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Vitamin B-12 deficiency is a major public health problem affecting individuals across the lifespan, with known hematological, neurological, and obstetric consequences. Emerging evidence suggests that vitamin B-12 may have an important role in other aspects of human health, including the composition and function of the gastrointestinal (gut) microbiome. Vitamin B-12 is synthesized and utilized by bacteria in the human gut microbiome and is required for over a dozen enzymes in bacteria, compared to only two in humans. However, the impact of vitamin B-12 on the gut microbiome has not been established. This systematic review was conducted to examine the evidence that links vitamin B-12 and the gut microbiome. A structured search strategy was used to identify in vitro, animal, and human studies that assessed vitamin B-12 status, dietary intake, or supplementation, and the gut microbiome using culture-independent techniques. A total of 22 studies (3 in vitro, 8 animal, 11 human observational studies) were included. Nineteen studies reported vitamin B-12 intake, status, or supplementation was associated with gut microbiome outcomes, including beta-diversity, alpha-diversity, relative abundance of bacteria, functional capacity, or short chain fatty acid production. Evidence suggests vitamin B-12 may be associated with changes in bacterial abundance. While results from in vitro studies suggest vitamin B-12 may increase alpha-diversity and shift gut microbiome composition (beta-diversity), findings from animal studies and observational human studies were heterogeneous. Based on evidence from in vitro and animal studies, microbiome outcomes may differ by cobalamin form and co-intervention. To date, few prospective observational studies and no randomized trials have been conducted to examine the effects of vitamin B-12 on the human gut microbiome. The impact of vitamin B-12 on the gut microbiome needs to be elucidated to inform screening and public health interventions. Statement of significance: Vitamin B-12 is synthesized and utilized by bacteria in the human gut microbiome and is required by over a dozen enzymes in bacteria. However, to date, no systematic reviews have been conducted to evaluate the impact of vitamin B-12 on the gut microbiome, or its implications for human health.

Liquid hog manure (LHM) and solid cattle manure (SCM) co-applied with biochar could beneficially influence soil nitrogen (N) and phosphorus (P) dynamics. A split-plot design was used at two sites (Brown and Black) in Saskatchewan to assess LHM and SCM (100 kg N·ha−1) applied alone or in combination with biochars (8 Mg carbon·ha−1) produced using slow or fast pyrolysis. Crop growth and nutrient uptake, along with fertilizer 15N recovery, were followed over four years in a cereal–oilseed rotation. Crop growth on the Brown soil was more responsive to the treatments than the Black soil, reflecting lower fertility of the Brown soil. The manure and biochar, applied alone or in combination, had little impact on available soil P. Manure and biochar effects on crop growth and nutrient uptake were attributed to temporal effects on soil N immobilization–mineralization influencing plant-available soil N. A negative impact of the fast pyrolysis biochar on growth and plant uptake was observed and attributable to its greater labile-carbon content, which likely promoted soil N immobilization. Synergism observed between SCM and the slow pyrolysis biochar may reflect enhanced net SCM-N mineralization and increased water-holding capacity. The majority (55%–80%) of plant 15N recovery occurred during the first year, with 86% of fertilizer 15N conserved within the soil–plant system after four years. Greater (40%) plant 15N recovery without biochar addition, coupled with increased (38%) soil 15N recovery with added biochar, suggests biochar-related immobilization and (or) sorption in the biochar-amended soils.

Termination by tillage is one strategy used for regenerating pasture stands. Yet, research gaps exist on how tillage affects carbon (C) and nitrogen (N) forms and amounts in western Canadian soils. We measured total soil organic C (SOC), dissolved organic C (DOC), total dissolved N (TDN), light fraction organic C (LFOC) and light fraction organic N (LFON), microbial biomass C (MBC) and microbial biomass N (MBN), and inorganic N as indicators of soil organic matter (SOM) dynamics. After tillage termination in fall 2018, we sampled soils (0–10 cm; 0–15 cm) under three legume species (alfalfa, cicer milkvetch, and sainfoin) three times (spring, summer, and fall of 2019) across three landscape positions. Legume species did not affect the measured parameters. Over time, tillage affected DOC, TDN, and inorganic N. Averaged across three pasture legumes and three landscape positions, tillage increased DOC 29% by summer. Fall-applied tillage led to 59% and 33% higher TDN in the succeeding summer and fall. Inorganic N increased by 14% and 40% across landscape positions and sampling after tillage. Averaged across landscape positions, MBC decreased by 31% from spring to summer and increased by 51% from summer to fall. However, MBN increased by 53% and decreased by 5% within the same period. The seasonal fluctuations in MBC and MBN reflected variations in moisture, temperature, and substrate quality. Total SOC, LFOC, and LFON increased on the upper slopes and fall sampling time. Although single intensive tillage did not affect total SOC, several tillage operations could accelerate SOM loss and reduced total C storage over time.

Cover crops have the potential to immobilize nitrogen (N) that would otherwise be lost before or after the main crop production, leading to improved N management. However, information on how cover crops influence N management in intensive vegetable cropping systems is scarce. This study aimed to determine how an overwintering rye cover crop impacts crop yield and N cycling, for three common prairie vegetable crops. From 2017 to 2019, a broccoli – sweet corn – root crop sequence was tested (in which all crops of rotation were present each year), with each crop type receiving five N fertilizer treatments, ranging from 0 to 300 kg N·ha−1. After harvest each year, sub-plots were established with vs. without a rye cover crop, and the effect on vegetable yield, soil inorganic N, and N use efficiency (NUE) was followed into the subsequent growing season. In most cases, the cover crop increased vegetable crop productivity and N content in the subsequent growing season. The cover crop also lowered soil inorganic N levels at vegetable planting but increased levels at harvest. Vegetable crop NUE indices were frequently improved with vs. without the cover crop. As for the N fertilizer response, increasing N fertilizer rate did not continually increase vegetable crop productivity and N content. Higher N fertilizer rates increased soil inorganic N levels at vegetable planting and harvest, and often lowered vegetable crop NUE indices. These results demonstrate the importance of adjusting soil N levels to better align with crop needs — and that including a rye cover crop in the vegetable rotation is one method of doing so.

