The Use of Baculovirus-Mediated Gene Expression in Mammalian Cells for Recombinant Protein Production.

Baculovirus-mediated gene expression in mammalian cells, BacMam, is a useful alternative to transient transfection for recombinant protein production in various types of mammalian cell lines. We decided to establish BacMam in our lab in order to streamline our workflows for gene expression in insect and mammalian cells, as it is straightforward to parallelize the baculovirus generation for both types of eukaryotic cells. This chapter provides a step-by-step description of the protocols we use for the generation of the recombinant BacMam viruses, the transduction of mammalian cell cultures, and optimization of the protein production conditions through small-scale expression and purification tests.

Estimating the cumulative incidence of SARS-CoV-2 infection in Costa Rica: modelling seroprevalence data in a population-based cohort.

Background: The true incidence of SARS-CoV-2 infection in Costa Rica was likely much higher than officially reported, because infection is often associated with mild symptoms and testing was limited by official guidelines and socio-economic factors.

Methods: Using serology to define natural infection, we developed a statistical model to estimate the true cumulative incidence of SARS-CoV-2 in Costa Rica early in the pandemic. We estimated seroprevalence from 2223 blood samples collected from November 2020 to October 2021 from 1976 population-based controls from the RESPIRA study.

Peptidylarginine deiminase 2 regulates expression of DGCR8 affecting miRNA biogenesis in gonadotrope cells.

In Brief: DGCR8 microprocessor complex, which is important for miRNA biogenesis, is regulated by peptidylarginine deiminase 2 and expression fluctuates in gonadotrope cells across the mouse estrous cycle.

Abstract: Canonical miRNA biogenesis requires DGCR8 microprocessor complex subunit, which helps cleave pri-miRNAs into pre-miRNAs. Previous studies found that inhibiting peptidylarginine deiminase (PAD) enzyme activity results in increased DGCR8 expression.

Conservation of vaccine antigen sequences encoded by sequenced strains of Streptococcus equi subsp. equi.

Background: Streptococcus equi subspecies equi (S equi) is the cause of Strangles, one of the most prevalent diseases of horses worldwide. Variation within the immunodominant SeM protein has been documented, but a new eight-component fusion protein vaccine, Strangvac, does not contain live S equi or SeM and conservation of the antigens it contains have not been reported.

Objective: To define the diversity of the eight Strangvac antigens across a diverse S equi population.

Protein abundance and folding rather than the redox state of Kelch13 determine the artemisinin susceptibility of Plasmodium falciparum.

Decreased susceptibilities of the human malaria parasite Plasmodium falciparum towards the endoperoxide antimalarial artemisinin are linked to mutations of residue C580 of PfKelch13, a homologue of the redox sensor Keap1 and other vertebrate BTB-Kelch proteins. Here, we addressed whether mutations alter the artemisinin susceptibility by modifying the redox properties of PfKelch13 or by compromising its native fold or abundance. Using selection-linked integration and the glmS ribozyme, efficient down-regulation of PfKelch13 resulted in ring-stage survival rates around 40%.

The impact of demonstration plots on improved agricultural input purchase in Tanzania: Implications for policy and practice.

In this paper, the authors use survey data from over 800 households to examine the impact of demonstration plots and associated activities (distribution of small packs of agricultural inputs) on smallholder farmers' decisions to buy agricultural inputs in Tanzania. Using propensity score matching and inverse probability-weighted adjustment models, the authors estimated the effect of access to demonstration plots alone and demonstration plots combined with small packs of agricultural inputs on a household's decision to purchase improved inputs. The results indicate that access to demonstration plots and demonstration plots with small packs increased the probability of purchasing improved inputs by 13-17 percentage points.

Intramuscular vaccination with Strangvac is safe and induces protection against equine strangles caused by Streptococcus equi.

The equine disease strangles, caused by Streptococcus equi, remains a major cause of welfare and economic cost to the global horse industry. Here we report the safety, immunogenicity and efficacy of a novel multi-component chimeric fusion protein vaccine, called Strangvac, when administered to ponies via the intramuscular route. Across the four studies, Strangvac was safe and induced robust antibody responses towards the vaccine components in blood serum and the nasopharynx, which were boosted by revaccination up to 12 months after a primary course of 2 vaccinations 4 weeks apart.

