Newly emerged enterovirus-A71 C4 sublineage may be more virulent than B5 in the 2015-2016 hand-foot-and-mouth disease outbreak in northern Vietnam.

Enterovirus-A71 (EV-A71) is a common cause of hand-foot-and-mouth disease (HFMD) and, rarely, causes severe neurological disease. This study aimed to elucidate the epidemiological and genetic characteristics and virulence of EV-A71 strains isolated from children diagnosed with HFMD. Rectal and throat swabs were collected from 488 children with HFMD in Hanoi, Vietnam, in 2015-2016. From 391 EV-positive patients, 15 EVs, including coxsackievirus A6 (CV-A6; 47.1%) and EV-A71 (32.5%, n = 127), were identified. Of the 127 EV-A71 strains, 117 (92.1%) were the B5 subgenotype and 10 (7.9%) were the C4 subgenotype. A whole-genome analysis of EV-A71 strains showed that seven of the eight C4a strains isolated in 2016 formed a new lineage, including two possible recombinants between EV-A71 C4 and CV-A8. The proportion of inpatients among C4-infected children was higher than among B5-infected children (80.0% vs. 27.4%; P = 0.002). The virulence of EV-A71 strains was examined in human scavenger receptor class B2 (hSCARB2)-transgenic mice, and EV-A71 C4 strains exhibited higher mortality than B5 strains (80.0% vs. 30.0%, P = 0.0001). Thus, a new EV-A71 C4a-lineage, including two possible recombinants between EV-A71 C4 and CV-A8, appeared in 2016 in Vietnam. The EV-A71 C4 subgenotype may be more virulent than the B5 subgenotype.

Role of GAP-43 in sequestering phosphatidylinositol 4,5-bisphosphate to Raft bilayers.

The lipid phosphatidylinositol 4,5-bisphosphate (PIP(2)) is critical for a number of physiological functions, and its presence in membrane microdomains (rafts) appears to be important for several of these spatially localized events. However, lipids like PIP(2) that contain polyunsaturated hydrocarbon chains are usually excluded from rafts, which are enriched in phospholipids (such as sphingomyelin) containing saturated or monounsaturated chains. Here we tested a mechanism by which multivalent PIP(2) molecules could be transferred into rafts through electrostatic interactions with polybasic cytoplasmic proteins, such as GAP-43, which bind to rafts via their acylated N-termini. We analyzed the interactions between lipid membranes containing raft microdomains and a peptide (GAP-43P) containing the linked N-terminus and the basic effector domain of GAP-43. In the absence or presence of nonacylated GAP-43P, PIP(2) was found primarily in detergent-soluble membranes thought to correspond to nonraft microdomains. However, when GAP-43P was acylated by palmitoyl coenzyme A, both the peptide and PIP(2) were greatly enriched in detergent-resistant membranes that correspond to rafts; acylation of GAP-43P changed the free energy of transfer of PIP(2) from detergent-soluble membranes to detergent-resistant membranes by -1.3 kcal/mol. Confocal microscopy of intact giant unilamellar vesicles verified that in the absence of GAP-43P PIP(2) was in nonraft microdomains, whereas acylated GAP-43P laterally sequestered PIP(2) into rafts. These data indicate that sequestration of PIP(2) to raft microdomains could involve interactions with acylated basic proteins such as GAP-43.

Lab-scale evaluation of aerated burial concept for treatment and emergency disposal of infectious animal carcasses.

