SARS-CoV-2 papain-like protease plays multiple roles in regulating cellular proteins in the endoplasmic reticulum.

Nsp3s are the largest nonstructural proteins of coronaviruses. These transmembrane proteins include papain-like proteases (PLpro) that play essential roles in cleaving viral polyproteins into their mature units. The PLpro of SARS-CoV viruses also have deubiquitinating and deISGylating activities. As Nsp3 is an endoplasmic reticulum (ER)-localized protein, we asked if the deubiquitinating activity of SARS-CoV-2 PLpro affects proteins that are substrates for ER-associated degradation (ERAD). Using full-length Nsp3 as well as a truncated transmembrane form we interrogated, by coexpression, three potential ERAD substrates, all of which play roles in regulating lipid biosynthesis. Transmembrane PLpro increases the level of INSIG-1 and decreases its ubiquitination. However, different effects were seen with SREBP-1 and SREBP-2. Transmembrane PLpro cleaves SREBP-1 at three sites, including two noncanonical sites in the N-terminal half of the protein, resulting in a decrease in precursors of the active transcription factor. Conversely, cleavage of SREBP-2 occurs at a single canonical site that disrupts a C-terminal degron, resulting in increased SREBP-2 levels. When this site is mutated and the degron can no longer be interrupted, SREBP-2 is still stabilized by transmembrane PLpro, which correlates with a decrease in SREBP-2 ubiquitination. All of these observations are dependent on PLpro catalytic activity. Our findings demonstrate that, when anchored to the ER membrane, SARS-CoV-2 Nsp3 PLpro can function as a deubiquitinating enzyme to stabilize ERAD substrates. Additionally, SARS-CoV-2 Nsp3 PLpro can cleave ER-resident proteins, including at sites that could escape analyses based on the established consensus sequence.

A structurally conserved site in AUP1 binds the E2 enzyme UBE2G2 and is essential for ER-associated degradation.

Endoplasmic reticulum-associated degradation (ERAD) is a protein quality control pathway of fundamental importance to cellular homeostasis. Although multiple ERAD pathways exist for targeting topologically distinct substrates, all pathways require substrate ubiquitination. Here, we characterize a key role for the UBE2G2 Binding Region (G2BR) of the ERAD accessory protein ancient ubiquitous protein 1 (AUP1) in ERAD pathways. This 27-amino acid (aa) region of AUP1 binds with high specificity and low nanomolar affinity to the backside of the ERAD ubiquitin-conjugating enzyme (E2) UBE2G2. The structure of the AUP1 G2BR (G2BRAUP1) in complex with UBE2G2 reveals an interface that includes a network of salt bridges, hydrogen bonds, and hydrophobic interactions essential for AUP1 function in cells. The G2BRAUP1 shares significant structural conservation with the G2BR found in the E3 ubiquitin ligase gp78 and in vitro can similarly allosterically activate ubiquitination in conjunction with ERAD E3s. In cells, AUP1 is uniquely required to maintain normal levels of UBE2G2; this is due to G2BRAUP1 binding to the E2 and preventing its rapid degradation. In addition, the G2BRAUP1 is required for both ER membrane recruitment of UBE2G2 and for its activation at the ER membrane. Thus, by binding to the backside of a critical ERAD E2, G2BRAUP1 plays multiple critical roles in ERAD.

Citizen science and online data: Opportunities and challenges for snake ecology and action against snakebite.

The secretive behavior and life history of snakes makes studying their biology, distribution, and the epidemiology of venomous snakebite challenging. One of the most useful, most versatile, and easiest to collect types of biological data are photographs, particularly those that are connected with geographic location and date-time metadata. Photos verify occurrence records, provide data on phenotypes and ecology, and are often used to illustrate new species descriptions, field guides and identification keys, as well as in training humans and computer vision algorithms to identify snakes. We scoured eleven online and two offline sources of snake photos in an attempt to collect as many photos of as many snake species as possible, and attempt to explain some of the inter-species variation in photograph quantity among global regions and taxonomic groups, and with regard to medical importance, human population density, and range size. We collected a total of 725,565 photos-between 1 and 48,696 photos of 3098 of the world's 3879 snake species (79.9%), leaving 781 "most wanted" species with no photos (20.1% of all currently-described species as of the December 2020 release of The Reptile Database). We provide a list of most wanted species sortable by family, continent, authority, and medical importance, and encourage snake photographers worldwide to submit photos and associated metadata, particularly of "missing" species, to the most permanent and useful online archives: The Reptile Database, iNaturalist, and HerpMapper.