On-line electrolytic dissolution for soluble aluminium determination in high-silicon steel/electrical sheet by ED-FIA-ICP-OES.

Soluble aluminium (Al(sl)) in high silicon alloys is determined by electrolytic dissolution flow injection analysis (ED-FIA) and inductive coupled plasma with optical emission spectrometry (ICP-OES). Skipping the time consuming sample preparation known from acid hydrolysis reduces the required time per determination from 15 to 4 min. A new calculation procedure is used to compensate the discrepancy in matters of dissolution between acid hydrolysis and electrolytic dissolution.

Strangvac: A recombinant fusion protein vaccine that protects against strangles, caused by Streptococcus equi.

The host-restricted pathogen Streptococcus equi causes strangles in the horse, which is characterised by abscessation of the lymph nodes of the head and neck. The disease is endemic throughout the world causing considerable welfare and economic cost to the horse industry. Here we report the results of three studies where ponies were vaccinated with combinations of recombinant fusion proteins to optimise vaccine production and the level of protection conferred.

Correction: A single-source precursor route to anisotropic halogen-doped zinc oxide particles as a promising candidate for new transparent conducting oxide materials.

[This corrects the article DOI: 10.3762/bjnano.6.

A single-source precursor route to anisotropic halogen-doped zinc oxide particles as a promising candidate for new transparent conducting oxide materials.

Numerous applications in optoelectronics require electrically conducting materials with high optical transparency over the entire visible light range. A solid solution of indium oxide and substantial amounts of tin oxide for electronic doping (ITO) is currently the most prominent example for the class of so-called TCOs (transparent conducting oxides). Due to the limited, natural occurrence of indium and its steadily increasing price, it is highly desired to identify materials alternatives containing highly abundant chemical elements.

Two-colour high-speed asynchronous optical sampling based on offset-stabilized Yb:KYW and Ti:sapphire oscillators.

We present a high-speed asynchronous optical sampling system, based on two different Kerr-lens mode-locked lasers with a GHz repetition rate: An Yb:KYW oscillator and a Ti:sapphire oscillator are synchronized in a master-slave configuration at a repetition rate offset of a few kHz. This system enables two-colour pump-probe measurements with resulting noise floors below 10⁻⁶ at a data aquisition time of 5 seconds. The measured temporal resolution within the 1 ns time window is below 350 fs, including a timing jitter of less than 50 fs.

The role of 2,6-diaminopyridine ligands in the isolation of an unprecedented, low-valent tin complex.

Stabilization of the central atom in an oxidation state of zero through coordination of neutral ligands is a common bonding motif in transition-metal chemistry. However, the stabilization of main-group elements in an oxidation state of zero by neutral ligands is rare. Herein, we report that the transamination reaction of the DAMPY ligand system (DAMPY=2,6-[ArNH-CH2 ]2 (NC5 H3 ) (Ar=C6 H3 -2,6-iPr2 )) with Sn[N(SiMe3 )2 ]2 produces the DIMPYSn complex (DIMPY=(2,6-[ArNCH]2 (NC5 H3 )) with the Sn atom in a formal oxidation state of zero.

Staphylococcus aureus subsp. anaerobius strain ST1464 genome sequence.

Staphylococcus aureus subsp. anaerobius is responsible for Morel's disease in animals and a cause of abscess in humans. It is characterized by a microaerophilic growth, contrary to the other strains of S.

Homogenates of skeletal muscle injected with snake venom inhibit myogenic differentiation in cell culture.

Introduction: Viperid snakebite envenomings are characterized by muscle necrosis and a deficient regenerative response.

Methods: Homogenates from gastrocnemius muscles of mice injected with the venom of the snake Bothrops asper or with 2 tissue-damaging toxins were added to cultures of C2C12 myogenic cells. Myoblasts proliferation and fusion were assessed.

Protective immunization against Staphylococcus aureus infection in a novel experimental wound model in mice.

A novel murine experimental wound infection model was used to assess the efficacy of multi-component immunization against Staphylococcus aureus infection. Necrotic lesions were induced in mice with venom from Bothrops asper and infected with a low inoculum, 1 × 10(2) CFU. The wound infection model therefore more resembles a clinical case of S.

