Search

Abstract Ultrastructure of human brain tissue has traditionally been examined using electron microscopy (EM) following fixation, staining, and sectioning, which limit resolution and introduce artifacts. Alternatively, cryo-electron tomography (cryo-ET) allows higher resolution imaging of unfixed cellular samples while preserving architecture, but it requires samples to be vitreous and thin enough for transmission EM. Due to these requirements, cryo-ET has yet to be employed to investigate unfixed, never previously frozen human brain tissue. Here we present a method for generating lamellae in human brain tissue obtained at time of autopsy that can be imaged via cryo-ET. We vitrify the tissue via plunge-freezing and use xenon plasma focused ion beam (FIB) milling to generate lamellae directly on-grid at variable depth inside the tissue. Lamellae generated in Alzheimer’s disease brain tissue reveal intact subcellular structures including components of autophagy and potential pathologic tau fibrils. Furthermore, we reveal intact compact myelin and functional cytoplasmic expansions. These images indicate that plasma FIB milling with cryo-ET may be used to elucidate nanoscale structures within the human brain.

Relative to soluble N sources, enhanced-efficiency fertilizers (EEFs) support steady turfgrass growth and dense canopy quality while abating N loss as nitrate, ammonia, and/or N2O from turfgrass systems. Modern EEFs provide turfgrass managers greater operational effect and versatility in their nutrient management efforts and compel field characterization of their temporal response. Likewise, field confirmation of commercial EEF nutrient recovery helps stakeholders select the appropriate EEF for their specific application. Our research objective was to quantify the temporal response of Kentucky bluegrass growth/yield, canopy density and color, and fertilizer N recovery to a practical application of conventional urea or an enhanced-efficiency granular fertilizer. In May 2014 and June 2018, Kentucky bluegrass plots were fertilized by granules of conventional urea, N-(n-butyl) thiophosphoric triamide (NBPT)-, and dicyandiamide (DCD)-stabilized urea, or polymer-/sulfur-coated urea (PSCU) at a N rate of 43.9 kg ha−1 (0.9 lbs/1000 sq. ft.). The dependent variable response over the two growing seasons was highly affected by efficiency enhancement. Following the repeated 16.5-week evaluations, the mean percent of fertilizer N recovered from conventional urea, stabilized urea, and PSCU totaled 57.5, 68.4, and 89.1%, respectively. In the 23 to 51 days from treatment (DFT), recovery of PSCU-N significantly exceeded that from conventional or stabilized urea.

Introduction: The growing ability to provide online services has enabled the proliferation of exercise-based telehealth interventions; however, adoption in older adults may be impacted by low digital literacy and "technophobia." Objectives: The aim of this study was to explore the experience of community and aged-care dwelling older adults following exercise-based telehealth services to provide insights that could guide future telehealth exercise delivery. Design: Semi-structured interviews for qualitative analysis. Methods: Participants (age ≥60) who had completed at least one online exercise session from a registered health professional were recruited through a combination of purposeful and snowball sampling methods via their exercise facility or provider. A semi-structured interview guide was used by 2 interviewers to investigate participants' experiences and a "critical friends" approach used to identify common themes. Results: Thirteen interviews with 21 participants were conducted from 2 different facilities. Analysis identified meaning units within 3 themes and subthemes. Technology subthemes related to digital confidence prior to telehealth and changes during interventions, as well as the usability of technology for telehealth. Clinical practice subthemes described the different motivations to exercise, perceived benefits of telehealth, important implications for practitioners, and perceptions of safety. The social connection theme related to the social benefits of telehealth. Conclusions: Older adults in our sample were technologically confident and capable of performing exercise sessions delivered via telehealth. They notice benefits from this form of exercise delivery however, prefer face-to-face exercise delivery. [ABSTRACT FROM AUTHOR]

