Successful synthesis of a glial-specific blood-brain barrier shuttle peptide following a fragment condensation approach on a solid-phase resin.

Successful manual synthesis of the TD2.2 peptide acting as a blood-brain barrier shuttle was achieved. TD2.2 was successfully synthesised by sequential condensation of four protected peptide fragments on solid-phase settings, after several unsuccessful attempts using the stepwise approach. These fragments were chosen to minimise the number of demanding amino acids (in terms of coupling, Fmoc removal) in each fragment that are expected to hamper the overall synthetic process. Thus, the hydrophobic amino acids as well as Arg(Pbf) were strategically spread over multiple fragments rather than having them congested in one fragment. This study shows how a peptide that shows big challenges in the synthesis using the common stepwise elongation methodology can be synthesised with an acceptable purity. It also emphasises that choosing the right fragment with certain amino acid constituents is key for a successful synthesis. It is worth highlighting that lower amounts of reagents were required to synthesise the final peptide with an identical purity to that obtained by the automatic synthesiser.

Prediction of peptides retention behavior in reversed-phase liquid chromatography based on their hydrophobicity.

Hydrophobicity is an important physicochemical property of peptides and proteins. It is responsible for their conformational changes, stability, as well as various chemical intramolecular and intermolecular interactions. Enormous efforts have been invested to study the extent of hydrophobicity and how it could influence various biological processes, in addition to its crucial role in the separation and purification endeavor as well. Here, we have reviewed various studies that were carried out to determine the hydrophobicity starting from (i) simple amino acids solubility behavior, (ii) experimental approach that was undertaken in the reversed-phase liquid chromatography mode, and ending with (iii) some examples of more advanced computational and machine learning models.

Increasing the Diameter of Vertically Aligned, Hexagonally Ordered Pores in Mesoporous Silica Thin Films.

The variation in pore size in mesoporous films produced by electrochemically assisted self-assembly (EASA) with the surfactant chain length is described. EASA produces a hexagonal array of pores perpendicular to the substrate surface by using an applied potential to organize cationic surfactants and the resultant current to drive condensation in a silica sol. Here, we show that a range of pore sizes between 2 and 5 nm in diameter is available with surfactants of the form [Me3NCnH2n+1]Br, with alkyl chain lengths between C14 and C24. The film quality, pore order, pore size, and pore accessibility are probed with a range of techniques.

N95 respirators: quantitative fit test pass rates and usability and comfort assessment by health care workers.

OBJECTIVES: To compare the performance of four N95 respirator types with respect to quantitative fit test pass rate and health care worker-rated usability and comfort. DESIGN, SETTING, PARTICIPANTS: Health care workers who participated in the respiratory protection program at the Royal Melbourne Hospital, 1 October 2020 - 31 May 2021. Participants underwent quantitative N95 respirator fit testing (at least three of four types: semi-rigid cup, flat-fold cup, duckbill, and three-panel flat-fold types), and were invited to complete an online usability and comfort assessment for respirators for which their fit test results were passes. MAIN OUTCOME MEASURES: Fit test pass rate, and user-rated overall comfort and assessment ratings (five-point Likert scales), by N95 respirator type. RESULTS: A total of 2161 health care workers underwent quantitative fit testing (women, 1586 [73.4%]; nurses, 1271 [58.8%]). The overall fit test pass rates were 65.0% for the semi-rigid cup respirators (1029/1583 tests), 32.4% for the flat-fold respirator (660/2035 tests), 59.2% for the duckbill respirators (2005/3387 tests), and 96.4% for the three-panel flat-fold respirator (1876/1946 tests). 378 health care workers completed the comfort and usability survey. Overall comfort and assessment ratings each differed by respirator group (P < 0.001); the median overall comfort (4; IQR, 3-4) and overall assessment values (4; IQR, 3-5) were highest for the three-panel flat-fold respirator and lowest for the semi-rigid cup respirators (comfort: 2 [IQR, 1-3]; assessment: 2 [IQR, 2-3]). CONCLUSIONS: The three-panel flat-fold N95 respirator outperformed the three alternative types with regard to fit test pass rate and user-rated comfort and usability. To maximise respiratory protection for health care workers, these factors should be considered when making respirator procurement decisions.