Oligomeric complexes formed by Redß single strand annealing protein in its different DNA bound states.

Redß is a single strand annealing protein from bacteriophage λ that binds loosely to ssDNA, not at all to pre-formed dsDNA, but tightly to a duplex intermediate of annealing. As viewed by electron microscopy, Redß forms oligomeric rings on ssDNA substrate, and helical filaments on the annealed duplex intermediate. However, it is not clear if these are the functional forms of the protein in vivo. We have used size-exclusion chromatography coupled with multi-angle light scattering, analytical ultracentrifugation and native mass spectrometry (nMS) to characterize the size of the oligomers formed by Redß in its different DNA-bound states. The nMS data, which resolve species with the highest resolution, reveal that Redß forms an oligomer of 12 subunits in the absence of DNA, complexes ranging from 4 to 14 subunits on 38-mer ssDNA, and a much more distinct and stable complex of 11 subunits on 38-mer annealed duplex. We also measure the concentration of Redß in cells active for recombination and find it to range from 7 to 27 µM. Collectively, these data provide new insights into the dynamic nature of the complex on ssDNA, and the more stable and defined complex on annealed duplex.

Citizen science and online data: opportunities and challenges for snake ecology and action against snakebite

The secretive behavior and life history of snakes makes studying their biology, distribution, and the epidemiology of venomous snakebite challenging. One of the most useful, most versatile, and easiest to collect types of biological data are photographs, particularly those that are connected with geographic location and date-time metadata. Photos verify occurrence records, provide data on phenotypes and ecology, and are often used to illustrate new species descriptions, field guides and identification keys, as well as in training humans and computer vision algorithms to identify snakes. We scoured eleven online and two offline sources of snake photos in an attempt to collect as many photos of as many snake species as possible, and attempt to explain some of the inter-species variation in photograph quantity among global regions and taxonomic groups, and with regard to medical importance, human population density, and range size. We collected a total of 725,565 photos—between 1 and 48,696 photos of 3098 of the world's 3879 snake species (79.9%), leaving 781 “most wanted” species with no photos (20.1% of all currently-described species as of the December 2020 release of The Reptile Database). We provide a list of most wanted species sortable by family, continent, authority, and medical importance, and encourage snake photographers worldwide to submit photos and associated metadata, particularly of “missing” species, to the most permanent and useful online archives: The Reptile Database, iNaturalist, and HerpMapper.

Is Routine Preprocedural Bloodwork Needed for Elective Central Venous Access Device Removals in Children without Bleeding Dyscrasias?

PURPOSE: To assess the utility of routine preprocedural bloodwork during elective removal of central venous access devices (CVADs) with respect to bleeding complications. MATERIALS AND METHODS: Patients who underwent removal of a CVAD (tunneled central venous catheter [CVC] or port) by the interventional radiology service between January 2009 and December 2013 were retrospectively reviewed. Removals for infection or malfunction, without preprocedural bloodwork, with another concurrent procedure at the time of CVAD removal, or in patients with a bleeding dyscrasia were excluded. Peripherally inserted central catheter removals and temporary CVAD removals were also excluded. Routine preprocedural bloodwork included hemoglobin, platelet count, partial thromboplastin time, and International Normalized Ratio. Postprocedural complications were classified according to the Society of Interventional Radiology clinical practice guidelines. RESULTS: There were 802 CVAD removals in 777 patients (351 female, 426 male). Average patient age was 8.6 years (range, 5 wk to 19 y). In total, 246 permanent CVCs and 556 ports were removed. A total of 802 cases had preprocedural bloodwork. Of the 49 patients who had a bleeding complication after the procedure (6.1%; 49 of 802), 44 had normal findings on preprocedural bloodwork and 5 had abnormal findings. There was no statistically significant difference in bleeding complications between those with normal and abnormal bloodwork results (P = .7740). CONCLUSIONS: Routine bloodwork is not necessary before elective CVAD removal in children without a bleeding dyscrasia. Most children have normal findings on preprocedural bloodwork, and the incidence of postprocedural bleeding is low and not determined by bloodwork results.