Nearly 55,000 outbreaks of animal disease were reported to the World Animal Health Information Database between 2005 and 2016. To suppress the spread of disease, large numbers of animal mortalities often must be disposed of quickly and are frequently buried on the farm where they were raised. While this method of emergency disposal is fast and relatively inexpensive, it also can have undesirable and lasting impacts (slow decay, concerns about groundwater contamination, pathogens re-emergence, and odor). Following the 2010 foot-and-mouth disease outbreak, the Republic of Korea's National Institute of Animal Science funded research on selected burial alternatives or modifications believed to have potential to reduce undesirable impacts of burial. One such modification involves the injection of air into the liquid degradation products from the 60-70% water from decomposing carcasses in lined burial trenches. Prior to prototype development in the field, a laboratory-scale study of aerated decomposition (AeD) of poultry carcasses was conducted to quantify improvements in time of carcass decomposition, reduction of potential groundwater pollutants in the liquid products of decomposition (since trench liners may ultimately leak), and reduction of odorous VOCs emitted during decomposition. Headspace gases also were monitored to determine the potential for using gaseous biomarkers in the aerated burial trench exhaust stream to monitor completion of the decomposition. Results of the lab-scale experiments show that the mass of chicken carcasses was reduced by 95.0 ±â€¯0.9% within 3 months at mesophilic temperatures (vs. negligible reduction via mesophilic anaerobic digestion typical of trench burial) with concomitant reduction of biochemical oxygen demand (BOD; 99%), volatile suspended solids (VSS; 99%), total suspended solids (TSS; 99%), and total ammonia nitrogen (TAN; 98%) in the liquid digestate. At week #7 BOD and TSS in digestate met the U.S. EPA standards for treated wastewater discharge to surface water. Salmonella and Staphylococcus were inactivated by the AeD process after week #1 and #3, respectively. Five gaseous biomarkers: pyrimidine; p-cresol; phenol; dimethyl disulfide; and dimethyl trisulfide; were identified and correlated with digestate quality. Phenol was the best predictor of TAN (R = 0.96), BOD (R = 0.92), and dissolved oxygen (DO) (R = -0.91). Phenol was also the best predictor populations of Salmonella (R = 0.95) and aerobes (R = 0.88). P-cresol was the best predictor for anaerobes (R = 0.88). The off-gas from AeD will require biofiltration or other odor control measures for a much shorter time than anaerobic decomposition. The lab-scale studies indicate that AeD burial has the potential to make burial a faster, safer, and more environmentally friendly method for emergency disposal and treatment of infectious animal carcasses and that this method should be further developed via prototype-scale field studies.

Oropharyngeal Necrobacillosis with Septic Thrombophlebitis and Pulmonary Embolic Abscesses: Lemierre's Syndrome in a New Zealand White Rabbit.

Lemierre's syndrome, oropharyngeal infections induced by anaerobic bacteria, leading to fatal septic thrombophlebitis of the internal jugular vein and pulmonary embolic abscesses in humans, was diagnosed in a 6-month-old, male, New Zealand White rabbit. After acute onset of anorexia, lethargy, and depression, the rabbit died suddenly despite emergency clinical care. Necropsy revealed swelling, necrosis, and abscess in the soft tissues around the left caudal mandibular ramus, oral mucosa, and molar teeth, with systemic embolic abscesses and necrosis, especially in the jugular vein, lungs, and brain. Histologic examination revealed necrosis and embolic abscesses with filamentous bacteria in the mandibular soft tissues, salivary gland, jugular veins, alveolar bone and marrow, periodontal tissues and dental pulp, oral and pharyngeal mucosa, lungs, brain, liver, myocardium, meninges, and small intestine. Bacterial culture of the mandibular abscess and heart blood yielded Fusobacterium necrophorum.

Polymorphism of highly cross-linked F-actin networks: probing multiple length scales.

The assembly properties of F-actin filaments in the presence of different biological cross-linker concentrations and types have been investigated using a combined approach of fluorescence confocal microscopy and coarse-grained molecular dynamics simulation. In particular for highly cross-linked regimes, new network morphologies are observed. Complex network formation and the details of the resulting structure are strongly dependent on the ratio of cross-linkers to actin monomers and cross-linker shape but only weakly dependent on overall actin concentration and filament length. The work presented here may help to provide some fundamental understanding of how excessive cross-linkers interact with the actin filament solution, creating different structures in the cell under high cross-linker concentrations. F-actin is not only of biological importance but also, as an example of a semiflexible polymer, has attracted significant interest in its physical behavior. In combination with different cross-linkers semiflexible filaments may provide new routes to bio-materials development and act as the inspiration for new hierarchical network-based materials.