A longitudinal study of the psychological impact of the COVID-19 pandemic on frontline perioperative healthcare staff in an Australian tertiary public hospital.

OBJECTIVE: This longitudinal study examined changes in psychological outcomes of perioperative frontline healthcare workers at one of Australia's most COVID-19 affected hospitals, following the surge and decline of a pandemic wave. METHOD: A single-centred longitudinal online survey was conducted between 26 May and 17 November 2020. Recruitment was via poster advertisement and email invitation. The survey was sent out every 4 weeks, resulting in seven time-points. RESULTS: In total, 385 survey results were analysed from 193 staff (about 64% response rate), 72 (37%) of whom completed the survey more than once. The prevalence of moderate-to-severe anxiety and depressive symptoms peaked at 27% and 25%, respectively, during the pandemic surge. Up to 35% displayed post-traumatic stress disorder (PTSD) symptoms. Although not statistically significant, the trend of depressive and PTSD symptoms worsened over time, especially among females and anaesthetic/surgical trainees, despite subsidence of the pandemic curve. Technicians and anaesthetic/scrub nurses were the at-risk groups with worst psychological outcomes. CONCLUSION: We found persistent mental health impacts on frontline perioperative HCWs despite subsidence of the pandemic wave. Further research is needed to determine the extent and trajectory of such impacts with larger sample sizes to determine generalisability to frontline HCWs in general.

Identification of the preferential CO and SO2 adsorption sites within NOTT-401.

NOTT-401 was found to be a highly stable adsorbent for SO2 and CO with excellent cyclability and a straightforward regeneration at room temperature. Moreover, the preferential CO binding sites within the MOF material have been identified by experimental in situ DRIFT spectroscopy coupled with DFT and QTAIM calculations. Such preferential CO adsorption sites were correlated to identify the most significant SO2 interactions within NOTT-401. This study sheds light on the role of the thiophene and hydroxo functionality, for a MOF material, in the binding of SO2 or CO.

On the Origins of Some Spectroscopic Properties of "Purple Iron" (the Tetraoxoferrate(VI) Ion) and Its Pourbaix Safe-Space.

In this work, the authors attempt to interpret the visible, infrared and Raman spectra of ferrate(VI) by means of theoretical physical-inorganic chemistry and historical highlights in this field of interest. In addition, the sacrificial decomposition of ferrate(VI) during water treatment will also be discussed together with a brief mention of how Rayleigh scattering caused by the decomposition of FeVIO42- may render absorbance readings erroneous. This work is not a compendium of all the instrumental methods of analysis which have been deployed to identify ferrate(VI) or to study its plethora of reactions, but mention will be made of the relevant techniques (e.g., Mössbauer Spectroscopy amongst others) which support and advance this overall discourse at appropriate junctures, without undue elaboration on the foundational physics of these techniques.

A Robust and Biocompatible Bismuth Ellagate MOF Synthesized Under Green Ambient Conditions.

The first bioinspired microporous metal-organic framework (MOF) synthesized using ellagic acid, a common natural antioxidant and polyphenol building unit, is presented. Bi2O(H2O)2(C14H2O8)·nH2O (SU-101) was inspired by bismuth phenolate metallodrugs, and could be synthesized entirely from nonhazardous or edible reagents under ambient aqueous conditions, enabling simple scale-up. Reagent-grade and affordable dietary supplement-grade ellagic acid was sourced from tree bark and pomegranate hulls, respectively. Biocompatibility and colloidal stability were confirmed by in vitro assays. The material exhibits remarkable chemical stability for a bioinspired MOF (pH = 2-14, hydrothermal conditions, heated organic solvents, biological media, SO2 and H2S), attributed to the strongly chelating phenolates. A total H2S uptake of 15.95 mmol g-1 was recorded, representing one of the highest H2S capacities for a MOF, where polysulfides are formed inside the pores of the material. Phenolic phytochemicals remain largely unexplored as linkers for MOF synthesis, opening new avenues to design stable, eco-friendly, scalable, and low-cost MOFs for diverse applications, including drug delivery.