Arsenic, Cadmium, Lead, and Mercury Concentrations in the Livers of Free-Ranging Common Garter Snakes (Thamnophis sirtalis) from Minnesota, USA.

Livers of 25 common garter snakes (Thamnophis sirtalis) from Minnesota (2015-16) were analyzed for heavy metals by inductively coupled plasma atomic emission spectroscopy. Dry weight mean concentrations (ranges µg/g) were arsenic, 118.94 (0.26-1245.56); cadmium, 1.44 (0.15-7.59); lead, 0.21 (<0.10-0.78); and mercury, 0.30 (<0.5-2.25).

Work function and temperature dependence of electron tunneling through an N-type perylene diimide molecular junction with isocyanide surface linkers.

Conducting probe atomic force microscopy (CP-AFM) was employed to examine electron tunneling in self-assembled monolayer (SAM) junctions. A 2.3 nm long perylene tetracarboxylic acid diimide (PDI) acceptor molecule equipped with isocyanide linker groups was synthesized, adsorbed onto Ag, Au and Pt substrates, and the current-voltage (I-V) properties were measured by CP-AFM. The dependence of the low-bias resistance (R) on contact work function indicates that transport is LUMO-assisted ('n-type behavior'). A single-level tunneling model combined with transition voltage spectroscopy (TVS) was employed to analyze the experimental I-V curves and to extract the effective LUMO position εl = ELUMO - EF and the effective electronic coupling (Γ) between the PDI redox core and the contacts. This analysis revealed a strong Fermi level (EF) pinning effect in all the junctions, likely due to interface dipoles that significantly increased with increasing contact work function, as revealed by scanning Kelvin probe microscopy (SKPM). Furthermore, the temperature (T) dependence of R was found to be substantial. For Pt/Pt junctions, R varied more than two orders of magnitude in the range 248 K < T < 338 K. Importantly, the R(T) data are consistent with a single step electron tunneling mechanism and allow independent determination of εl, giving values compatible with estimates of εl based on analysis of the full I-V data. Theoretical analysis revealed a general criterion to unambiguously rule out a two-step transport mechanism: namely, if measured resistance data exhibit a pronounced Arrhenius-type temperature dependence, a two-step electron transfer scenario should be excluded in cases where the activation energy depends on contact metallurgy. Overall, our results indicate (1) the generality of the Fermi level pinning phenomenon in molecular junctions, (2) the utility of employing the single level tunneling model for determining essential electronic structure parameters (εl and Γ), and (3) the importance of changing the nature of the contacts to verify transport mechanisms.

Genome Sequences of Four Subcluster L2 Mycobacterium Phages, Finemlucis, Miley16, Wilder, and Zakai.

Four subcluster L2 mycobacteriophages, Finemlucis, Miley16, Wilder, and Zakai, that infect Mycobacterium smegmatis mc2155 were isolated. The four phages are closely related to each other and code for 12 to 14 tRNAs and 130 to 132 putative protein-coding genes, including tyrosine integrases, cro, immunity repressors, and excise genes involved in the establishment of lysogeny.

Exceptionally Small Statistical Variations in the Transport Properties of Metal-Molecule-Metal Junctions Composed of 80 Oligophenylene Dithiol Molecules.