A change in facility for the synthesis of the step 2 intermediate for talazoparib tosylate (1·TsOH) required the development of an alternative, intrinsically safer process. Rapid route scouting and...

We present 5--14~$\mu$m spectra at two different positions across the Orion Bar photodissociation region (PDR) obtained with the Infrared Spectrograph onboard the Spitzer Space Telescope and 3.3~$\mu$m PAH observations obtained with the Stratospheric Observatory for Infrared Astronomy (SOFIA). We aim to characterize emission from Polycyclic Aromatic Hydrocarbon (PAH), dust, atomic and molecular hydrogen, argon, sulfur, and neon as a function of distance from the primary illuminating source. We find that all the major PAH bands peak between the ionization front and the PDR front, as traced by H$_{2}$, while variations between these bands become more pronounced moving away from this peak into the face-on PDRs behind the PDR front and at the backside of the \HII\, region. While the relative PAH intensities are consistent with established PAH characteristics, we report unusual behaviours and attribute these to the PDR viewing angle and the strength of the FUV radiation field impinging on the PDRs. We determine the average PAH size which varies across the Orion Bar. We discuss subtle differences seen between the cationic PAH bands and highlight the photo-chemical evolution of carbonaceous species in this PDR environment. We find that PAHs are a good tracer of environmental properties such as the strength of the FUV radiation field and the PAH ionization parameter., Comment: 25 pages, 19 Figures. To be published in MNRAS

3,5-Diiodo-8-aryl-BODIPYs are prepared from the corresponding 3,5-dichloro- and 3,5-dibromo- congeners via a double aromatic Finkelstein reaction with NaI in refluxing propionitrile, in yields from 51 to 100%. Rate enhancement is observed for 3,5-dibromo-BODIPYs (ArBr > ArCl) suggesting a concerted SNAr mechanism for this transformation. Thus formed 3,5-diiodo-BODIPYs are examined as coupling partners in Migita-Kosugi-Stille, Mizoroki-Heck, Sonogashira, and Suzuki-Miyaura reactions, providing routes toward complex BODIPY architectures.

Chronic disruption of rhythms (CDR) impacts sleep and can result in circadian misalignment of physiological systems which, in turn, is associated with increased disease risk. Exposure to repeated or severe stressors also disturbs sleep and diurnal rhythms. Prebiotic nutrients produce favorable changes in gut microbial ecology, the gut metabolome, and reduce several negative impacts of acute severe stressor exposure, including disturbed sleep, core body temperature rhythmicity, and gut microbial dysbiosis. In light of previous compelling evidence that prebiotic diet broadly reduces negative impacts of acute, severe stressors, we hypothesize that prebiotic diet will also effectively mitigate the negative impacts of chronic disruption of circadian rhythms on physiology and sleep/wake behavior. Male, Sprague Dawley rats were fed diets enriched in prebiotic substrates or calorically matched control chow. After 5 weeks on diet, rats were exposed to CDR (12 h light/dark reversal, weekly for 8 weeks) or remained on undisturbed normal light/dark cycles (NLD). Sleep EEG, core body temperature, and locomotor activity were recorded via biotelemetry in freely moving rats. Fecal samples were collected on experimental days –33, 0 (day of onset of CDR), and 42. Taxonomic identification and relative abundances of gut microbes were measured in fecal samples using 16S rRNA gene sequencing and shotgun metagenomics. Fecal primary, bacterially modified secondary, and conjugated bile acids were measured using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Prebiotic diet produced rapid and stable increases in the relative abundances of Parabacteroides distasonis and Ruminiclostridium 5. Shotgun metagenomics analyses confirmed reliable increases in relative abundances of Parabacteroides distasonis and Clostridium leptum, a member of the Ruminiclostridium genus. Prebiotic diet also modified fecal bile acid profiles; and based on correlational and step-wise regression analyses, Parabacteroides distasonis and Ruminiclostridium 5 were positively associated with each other and negatively associated with secondary and conjugated bile acids. Prebiotic diet, but not CDR, impacted beta diversity. Measures of alpha diversity evenness were decreased by CDR and prebiotic diet prevented that effect. Rats exposed to CDR while eating prebiotic, compared to control diet, more quickly realigned NREM sleep and core body temperature (ClockLab) diurnal rhythms to the altered light/dark cycle. Finally, both cholic acid and Ruminiclostridium 5 prior to CDR were associated with time to realign CBT rhythms to the new light/dark cycle after CDR; whereas both Ruminiclostridium 5 and taurocholic acid prior to CDR were associated with NREM sleep recovery after CDR. These results support our hypothesis and suggest that ingestion of prebiotic substrates is an effective strategy to increase the relative abundance of health promoting microbes, alter the fecal bile acid profile, and facilitate the recovery and realignment of sleep and diurnal rhythms after circadian disruption.

