Crystal structure of the 3C protease from South African Territories type 2 foot-and-mouth disease virus

The replication of foot-and-mouth disease virus (FMDV) is dependent on the virus-encoded 3C protease (3Cpro). As in other picornaviruses, 3Cpro performs most of the proteolytic processing of the polyprotein expressed from the single open reading frame in the RNA genome of the virus. Previous work revealed that the 3Cpro from serotype A – one of the seven serotypes of FMDV – adopts a trypsin-like fold. Phylogenetically the FMDV serotypes are grouped into two clusters, with O, A, C, and Asia 1 in one, and the three South African Territories serotypes, (SAT-1, SAT-2 and SAT-3) in another. We report here the cloning, expression and purification of 3C proteases from four SAT serotype viruses (SAT2/GHA/8/91, SAT1/NIG/5/81, SAT1/UGA/1/97, and SAT2/ZIM/7/83) and the crystal structure at 3.2 Å resolution of 3Cpro from SAT2/GHA/8/91)

Differential leukocyte count method for bovine low somatic cell count milk

Whereas many differential leukocyte count methods for high somatic cell count (SCC) milk from mastitic cows are available, only a few have been developed for low SCC milk. We have developed a flow cytometric differential leukocyte count method for low SCC milk. The procedure consists of 1) 1.5 ml of diluted milk sample (30%, vol/vol dilution with PBS), 2) centrifugation, 3) leukocyte labeling with SYTO 13 and 4) flow cytometric analysis. Four major leukocyte populations can be clearly identified in the green fluorescence-side scatter dot plot: lymphocytes and monocytes (LM), polymorphonuclear neutrophils (PMN), mature macrophages (MO), and cells with apoptotic features based on chromatin condensation and nuclear fragmentation. The optimal processing temperature was 20degreesC. Significant differences among samples with similar differential leukocyte counts were found. Storage of milk samples during 2 d at 7degreesC had no effect on differential leukocyte count. Using the new method, differential leukocyte count was performed in low SCC milk samples from cows in early, mid, and late lactation. In accordance with previous studies, PMN and M P percentages were lower and LM percentages were higher in early lactation than in the other stages of lactation. The percentage of cells with apoptotic features was higher in early lactation than in mid and late lactation. In conclusion, a rapid, simple, accurate, and reproducible standard procedure was developed to determine the differential leukocyte count (MO, PMN, LM, and cells with apoptotic features) of bovine low SCC milk

Bubble size prediction in co-flowing streams

In this paper, the size of bubbles formed through the breakup of a gaseous jet in a co-axial microfluidic device is derived. The gaseous jet surrounded by a co-flowing liquid stream breaks up into monodisperse microbubbles and the size of the bubbles is determined by the radius of the inner gas jet and the bubble formation frequency. We obtain the radius of the gas jet by solving the Navier-Stokes equations for low Reynolds number flows and by minimization of the dissipation energy. The prediction of the bubble size is based on the system's control parameters only, i.e. the inner gas flow rate $Q_i$, the outer liquid flow rate $Q_o$, and the tube radius $R$. For a very low gas-to-liquid flow rate ratio ($Q_i / Q_o \rightarrow 0$) the bubble radius scales as $r_b / R \propto \sqrt{Q_i / Q_o}$, independently of the inner to outer viscosity ratio $\eta_i/\eta_o$ and of the type of the velocity profile in the gas, which can be either flat or parabolic, depending on whether high-molecular-weight surfactants cover the gas-liquid interface or not. However, in the case in which the gas velocity profiles are parabolic and the viscosity ratio is sufficiently low, i.e. $\eta_i/\eta_o \ll 1$, the bubble diameter scales as $r_b \propto (Q_i/Q_o)^\beta$, with $\beta$ smaller than 1/2

Cyclic thermo-mechanical material modelling and testing of 316 stainless steel

A programme of cyclic mechanical testing of a 316 stainless steel, at temperatures of up to 600 °C under isothermal conditions, for the identification of material constitutive constants, has been carried out using a thermo-mechanical fatigue test machine (with induction coil heating). The constitutive model adopted is a modified Chaboche unified viscoplasticity model, which can deal with both cyclic effects, such as combined isotropic and kinematic hardening, and rate-dependent effects, associated with viscoplasticity. The characterisation of 316 stainless steel is presented and compared with results from tests consisting of cyclic isothermal, as well as in-phase and out-of-phase thermo-mechanical fatigue conditions, using interpolation between the isothermal material constants to predict the material behaviour under anisothermal conditions

Haemangiosarcoma in a captive Asiatic lion (Panthera leo persica)

A 2.7-year-old male captive Asiatic lion (Panthera leo persica) died unexpectedly without preceding symptoms. Gross necropsy revealed liver and lung tumours, which proved to be haemangiosarcomas by histopathology. Some of the liver tumours were ruptured, leading to massive intra-abdominal haemorrhage and death. Haemangiosarcomas are rare in domestic and exotic felids, occurring in skin, thoracic-abdominal cavity and bones. Although these tumours mainly appear to be occurring in older cats, they are sometimes observed in younger animals, as in the present case. This is the first description of haemangiosarcoma in a young Asiatic lion

LPS resistance of SPRET/Ei mice is mediated by Gilz, encoded by the Tsc22d3 gene on the X chromosome

Natural variation for LPS-induced lethal inflammation in mice is useful for identifying new genes that regulate sepsis, which could form the basis for novel therapies for systemic inflammation in humans. Here we report that LPS resistance of the inbred mouse strain SPRET/Ei, previously reported to depend on the glucocorticoid receptor (GR), maps to the distal region of the X-chromosome. The GR-inducible gene Tsc22d3, encoding the protein Gilz and located in the critical region on the X-chromosome, showed a higher expressed SPRET/Ei allele, regulated in cis. Higher Gilz levels were causally related to reduced inflammation, as shown with knockdown and overexpression studies in macrophages. Transient overexpression of Gilz by hydrodynamic plasmid injection confirmed that Gilz protects mice against endotoxemia Our data strongly suggest that Gilz is responsible for the LPS resistance of SPRET/Ei mice and that it could become a treatment option for sepsis

Wnt/Β-Catenin and Sex Hormone Signaling In Endometrial Homeostasis and Cancer

A delicate balance between estrogen and progestagen signaling underlies proper functioning of the female reproductive tract and, in particular, the monthly re- and degenerative phases characteristic of the menstrual cycle. Here, we propose that the canonical Wnt/β-catenin signaling pathway may underlie this finely tuned hormonal equilibrium in endometrial homeostasis and, upon its constitutive activation, lead to neoplastic transformation of the endometrium. During the menstrual cycle, estradiol will enhance Wnt/β-catenin signaling in the proliferative phase, while progesterone inhibits Wnt/β-catenin signaling, thus restraining estrogens' proliferative actions, during the secretory phase. In case of enhanced or unopposed estrogen signaling, constitutive activation of Wnt/β-catenin signaling will trigger endometrial hyperplasia, which may develop further into endometrial cancer