Strong stochastic fluctuations witnessed as very broad resistance (R) histograms with widths comparable to or even larger than the most probable values characterize many measurements in the field of molecular electronics, particularly those measurements based on single molecule junctions at room temperature. Here we show that molecular junctions containing 80 oligophenylene dithiol molecules (OPDn, 1 ≤ n ≤ 4) connected in parallel display small relative statistical deviations-δR/R ≈ 25% after only ∼200 independent measurements-and we analyze the sources of these deviations quantitatively. The junctions are made by conducting probe atomic force microscopy (CP-AFM) in which an Au-coated tip contacts a self-assembled monolayer (SAM) of OPDs on Au. Using contact mechanics and direct measurements of the molecular surface coverage, the tip radius, tip-SAM adhesion force (F), and sample elastic modulus (E), we find that the tip-SAM contact area is approximately 25 nm2, corresponding to about 80 molecules in the junction. Supplementing this information with I-V data and an analytic transport model, we are able to quantitatively describe the sources of deviations δR in R: namely, δN (deviations in the number of molecules in the junction), δε (deviations in energetic position of the dominant molecular orbital), and Î´Γ (deviations in molecule-electrode coupling). Our main results are (1) direct determination of N; (2) demonstration that δN/N for CP-AFM junctions is remarkably small (≤2%) and that the largest contributions to δR are δε and δΓ; (3) demonstration that δR/R after only ∼200 measurements is substantially smaller than most reports based on >1000 measurements for single molecule break junctions. Overall, these results highlight the excellent reproducibility of junctions composed of tens of parallel molecules, which may be important for continued efforts to build robust molecular devices.

Effect of Heteroatom Substitution on Transport in Alkanedithiol-Based Molecular Tunnel Junctions: Evidence for Universal Behavior.

The transport properties of molecular junctions based on alkanedithiols with three different methylene chain lengths were compared with junctions based on similar chains wherein every third -CH2- was replaced with O or S, that is, following the general formula HS(CH2CH2X)nCH2CH2SH, where X = CH2, O, or S and n = 1, 2, or 3. Conducting probe atomic force microscopy revealed that the low bias resistance of the chains increased upon substitution in the order CH2 < O < S. This change in resistance is ascribed to the observed identical trend in contact resistance, Rc, whereas the exponential prefactor ß (length sensitivity) was essentially the same for all chains. Using an established, analytical single-level model, we computed the effective energy offset εh (i.e., Fermi level relative to the effective HOMO level) and the electronic coupling strength Γ from the current-voltage (I-V) data. The εh values were only weakly affected by heteroatom substitution, whereas the interface coupling strength Γ varied by over an order of magnitude. Consequently, we ascribe the strong variation in Rc to the systematic change in Γ. Quantum chemical calculations reveal that the HOMO density shifts from the terminal SH groups for the alkanedithiols to the heteroatoms in the substituted chains, which provides a plausible explanation for the marked decrease in Γ for the dithiols with electron-rich heteroatoms. The results indicate that the electronic coupling and thus the resistance of alkanedithiols can be tuned by substitution of even a single atom in the middle of the molecule. Importantly, when appropriately normalized, the experimental I-V curves were accurately simulated over the full bias range (±1.5 V) using the single-level model with no adjustable parameters. The data could be collapsed to a single universal curve predicted by the model, providing clear evidence that the essential physics is captured by this analytical approach and supporting its utility for molecular electronics.

Charge Transport in 4 nm Molecular Wires with Interrupted Conjugation: Combined Experimental and Computational Evidence for Thermally Assisted Polaron Tunneling.

We report the synthesis, transport measurements, and electronic structure of conjugation-broken oligophenyleneimine (CB-OPI 6) molecular wires with lengths of ∼4 nm. The wires were grown from Au surfaces using stepwise aryl imine condensation reactions between 1,4-diaminobenzene and terephthalaldehyde (1,4-benzenedicarbaldehyde). Saturated spacers (conjugation breakers) were introduced into the molecular backbone by replacing the aromatic diamine with trans-1,4-diaminocyclohexane at specific steps during the growth processes. FT-IR and ellipsometry were used to follow the imination reactions on Au surfaces. Surface coverages (∼4 molecules/nm(2)) and electronic structures of the wires were determined by cyclic voltammetry and UV-vis spectroscopy, respectively. The current-voltage (I-V) characteristics of the wires were acquired using conducting probe atomic force microscopy (CP-AFM) in which an Au-coated AFM probe was brought into contact with the wires to form metal-molecule-metal junctions with contact areas of ∼50 nm(2). The low bias resistance increased with the number of saturated spacers, but was not sensitive to the position of the spacer within the wire. Temperature dependent measurements of resistance were consistent with a localized charge (polaron) hopping mechanism in all of the wires. Activation energies were in the range of 0.18-0.26 eV (4.2-6.0 kcal/mol) with the highest belonging to the fully conjugated OPI 6 wire and the lowest to the CB3,5-OPI 6 wire (the wire with two saturated spacers). For the two other wires with a single conjugation breaker, CB3-OPI 6 and CB5-OPI 6, activation energies of 0.20 eV (4.6 kcal/mol) and 0.21 eV (4.8 kcal/mol) were found, respectively. Computational studies using density functional theory confirmed the polaronic nature of charge carriers but predicted that the semiclassical activation energy of hopping should be higher for CB-OPI molecular wires than for the OPI 6 wire. To reconcile the experimental and computational results, we propose that the transport mechanism is thermally assisted polaron tunneling in the case of CB-OPI wires, which is consistent with their increased resistance.