Objective Calcium is an essential mineral involved in the functioning of nearly every human cell. Calcium levels are regulated by dietary absorption, vitamin D status, and parathyroid hormone (PTH). This report describes a patient in whom childhood bowel resection and partial gastrectomy resulted in malabsorptive hypocalcemia in adulthood. Case Report A 21-year-old man presented with syncope and a fall resulting in a right femoral neck fracture. His medical history included small bowel obstructions at age 9 requiring bowel resection, and at age 12 with gastric perforation and partial gastrectomy. Laboratory values showed calcium level of 4.9 mg/dL (8.9-10.3 mg/dL). PTH level was 273 pg/mL (12.0-88.0 pg/mL), 25-hydroxy-vitamin D was 28 ng/dL (30-100 ng/mL), and 1,25-dihydroxy-vitamin D was 54 pg/dL (18-72 pg/mL). Furthermore, magnesium and phosphorus levels were 2.1 mg/dL (1.5-2.6 mg/dL) and 4.4 mg/dL (2.4-4.7 mg/dL), respectively. Calcium levels improved to 9.5 mg/dL on 10% calcium gluconate drip but could not be maintained above 7 mg/dL on oral calcium carbonate supplementation, despite doses as high as 3750 mg three times daily with calcitriol 0.75 mcg twice daily. After switching from calcium carbonate to calcium citrate 3500 mg three times daily, the calcium level improved and was maintained between 8.3 and 9.0 mg/dL. Discussion High calcium needs, other nutrient deficiencies, and response to calcium citrate versus calcium carbonate suggest malabsorption from achlorhydria and small bowel resection. Conclusion This case emphasizes the gastrointestinal physiology in calcium homeostasis and highlights the recognition of hypocalcemia as a complication of gastric and bowel resection.

Purpose A major difficulty in monoclonal antibody (mAb) therapeutic development is product aggregation. In this study, intermolecular isopeptide bonds in mAb aggregates were characterized for the first time. We aim to propose a mechanism of covalent aggregation in a model antibody using stressed studies at raised temperatures to aid in the understanding of mAb aggregation pathways. Methods Aggregate fractions were generated using raised temperature and were purified using size-exclusion chromatography (SEC). The fractions were tryptically digested and characterized using liquid chromatography hyphenated to tandem mass-spectrometry (LC–MS/MS). Results An increased amount of clipping between aspartic acid and proline in a solvent accessible loop in the constant heavy 2 (CH2) domain of the mAb was observed under these conditions. Detailed peptide mapping revealed 14 isopeptide bonds between aspartic acid at that cleavage site and lysine residues on adjacent antibodies. Two additional isopeptide bonds were identified between the mAb HC N-terminal glutamic acid or a separate aspartic acid to lysine residues on adjacent antibodies. Conclusions Inter-protein isopeptide bonds between the side chains of acidic amino acids (aspartate and glutamate) and lysine were characterized for the first time in mAb aggregates. A chemical mechanism was presented whereby spontaneous isopeptide bond formation could be facilitated via either the aspartic acid side chain or C-terminus.

In 2017, the Nuclear Forensics International Technical Working Group organized their fifth Collaborative Materials Exercise (CMX-5). The exercise samples were two uranium dioxide fuel pellets manufactured from the same starting materials by different processes to have similar bulk isotopic composition, but different spatial uranium isotopic distributions. Sets of identical materials were sent to all participating laboratories, who then utilized their existing nuclear forensic capabilities to independently analyse fuel pellets and identify similarities and differences of the materials’ characteristics. Here we present and compare the ability of different analytical techniques to spatially resolve uranium isotopic heterogeneity in the uranium oxide fuel pellets.

For years, organophosphorus poisoning has been a major concern of health problems throughout the world. An estimated 200,000 acute pesticide poisoning deaths occur each year, many in developing countries. Apart from the agricultural pesticide poisoning, terrorists have used these organophosphorus compounds to attack civilian populations in some countries. Recent misuses of sarin in the Syrian conflict had been reported in 2018. Since the 1950s, the therapy to overcome this health problem is to utilize a reactivator to reactivate the inhibited acetylcholinesterase by these organophosphorus compounds. However, many questions remain unanswered regarding the efficacy and toxicity of this reactivator. Pralidoxime, MMB-4, TMB-4, obidoxime, and HI-6 are the examples of the established oximes, yet they are of insufficient effectiveness in some poisonings and only a limited spectrum of the different nerve agents and pesticides are being covered. Alternatively, an option in the treatment of organophosphorus poisoning that has been explored is through the use of enzyme therapy. Organophosphorus hydrolases are a group of enzymes that look promising for detoxifying organophosphorus compounds and have recently gained much interest. These enzymes have demonstrated remarkable protective and antidotal value against some different organophosphorus compounds in vivo in animal models. Apart from that, enzyme treatments have also been applied for decontamination purposes. In this review, the restrictions and obstacles in the therapeutic development of oximes, along with the new strategies to overcome the problems, are discussed. The emerging interest in enzyme treatment with its advantages and disadvantages is described as well.

Gene therapy continues to be a promising contender for the treatment of cystic fibrosis (CF) airway disease. We have previously demonstrated that airway conditioning with lysophosphatidylcholine (L...

A variety of normal operation and accident scenarios can generate thermal stresses large enough to cause cracking in ceramic fuel pellets. Cracking in fuel pellets can lead to reduced heat removal,...

