A rapid influenza diagnostic test based on detection of viral neuraminidase activity

Current methods used for diagnosis of acute infection of pathogens rely on detection of nucleic acids, antigens, or certain classes of antibodies such as IgM. Here we report a virus enzyme assay as an alternative to these methods for detection of acute viral infection. In this method, we used a luciferin derivative as the substrate for detection of the enzyme activity of influenza viral neuraminidase as a means for diagnosis of influenza. The resulting commercial test, the qFLU Dx Test, uses a different supply chain that does not compete with those for the current tests. The assay reagents were formulated as a master mix that accommodated both the neuraminidase and luciferase reactions, thereby enabling rapid and prolonged production of stable light signal in the presence of influenza virus in the sample. The assay was evaluated using depository throat swab specimens. As expected, the assay exhibited similar detection rates for all influenza types and subtypes except for A(H7N9), which exhibited lower detection rate due to lower viral titer in the specimens. When throat swab specimens were diluted with the sample buffer of the test kit and tested with the qFLU Dx Test. The sensitivity and specificity were 82.41% (95% confidence interval: 79.66–85.84%) and 95.39% (95% confidence interval: 94.32–96.46%), respectively, for these diluted specimens in comparison to a real-time polymerase chain reaction assay. The uniqueness of the qFLU Dx Test as an enzymatic assay makes it highly complementary with currently available methods.

Alterations in Cortisol and Interleukin-6 Secretion in Patients With COVID-19 Suggestive of Neuroendocrine-Immune Adaptations

Purpose: The beneficial effect of glucocorticoids in coronavirus disease (COVID-19) is established, but whether adrenal cortisol secretion is impaired in COVID-19 is not fully elucidated. In this case-control study we investigated the diurnal free bioavailable salivary cortisol secretion in COVID-19 patients.Methods: Fifty-two consecutive COVID-19 patients -before dexamethasone treatment- recruited between April 15th -June15th-2021, (NCT04988269) at Laikon Athens University-Hospital, and 33 healthy age- and sex-matched controls were included. Diurnal salivary cortisol (8am, 12, 6, and 10pm), plasma adrenocorticotropin (ACTH) and aldosterone, and serum interleukin-6 (IL-6) and C-reactive protein (CRP) levels were assessed. Diurnal salivary dehydroepiandrosterone (DHEA) and IL-6 were also assessed in subgroups of patients.ResultsMedian CRP and IL-6 measurements were about 6-fold higher in patients than controls (both p<0.001) Morning salivary cortisol levels did not differ between the two groups, but patients exhibited higher median levels of evening and nocturnal salivary cortisol compared to controls [0.391(0.054, 0,663) vs. 0.081(0.054, 0.243)μg/dl, p<0.001 and 0.183(0.090, 0.834) vs. 0.054(0.054, 0.332)μg/dl, p<0.001, respectively], resulting in higher time-integrated area under the curve (AUC) (4.81±2.46 vs. 2.75±0.810, respectively, p<0.001). Circulating ACTH, DHEA, and aldosterone levels were similar in patients and controls. Serum IL-6, but not ACTH levels, WAS strongly correlated with nocturnal cortisol salivary levels (rho=0.555, p<0.001) in patients.ConclusionIncreased evening and nocturnal but not morning cortisol secretion occur in even clinically mild COVID-19. In the context of acute viral infection (Covid-19), IL-6 may partially replace ACTH as a stimulus of the glucocorticoid-secreting adrenal zona-fasciculata without influencing the secretion of DHEA and aldosterone.

A PROSPECTIVE STUDY ON VARIOUS SEROLOGICAL MARKERS AND PLATELET COUNT IN DENGUE INFECTION IN AND AROUND KANCHEEPURAM.

INTRODUCTION: Dengue is an acute viral infection with potential fatal complications, presenting with non-specic fever that mimics other febrile illness. Specic antibody detection has been the mainstay of diagnosis which is prone for false positive and negative reactions. The newer parameter NS1 appears to be highly specic and reliable for diagnosis. Therefore, we tried to evaluate the association of platelet counts against NS1 and IgM / IgG in dengue infections. OBJECTIVE: To study the association of thrombocytopenia with NS1 antigen and specic antibodies in dengue infection in kancheepuram population. MATERIALS AND METHODS: Serum samples from clinically suspected dengue cases were tested for NS1,IgM,IgG by immunochromatography – based test. Platelet counts were obtained from all positive cases. Test results of dengue-specic parameters and platelet counts were compared. RESULTS: Of the 612 samples tested, 150 were positive for one or more dengue serological markers. Of the 150, 27(18%) were positive for NS1only, 45(30%)were positive for IgM, 54(36%) were positive for IgG only. Of the 150, 39(26%) showed platelet count less than 1 lakh. CONCLUSION: Inclusion of NS1 in the diagnosis of dengue increases the detection and specicity rate. Thus correlation of platelet count, NS1,IgM and ,IgG gives detailed picture of the stage and prognosis of the disease

Acute HIV-1 and SARS-CoV-2 Infections Share Slan+ Monocyte Depletion—Evidence from an Hyperacute HIV-1 Case Report

