IgG antibody production and persistence to 6 months following SARS-CoV-2 vaccination: A Northern Ireland observational study.

BACKGROUND: This study evaluates spike protein IgG antibody response following Oxford-AstraZeneca COVID-19 vaccination using the AbC-19™ lateral flow device. METHODS: Plasma samples were collected from n = 111 individuals from Northern Ireland. The majority were >50 years old and/or clinically vulnerable. Samples were taken at five timepoints from pre-vaccination until 6-months post-first dose. RESULTS: 20.3% of participants had detectable IgG responses pre-vaccination, indicating prior COVID-19. Antibodies were detected in 86.9% of participants three weeks after the first vaccine dose, falling to 74.7% immediately prior to the second dose, and rising to 99% three weeks post-second vaccine. At 6-months post-first dose, this decreased to 90.5%. At all timepoints, previously infected participants had significantly higher antibody levels than those not previously infected. CONCLUSION: This study demonstrates that strong anti-spike protein antibody responses are evoked in almost all individuals that receive two doses of Oxford-AstraZeneca vaccine, and which largely persist beyond six months after first vaccination.

Synthetic biology open language visual (SBOL Visual) version 2.3.

People who are engineering biological organisms often find it useful to communicate in diagrams, both about the structure of the nucleic acid sequences that they are engineering and about the functional relationships between sequence features and other molecular species. Some typical practices and conventions have begun to emerge for such diagrams. The Synthetic Biology Open Language Visual (SBOL Visual) has been developed as a standard for organizing and systematizing such conventions in order to produce a coherent language for expressing the structure and function of genetic designs. This document details version 2.3 of SBOL Visual, which builds on the prior SBOL Visual 2.2 in several ways. First, the specification now includes higher-level "interactions with interactions," such as an inducer molecule stimulating a repression interaction. Second, binding with a nucleic acid backbone can be shown by overlapping glyphs, as with other molecular complexes. Finally, a new "unspecified interaction" glyph is added for visualizing interactions whose nature is unknown, the "insulator" glyph is deprecated in favor of a new "inert DNA spacer" glyph, and the polypeptide region glyph is recommended for showing 2A sequences.

Nanophotonic-Carbohydrate Lab-on-a-Microneedle for Rapid Detection of Human Cystatin C in Finger-Prick Blood.

Miniaturized total analysis systems, for the rapid detection of disease biomarkers, with features including high biomarker sensitivity, selectivity, biocompatibility, and disposability, all at low cost are of profound importance in the healthcare sector. Within this frame of reference, we developed a lab-on-a-carbohydrate-microneedle biodevice by integrating localized surface plasmon resonance (LSPR) paper-based substrates with biocompatible microneedles of high aspect ratio (>60:1 length:width). These microneedles are completely fabricated with carbohydrate (maltose) and further coated with poly lactic-co-glycolic acid (PLGA), which together serves the purpose of fluid channels. The porous nature of PLGA, in addition to drawing blood by capillary action, filters out the whole blood, allowing only the blood plasma to reach the biorecognition layer of the developed biodevice. While the use of maltose provides biocompatibility to the microneedle, the axial compression and transverse load analysis revealed desired mechanical strength of the microneedle, with mechanical failure occurring at 11N and 9 N respectively for the compressive and transverse load. For a proof-of-principle demonstration, the developed biodevice is validated for its operational features by direct detection of cystatin C in finger-prick blood and up to a concentration of 0.01 µg/mL in buffered conditions using the LSPR technique. Furthermore, by changing the biorecognition layer, the use of the developed needle can be extended to other disease biomarkers, and therefore the innovation presented in this work represents a hallmark in the state of the art of lab-on-a-chip biodevices.

Synthetic biology open language visual (SBOL visual) version 2.2.