Prehospital Naloxone and Emergency Department Adverse Events: A Dose-Dependent Relationship.

BACKGROUND: The purpose of this study was to evaluate prehospital and emergency department (ED) interventions and outcomes of patients who received prehospital naloxone for a suspected opioid overdose. OBJECTIVES: The primary objective was to evaluate if the individual dose, individual route, total dose, number of prehospital naloxone administrations, or occurrence of a prehospital adverse event (AE) were associated with the occurrence of AEs in the ED. Secondary objectives included a subset analysis of patients who received additional naloxone while in the ED, or were admitted to an intensive care or step-down unit (ICU). METHODS: This was a retrospective, observational chart review of adult patients who received prehospital naloxone and were transported by ambulance to a suburban academic tertiary care center between 2014 and 2017. Descriptive, univariate, and multivariate statistics were used, with p < 0.05 indicating significance. RESULTS: There were 513 patients included in the analysis, with a median age of 29 years, and median total prehospital naloxone dose of 2 mg. An increasing number of prehospital naloxone doses, an occurrence of a prehospital AE, and a route of administration other than intranasally for the first dose of prehospital naloxone were significantly associated with an increased likelihood of an ED AE. Patients who received < 2 mg of prehospital naloxone had the least likelihood of being admitted to an ICU, whereas patients who received at least 6 mg had a dramatically increased likelihood of ICU admission. CONCLUSIONS: Our results suggest that an increasing number of prehospital naloxone doses was significantly associated with an increased likelihood of an ED adverse event.

Mapping the mAb Aggregation Propensity Using Self-Interaction Chromatography as a Screening Tool.

The osmotic second virial coefficient ( B2), which describes protein-protein molecular interactions in solution, was determined using self-interaction chromatography (SIC) for an IgG1-type mAb across a wide range of solution conditions. These data were compared with its time dependent aggregation behavior, as determined using size-exclusion chromatography (SEC), and its temperature dependent aggregation behavior using dynamic light scattering (DLS) over a four-week period (SEC) or overnight (DLS). DLS and SEC gave consistent data on aggregation behavior, which correlated well with experimental B2 trends across the wide pH (4-9) and NaCl concentration (0-1.0 M) ranges studied. The IgG aggregated at pH 4 for 0.5-1.0 M NaCl concentrations and for 0 M NaCl concentrations at pH 8. Best stability against aggregation was exhibited for the pH range from 5 to 8 at 0.8-1.0 M NaCl. SIC data were able to be classified within the one-day solution conditions for aggregation, which were not identified for 2-3 weeks in the accelerated SEC stability study. The ability of SIC to provide such data rapidly reflects the fundamentally thermodynamic nature of this parameter and of the aggregation process itself. Proteins with attractive protein-protein interactions and negative B2 coefficients in the range -3 to -6 clearly exhibit aggregation behavior, while B2 values in the range 0 to 2 showed good stability toward aggregation. SIC allows the rapid screening of solution conditions for which mAbs will exhibit stability to aggregation while requiring 90% less time and material compared with that required for a conventional SEC aggregation study.

Comparative Plasma and Cerebrospinal Fluid Pharmacokinetics of Paracetamol After Intravenous and Oral Administration.