Purpose To investigate corneal stiffness parameters (SPs) as predictors of future progression risk in glaucoma suspect eyes. Design Prospective, longitudinal study. Participants Three hundred seventy-one eyes from 228 primary open-angle glaucoma suspects, based on optic disc appearance, with normal baseline Humphrey Visual Field (HVF; Carl Zeiss Meditec) results. Methods Baseline corneal SPs were measured using Corvis ST (Oculus Optikgerate GmbH). Participants were followed up every 6 months with clinical examination, HVF testing, and OCT. The baseline SP at first applanation (SP-A1) and highest concavity predicted the prospective outcome measures. Main Outcome Measures Structural progression was measured by the OCT rate of thinning of the retinal nerve fiber layer (RNFL) and ganglion cell–inner plexiform layer (GCIPL). Functional progression was assessed by permutation analysis of pointwise linear regression criteria on HVF testing. Results Stiffness parameters correlated positively with central corneal thickness (CCT), which was adjusted for in all analyses. A higher SP-A1, suggestive of a stiffer cornea, was associated with a faster rate of RNFL thinning (P Conclusions Glaucoma suspect eyes with higher corneal SPs and lower CCT, suggestive of thin and stiff corneas, are at greater risk of progression. Corneal SPs seem to act synergistically with CCT as risk factors for glaucoma progression.

The ages of sedimentation and copper-silver mineralization in the late Meso- to Neoproterozoic Kalahari Copperbelt in Botswana, an economically significant copper province, have previously been poorly constrained within an ~600 m.y. period that spans the Neoproterozoic from the assembly and breakup of Rodinia to the assembly of Gondwana. Rhenium-osmium geochronology of molybdenite and copper sulfide minerals and U-Th-Pb laser ablation split-stream inductively coupled plasma-mass spectrometry (LASS ICP-MS) analysis of xenotime grains are utilized to provide absolute and relative age data on the host rocks and mineralizing events within the Ghanzi Ridge region of the Kalahari Copperbelt. The data reveal a prolonged history of events, which is partially comparable with depositional and mineralizing events in the neighboring Central African Copperbelt. Abundant disseminated molybdenite is located within a shale layer near the base of the Proterozoic D’Kar Formation at the Northeast Mango Two deposit. Unusual molybdenite textures suggest organic matter may have been a precursor. Two molybdenite separates from a small calcite-molybdenite stringer in a wall-rock fragment that is enclosed within an epigenetic quartz-calcite-chalcopyrite vein with ill-defined and mismatched margins yielded Re-Os ages of 981 ± 3 and 981 ± 7 Ma. These ages indicate an early hydrothermal mineralizing event in the basin. A xenotime inclusion intergrown with molybdenite and chalcopyrite within the epigenetic vein yielded a younger U-Th-Pb age of 538 ± 8 Ma, suggesting two mineralizing events are preserved in a complex 6-cm-wide vein. Based on vein texture and alteration, the ages represent an ~981 Ma calcite-molybdenite mineralization event overprinted by an ~538 Ma quartz-chalcopyrite-molybdenite mineralization event, perhaps during reopening of the vein. Re-Os and U-Th-Pb geochronology were utilized at the Zone 6 deposit on minerals associated with a hydrothermal quartz-calcite-chalcocite-idaite-bornite vein. Several authigenic xenotime grains that occur along the margin of the vein yielded three concordant U-Th-Pb ages that indicate xenotime growth at ~950 to 925 Ma while other xenotime grains in a similar position yielded mostly discordant data, suggesting disturbance of the isotopic system in the xenotime grains. A coprecipitated chalcocite-idaite mixture within the hydrothermal vein produced an Re-Os age of 549.0 ± 11.2 Ma. Re-Os analysis obtained from a coprecipitated molybdenite-bornite mixture at the Northeast Fold deposit yielded an age of 515.9 ± 2 Ma. Together, the earliest Neoproterozoic Re-Os molybdenite and U-Th-Pb xenotime ages provide both a minimum depositional age constraint for the lowermost D’Kar Formation and clear evidence that diagenetic hydrothermal mineralizing events took place within the Ghanzi basin. The timing of this mineralizing event corresponds with a poorly documented regional thermal event that affected the northern margin of the Kalahari craton during the final stages of the assembly of Rodinia at ~980 Ma. The lower to middle Ghanzi Group of the Kalahari Copperbelt is at least 100 m.y. older than the host rocks within the neighboring Central African Copperbelt, which are associated with the breakup of Rodinia. The latest Neoproterozoic to Cambrian Re-Os and U-Th-Pb ages indicate that hydrothermal copper-silver mineralizing events occurred during the Pan-African (~600–480 Ma) fold-thrust evolution of the Ghanzi-Chobe zone and were broadly synchronous with widespread epigenetic hydrothermal copper-cobalt mineralizing events in the adjacent Central African Copperbelt.

Synthetically straightforward pi-extension of helically chiral N,N,O,O-boron chelated borondipyrromethenes (BODIPYs) has been achieved by incorporation of naphthyl rings into the BODIPY 3- and 5-positions. The effect of this extension on the photophysical and chiroptical properties is strongly dependent on the regiochemistry of the naphthyl substituents (4 exo, 4 lin, 4 endo). The most helicene-like regioisomer 4 endo is a near infra-red (NIR) emitter (lambda(em)(max)=751 nm) with an emission band between 675-875 nm, displaying circularly polarised luminescence (CPL) with a luminescence dissymmetry factor, vertical bar g(lum)vertical bar=5.7x10(-3) and CPL brightness, B-CPL=27, rivalling the best NIR CPL-emitting small organic molecules reported to date.