Monocytes are key modulators in acute viral infections, determining both inflammation and development of specific B- and T-cell responses. Recently, these cells were shown to be associated to different SARS-CoV-2 infection outcome. However, their role in acute HIV-1 infection remains unclear. We had the opportunity to evaluate the mononuclear cell compartment in an early hyper-acute HIV-1 patient in comparison with an untreated chronic HIV-1 and a cohort of SARS-CoV-2 infected patients, by high dimensional flow cytometry using an unsupervised approach. A distinct polarization of the monocyte phenotype was observed in the two viral infections, with maintenance of pro-inflammatory M1-like profile in HIV-1, in contrast to the M2-like immunosuppressive shift in SARS-CoV-2. Noticeably, both acute infections had reduced CD14low/-CD16+ non-classical monocytes, with depletion of the population expressing Slan (6-sulfo LacNac), which is thought to contribute to immune surveillance through pro-inflammatory properties. This depletion indicates a potential role of these cells in acute viral infection, which has not previously been explored. The inflammatory state accompanied by the depletion of Slan+ monocytes may provide new insights on the critical events that determine the rate of viral set-point in acute HIV-1 infection and subsequent impact on transmission and reservoir establishment.

Acute sinusitis in daily clinical practice

Understanding the appropriate use of diagnostics and treatment in acute rhinosinusitis is of immense importance given the high prevalence of this disease in the general population. The ability to differentiate between the principal phenotypes of acute sinusitis, namely acute viral infection (cold), acute post-viral sinusitis and acute bacterial sinusitis, determines the future management and is fundamental to providing rational therapeutic recommendations – especially as regards antibiotic treatment, which is very often overused in acute sinusitis even though bacterial phenotypes only account for 0.5–2% of all cases of the disease. The latest therapeutic recommendations contained in the EPOS2020 position paper introduce a system based on integrated care pathways (ICPs), which comprise pharmacy-supported self-care and e-health as the first level, followed by primary care as the second, with specialist care being reserved for patients who develop a more severe course of the disease, have suspected complications or suffer from recurrent acute sinusitis. Management of acute sinusitis is primarily based on symptomatic treatment modalities, with phytotherapeutic support, as well as on antiinflammatory treatment, while antibiotic therapy is used in very specific and limited indications. Complications are relatively rare in acute sinusitis and they are not considered to be associated with antibiotic intake. Considering the high prevalence of acute forms of sinusitis, their significant impact on quality of life and high direct and indirect costs of treatment, the right diagnosis and management, without unnecessary escalation of therapy, can substantially translate into a number of public health benefits.

Ddb1 Is Essential for the Expansion of CD4+ Helper T Cells by Regulating Cell Cycle Progression and Cell Death

Follicular helper T (TFH) cells are specialized CD4+ helper T cells that provide help to B cells in humoral immunity. However, the molecular mechanism underlying generation of TFH cells is incompletely understood. Here, we reported that Damage-specific DNA binding protein 1 (Ddb1) was required for expansion of CD4+ helper T cells including TFH and Th1 cells, germinal center response, and antibody response to acute viral infection. Ddb1 deficiency in activated CD4+ T cells resulted in cell cycle arrest at G2-M phase and increased cell death, due to accumulation of DNA damage and hyperactivation of ATM/ATR-Chk1 signaling. Moreover, mice with deletion of both Cul4a and Cul4b in activated CD4+ T cells phenocopied Ddb1-deficient mice, suggesting that E3 ligase-dependent function of Ddb1 was crucial for genome maintenance and helper T-cell generation. Therefore, our results indicate that Ddb1 is an essential positive regulator in the expansion of CD4+ helper T cells.

Pharmacological treatment of the patients with croup

Background: Croup is a respiratory illness usually caused by acute viral infection of the larynx, trachea, and bronchi, and characterized by the abrupt onset of a barking cough, inspiratory stridor, hoarseness, and respiratory distress due to upper airway obstruction. Croup commonly affects children younger than 6 years of age, with peak incidence between 7 and 36 months. Although the disease is usually self-limited, it may occasionally become life threatening, and can, on rare occasion, lead to respiratory failure.Current Concepts: Treatment of viral croup depends on the severity of symptoms as denoted by Westley croup score (i.e., mild, moderate, or severe). A single dose of oral or intramuscular dexamethasone (0.15-0.6 mg/kg) is the mainstay of treatment for viral croup, irrespective of severity. A single dose of nebulized budesonide (2 mg) is equally effective as systemically administered dexamethasone, and is considered when a patient is unable to take a medicine orally. Nebulized L-epinephrine (1:1,000, 3-5 mL) causes vasoconstriction in the mucosa, rapidly reducing airway edema. Addition of nebulized L-epinephrine is indicated in the patients with croup of at least moderate severity, displaying chest retraction and signs of labored breathing.Discussion and Conclusion: The most effective pharmacological treatments for patients with viral croup are oral or intramuscular dexamethasone, and nebulized L-epinephrine. Especially, corticosteroids can significantly decrease the intensity of croup symptoms and reduce hospital admissions, return visits to emergency department and length of stay in the hospital.