BACKGROUND: We compared plasma and cerebrospinal fluid (CSF) pharmacokinetics of paracetamol after intravenous (IV) and oral administration to determine dosing regimens that optimize CSF concentrations. METHODS: Twenty-one adult patients were assigned randomly to 1 g IV, 1 g oral or 1.5 g oral paracetamol. An IV cannula and lumbar intrathecal catheter were used to sample venous blood and CSF, respectively, over 6 hours. The plasma and CSF maximum concentrations (Cmax), times to maximum concentrations (Tmax), and area under the plasma and CSF concentration-time curves (AUCs) were calculated using noncompartmental techniques. Significance was defined by P < .0167 (Bonferroni correction for 3 comparisons for each parameter). Probability (X < Y) (p″) with Bonferroni corrected 95% confidence intervals (CIs) were calculated (CIs including 0.5 meet the null hypothesis). Results are presented as median (range) or p″ (CI). P values are listed as 1 g IV vs 1 g orally, 1 g IV vs 1.5 g orally and 1 g orally vs 1.5 g orally, respectively. RESULTS: Wide variation in measured paracetamol concentrations was observed, especially in the oral groups. The median plasma Cmax in the 1 g IV group was significantly greater than the oral groups. In contrast, the median CSF Cmax was not different between groups. The median plasma Tmax in the 1 g IV group was 105 and 75 minutes earlier than in the 1 and 1.5 g oral groups. The median CSF Tmax was not significantly different between groups. The median plasma AUC (total) was not significantly different between groups; however, in the first hour, the median plasma AUC was significantly greater in the IV group than in the oral groups. In the second hour, there was no difference between groups. The median CSF AUC (total) did not significantly differ between groups; however, in the first hour, the median CSF AUC was significantly greater in the IV compared with the orally groups. In the second hour, there was no difference between groups. Our analysis indicated that the median Cmax, Tmax, and AUC values lacked precision because of small sample sizes. CONCLUSIONS: Peak plasma concentrations were greater and reached earlier after IV than oral dosing. Evidence for differences in CSF Cmax and Tmax was lacking because of the small size of this study.

A new method to determine dispersive surface energy site distributions by inverse gas chromatography.

A computational model to predict the relative energy site contributions of a heterogeneous material from data collected by finite dilution-inverse gas chromatography (FD-IGC) is presented in this work. The methodology employed a multisolvent system site filling model utilizing Boltzmann statistics, expanding on previous efforts to calculate "experienced energies" at varying coverage, yielding a retention volume distribution allowing calculation of a surface free energy distribution. Surface free energy distributions were experimentally measured for racemic ibuprofen and ß-mannitol powders, the energies of each were found in the ranges 43-52 and 40-55 mJ/m(2), respectively, over a surface coverage range of 0-8%. The computed contributions to surface energy values were found to match closely with data collected on macroscopic crystals by alternative techniques (±<1.5 mJ/m(2)).

Magnetic resonance imaging respirators: a randomized crossover trial to assess respiratory protection, usability, and comfort.

Objective: Many available facepiece filtering respirators contain ferromagnetic components, which may cause significant problems in the magnetic resonance imaging (MRI) environment. We conducted a randomized crossover trial to assess the effectiveness, usability, and comfort of 3 types of respirators, judged to be "conditionally MRI safe" with an aluminum nosepiece (Halyard 46727 duckbill-type respirators and Care Essentials MSK-002 bifold cup-type respirators) or "MRI safe" completely metal free (Eagle AG2200 semirigid cup-type respirators). Design and setting: We recruited 120 participants to undergo a quantitative fit test (QNFT) on each of the 3 respirators in a randomized order. Participants then completed a usability and comfort assessment of each respirator. Results: There were significant differences in the QNFT pass rates (51% for Halyard 46727, 73% for Care Essentials MSK-002, and 86% for Eagle AG2200, P < .001). The first-time fit test pass rate and overall fit factor were significantly higher for Eagle AG2200 compared with the other 2 respirators. Eagle AG2200 scored the lowest ratings in the ease of use and overall comfort. There were no significant differences in other modalities, including the seal rating, breathability, firmness, and overall assessment. Conclusions: Our study supports the utility of the Eagle AG2200 and Care Essentials MSK-002 respirators for healthcare professionals working in an MRI environment, based on their high QNFT pass rates and reasonably good overall usability and comfort scores. Eagle AG2200 is unique because of its metal-free construction. However, its comparatively lower usability and comfort ratings raise questions about practicality, which may be improved by greater user training.