Nineteen host plant volatiles (HPVs) were screened for attractivity to adult codling moth Cydia pomonella (L.) as a fourth component of core blends (3K) including (E,Z)-2,4-ethyl decadienoate, (E)-4,8-dimethyl-1,3,7-nonatriene and acetic acid. Each new quaternary combination was compared with a previously reported attractive bisexual lure (4K), consisting of the 3K blend plus 6-ethenyl-2,2,6-trimethyloxan-3-ol (pyranoid linalool oxide, pyrLOX). All lure evaluations were conducted in apple, Malus domestica (Borkhausen). Several compounds were found to significantly lower total and/or female catches when added to the 3K blend, including (Z)-3-hexenol, (E)-2-hexanal and hexyl butanoate (female and total moths), and (Z)-3-hexenyl acetate and linalool (female moths). Other compounds when added to the 3K blend did not increase or decrease moth catches, including methyl salicylate, (E)-beta-ocimene, limonene, beta-caryophyllene, butyl hexanoate, farnesol, terpineol, terpinen-4-ol and alpha-pinene. A few added compounds significantly increased moth catches compared with the 3K blend, including beta-pinene (male moths), (Z)-jasmone (male and total moths), (E)-beta-farnesene and beta-myrcene (female and total moths), and (E,E)-alpha-farnesene (male, female, and total moths). In addition, each of these five compounds when added to the 3K core blend performed similarly to the 4K lure (male, females, and total moths). Further studies should expand these results through tests of these and other new blends with a range of component ratios and total loading amounts. Field trials should also be replicated within all host crops of codling moth and across major geographical production regions.

Abstract The iridium-catalyzed C-H borylation of diethyl phenylphosphonate results in nonselective mono and bisborylation to afford a near statistical mixture of 3-, 3,5- and 4-boryl substituted aryl phosphonates whereas 3-substituted aryl phosphonates undergo highly regioselective C-H borylation to afford the corresponding meta-phosphonate substituted arylboronic esters as the sole product; the resulting boronic esters were used as nucleophilic reagents in a subsequent palladium-catalyzed Suzuki–Miyaura cross-coupling to generate a range of biarylmonophosphonates. Gratifyingly, the Suzuki–Miyaura cross-coupling can be conducted without purifying the boronic ester which greatly simplifies the synthetic procedure. Graphical Abstract

A simple, low cost and rapid analytical method for determination of HCl concentration after being treated with cellulose and cellulose nanofibers (CNF) is developed. This method is based on color intensity after the HCl solution is doped with sodium iodide (NaI). The color of HCl solution changes from colorless to yellow. The intensity of the color is measured by UV – Visible spectroscopy. The UV-Visible spectra of 0.15 M HCl treated with cellulose and cellulose nanofibers is reduced from its initial concentration. The CNF absorption capacity is higher as compared to cellulose. FT-IR analysis showed that there is interaction between C-H group from the CNF backboned and chloride ion from HCl solution.

1. Neonicotinoid insecticides are the most widely used class of insecticides in the world and can have both lethal and sub-lethal effects on non-target organisms in agricultural areas. Monarch butterflies (Danaus plexippus) have experienced dramatic declines in recent decades and, given that a large proportion of milkweed on the landscape grows in agricultural areas, there is concern about the negative effects of neonicotinoids on this non-target insect. 2. In the field, we exposed common milkweed (Asclepias syriaca), an obligate host plant of monarch butterflies, to agriculturally-realistic levels of clothianidin, a widely used neonicotinoid insecticide. We tested whether this treatment influenced the number of eggs laid and larval survival over two years. 3. Milkweeds were transplanted into 60 experimental plots alongside a corn crop planted with a clothianidin seed-coat and 60 control plots alongside an untreated corn crop. The number of eggs, larvae at each stage (first-fifth instar), and presence of other arthropods were recorded weekly from June to the end of August and survival from egg to fifth instar was estimated using a Bayesian state-space statistical model. 4. We counted more eggs in treated plots compared to control plots, suggesting a preference for treated milkweed. The number of plots with arthropods did not differ between treatments, but within treated plots, there was a greater decrease in the number of arthropods throughout the season. There was no evidence that monarchs selected plots with fewer arthropods for oviposition. Larval survival was lower in clothianidin-treated plots compared to control plots. 5. Our results suggest milkweed near clothianidin-treated crops can reduce larval survival of monarch butterflies. While we provide some evidence that clothianidin could also act as an ecological trap for this species, further work is needed to identify additional components of fitness, including individual egg-laying rates and survival beyond the pupal stage. Our findings add to a growing body of evidence that neonicotinoids can negatively affect non-target organisms.

Therapeutic development of histone deacetylase inhibitors (HDACi) has been hampered by a number of barriers to drug delivery, including poor solubility and inadequate tissue penetration. Nanoparticle encapsulation could be one approach to improve the delivery of HDACi to target tissues; however, effective and generalizable loading of HDACi within nanoparticle systems remains a long-term challenge. We hypothesized that the common terminally ionizable moiety on many HDACi molecules could be capitalized upon for loading in polymeric nanoparticles. Here, we describe the simple, efficient formulation of a novel library of β-cyclodextrin-poly (β-amino ester) networks (CDN) to achieve this goal. We observed that network architecture was a critical determinant of CDN encapsulation of candidate molecules, with a more hydrophobic core enabling effective self-assembly and a PEGylated surface enabling high loading (up to ∼30% w/w), effective self-assembly of the nanoparticle, and slow release of drug into aqueous media (up to 24 days) for the model HDACi panobinostat. We next constructed a library of CDNs to encapsulate various small, hydrophobic, terminally ionizable molecules (panobinostat, quisinostat, dacinostat, givinostat, bortezomib, camptothecin, nile red, and cytarabine), which yielded important insights into the structural requirements for effective drug loading and CDN self-assembly. Optimized CDN nanoparticles were taken up by GL261 cells in culture and a released panobinostat was confirmed to be bioactive. Panobinostat-loaded CDNs were next administered by convection-enhanced delivery (CED) to mice bearing intracranial GL261 tumors. These studies confirm that CDN encapsulation enables a higher deliverable dose of drug to effectively slow tumor growth. Matrix-assisted laser desorption/ionization (MALDI) analysis on tissue sections confirms higher exposure of tumor to drug, which likely accounts for the therapeutic effects. Taken in sum, these studies present a novel nanocarrier platform for encapsulation of HDACi via both ionic and hydrophobic interactions, which is an important step toward better treatment of disease via HDACi therapy.