Domain Structure of the Redß Single-Strand Annealing Protein: the C-terminal Domain is Required for Fine-Tuning DNA-binding Properties, Interaction with the Exonuclease Partner, and Recombination in vivo.

Redß is a component of the Red recombination system of bacteriophage λ that promotes a single strand annealing (SSA) reaction to generate end-to-end concatemers of the phage genome for packaging. Redß interacts with λ exonuclease (λexo), the other component of the Red system, to form a "synaptosome" complex that somehow integrates the end resection and annealing steps of the reaction. Previous work using limited proteolysis and chemical modification revealed that Redß consists of an N-terminal DNA binding domain, residues 1-177, and a flexible C-terminal "tail", residues 178-261. Here, we quantitatively compare the binding of the full-length protein (Redß(FL)) and the N-terminal domain (Redß(177)) to different lengths of ssDNA substrate and annealed duplex product. We find that in general, Redß(FL) binds more tightly to annealed duplex product than to ssDNA substrate, while Redß(177) binds more tightly to ssDNA. In addition, the C-terminal region of Redß corresponding to residues 182-261 was purified and found to fold into an α-helical domain that is required for the interaction with λexo to form the synaptosome complex. Deletion analysis of Redß revealed that removal of just eleven residues from the C-terminus disrupts the interaction with λexo as well as ssDNA and dsDNA recombination in vivo. By contrast, the determinants for self-oligomerization of Redß appear to reside solely within the N-terminal domain. The subtle but significant differences in the relative binding of Redß(FL) and Redß(177) to ssDNA substrate and annealed duplex product may be important for Redß to function as a SSA protein in vivo.

Snake fungal disease: an emerging threat to wild snakes.

Since 2006, there has been a marked increase in the number of reports of severe and often fatal fungal skin infections in wild snakes in the eastern USA. The emerging condition, referred to as snake fungal disease (SFD), was initially documented in rattlesnakes, where the infections were believed to pose a risk to the viability of affected populations. The disease is caused by Ophidiomyces ophiodiicola, a fungus recently split from a complex of fungi long referred to as the Chrysosporium anamorph of Nannizziopsis vriesii (CANV). Here we review the current state of knowledge about O. ophiodiicola and SFD. In addition, we provide original findings which demonstrate that O. ophiodiicola is widely distributed in eastern North America, has a broad host range, is the predominant cause of fungal skin infections in wild snakes and often causes mild infections in snakes emerging from hibernation. This new information, together with what is already available in the scientific literature, advances our knowledge of the cause, pathogenesis and ecology of SFD. However, additional research is necessary to elucidate the factors driving the emergence of this disease and develop strategies to mitigate its impacts.This article is part of the themed issue 'Tackling emerging fungal threats to animal health, food security and ecosystem resilience'.

Length-Dependent Nanotransport and Charge Hopping Bottlenecks in Long Thiophene-Containing π-Conjugated Molecular Wires.