Prospective comprehensive evaluation of an elastic-band beard cover for filtering facepiece respirators in healthcare workers.

OBJECTIVE: To undertake a healthcare-based multimodal evaluation of the combination of filtering facepiece respirator (FFR) with the elastic-band beard cover technique, including quantitative fit test (QNFT) results, skills assessment, and usability assessment. DESIGN AND SETTING: We conducted a prospective study through the Respiratory Protection Program at the Royal Melbourne Hospital from May 2022 to January 2023. PARTICIPANTS: Healthcare workers who required respiratory protection and could not shave for religious, cultural, or medical reasons. INTERVENTION: Online education and personal face-to-face training on the use of FFR with the elastic-band beard cover technique. RESULTS: Among 87 participants (median beard length 38 mm; interquartile range [IQR], 20-80), 86 (99%) passed 3 QNFTs consecutively with the elastic-band beard cover under a Trident P2 respirator and 68 (78%) passed 3 QNFTs consecutively with a 3M 1870+ Aura respirator. The first QNFT pass rate and the overall fit factors were significantly higher when using the technique than without the elastic-band beard cover. Most participants displayed a high skill level in their donning, doffing, and user seal-check techniques. Of 87 participants, 83 (95%) completed the usability assessment. The overall ease of use, comfort, and overall assessment were rated highly. CONCLUSIONS: The elastic-band beard cover technique can provide safe and effective respiratory protection for bearded healthcare workers. The technique was easily taught, comfortable, well tolerated and accepted by healthcare workers, potentially allowing them full participation in the workforce during pandemics with airborne transmission. We recommend further research and evaluation of this technique in a broader health workforce.

Development of a rapid, multi-organisational, multi-modal assessment of a newly available disposable respirator.

BACKGROUND: A rapid large-scale evaluation of a newly available duckbill style P2/N95 respirator, the Care Essentials (CE) MSK-003, was required to determine its suitability for deployment into the Victorian healthcare service. The aims of this study were to assess the feasibility of establishing a rapid, multi-organisational and multi-modal evaluation of the respirator, and to investigate whether this respirator would meet the needs of healthcare workers. METHODS: The evaluation was a collaboration among three healthcare organisations - two tertiary hospitals in metropolitan Melbourne and a rural-based hospital. Participants were healthcare workers undertaking their routine fit tests. They were required to complete quantitative fit testing and a usability assessment survey on the CE MSK-003 respirator. The a priori performance criteria were set as fit test pass rate of >70%, plus satisfactory subjective overall comfort and performance assessments, defined as a rating of adequate, good, or very good in >90% of the cohort. RESULTS: A total of 1070 participants completed the multi-modal assessment within a month. Seventy-eight percent of participants passed their quantitative fit test. Over 90% of survey respondents reported that the CE MSK-003 was adequate, good or very good in terms of its overall comfort and performance assessments. CONCLUSION: We demonstrated that a multi-modal evaluation of a new respirator can be rapidly conducted with a high level of participation in a controlled, consistent manner across multiple organisations. The evaluation results of the CE MSK-003 respirator exceeded our predetermined (a priori) minimal criteria, making it suitable for broad distribution to healthcare organisations.

Nitrite regulates hypoxic vasodilation via myoglobin-dependent nitric oxide generation.

