Illumination of Parainfluenza Virus Infection and Transmission in Living Animals Reveals a Tissue-Specific Dichotomy

The parainfluenza viruses (PIVs) are highly contagious respiratory paramyxoviruses and a leading cause of lower respiratory tract (LRT) disease. Since no vaccines or antivirals exist, non-pharmaceutical interventions are the only means of control for these pathogens. Here we used bioluminescence imaging to visualize the spatial and temporal progression of murine PIV1 (Sendai virus) infection in living mice after intranasal inoculation or exposure by contact. A non-attenuated luciferase reporter virus (rSeV-luc(M-F*)) that expressed high levels of luciferase yet was phenotypically similar to wild-type Sendai virus in vitro and in vivo was generated to allow visualization. After direct intranasal inoculation, we unexpectedly observed that the upper respiratory tract (URT) and trachea supported robust infection under conditions that result in little infection or pathology in the lungs including a low inoculum of virus, an attenuated virus, and strains of mice genetically resistant to lung infection. The high permissivity of the URT and trachea to infection resulted in 100% transmission to naïve contact recipients, even after low-dose (70 PFU) inoculation of genetically resistant BALB/c donor mice. The timing of transmission was consistent with the timing of high viral titers in the URT and trachea of donor animals but was independent of the levels of infection in the lungs of donors. The data therefore reveals a disconnect between transmissibility, which is associated with infection in the URT, and pathogenesis, which arises from infection in the lungs and the immune response. Natural infection after transmission was universally robust in the URT and trachea yet limited in the lungs, inducing protective immunity without weight loss even in genetically susceptible 129/SvJ mice. Overall, these results reveal a dichotomy between PIV infection in the URT and trachea versus the lungs and define a new model for studies of pathogenesis, development of live virus vaccines, and testing of antiviral therapies

Influence of polymer composition on the sensitivity towards nitrite and nitric oxide of colorimetric disposable test strips

The influence of polymer composition on the sensitivity towards nitrite (NO 2 − ) and nitric oxide (NO) of a series of 19 polymeric hydrogel films has been stud- ied. The polymers, based on the hydrophilic monomer 2- hydroxyethylmethacrylate (HEMA), are able to encapsu- late the colorimetric indicator 1,2-diaminoanthraquinone (DAQ) and to respond to NO 2 − and NO by visual chang- es. In the case of nitrite, the calculated limits of detection (LOD) for two of the polymeric sensors (10 μ M) are very close to the sensitivity estimated for free DAQ in solution (LOD 5 μ M), but with the advantage of a solid supported sensor with the format of a disposable test- strip made with affordable starting chemicals. The re- sults are interpreted taking into account the nature and proportions of monomers and cross-linkers used for the synthesis of polymers. Key factors for obtaining sensi- tive materials are the hydro philic character of the film along with the utilization of low levels of cross-linker and the use of an acidic monomer, like acrylic acid, as a building block.Financial support from the Spanish MINECO (CTQ2015-68429-R) and Fundació Caixa Castelló-UJI (P1·1B2015-76) is acknowledged. V. F. thanks the financial support from UJI (predoctoral fellowship). We thank SCIC/UJI for technical assistance

Pre-analytical issues in the haemostasis laboratory: guidance for the clinical laboratories

Ensuring quality has become a daily requirement in laboratories. In haemostasis, even more than in other disciplines of biology, quality is determined by a pre-analytical step that encompasses all procedures, starting with the formulation of the medical question, and includes patient preparation, sample collection, handling,transportation, processing, and storage until time of analysis. This step, based on a variety of manual activities, is the most vulnerable part of the total testing process and is a major component of the reliability and validity of results in haemostasis and constitutes the most important source of erroneous or un-interpretable results. Pre-analytical errors may occur throughout the testing process and arise from unsuitable, inappropriate or wrongly handled procedures. Problems may arise during the collection of blood specimens such as misidentification of the sample, use of inadequate devices or needles, incorrect order of draw, prolonged tourniquet placing, unsuccessful attempts to locate the vein, incorrect use of additive tubes, collection of unsuitable samples for quality or quantity, inappropriate mixing of a sample, etc. Some factors can alter the result of a sample constituent after collection during transportation, preparation and storage. Laboratory errors can often have serious adverse consequences. Lack of standardized procedures for sample collection accounts for most of the errors encountered within the total testing process. They can also have clinical consequences as well as a significant impact on patient care, especially those related to specialized tests as these are often considered as “diagnostic”. Controlling pre-analytical variables is critical since this has a direct influence on the quality of results and on their clinical reliability. The accurate standardization of the pre-analytical phase is of pivotal importance for achieving reliable results of coagulation tests and should reduce the side effects of the influence factors. This review is a summary of the most important recommendations regarding the importance of pre-analytical factors for coagulation testing and should be a tool to increase awareness about the importance of pre-analytical factors for coagulation testing