Self-assembled conjugated molecular wires containing thiophene up to 6 nm in length were grown layer-by-layer using click chemistry. Reflection-absorption infrared spectroscopy, ellipsometry and X-ray photoelectron spectroscopy were used to follow the stepwise growth. The electronic structure of the conjugated wires was studied with cyclic voltammetry and UV-vis spectroscopy as well as computationally with density functional theory (DFT). The current-voltage curves (±1 V) of the conjugated molecular wires were measured with conducting probe atomic force microscopy (CP-AFM) in which the molecular wire film bound to a gold substrate was contacted with a conductive AFM probe. By systematically measuring the low bias junction resistance as a function of length for molecules 1-4 nm long, we extracted the structure dependent tunneling attenuation factor (ß) of 3.4 nm(-1) and a contact resistance of 220 kΩ. The crossover from tunneling to hopping transport was observed at a molecular length of 4-5 nm with an activation energy of 0.35 eV extracted from Arrhenius plots of resistance versus temperature. DFT calculations revealed localizations of spin densities (polarons) on molecular wire radical cations. The calculations were employed to gauge transition state energies for hopping of polarons along wire segments. Individual estimated transition state energies were 0.2-0.4 eV, in good agreement with the experimental activation energy. The transition states correspond to flattening of dihedral angles about specific imine bonds. These results open up possibilities to further explore the influence of molecular architecture on hopping transport in molecular junctions, and highlight the utility of DFT to understand charge localization and associated hopping-based transport.

A Structure-Activity Analysis for Probing the Mechanism of Processive Double-Stranded DNA Digestion by λ Exonuclease Trimers.

λ exonuclease (λexo) is an ATP-independent 5'-to-3' exonuclease that binds to double-stranded DNA (dsDNA) ends and processively digests the 5'-strand into mononucleotides. The crystal structure of λexo revealed that the enzyme forms a ring-shaped homotrimer with a central funnel-shaped channel for tracking along the DNA. On the basis of this structure, it was proposed that dsDNA enters the open end of the channel, the 5'-strand is digested at one of the three active sites, and the 3'-strand passes through the narrow end of the channel to emerge out the back. This model was largely confirmed by the structure of the λexo-DNA complex, which further revealed that the enzyme unwinds the DNA by 2 bp prior to cleavage, to thread the 5'-end of the DNA into the active site. On the basis of this structure, an "electrostatic ratchet" model was proposed, in which the enzyme uses a hydrophobic wedge to insert into the base pairs to unwind the DNA, a two-metal mechanism for nucleotide hydrolysis, a positively charged pocket to bind to the terminal 5'-phosphate generated after each round of cleavage, and an arginine residue (Arg-45) to bind to the minor groove of the downstream end of the DNA. To test this model, in this study we have determined the effects of 11 structure-based mutations in λexo on DNA binding and exonuclease activities in vitro, and on DNA recombination in vivo. The results are largely consistent with the model for the mechanism that was proposed on the basis of the structure and provide new insights into the roles of particular residues of the protein in promoting the reaction. In particular, a key role for Arg-45 in DNA binding is revealed.

Experimental and Theoretical Analysis of Nanotransport in Oligophenylene Dithiol Junctions as a Function of Molecular Length and Contact Work Function.

We report the results of an extensive investigation of metal-molecule-metal tunnel junctions based on oligophenylene dithiols (OPDs) bound to several types of electrodes (M1-S-(C6H4)n-S-M2, with 1 ≤ n ≤ 4 and M1,2 = Ag, Au, Pt) to examine the impact of molecular length (n) and metal work function (Φ) on junction properties. Our investigation includes (1) measurements by scanning Kelvin probe microscopy of electrode work function changes (ΔΦ = ΦSAM - Φ) caused by chemisorption of OPD self-assembled monolayers (SAMs), (2) measurements of junction current-voltage (I-V) characteristics by conducting probe atomic force microscopy in the linear and nonlinear bias ranges, and (3) direct quantitative analysis of the full I-V curves. Further, we employ transition voltage spectroscopy (TVS) to estimate the energetic alignment εh = EF - EHOMO of the dominant molecular orbital (HOMO) relative to the Fermi energy EF of the junction. Where photoelectron spectroscopy data are available, the εh values agree very well with those determined by TVS. Using a single-level model, which we justify via ab initio quantum chemical calculations at post-density functional theory level and additional UV-visible absorption measurements, we are able to quantitatively reproduce the I-V measurements in the whole bias range investigated (∼1.0-1.5 V) and to understand the behavior of εh and Γ (contact coupling strength) extracted from experiment. We find that Fermi level pinning induced by the strong dipole of the metal-S bond causes a significant shift of the HOMO energy of an adsorbed molecule, resulting in εh exhibiting a weak dependence with the work function Φ. Both of these parameters play a key role in determining the tunneling attenuation factor (ß) and junction resistance (R). Correlation among Φ, ΔΦ, R, transition voltage (Vt), and εh and accurate simulation provide a remarkably complete picture of tunneling transport in these prototypical molecular junctions.