People who are engineering biological organisms often find it useful to communicate in diagrams, both about the structure of the nucleic acid sequences that they are engineering and about the functional relationships between sequence features and other molecular species. Some typical practices and conventions have begun to emerge for such diagrams. The Synthetic Biology Open Language Visual (SBOL Visual) has been developed as a standard for organizing and systematizing such conventions in order to produce a coherent language for expressing the structure and function of genetic designs. This document details version 2.2 of SBOL Visual, which builds on the prior SBOL Visual 2.1 in several ways. First, the grounding of molecular species glyphs is changed from BioPAX to SBO, aligning with the use of SBO terms for interaction glyphs. Second, new glyphs are added for proteins, introns, and polypeptide regions (e. g., protein domains), the prior recommended macromolecule glyph is deprecated in favor of its alternative, and small polygons are introduced as alternative glyphs for simple chemicals.

Rapid removal of lead(II) ions from water using iron oxide-tea waste nanocomposite - a kinetic study.

Lead (Pb) ions are a major concern to the environment and human health as they are contemplated cumulative poisons. In this study, facile synthesis of magnetic iron oxide-tea waste nanocomposite is reported for adsorptive removal of lead ions from aqueous solutions and easy magnetic separation of the adsorbent afterwards. The samples were characterised by scanning electron microscopy, Fourier transform-infrared spectroscopy, X-ray diffraction, and Braunner-Emmet-Teller nitrogen adsorption study. Adsorptive removal of Pb(II) ions from aqueous solution was followed by ultraviolet-visible (UV-Vis) spectrophotometry. About 95% Pb(II) ion removal is achieved with the magnetic tea waste within 10 min. A coefficient of regression R2 ≃ 0.99 and adsorption density of 18.83 mg g-1 was found when Pb(II) ions were removed from aqueous solution using magnetic tea waste. The removal of Pb(II) ions follows the pseudo-second-order rate kinetics. External mass transfer principally regulates the rate-limiting phenomena of adsorption of Pb(II) ions on iron oxide-tea waste surface. The results strongly imply that magnetic tea waste has promising potential as an economic and excellent adsorbent for the removal of Pb(II) from water.

Alternative strategies for the treatment of classical congenital adrenal hyperplasia: pitfalls and promises.

Despite decades of different treatment algorithms, the management of congenital adrenal hyperplasia (CAH) remains clinically challenging. This is due to the inherent difficulty of suppressing adrenal androgen production using near physiological dosing of glucocorticoids (GC). As a result, alternating cycles of androgen versus GC excess can occur and may lead to short stature, obesity, virilization, and alterations in puberty. Novel therapeutic alternatives, including new and more physiological means of GC delivery, inhibitors at the level of CRH or ACTH secretion and/or action, as well as "rescue strategies", such as GnRH analogs, anti-androgens, aromatase inhibitors, and estrogen receptor blockers, are available; many of these agents, however, still require active investigation in CAH. Bilateral adrenalectomy is effective but it is also still an experimental approach. Gene therapy and stem cells, to provide functional adrenal cortical tissue, are at preclinical stage but provide exciting avenues for a potential cure for CAH.

An allosteric modulator of alpha7 nicotinic receptors, N-(5-Chloro-2,4-dimethoxyphenyl)-N'-(5-methyl-3-isoxazolyl)-urea (PNU-120596), causes conformational changes in the extracellular ligand binding domain similar to those caused by acetylcholine.

Nicotinic acetylcholine receptors are implicated in several neuropsychiatric disorders, including nicotine addiction, Alzheimer's, schizophrenia, and depression. Therefore, they represent a critical molecular target for drug development and targeted therapeutic intervention. Understanding the molecular mechanisms by which allosteric modulators enhance activation of these receptors is crucial to the development of new drugs. We used the substituted cysteine accessibility method to study conformational changes induced by the positive allosteric modulator N-(5-chloro-2,4-dimethoxyphenyl)-N'-(5-methyl-3-isoxazolyl)-urea (PNU-120596) in the extracellular ligand binding domain of alpha7 nicotinic receptors carrying the L247T mutation. PNU-120596 caused changes in cysteine accessibility at the inner beta sheet, transition zone, and agonist binding site. These changes in accessibility are similar to but not identical to those caused by ACh alone. In particular, PNU-120596 induced changes in MTSEA accessibility at N170C (in the transition zone) that were substantially different from those evoked by acetylcholine (ACh). We found that PNU-120596 induced changes at position E172C in the absence of allosteric modulation. We identified a cysteine mutation of the agonist binding site (W148C) that exhibited an unexpected phenotype in which PNU-120596 acts as a full agonist. In this mutant, ACh-evoked currents were more sensitive to thiol modification than PNU-evoked currents, suggesting that PNU-120596 does not bind at unoccupied agonist-binding sites. Our results provide evidence that binding sites for PNU-120596 are not in the agonist-binding sites and demonstrate that positive allosteric modulators such as PNU-120596 enhance agonist-evoked gating of nicotinic receptors by eliciting conformational effects that are similar but nonidentical to the gating conformations promoted by ACh.