Orbital frequency analyzer (OFA) is one of the electrostatic ion trap mass analysers for FTMS, which offers ultrahigh mass resolution and high ion charge capacity. In order to analyze multiply charged proteins and other large biological particles by means of charge detection mass spectrometry, a data processing algorithm was created to suit the image charge signal of nonharmonic waveform nature. The algorithm is capable of detecting collisions between ions and residual gas molecules, to determine lifetime of ions, and to evaluate the charge and mass values for ions having lifetime above a threshold. With the filtering of the lifetime and charge value, the chemical noise from small molecules and protein fragments can be eliminated in the reconstructed spectrum, facilitating measurement of protein content at a very low concentration, down to tens of nanomolars. The standard deviation of charge measurement is between 1.1 to 1.8 e for ions with oscillation lifetimes from 1 to 0.4 s, and this in turn determines the CDMS spectrum mass peak width. It has been found that the lower voltage setting of the OFA results in a larger population of ions surviving for longer times and thus produces narrower mass peak width. While OFA is able to run multiplexed CDMS without ion motion interference, coexisting ions of the same or very close m/z can cause interference between their image charge signals, which increases the error in charge determination.

The occurrence of aerobic biodegradation in the vadose zone between a subsurface source and a building foundation can all‐but eliminate the risks from methane and petroleum vapor intrusion (PVI). Understanding oxygen availability and the factors that affect it (e.g., building sizes and their distribution) are therefore critical. Uncovered ground surfaces allow oxygen access to the subsurface to actively biodegrade hydrocarbons (inclusive of methane). Buildings can reduce the net flux of oxygen into the subsurface and so reduce degradation rates. Here we determine when PVI and methane risk is negligible and/or extinguished; defined by when oxygen is present across the entire sub‐slab region of existing or planned slab‐on‐ground buildings. We consider all building slab sizes, all depths to vapor sources and the effect of spacings between buildings on the availability of oxygen in the subsurface. The latter becomes critical where buildings are in close proximity or when increased building density is planned. Conservative assumptions enable simple, rapid and confident screening should sites and building designs comply to model assumptions. We do not model the aboveground “building” processes (e.g., air exchange), and assume the slab‐on‐ground seals the ground surface so that biodegradation of hydrocarbons is minimized under the built structure (i.e., the assessment remains conservative). Two graphs represent the entirety of the outcomes that allow simple screening of hydrocarbon vapors based only on the depth to the source of vapors below ground, the concentration of vapors within the source, the width of the slab‐on‐ground building, and the gap between buildings; all independent of soil type. Rectangular, square, and circular buildings are considered. Comparison with field sites and example applications are provided, along with a simple 8‐step screening guide set in the context of existing guidance on PVI assessment.

BACKGROUND Ocular coherence tomography angiography (OCTA) is available in varying size and resolution. We sought to characterise associations of cardiometabolic factors with retinal microvascular changes using 3 × 3, 6 × 6 and 8 × 8-mm OCTA scans to determine differences in detection with varying scan size. METHODS Cross-sectional study of 247 cardiovascular patients from a single-centre tertiary-care hospital. Demographic, comorbidity and medication data were obtained. Patients underwent 3 × 3, 6 × 6 and 8 × 8-mm macula OCTA scanning using Carl Zeiss CIRRUS HD-OCT Model 5000. Angioplex and AngioTool software was used to quantify vascular parameters in the superficial capillary plexus. RESULTS Increasing age, hypertension, dyslipidaemia, diabetes, chronic kidney disease, coronary artery disease and peripheral vascular disease were associated with reductions in vessel density, vessel perfusion, average vessel length and/or junction density in 3 × 3-mm OCTA (P

Besides the ecotoxicological consequences of microplastics and associated chemicals, the association of microbes on plastics has greater environmental implications as microplastics may select for unique microbiome participating in environmentally significant functions. Despite this, the functional potential of the microbiome associated with different types of plastics is understudied. Here, we investigate the interaction between plastic and marine biofilm-forming microorganisms through a whole-genome sequencing approach on four types of microplastics incubated in the marine environment. Taxonomic analysis suggested that the microplastic surfaces exhibit unique microbial profiles and niche partitioning among the substrates. In particular, the abundance of Vibrio alginolyticus and Vibrio campbellii suggested that microplastic pollution may pose a potential risk to the marine food chain and negatively impact aquaculture industries. Microbial genera involved in xenobiotic compound degradation, carbon cycling, and genes associated with the type IV secretion system, conjugal transfer protein TraG, plant-pathogen interaction, CusA/CzcA family heavy metal efflux transfer proteins, and TolC family proteins were significantly enriched on all the substrates, indicating the variety of processes operated by the plastic-microbiome. The present study gives a detailed characterization of the rapidly altering microbial composition and gene pools on plastics and adds new knowledge surrounding the environmental ramifications of marine plastic pollution.

The correct choice of formulation buffer is a critical aspect of drug development and is chosen primarily to improve the stability of a protein therapeutic and protect against degradation. Amino acids are frequently incorporated into formulation buffers. In this study we have identified and characterized light induced cross-links between the side chain of histidine residues in an IgG4 monoclonal antibody and different amino acids commonly used in formulation buffers. These reactions have the potential to impact the overall product quality of the drug. The structure of each cross-link identified was elucidated using high performance liquid chromatography (HPLC) hyphenated to tandem mass spectrometry (MS/MS) with higher energy collisional dissociation (HCD). Furthermore, we speculate on the role of amino acids in formulation buffers and their influence on mAb stability. We theorize that whilst the adduction of formulation buffer amino acids could have a negative impact on product quality, it may protect against other pathways of photo-degradation.