Absorbable magnesium-based stent: physiological factors to consider for in vitro degradation assessments.

Absorbable metals have been widely tested in various in vitro settings using cells to evaluate their possible suitability as an implant material. However, there exists a gap between in vivo and in vitro test results for absorbable materials. A lot of traditional in vitro assessments for permanent materials are no longer applicable to absorbable metallic implants. A key step is to identify and test the relevant microenvironment and parameters in test systems, which should be adapted according to the specific application. New test methods are necessary to reduce the difference between in vivo and in vitro test results and provide more accurate information to better understand absorbable metallic implants. In this investigative review, we strive to summarize the latest test methods for characterizing absorbable magnesium-based stent for bioabsorption/biodegradation behavior in the mimicking vascular environments. Also, this article comprehensively discusses the direction of test standardization for absorbable stents to paint a more accurate picture of the in vivo condition around implants to determine the most important parameters and their dynamic interactions.

The effects of whole grain high-amylose maize flour as a source of resistant starch on blood glucose, satiety, and food intake in young men.

The objective of this study was to determine the dose response effect of whole grain high-amylose maize (HAM) flour as a source of resistant starch (RS) on blood glucose, appetite and short-term food intake. In a repeated-measures crossover trial, healthy men (n = 30, 22.9 ± 0.6 y, BMI of 22.6 ± 0.3 kg/m(2)) were randomly assigned to consume 1 of 3 cookies once a week for 3 wk. Cookies were control (100% wheat flour), low-dose (63% wheat flour,37% HAM flour), and high-dose (33% wheat flour, 67% HAM flour) providing 53.5, 43.5, and 36.3 g of available carbohydrate, respectively. Ad libitum food intake was measured 120 min at a pizza meal, blood glucose and subjective appetite were measured after consumption of the cookie (0 to 120 min) and after the pizza meal (140 to 200 min). Blood glucose concentrations were lower at 30 and 45 min after high-dose treatment, and at 120 min after both high- and low-dose treatments compared to control (P < 0.05). Blood glucose AUC before the pizza meal (0 to 120 min) was 44% and 14% lower, and higher by 43% and 41% after the pizza meal (140 to 200 min) compared with control. Yet despite the higher response following the meal, cumulative AUC (0 to 200 min) was still 22% lower after the high-dose treatment (P < 0.05). All treatments equally suppressed subjective appetite and there was no effect on food intake. In conclusion, HAM flour as a source of RS and incorporated into a cookie was associated with better glycemic control in young men.

The acute effect of commercially available pulse powders on postprandial glycaemic response in healthy young men.

Whole pulses (beans, peas, chickpeas and lentils) elicit low postprandial blood glucose (BG) responses in adults; however, their consumption in North America is low. One potential strategy to increase the dietary intake of pulses is the utilisation of commercial pulse powders in food products; however, it is unclear whether they retain the biological benefits observed with whole pulses. Therefore, the present study examined the effects of commercially prepared pulse powders on BG response before and after a subsequent meal in healthy young men. Overall, three randomised, within-subject experiments were conducted. In each experiment, participants received whole, puréed and powdered pulses (navy beans in Expt 1; lentils in Expt 2; chickpeas in Expt 3) and whole-wheat flour as the control. All treatments were controlled for available carbohydrate content. A fixed-energy pizza meal (50·2 kJ/kg body weight) was provided at 120 min. BG concentration was measured before (0-120 min) and after (140-200 min) the pizza meal. BG concentration peaked at 30 min in all experiments, and pulse forms did not predict their effect on BG response. Compared with the whole-wheat flour control, navy bean treatments lowered peak BG concentrations (Expt 1, P< 0.05), but not the mean BG concentration over 120 min. The mean BG concentration was lower for all lentil (Expt 2, P= 0.008) and chickpea (Expt 3, P= 0.002) treatments over 120 min. Processing pulses to powdered form does not eliminate the benefits of whole pulses on BG response, lending support to the use of pulse powders as value-added food ingredients to moderate postprandial glycaemic response.