Early detection of Ebola virus proteins in peripheral blood mononuclear cells from infected mice.

BACKGROUND: Detection of viral ribo-nucleic acid (RNA) via real-time polymerase chain reaction (RT-PCR) is the gold standard for the detection of Ebola virus (EBOV) during acute infection. However, the earliest window for viral RNA detection in blood samples is 48-72 h post-onset of symptoms. Therefore, efforts to develop additional orthogonal assays using complementary immunological and serological technologies are still needed to provide simplified methodology for field diagnostics. Furthermore, unlike RT-PCR tests, immunoassays that target viral proteins and/or early host responses are less susceptible to sequence erosion due to viral genetic drift. Although virus is shed into the bloodstream from infected cells, the wide dynamic range of proteins in blood plasma makes this a difficult sample matrix for the detection of low-abundant viral proteins. We hypothesized that the isolation of peripheral blood mononuclear cells (PBMCs), which are the first cellular targets of the Ebola virus (EBOV), may provide an enriched source of viral proteins. METHODS: A mouse infection model that employs a mouse-adapted EBOV (MaEBOV) was chosen as a proof-of-principal experimental paradigm to determine if viral proteins present in PBMCs can help diagnose EBOV infection pre-symptomatically. We employed a liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) platform to provide both high sensitivity and specificity for the detection and relative quantitation of viral proteins in PBMCs collected during MaEBOV infection. Blood samples pooled from animals at the post-infection time-points were used to determine the viral load by RT-PCR and purify PBMCs. RESULTS: Using quantitative LC-MS/MS, we detected two EBOV proteins (vp40 and nucleoprotein) in samples collected on Day 2 post-infection, which was also the first day of detectable viremia via RT-PCR. These results were confirmed via western blot which was performed on identical PBMC lysates from each post-infection time point. CONCLUSIONS: While mass spectrometry is not currently amenable to field diagnostics, these results suggest that viral protein enrichment in PBMCs in tandem with highly sensitive immunoassays platforms, could lead to the development of a rapid, high-throughput diagnostic platform for pre-symptomatic detection of EBOV infection.

Concussion Care in Teenage Athletes.

Concussion is a form of traumatic brain injury that occurs commonly in pediatric patients. One group at higher risk for concussion is adolescent and teenage athletes. Athletes may sustain one or more concussions throughout their athletic careers, and these injuries may lead to significant morbidities for children. Although most concussion symptoms will resolve in less than 1 month, there is a subset of patients for whom symptoms may persist. Given the increased interest in preventing long-term sequelae related to concussion, legislation has been passed across the United States that aims to identify concussion promptly and remove participants from play after an injury has occurred. Care and treatment for concussion is evolving, with newer recommendations stating that complete or extreme rest may be unnecessary during the recovery period. Pediatricians and those who interact with young athletes should be aware of the guidelines surrounding return-to-play and return-to-learn in their geographic area of practice. [Pediatr Ann. 2024;53(2):e39-e41.].

Evaluation and Management of Blunt Head Trauma in the Pediatric Clinic.

Head injuries, and specifically blunt head trauma, are common among pediatric patients of all ages. Patients may present to their primary care provider, to urgent care, or to the emergency department after head trauma. Such injuries may occur as a result of a variety of mechanisms, including falls, motor vehicle collisions, or sports injuries. Clinical decision rules exist to help guide the clinician in the initial evaluation of head injury and in determining when head imaging may be indicated. One such guideline that is widely used in the United States is known as the PECARN (Pediatric Emergency Care Applied Research Network) criteria. Pediatricians should also evaluate for the presence of symptoms consistent with concussion that may occur as a result of blunt head trauma and be familiar with the management and sequelae of concussion and head injuries. [Pediatr Ann. 2023;52(8):e279-e281.].

"The Fever Is Still There!": Workup and Management of Prolonged Fever.

Fever is one of the most common chief complaints that brings pediatric patients to seek medical care. Although fever is, in most cases, a physiologic response to a pathogen that has alerted the immune system, prolonged fever can be challenging to work up for the treating pediatrician. In addition to assessing causes of fever, pediatricians must also address the fears that many caregivers may have surrounding elevations in body temperature and provide education as to when fever becomes a concern. Fever can be classified by its duration as well as the presence or absence of associated symptoms. Fever without a source is defined as a fever that has been present for 1 week without a clear cause. Fever of unknown origin is a fever that has been present daily for 8 days or more without an apparent source. This article will walk through considerations for a clinician evaluating a pediatric patient with prolonged fever. [Pediatr Ann. 2023;52(4):e124-e126.].

Updates on the Evaluation and Management of Febrile Neonates.

In August 2021, the American Academy of Pediatrics published a guideline, the first of its kind, to synthesize decades of research on the care of febrile neonates. The guideline is not meant to dictate one particular course of treatment for every clinical scenario, but rather to provide evidence-based recommendations to guide care in collaboration with the patient's caretaker(s). The guideline considers evolving bacteriology, costs of care, advances in laboratory testing and inflammatory markers, and known risks related to the hospitalization of young infants. The guideline highlights 21 key action statements to guide practitioners in the care of neonates presenting with fever, with the aim of optimizing outcomes while reducing risk and unnecessary costs. [Pediatr Ann. 2022;51(2):e49-e51.].

Pulse Oximetry Screening for Congenital Heart Defects in the Newborn Nursery: A Review for the General Pediatrician.

Congenital heart defects are a heterogenous group of disorders that may present in varied ways early in life. Of particular importance within the category of congenital heart defects are "critical" congenital heart defects, which require surgery, interventional catheterization, or prostaglandin dependence before intervention by age 1 month or 1 year, depending on which definition is used. Because of the time-sensitive element of identifying such lesions, the question is raised: how can children affected by critical congenital heart defects best be identified? One strategy that has been used since the mid-1990s is a pulse oximetry screening assessment, which detects such abnormalities in neonates and has led to improved child health outcomes across the United States and the world. This article will serve as a review of the critical congenital heart defect pulse oximetry screening tool for general pediatricians and those who care for infants. [Pediatr Ann. 2022;51(11):e411-e413.].

Social Media, Adolescents, and the Role of the Pediatrician: Exploring Potential Benefits and Harms in Today's Digital Landscape.

The use of technology among children, adolescents, and adults has increased dramatically in the last 15 to 20 years. Along with the presence of technology has come increased exposure and access to social media, particularly for adolescents and teenagers. Social media may include any form of electronic communication where users share information and build communities; some examples of commonly used social media sites include YouTube, TikTok, Snapchat, Instagram, and Facebook. With the near-ubiquitous presence of smartphones and tablets, most adolescents and teens are using devices and associated social media sites. There are advantages and disadvantages to the use of social media, and pediatricians should be aware of potential risks as well as ways that social media use may be beneficial to teens. Communication with caregivers and patients about technology and social media use is an important piece of anticipatory guidance during health care visits, and there are multiple ways that pediatricians can facilitate healthy usage of social media with their patients. [Pediatr Ann. 2022;51(8):e300-e303.].

Effects of the COVID-19 Pandemic on Well-Child Care and Recommendations for Remediation.

The coronavirus disease 2019 pandemic has touched nearly every aspect of medical care in the United States, including the provision of well-child care to pediatric patients. According to the American Academy of Pediatrics, well-child visits are a crucial component of ensuring the optimal health and well-being of children of all ages.1 The pandemic has generated challenges to providing effective and timely well-child visits, which has led to effects at the population level, the practice level, for special patient populations, and for individual patients and families. This article aims to outline these effects and identify solutions and remedies for pediatric providers. [Pediatr Ann. 2021;50(12):e488-e493.].