The physical basis of phase separation is thought to consist of the same types of bonds that specify conventional macromolecular interactions yet is unsatisfyingly often referred to as ‘fuzzy’. Gaining clarity on the biogenesis of membraneless cellular compartments is one of the most demanding challenges in biology. Here, we focus on the chromosome passenger complex (CPC), that forms a chromatin body that regulates chromosome segregation in mitosis. Within the three regulatory subunits of the CPC implicated in phase separation – a heterotrimer of INCENP, Survivin, and Borealin – we identify the contact regions formed upon droplet formation using hydrogen/deuterium exchange mass spectrometry (HXMS). These contact regions correspond to some of the interfaces seen between individual heterotrimers within the crystal lattice they form. A major contribution comes from specific electrostatic interactions that can be broken and reversed through initial and compensatory mutagenesis, respectively. Our findings reveal structural insight for interactions driving liquid-liquid demixing of the CPC. Moreover, we establish HXMS as an approach to define the structural basis for phase separation.

This study examined the challenges associated with knowledge management implementation and academic staff retention in selected tertiary institutions in South East, Nigeria. The research design adopted was a cross-sectional survey design. The main instrument used for data collection was questionnaire. The population consisted of 7,423 academic staff of the 10 randomly selected institutions in the South East Nigeria. A total sample size of 555 was drawn from the population. The instrument was checked for reliability using Cronbach method and the reliability co-efficient result of 0.915 showed that the instrument had high degree of item reliability. The hypothesis formulated was tested using Friedman chi-square statistics. The findings indicate that there is positive significant challenge in knowledge management implementation and academic staff retention in selected tertiary institution. The study concluded that lack of Knowledge Management implementation has posed a great challenge in academic staff retention in tertiary institutions. It was advised that tertiary institutions in Nigeria should adequately manage the process of knowledge acquisition, sharing and development so as to enjoy a Stella performance.

Nodular regenerative hyperplasia (NRH) is a primary disease of the liver that may cause noncirrhotic portal hypertension. Common causes include autoimmune, hematologic, immune deficiency, and myeloproliferative disorders. Given the limited data regarding the development of NRH in contemporary immunosuppressive protocols and the occurrence of NRH post-liver transplantation, we systematically reviewed NRH as it pertains to liver transplantation. We performed a comprehensive search for NRH and transplantation. Nineteen studies were identified with relevant data for NRH as an indication for a liver transplant. Thirteen studies were identified with relevant data pertaining to NRH development after liver transplant. Pooled analysis revealed 0.9% of liver transplant recipients had NRH. A total of 113 patients identified with NRH underwent liver transplantation. Most series report transplants done after the failure of endoscopic banding and TIPS management of portal hypertension. Reported 5-year graft and patient survival ranged from 73%–78% and 73%–90%. The pooled incidence of NRH after liver transplant for all indications was 2.9% and caused complications of portal hypertension. Complications related to portal hypertension secondary to NRH are a rare indication for a liver transplant. NRH can develop at any time after liver transplantation often without an identifiable cause, which may lead to portal hypertension requiring treatment or even re-transplantation.

Abstract This investigation aimed to quantitatively measure the changes in inflammation of subjects with healthy and unhealthy gums during a period of induced gingivitis. A total of 30 subjects (15 healthy, 15 with gum inflammation) were enlisted and given oral exams by a dental hygienist. Baseline measurements were acquired before a 3-week period of oral hygiene abstinence. The lobene modified gingival index scoring was used for inflammation scoring and hyperspectral spatial frequency domain imaging was used to quantitatively measure oxy- and deoxygenated blood volume fraction at two time points: at Baseline and after 3 weeks of oral hygiene abstinence. We found that abstaining from oral hygiene causes a near proportional increase in oxygenated and deoxygenated blood volume fraction for healthy individuals. For individuals who started the study with mild to moderate gingivitis, increases in blood volume were mainly due to deoxygenated blood.

CENP-A is the histone H3 variant necessary to specify the location of all eukaryotic centromeres via its CENP-A targeting domain and either one of its terminal regions. In humans, several post-translational modifications occur on CENP-A, but their role in centromere function remains controversial. One of these modifications of CENP-A, phosphorylation on serine 7, has been proposed to control centromere assembly and function. Here, using gene targeting at both endogenous CENP-A alleles and gene replacement in human cells, we demonstrate that a CENP-A variant that cannot be phosphorylated at serine 7 maintains correct CENP-C recruitment, faithful chromosome segregation and long-term cell viability. Thus, we conclude that phosphorylation of CENP-A on serine 7 is dispensable to maintain correct centromere dynamics and function.