Antiphagocytic function of an IgG glycosyl hydrolase from Streptococcus equi subsp. equi and its use as a vaccine component.

EndoSe from Streptococcus equi subsp. equi is an enzyme hydrolyzing glycosyl groups on IgG, analogous to EndoS from Streptococcus pyogenes. We here show that the activity of EndoSe leads to an antiphagocytic function and may thus be a contributory factor to immune evasion of S.

Determination of diffusible and total hydrogen concentration in coated and uncoated steel using melt and solid extraction techniques: Part I.

It is essentially to know the bulk hydrogen concentration in various types of steel because it indicates the amount of hydrogen that can be trapped by the different alloys of steel. This information leads to more knowledge about the interactions of steel alloys with hydrogen containing environment and stability of the steel material during usage. To get this information precise analytical methods are necessary.

Levels of antibody against 11 Staphylococcus aureus antigens in a healthy population.

Serum samples from 151 healthy individuals aged from 15 to 89 years were investigated by enzyme-linked immunosorbent assay (ELISA) for IgG levels against 11 different purified antigens from Staphylococcus aureus. Surface antigens, such as teichoic acid, clumping factors A and B, and bone sialoprotein binding protein, and extracellular proteins, such as alpha-toxin, lipase, enterotoxin A, toxic shock syndrome toxin, scalded-skin syndrome toxin, fibrinogen binding protein, and extracellular adherence protein, were used. The IgG values were analyzed in relation to the state of nasal carriage at the time of sampling.

Ovine clone ST1464: a predominant genotype of Staphylococcus aureus subsp. anaerobius isolated from sheep in Sudan.

Background: The aim of the present study was to examine the phenotypic and genotypic relatedness of 17 Staphylococcus aureus subsp. anaerobius isolates recovered from sheep abscesses in Khartoum state, Sudan, during the period 2007-2008.

Methodology: This sample was characterised using antibiogram typing, biochemical typing with the commercial PhenePlate system (PhP-CS) and multilocus sequence typing (MLST).

Extracellular adherence protein (Eap) from Staphylococcus aureus does not function as a superantigen.

Extracellular adherence protein (Eap) from Staphylococcus aureus has been reported to have strong anti-inflammatory properties, which make Eap a potential anti-inflammatory agent. However, Eap has also been demonstrated to trigger T-cell activation and to share structural homology with superantigens. In this study, we focused on whether Eap fulfilled the definition criteria for a superantigen.

Getting to grips with strangles: an effective multi-component recombinant vaccine for the protection of horses from Streptococcus equi infection.

Streptococcus equi subspecies equi (S. equi) is a clonal, equine host-adapted pathogen of global importance that causes a suppurative lymphodendopathy of the head and neck, more commonly known as Strangles. The disease is highly prevalent, can be severe and is highly contagious.

Two novel IgG endopeptidases of Streptococcus equi.

Streptococcus equi ssp. equi causes strangles, a highly contagious and serious disease in the upper respiratory tract of horses. Streptococcus equi ssp.

Staphylococcus epidermidis isolated from newborn infants express pilus-like structures and are inhibited by the cathelicidin-derived antimicrobial peptide LL37.

Coagulase-negative staphylococci and its subtype Staphylococcus epidermidis are major indigenous Gram-positive inhabitants of the human skin. Colonization occurs in direct connection with birth and terrestrial adaptation. This study focuses on factors that may influence skin colonization of the newborn infant that relates to the immune status of both the bacteria and the host.

More than one tandem repeat domain of the extracellular adherence protein of Staphylococcus aureus is required for aggregation, adherence, and host cell invasion but not for leukocyte activation.

The extracellular adherence protein (Eap) is a multifunctional Staphylococcus aureus protein and broad-spectrum adhesin for several host matrix and plasma proteins. We investigated the interactions of full-length Eap and five recombinant tandem repeat domains with host proteins by use of surface plasmon resonance (BIAcore) and ligand overlay assays. In addition, agglutination and host cell interaction, namely, adherence, invasion, and stimulation of proliferation, were determined.