Conformational changes in alpha 7 acetylcholine receptors underlying allosteric modulation by divalent cations.

Allosteric modulation of membrane receptors is a widespread mechanism by which endogenous and exogenous agents regulate receptor function. For example, several members of the nicotinic receptor family are modulated by physiological concentrations of extracellular calcium ions. In this paper, we examined conformational changes underlying this modulation and compare these with changes evoked by ACh. Two sets of residues in the alpha 7 acetylcholine receptor extracellular domain were mutated to cysteine and analyzed by measuring the rates of modification by the thiol-specific reagent 2-aminoethylmethane thiosulfonate. Using Ba2+ as a surrogate for Ca2+, we found a divalent-dependent decrease the modification rates of cysteine substitutions at M37 and M40, residues at which rates were also slowed by ACh. In contrast, Ba2+ had no significant effect at N52C, a residue where ACh increased the rate of modification. Thus divalent modulators cause some but not all of the conformational effects elicited by agonist. Cysteine substitution of either of two glutamates (E44 or E172), thought to participate in the divalent cation binding site, caused a loss of allosteric modulation, yet Ba2+ still had a significant effect on modification rates of these residues. In addition, the effect of Ba2+ at these residues did not appear to be due to direct occlusion. Our data demonstrate that modulation by divalent cations involves substantial conformational changes in the receptor extracellular domain. Our evidence also suggests the modulation occurs via a binding site distinct from one which includes either (or both) of the conserved glutamates at E44 or E172.

Cure of Acanthamoeba cerebral abscess in a liver transplant patient.

Acanthamoeba-related cerebral abscess and encephalitis are rare but usually fatal, being caused by free-living amoebic infections usually occurring in immunocompromised patients. In patients receiving transplants, a literature review showed that the infection is universally fatal. The diagnosis is often missed despite appropriate investigations including lumbar puncture, computerized tomography, and brain biopsy. We present the first reported liver transplant patient with Acanthamoeba cerebral abscess. The diagnosis was made in brain tissue removed at decompressive frontal lobectomy. He was successfully treated with a 3-month course of co-trimoxazole and rifampicin. There was no recurrence of the disease after 11 years of follow-up.

Agonist-driven conformational changes in the inner beta-sheet of alpha7 nicotinic receptors.

Cys-loop ligand-gated ion channels assemble as pentameric proteins, and each monomer contributes two structural elements: an extracellular ligand-binding domain (LBD) and a transmembrane ion channel domain. Models of receptor activation include rotational movements of subunits leading to opening of the ion channel. We tested this idea using substituted cysteine accessibility to track conformational changes in the inner beta sheet of the LBD. Using a nondesensitizing chick alpha7 background (L(247)T), we constructed 18 consecutive cysteine replacement mutants (Leu(36) to Ile(53)) and tested each for expression of acetylcholine (ACh)-evoked currents and functional sensitivity to thiol modification. We measured rates of modification in the presence and absence of ACh to identify conformational changes associated with receptor activation. Resting modification rates of eight substituted cysteines in the beta1 and beta2 strands and the sequence between them (loop 2) varied over several orders of magnitude, suggesting substantial differences in the accessibility or electrostatic environment of individual side chains. These differences were in general agreement with structural models of the LBD. Eight of 18 cysteine replacements displayed ACh-dependent changes in modification rates, indicating a change in the accessibility or electrostatic environment of the introduced cysteine during activation. We were surprised that the effects of agonist exposure were difficult to reconcile with rotational models of activation. Acetylcholine reduced the modification rate of M(40)C but increased it at N(52)C despite the close physical proximity of these residues. Our results suggest that models that depend strictly on rigid-body rotation of the LBD may provide an incomplete description of receptor activation.

