol Sensor

In Pichia pastoris, alcohol oxidase (AOX) is the first enzyme in the methanol utilization pathway. This enzyme is encoded by the AOXI gene. If exposed to an environment containing both methanol and ethanol, P. pastoris preferentially metabolizes ethanol. The production of the AOX enzyme is subject to the concentration of ethanol. This diauxic metabolism may be utilized as an ethanol sensor. When the AOXI promoter is fused with a gene encoding a fluorescent protein, the activation of the AOXI promoter may be detected by direct observation of fluorescence. Our project is the development of a device containing the AOXI promoter fused with a fluorescent protein gene to create an inexpensive ethanol sensor for a variety of applications. The concentration of ethanol in the environment may be deduced from the time period between exposure of the microorganism carrying the device to ethanol and methanol, until the detection of fluorescence

A Near-Miss Diagnosis of Necrotizing Breast Fasciitis Complicated by Atrial Fibrillation Secondary to Septic Focus: a Case Report and Brief Review of Literature

Necrotizing fasciitis of the breast is a rare, life-threatening entity characterized by a rapidly aggressive infection of the soft tissue. There are few literature reports on necrotizing fasciitis at the level of the breast tissue as the most common locations are within the abdominal wall or extremities, but this entity can lead to sepsis and systemic multiorgan failure if not adequately managed. Here, we report a case that highlights the course of a 68-year-old African American female with a past medical history of hypertension, hyperlipidemia, and poorly controlled diabetes mellitus, who presented with the complaint of a painful right breast abscess with intermittent, purulent drainage. An initial point-of-care ultrasound displayed an area of induration of the right breast as well as soft tissue edema with no identifiable fluid collection. A subsequent CT abdomen and pelvis was obtained given new onset abdominal pain, which demonstrated incidental findings of inflammatory changes and subcutaneous emphysema along with colonic diverticulosis. Surgical intervention was immediately sought for which she underwent debridement and exploration of the right breast with findings that were consistent with necrotizing transformation. The patient was sent back to the OR for an additional surgical debridement the next day. Of note, the patient had post-op atrial fibrillation with rapid ventricular response and had to be admitted to the ICU for conversion to sinus rhythm. She returned to sinus rhythm and was transferred back to medicine before application of a negative pressure wound dressing on discharge. The patient was transitioned from Enoxaparin to Apixaban for anticoagulation control in the setting of atrial fibrillation before being discharged to a Skilled Nursing Facility with long-term antibiotics. This case highlights the difficulty and significance in establishing a prompt diagnosis for necrotizing fasciitis

Tocilizumab for Severe COVID-19 Infection and Multisystem Inflammatory Syndrome in Adults and Children

Coronavirus disease 2019 (COVID-19) rapidly emerged as a global pandemic, placing imminent stress and burden on healthcare resources and workers worldwide. Many patients who present with a severe COVID-19 infection are at high risk of developing severe acute respiratory distress syndrome (ARDS), leading to a vast number of patients requiring mechanical ventilation and a high mortality rate. Similar to Middle East respiratory syndrome, COVID-19 demonstrates an initial viral replication phase that manifests as a variety of symptoms typically flu-like in nature, followed by a profound inflammatory response leading to rapid production of cytokines and uncontrolled inflammation. There have also been many cases of COVID-19 in pediatric patients presenting with elevated inflammatory markers and multisystem involvement labeled as a multisystem inflammatory syndrome (MIS-C) by the world health organization (WHO). The recent treatment of systemic inflammatory response to COVID-19 targets the secondary phase involving cytokine release syndrome. The detrimental effects of IL-6 can be profound and elevated levels are associated with a higher mortality rate and mechanical ventilation. Tocilizumab is an IL-6 inhibitor most widely investigated to target cytokine storm syndrome. Since June 2021, the FDA enacted an emergency use authorization for tocilizumab in the treatment of COVID-19. Several clinical trials have investigated tocilizumab combined with corticosteroids for treating severe ARDS associated with COVID-19. An increasing amount of evidence suggests that targeting the cytokine storm syndrome related to COVID-19 can lead to improved outcomes, especially in those patients requiring mechanical ventilation and with a critical illness. Additional studies are warranted to further look at the positive effects of tocilizumab in the COVID-19 population while additionally defining possible adverse effects

Neuroimaging in encephalitis:analysis of imaging findings and interobserver agreement

AimTo assess the role of imaging in the early management of encephalitis and the agreement on findings in a well-defined cohort of suspected encephalitis cases enrolled in the Prospective Aetiological Study of Encephalitis conducted by the Health Protection Agency (now incorporated into Public Health England).Materials and methodsEighty-five CT examinations from 68 patients and 101 MRI examinations from 80 patients with suspected encephalitis were independently rated by three neuroradiologists blinded to patient and clinical details. The level of agreement on the interpretation of images was measured using the kappa statistic. The sensitivity, specificity, and negative and positive predictive values of CT and MRI for herpes simplex virus (HSV) encephalitis and acute disseminated encephalomyelitis (ADEM) were estimated.ResultsThe kappa value for interobserver agreement on rating the scans as normal or abnormal was good (0.65) for CT and moderate (0.59) for MRI. Agreement for HSV encephalitis was very good for CT (0.87) and MRI (0.82), but only fair for ADEM (0.32 CT; 0.31 MRI). Similarly, the overall sensitivity of imaging for HSV encephalitis was ∼80% for both CT and MRI, whereas for ADEM it was 0% for CT and 20% for MRI. MRI specificity for HSV encephalitis between 3–10 days after symptom onset was 100%.ConclusionThere is a subjective component to scan interpretation that can have important implications for the clinical management of encephalitis cases. Neuroradiologists were good at diagnosing HSV encephalitis; however, agreement was worse for ADEM and other alternative aetiologies. Findings highlight the importance of a comprehensive and multidisciplinary approach to diagnosing the cause of encephalitis that takes into account individual clinical, microbiological, and radiological features of each patient