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Sulfated glycans are barely detectable in routine mass spectrometry (MS)-based glycomic analysis due to ion suppression by the significantly more abundant neutral glycans in the positive ion mode, and sialylated non-sulfated glycans in the negative ion mode, respectively. Nevertheless, the negative charge imparted by sulfate can be advantageous for selective detection in the negative ion mode if the sialic acids can first be neutralized. This is most conveniently achieved by a concerted sample preparation workflow in which permethylation is followed by solid phase fractionation to isolate the sulfated glycans prior to MS analysis. Importantly, we demonstrated that conventional NaOH/DMSO slurry permethylation method can retain the sulfates. Instead of extracting permethylated glycans into chloroform for sample clean-up, reverse phase C18 cartridge coupled with self-packed amine-tip or mixed mode weak anion exchange cartridge can be utilized to obtain in good yield the non-sulfated, mono-sulfated, and multiply sulfated permethylated glycans in separate fractions for sulfoglycomic analysis.Exploring the structure and function of protein complexes requires their isolation in the native state-a task that is made challenging when studying labile and/or low abundant complexes. The difficulties in preparing membrane-protein complexes are especially notorious. The cyanobacterium Synechocystis sp. PCC 6803 is a widely used model organism for the physiology of oxygenic phototrophs, and the biogenesis of membrane-bound photosynthetic complexes has traditionally been studied using this cyanobacterium. In a typical approach, the protein complexes are purified with a combination of His-affinity chromatography and a size-based fractionation method such as gradient ultracentrifugation and/or native electrophoresis. However, His-affinity purification harbors prominent contaminants and the levels of many proteins are too low for a feasible multi-step purification. Here, we have developed a purification method for the isolation of 3x FLAG-tagged proteins from the membrane and soluble fractions of Synechocystis. Soluble proteins or solubilized thylakoids are subjected to a single affinity purification step that utilizes the highly specific binding of FLAG-affinity resin. After an intensive wash, the captured proteins are released from the resin under native conditions using an excess of synthetic 3x FLAG peptide. The protocol allows fast isolation of low abundant protein complexes with a superb purity.Tick-host bloodmeal associations are important factors when characterizing risks of associated pathogen transmission and applying appropriate management strategies. Despite their biological importance, comparatively little is known about soft tick (Argasidae) host associations in the United States compared to hard ticks (Ixodidae). In this study, we evaluated a PCR and direct Sanger sequencing method for identifying the bloodmeal hosts of soft ticks. We collected 381 cave-associated Ornithodoros turicata near San Antonio, Texas, USA, and also utilized eight colony-reared specimens fed artificially on known host blood sources over 1.5 years ago. We correctly identified the vertebrate host bloodmeals of two colony-reared ticks (chicken and pig) up to 1,105 days post-feeding, and identified bloodmeal hosts from 19 out of 168 field-collected soft ticks, including raccoon (78.9%), black vulture (10.5%), Texas black rattlesnake (5.3%), and human (5.3%). Our results confirm the retention of vertebrate blood DNA in soft ticks and advance the knowledge of argasid host associations in cave-dwelling O. turicata.Mycobacterium avium subspecies paratuberculosis (MAP) is the aetiological agent of Johne's disease (JD), a chronic enteritis that causes major losses to the global livestock industry. Further, it has been associated with human Crohn's disease. Several strains of MAP have been identified, the two major groups being sheep strain MAP, which includes the Type I and Type III sub-lineages, and the cattle strain or Type II MAP lineage, of which bison strains are a sub-grouping. Major genotypic, phenotypic and pathogenic variations have been identified in prior comparisons, but the research has predominately focused on cattle strains of MAP. In countries where the sheep industries are more prevalent, however, such as Australia and New Zealand, ovine JD is a substantial burden. An information gap exists regarding the genomic differences between sheep strain sub-lineages and the relevance of Type I and Type III MAP in terms of epidemiology and/or pathogenicity. We therefore investigated sheep MAP isolates from Australilogical and virulence traits specific to sheep MAP. This knowledge will potentially contribute to improved vaccine development and control measures for these strains.The production of surplus male offspring illustrates a socioethical concern in the dairy industry. In this article, we highlight the animal health and welfare implications of production outputs for surplus dairy calves, namely veal production, dairy calf to beef production, and euthanasia. Moreover, we present a pilot study focus on exploring the perception of key industry actors within the dairy industry in Ireland regarding the use of sexed semen as a mitigation strategy to reduce the production of surplus male dairy calves. A pilot survey was completed by farmers (n = 6), veterinarians (n = 17), and dairy farm advisors (n = 11). https://www.selleckchem.com/products/FK-506-(Tacrolimus).html All the veterinarians, 80% of the farmers, and 62% of the advisors believed that the use of sexed semen had a positive influence on herd welfare. All participants identified the same barriers to the implementation of sexed semen lower conception rate, lower availability, and higher cost. The reviewed literature highlights the importance of tailored communication to support knowledge exchange between stakeholders and key industry actors such as dairy farmers, their veterinarians, and advisors. Research to understand stakeholders' perception is pivotal to address socioethical concerns such as the surplus male dairy calves.[This corrects the article DOI 10.3389/fcvm.2020.00116.].
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