High night temperatures (HNT) are shown to significantly reduce rice (Oryza sativa L.) yield and quality. A better understanding of the genetic architecture of HNT tolerance will help rice breeders to develop varieties adapted to future warmer climates. In this study, a diverse indica rice panel displayed a wide range of phenotypic variability in yield and quality traits under control night (24 °C) and higher night (29 °C) temperatures. Genome-wide association analysis revealed 38 genetic loci associated across treatments (18 for control and 20 for HNT). Nineteen loci were detected with the relative changes in the traits between control and HNT. Positive phenotypic correlations and co-located genetic loci with previously cloned grain size genes revealed common genetic regulation between control and HNT, particularly grain size. Network-based predictive models prioritized 20 causal genes at the genetic loci based on known gene/s expression under HNT in rice. Our study provides important insights for future candidate gene validation and molecular marker development to enhance HNT tolerance in rice. Integrated physiological, genomic, and gene network-informed approaches indicate that the candidate genes for stay-green trait may be relevant to minimizing HNT-induced yield and quality losses during grain filling in rice by optimizing source-sink relationships.

Funder: UCL Grand Challenges, Ethnographers frequently allude to alcoholism and related harms in Indigenous hunter-gatherer communities, but very few studies have quantified patterns of alcohol consumption or its health and social impacts. We present a case study of the Mbendjele BaYaka, a Congolese population undergoing socioeconomic transition. 83 adults answered questions about their frequency and quantity of alcohol consumption, underwent biometric measurements and reported whether they were currently experiencing a cough or diarrhoea; 56 participated in structured interviews about their experiences with alcohol. Based on WHO standards, we found 44.3% of the full sample, and 51.5% of drinkers (excluding abstainers), had a hazardous volume of alcohol consumption; and 35.1% of the full sample, and 40.9% of drinkers, engaged in heavy episodic drinking; consumption habits varied with sex and age. Total weekly consumption was a positive predictor of blood pressure and the likelihood of experiencing diarrhoea; associations with other biometric variables were not statistically significant. Interview responses indicated numerous other economic, mental and physical health harms of alcohol use, the prevalence of which demonstrate some variability between forest camps and permanent village settlements. These include high rates of drinking during pregnancy and breastfeeding (~40%); frequent alcohol-induced violence; and considerable exchange of foraged foods and engagement in exploitative labour activities to acquire alcohol or repay associated debts. Our findings demonstrate the prevalence of hazardous alcohol consumption among transitioning hunter-gatherers is higher than other segments of the Congolese population and indicate negative impacts on health and wellbeing, highlighting an urgent need for targeted public health interventions.

The discovery of the CRISPR-Cas9 protein has enabled facile genome editing in living cells and organisms. Structural and biochemical studies of Cas9 endonucleases have provided critical information about the core molecular requirements of the RNA-guided DNA cleavage reaction, but questions remain about how this bacterial protein navigates chromatin to identify DNA targets within living eukaryotic cells. In particular, the in vivo kinetics of on- versus off-target binding and Cas9 dependence on chromatin environment remain largely unknown. Here we present a single-molecule analysis of Cas9 searching in living mammalian cells. We provide evidence for a diffusion- dominated search mechanism and show that off-target binding at PAMs and short seed sequences is, on average, short-lived (milleseconds to seconds) by both single- molecule and bulk imaging techniques. The differential behavior of Cas9 in closed-off (heterochromatic) versus open (euchromatic) regions of the genome is also explored via heterochromatin protein 1 (HP1) staining and nuclear masking of single-particle trajectories. Comparative analysis of trajectories from these two regions suggests that Cas9 undersamples heterochromatin and moves more compactly within these regions. These data provide the first direct visualization of Cas9 searching in living cells and offer quantitative insights into how Cas9 navigates hierarchical organization of DNA within a eukaryotic nucleus. We additionally present mechanistic investigations of unrelated CRISPR-C2c2 proteins and identify two distinct enzymatic activities– pre- CRISPR RNA (pre-crRNA) processing and crRNA-stimulated general RNase activity. Collectively, this work expands our mechanistic understanding of CRISPR biology and highlights the utility of these enzymatically diverse proteins to be harnessed for biotechnological applications.

The biological activities of chemokine (C-C motif) ligand 2 (CCL2) are mediated via C-C chemokine receptor-2 (CCR2). Increased CCL2 level is associated with metastasis of many cancers. In this study, we investigated the role of the CCL2/CCR2 axis in the development of spontaneous intestinal tumorigenesis using the ApcMin/+ mouse model. Ablation of CCR2 in ApcMin/+ mice significantly increased the overall survival and reduced intestinal tumor burden. Immune cell analysis showed that CCR2-/- ApcMin/+ mice exhibited significant reduction in the myeloid cell population and increased IFNγ producing T cells both in spleen and mesenteric lymph nodes (mLN) compared to ApcMin/+ mice. The CCR2-/- ApcMin/+ tumors showed significantly reduced levels of IL-17 and IL-23 and increased IFNγ and Granzyme B compared to ApcMin/+ tumors. Transfer of CCR2+/+ ApcMin/+ CD4+ T cells into Rag2-/- mice led to development of colitis phenotype with increased CD4+ T cells hyper proliferation and IL-17 production. In contrast, adoptive transfer of CCR2-/- ApcMin/+ CD4+ T cells into Rag2-/- mice failed to enhance colonic inflammation or IL-17 production. These results a suggest novel additional role for CCR2, where it regulates migration of IL-17 producing cells mediating tumor-promoting inflammation in addition to its role in migration of tumor associated macrophages. This article is protected by copyright. All rights reserved.