Role of the outer beta-sheet in divalent cation modulation of alpha7 nicotinic receptors.

alpha-7 Nicotinic acetylcholine receptors (AChRs) exhibit a positive modulation by divalent cations similar to that observed in other AChRs. In the chick alpha7 AChR, this modulation involves a conserved glutamate in loop 9 (Glu172) that undergoes agonist-dependent movements during activation. From these observations, we hypothesized that movements of the nearby beta-sheet formed by the beta7, beta9, and beta10 strands may be involved in agonist activation and/or divalent modulation. To test this hypothesis, we examined functional properties of cysteine mutations of the beta7 and beta10 strands, alone or in pairs. We postulated that reduced flexibility or mobility of the beta7/beta9/beta10-sheet as a result of introduction of a disulfide bond between the beta strands would alter activation by agonists. Using a nondesensitizing alpha7 mutant background (L247T), we identified one mutant pair, K144C + T198C, that exhibited a unique characteristic: it was fully activated by divalent cations (Ca2+, Ba2+, or Sr2+) in the absence of acetylcholine (ACh). Divalent-evoked currents were blocked by the alpha7 antagonist methyllycaconitine and were abolished when Glu172 was mutated to glutamine. When the K144C + T198C pair was expressed in wild-type alpha7 receptors, activation required both ACh and divalent cations. We conclude that the introduction of a disulfide bond into beta7/beta9/beta10 lowers the energetic barrier between open and closed conformations, probably by reducing the torsional flexibility of the beta-sheet. In this setting, divalent cations, acting at the conserved glutamate in loop 9, act as full agonists or requisite coagonists.

Legislation for smoke-free workplaces and health of bar workers in Ireland: before and after study.

OBJECTIVES: To compare exposure to secondhand smoke and respiratory health in bar staff in the Republic of Ireland and Northern Ireland before and after the introduction of legislation for smoke-free workplaces in the Republic. DESIGN: Comparisons before and after the legislation in intervention and control regions. SETTING: Public houses in three areas in the Republic (intervention) and one area in Northern Ireland (control). PARTICIPANTS: 329 bar staff enrolled in baseline survey; 249 (76%) followed up one year later. Of these, 158 were non-smokers both at baseline and follow-up. MAIN OUTCOME MEASURES: Salivary cotinine concentration, self reported exposure to secondhand smoke, and respiratory and sensory irritation symptoms. RESULTS: In bar staff in the Republic who did not themselves smoke, salivary cotinine concentrations dropped by 80% after the smoke-free law (from median 29.0 nmol/l (95% confidence interval 18.2 to 43.2 nmol/l)) to 5.1 nmol/l (2.8 to 13.1 nmol/l) in contrast with a 20% decline in Northern Ireland over the same period (from median 25.3 nmol/l (10.4 to 59.2 nmol/l) to 20.4 nmol/l (13.2 to 33.8 nmol/l)). Changes in self reported exposure to secondhand smoke were consistent with the changes in cotinine concentrations. Reporting any respiratory symptom declined significantly in the Republic (down 16.7%, -26.1% to -7.3%) but not in Northern Ireland (0% difference, -32.7% to 32.7%). After adjustment for confounding, respiratory symptoms declined significantly more in the Republic than in Northern Ireland and the decline in cotinine concentration was twice as great. CONCLUSION: The smoke-free law in the Republic of Ireland protects non-smoking bar workers from exposure to secondhand smoke.

Agonist-induced conformational changes in the extracellular domain of alpha 7 nicotinic acetylcholine receptors.

