Profile of Membrane Cargo Trafficking Proteins and Transporters Expressed under N Source Derepressing Conditions in Aspergillus nidulans

Solute and ion transporters are proteins essential for cell nutrition, detoxification, signaling, homeostasis and drug resistance. Being polytopic transmembrane proteins, they are co-translationally inserted and folded into the endoplasmic reticulum (ER) of eukaryotic cells and subsequently sorted to their final membrane destination via vesicular secretion. During their trafficking and in response to physiological/stress signals or prolonged activity, transporters undergo multiple quality control processes and regulated turnover. Consequently, transporters interact dynamically and transiently with multiple proteins. To further dissect the trafficking and turnover mechanisms underlying transporter subcellular biology, we herein describe a novel mass spectrometry-based proteomic protocol adapted to conditions allowing for maximal identification of proteins related to N source uptake in A. nidulans. Our analysis led to identification of 5690 proteins, which to our knowledge constitutes the largest protein dataset identified by omics-based approaches in Aspergilli. Importantly, we detected possibly all major proteins involved in basic cellular functions, giving particular emphasis to factors essential for membrane cargo trafficking and turnover. Our protocol is easily reproducible and highly efficient for unearthing the full A. nidulans proteome. The protein list delivered herein will form the basis for downstream systematic approaches and identification of protein–protein interactions in living fungal cells.

Use of magnetic field in equine physiotherapy – preliminary study

This study aimed to determine the efficacy of magnetic field therapy in equine rehabilitation and the use of thermography as a diagnostic tool providing visualisation of the healing progress. The material for the study was a 23-year-old Mur-Insulan mare used for many years as a horse for riding lessons. Magnetic field therapy was applied every second day for 60 minutes. During the survey, blood samples were taken three times. Thermograms were taken before and after each magnetic therapy sessions. Magnetic field therapy had no negative effects on the mare’s condition, which was supported by haematological blood samples analysis. Thermograms showed increased temperature within the examined front limbs and trunk. During the sessions, the mare was calm and relaxed, which indicates that the application of a magnetic field had a calming and soothing effect on the horse's body. The conclusion was that magnetic field therapy may improve regeneration, provides proper cell nutrition, faster bone symphysis, analgesia and could decrease swelling.

Use of magnetic field in equine physiotherapy – preliminary study

This study aimed to determine the efficacy of magnetic field therapy in equine rehabilitation and the use of thermography as a diagnostic tool providing visualisation of the healing progress. The material for the study was a 23-year-old Mur-Insulan mare used for many years as a horse for riding lessons. Magnetic field therapy was applied every second day for 60 minutes. During the survey, blood samples were taken three times. Thermograms were taken before and after each magnetic therapy sessions. Magnetic field therapy had no negative effects on the mare’s condition, which was supported by haematological blood samples analysis. Thermograms showed increased temperature within the examined front limbs and trunk. During the sessions, the mare was calm and relaxed, which indicates that the application of a magnetic field had a calming and soothing effect on the horse's body. The conclusion was that magnetic field therapy may improve regeneration, provides proper cell nutrition, faster bone symphysis, analgesia and could decrease swelling.

Tales of tails in transporters

Cell nutrition, detoxification, signalling, homeostasis and response to drugs, processes related to cell growth, differentiation and survival are all mediated by plasma membrane (PM) proteins called transporters. Despite their distinct fine structures, mechanism of function, energetic requirements, kinetics and substrate specificities, all transporters are characterized by a main hydrophobic body embedded in the PM as a series of tightly packed, often intertwined, α-helices that traverse the lipid bilayer in a zigzag mode, connected with intracellular or extracellular loops and hydrophilic N- and C-termini. Whereas longstanding genetic, biochemical and biophysical evidence suggests that specific transmembrane segments, and also their connecting loops, are responsible for substrate recognition and transport dynamics, emerging evidence also reveals the functional importance of transporter N- and C-termini, in respect to transport catalysis, substrate specificity, subcellular expression, stability and signalling. This review highlights selected prototypic examples of transporters in which their termini play important roles in their functioning.

Characterization of the Opp Peptide Transporter ofCorynebacterium pseudotuberculosisand Its Role in Virulence and Pathogenicity

Despite the economic importance of caseous lymphadenitis (CLA), a chronic disease caused byCorynebacterium pseudotuberculosis, few genes related to the virulence of its etiologic agent have been characterized. The oligopeptide permease (Opp) transporters are located in the plasma membrane and have functions generally related to the uptake of peptides from the extracellular environment. These peptide transporters, in addition to having an important role in cell nutrition, also participate in the regulation of various processes involving intercellular signaling, including the control of the expression of virulence genes in pathogenic bacteria. To study the role of Opp inC. pseudotuberculosis, an OppD deficient strain was constructed via simple crossover with a nonreplicative plasmid carrying part of theoppDgene sequence. As occurred to the wild-type, the ΔoppDstrain showed impaired growth when exposed to the toxic glutathione peptide (GSH), indicating two possible scenarios: (i) that this component can be internalized by the bacterium through an Opp-independent pathway or (ii) that there is toxicity while the peptide is extracellular. Additionally, the ΔoppDmutant presented a reduced ability to adhere to and infect macrophages compared to the wild-type, although both strains exhibit the same potential to colonize spleens and cause injury and death to infected mice.

Teaching nutritional biochemistry: an experimental approach using yeast

In this report, we present a practical approach to teaching several topics in nutrition to science students at the high school and college freshmen levels. This approach uses baker's yeast ( Saccharomyces cerevisiae ) as a biological system model. The diameters of yeast colonies, which vary according to the nutrients present in the medium, can be observed, compared, and used to teach metabolic requirements. The experiments described in this report show simple macroscopic evidence of submicroscopic nutritional events. This can serve as a useful base for an analogy of heterotrophic human cell nutrition.

A New Fluorescence Photobleaching Method for Determining Solute Diffusive-Reactive Properties in Biological Tissues

Knowledge of solute transport in biological tissues is fundamental in order to understand cell nutrition and biosynthetic activity, and to design and fabricate tissue engineered constructs.