CBD-2115 was selected from a library of 148 compounds based on a pyridinyl-indole scaffold as a first-in-class 4R-tau radiotracer. In vitro binding assays showed [3H]CBD-2115 had a KD value of 6.9 nM and a nominal Bmax of 500 nM in 4R-tau expressing P301L transgenic mouse tissue. In binding assays with human brain tissue homogenates, [3H]CBD-2115 has a higher affinity (4.9 nM) for progressive supranuclear palsy specific 4R-tau deposits than [3H]flortaucipir (45 nM) or [3H]MK-6240 (>50 nM). [18F]CBD-2115 was reliably synthesized (3-11% radiochemical yield with molar activity of 27-111 GBq/μmol and >97% radiochemical purity). Dynamic PET imaging was conducted in mice, rats, and nonhuman primates, and all species showed initial brain uptake of 0.5-0.65 standardized uptake value with fast clearance from normal tissues. [3H]CBD-2115 could be a useful lead radioligand for further research in 4R-tauopathies, and PET radiotracer development will focus on improving brain uptake and binding affinity.

Biomechanical changes to the collagen fibrillar architecture in articular cartilage are believed to play a crucial role in enabling normal joint function. However, experimentally there is little quantitative knowledge about the structural response of the Type II collagen fibrils in cartilage to cyclic loading in situ, and the mechanisms that drive the ability of cartilage to withstand long-term repetitive loading. Here we utilize synchrotron small-angle X-ray scattering (SAXS) combined with in-situ cyclic loading of bovine articular cartilage explants to measure the fibrillar response in deep zone articular cartilage, in terms of orientation, fibrillar strain and inter-fibrillar variability in healthy cartilage and cartilage degraded by exposure to the pro-inflammatory cytokine IL-1β. We demonstrate that under repeated cyclic loading the fibrils reversibly change the width of the fibrillar orientation distribution whilst maintaining a largely consistent average direction of orientation. Specifically, the effect on the fibrillar network is a 3-dimensional conical orientation broadening around the normal to the joint surface, inferred by 3D reconstruction of X-ray scattering peak intensity distributions from the 2D pattern. Further, at the intrafibrillar level, this effect is coupled with reversible reduction in fibrillar pre-strain under compression, alongside increase in the variability of fibrillar pre-strain. In IL-1β degraded cartilage, the collagen rearrangement under cyclic loading is disrupted and associated with reduced tissue stiffness. These finding have implications as to how changes in local collagen nanomechanics might drive disease progression or vice versa in conditions such as osteoarthritis and provides a pathway to a mechanistic understanding of such diseases. Statement of significance Structural deterioration in biomechanically loaded musculoskeletal organs, e.g., joint osteoarthritis and back pain, are linked to breakdown and changes in their collagen-rich cartilaginous tissue matrix. A critical component enabling cartilage biomechanics is the ultrastructural collagen fibrillar network in cartilage. However, experimental probes of the dynamic structural response of cartilage collagen to biomechanical loads are limited. Here, we use X-ray scattering during cyclic loading (as during walking) on joint tissue to show that cartilage fibrils resist loading by a reversible, three-dimensional orientation broadening and disordering mechanism at the molecular level, and that inflammation reduces this functionality. Our results will help understand how changes to small-scale tissue mechanisms are linked to ageing and osteoarthritic progression, and development of biomaterials for joint replacements., Graphical abstract Image, graphical abstract

Acephate (Ac) is an organophosphate (OP) compound, which is able to inhibit the activity of acetylcholinesterase. Thus, the aim of this study was to optimize the detection of Ac using a thiolated acephate binding aptamer-citrate capped gold nanoparticle (TABA–Cit-AuNP) sensor that also incorporated an image processing technique. The effects of independent variables, such as the incubation period of TABA–Cit-AuNPs (3–24 h) for binding TABA to Cit-AuNPs, the concentration of phosphate buffer saline (PBS) (0.001–0.01 M), the concentration of thiolated acephate binding aptamer (TABA) (50–200 nM), and the concentration of magnesium sulphate (MgSO4) (1–300 mM) were investigated. A quadratic model was developed using a central composite design (CCD) from response surface methodology (RSM) to predict the sensing response to Ac. The optimum conditions such as the concentration of PBS (0.01 M), the concentration of TABA (200 nM), the incubation period of TABA–Cit-AuNPs (3 h), and the concentration of MgSO4 (1 mM) were used to produce a TABA–Cit-AuNPs sensor for the detection of Ac. Under optimal conditions, this sensor showed a detection ranging from 0.01 to 2.73 μM and a limit of detection (LOD) of 0.06 μM. Real sample analysis demonstrated this aptasensor as a good analytical method to detect Ac.

Disclosed in this communication is a thorough study on the dearomative addition of organomagnesium nucleophiles to N-alkyl pyridinium electrophiles. The regiochemical outcomes have observable and predictable trends associated with the substituent patterns on the pyridinium electrophile. Often, the substituent effects can be either additive, giving high selectivities, or ablative, giving competing outcomes. Additionally, the nature of the organometallic nucleophilic component was also investigated for its role in the regioselective outcome. The effects of either reactive component are important to both the overall reactivity and site of nucleophilic addition. The utility of these observed trends is demonstrated in a concise, dearomative synthesis of a tricyclic compound shown to have insecticidal activity.