The molecular mechanisms that couple agonist binding to the gating of Cys-loop ionotropic receptors are not well understood. The crystal structure of the acetylcholine (ACh) binding protein has provided insights into the structure of the extracellular domain of nicotinic receptors and a framework for testing mechanisms of activation. Key ligand binding residues are located at the C-terminal end of the beta9 strand. At the N-terminal end of this strand (loop 9) is a conserved glutamate [E172 in chick alpha7 nicotinic acetylcholine receptors (nAChRs)] that is important for modulating activation. We hypothesize that agonist binding induces the movement of loop 9. To test this, we used the substituted-cysteine accessibility method to examine agonist-dependent changes in the modification of cysteines introduced in loop 9 of L247T alpha7 nAChRs. In the absence of agonist, ACh-evoked responses of E172C/L247T alpha7 nAChRs were inhibited by 2-trimethylammonioethylmethane thiosulfonate (MTSET). Agonist coapplication with MTSET reduced the extent and rate of modification. The dose-dependence of ACh activation was nearly identical with that of ACh-dependent protection from modification. ACh increased the inhibition by methanethiosulfonate reagents of N170C and did not change inhibition of G171C receptors. The antagonist dihydro-beta-erythroidine did not mimic the effects of ACh. Combined with a structural model, the data suggest that receptor activation includes subunit rotation and/or intrasubunit conformational changes that move N170 to a more accessible position and E172 to a more protected position away from the vestibule. Thus, loop 9, located near the junction between the extracellular and transmembrane domains, participates in conformational changes triggered by ligand binding.

Glutamate 172, essential for modulation of L247T alpha7 ACh receptors by Ca2+, lines the extracellular vestibule.

Neuronal alpha7 nicotinic ACh receptors (nAChRs) are permeable to and modulated by Ca2+, Ba2+, and Sr2+. These permeant divalent cations interact with slowly desensitizing L247T alpha7 nAChRs to increase the potency and maximal efficacy of ACh, increase the efficacy of dihydro-beta-erythroidine (DHbetaE), and increase agonist-independent activity. Mutation of glutamate 172 (E172) to glutamine or cysteine eliminated these effects of permeant divalent cations. 2-(Trimethylammonium)ethyl methanethiosulfonate (MTSET), a cysteine-modifying reagent directed at water-accessible thiols, inhibited ACh-evoked currents of E172C/L247T alpha7 nAChRs by >90%, demonstrating that E172 was accessible to permeant ions. The data are consistent with a model of alpha7 receptors, derived from the crystal structure of the ACh binding protein (AChBP) from Lymnaea stagnalis, in which E172 projects toward the lumen of the extracellular vestibule. The observations that E172 was essential for divalent cation modulation of L247T alpha7 nAChRs and was accessible to permeating ions suggest that this residue participates in coupling ion permeation with modulation of receptor activity.

Physical activity and ethnic differences in hypertension prevalence in the United States.

BACKGROUND: In the United States, non-Hispanic blacks have higher rates of hypertension than other ethnic groups. In addition, they have higher rates of physical inactivity, a behavior linked to high blood pressure. We examined associations between ethnicity, leisure-time physical activity (LTPA), and hypertension prevalence in a representative sample of U.S. adults. METHODS: Using data on 16,246 adults in the third National Health and Nutrition Examination Survey, hypertension prevalence was determined for non-Hispanic white, non-Hispanic black, and Mexican Americans at various levels of LTPA (none, 0.1-4.9 bouts/week at any intensity, 5+ bouts/week of moderate-to-vigorous activity). Logistic regression was used to examine relationships between hypertension prevalence, race, LTPA, and other variables. RESULTS: Hypertension prevalence was significantly less in the most active group, compared with their sedentary peers (odds ratio = 0.73, CI 0.59 to 0.90). Blacks had an odds ratio for hypertension of 1.77 (CI 1.49 to 2.10) compared with non-Hispanic whites, after adjusting for gender, age, income, LTPA, smoking, BMI, salt intake, rural/urban dwelling, and alcohol intake. Mexican Americans had an adjusted odds ratio of 0.75 (CI 0.62 to 0.89), relative to non-Hispanic whites. CONCLUSION: Ethnicity and LTPA are both associated with hypertension prevalence after controlling for each other, as well as other confounders. Thus, race and physical activity are important independent contributors to hypertension prevalence.