BACKGROUND: Hypoxic vasodilation is a physiological response to low oxygen tension that increases blood supply to match metabolic demands. Although this response has been characterized for >100 years, the underlying hypoxic sensing and effector signaling mechanisms remain uncertain. We have shown that deoxygenated myoglobin in the heart can reduce nitrite to nitric oxide (NO·) and thereby contribute to cardiomyocyte NO· signaling during ischemia. On the basis of recent observations that myoglobin is expressed in the vasculature of hypoxia-tolerant fish, we hypothesized that endogenous nitrite may contribute to physiological hypoxic vasodilation via reactions with vascular myoglobin to form NO·. METHODS AND RESULTS: We show in the present study that myoglobin is expressed in vascular smooth muscle and contributes significantly to nitrite-dependent hypoxic vasodilation in vivo and ex vivo. The generation of NO· from nitrite reduction by deoxygenated myoglobin activates canonical soluble guanylate cyclase/cGMP signaling pathways. In vivo and ex vivo vasodilation responses, the reduction of nitrite to NO·, and the subsequent signal transduction mechanisms were all significantly impaired in mice without myoglobin. Hypoxic vasodilation studies in myoglobin and endothelial and inducible NO synthase knockout models suggest that only myoglobin contributes to systemic hypoxic vasodilatory responses in mice. CONCLUSIONS: Endogenous nitrite is a physiological effector of hypoxic vasodilation. Its reduction to NO· via the heme globin myoglobin enhances blood flow and matches O(2) supply to increased metabolic demands under hypoxic conditions.

Casey's conundrum, a review of the genus Semanotus Mulsant (Coleoptera: Cerambycidae: Cerambycinae: Callidiini) in North America.

The North American species of Semanotus Mulsant, 1839 are reviewed. Semanotus ligneus (Fabricius, 1787), Semanotus amplus amplus (Casey, 1912) new status, S. amplus basalis (Casey, 1924) new status, Semanotus amplus sequoiae Van Dyke, 1923 new status, Semanotus conformis Casey, 1924 new status, Semanotus terminatus Casey, 1912 new status, Semanotus amethystinus (LeConte, 1853), Semanotus juniperi (Fisher, 1915), and Semanotus litigiosus (Casey, 1891) are recognized as valid. A key to the North American fauna, as well as to Semanotus japonicus Lacordaire, 1869 and Semanotus australis Giesbert, 1993 is included. A morphometric study was conducted using 37 measured characters from adults of S. litigiosus. Data were examined using stepwise discriminant analysis to determine which characters aid in the diagnosis of taxa, and their amounts of resolving power using canonical variates analysis. Morphometrics revealed significant sexual dimorphism among taxa and suggested that three taxa were present, however, only two taxa could be reasonably separated using observable characters. Phylogenetic analysis using a hypothetical ancestor as out-group returned a single most parsimonious tree for North American Semanotus.

Quantitative fit-test concordance of a pair of similar-fit 3M Aura respirator models, 3M 9320A+ and 3M 1870+: A randomized crossover study.

OBJECTIVE: Some manufacturers provide information on similar-fit model pairings of filtering facepiece respirators (FFRs), suggesting that fit-test outcome of one model helps predict the other. This guidance may be useful during crisis capacity when FFR supplies and/or fit tests are constrained. The purpose of this study was to compare quantitative fit-test (QNFT) results and concordance between a pair of similar-fit 3M Aura FFRs: the 3M 9320A+ and 3M 1870+. METHODS: All participants completed online training and a QNFT with both respirators. The order of the respirator being examined first was randomly allocated. The outcomes included QNFT pass rate, concordance between the 2 models, overall and individual fit factors, and percentage of male and female participants who passed or failed the QNFT. RESULTS: We recruited 1,000 participants (668 females and 332 males). The QNFT pass rate, overall fit factors, and individual fit factors were significantly higher for the 3M 9320A+ than the 3M 1870+ FFR. The concordance between the models was "fair" (κ coefficient, 0.38). Male participants who passed a QNFT with either of the FFRs had 96% chance of passing the QNFT for the alternate model. Female participants who passed the 3M 1870+ had 97% chance of passing the QNFT for the 3M 9320A+ model. However, ∼1 in 12 females who passed the QNFT for the 3M 9320A+ failed the QNFT for the 3M 1870+ FFR. CONCLUSIONS: Similar-fit paired FFR models may provide a consequentially different level of respiratory protection, especially for women. Our findings are important for FFR stockpiling and fit-testing strategies, especially during crisis capacity.