Abstract Postoperative patients are at risk of life-threatening complications such as hemodynamic decompensation or arrhythmia. Automated detection of patients with such risks via a real-time clinical decision support system may provide opportunities for early and timely interventions that can significantly improve patient outcomes. We utilize multimodal features derived from digital signal processing techniques and tensor formation, as well as the electronic health record (EHR), to create machine learning models that predict the occurrence of several life-threatening complications up to 4 hours prior to the event. In order to ensure that our models are generalizable across different surgical cohorts, we trained the models on a cardiac surgery cohort and tested them on vascular and non-cardiac acute surgery cohorts. The best performing models achieved an area under the receiver operating characteristic curve (AUROC) of 0.94 on training and 0.94 and 0.82, respectively, on testing for the 0.5-hour interval. The AUROCs only slightly dropped to 0.93, 0.92, and 0.77, respectively, for the 4-hour interval. This study serves as a proof-of-concept that EHR data and physiologic waveform data can be combined to enable the early detection of postoperative deterioration events.

Aurora B, together with IN-box, the C-terminal part of INCENP, forms an enzymatic complex that ensures faithful cell division. The [Aurora B/IN-box] complex is activated by autophosphorylation in the Aurora B activation loop and in IN-box, but it is not clear how these phosphorylations activate the enzyme. We used a combination of experimental and computational studies to investigate the effects of phosphorylation on the molecular dynamics and structure of [Aurora B/IN-box]. In addition, we generated partially phosphorylated intermediates to analyze the contribution of each phosphorylation independently. We found that the dynamics of Aurora and IN-box are interconnected, and IN-box plays both positive and negative regulatory roles depending on the phosphorylation status of the enzyme complex. Phosphorylation in the activation loop of Aurora B occurs intramolecularly and prepares the enzyme complex for activation, but two phosphorylated sites are synergistically responsible for full enzyme activity.

Naturally occurring and structurally diverse alkene-containing substrates, terpenes, provided a platform for establishing chemo-, regio-, and diastereoselective reactivity in tantalum catalysed hydroaminoalkylation. Naturally derived 1,3-butadienes revealed the unique regio- and diastereoselective (Z)-1,4-addition products accessible from isoprene and β-myrcene by hydroaminoalkylation. Selective terpene functionalisation, within an industrially produced turpentine mixture, demonstrates functionalisation specificity of β-pinene and limonene. Lastly, sesquiterpene functionalisation using β-caryophyllene and humulene provide rare examples of trisubstituted alkene reactivity in hydroaminoalkylation, by leveraging strainrelease and stereoelectronic effects to control chemoselectivity. As a result of these reactivity studies using natural substrates, new tools for understanding alkene electronic, strain, and stereoelectronic effects on chemo- and diastereoselectivity outcomes have revealed new mechanistic insights into hydroaminoalkylation. [ABSTRACT FROM AUTHOR]

Relative to soluble N sources, enhanced-efficiency fertilizers (EEFs) support steady turfgrass growth and dense canopy quality while abating N loss as nitrate, ammonia, and/or N2O from turfgrass systems. Modern EEFs provide turfgrass managers greater operational effect and versatility in their nutrient management efforts and compel field characterization of their temporal response. Likewise, field confirmation of commercial EEF nutrient recovery helps stakeholders select the appropriate EEF for their specific application. Our research objective was to quantify the temporal response of Kentucky bluegrass growth/yield, canopy density and color, and fertilizer N recovery to a practical application of conventional urea or an enhanced-efficiency granular fertilizer. In May 2014 and June 2018, Kentucky bluegrass plots were fertilized by granules of conventional urea, N-(n-butyl) thiophosphoric triamide (NBPT)-, and dicyandiamide (DCD)-stabilized urea, or polymer-/sulfur-coated urea (PSCU) at a N rate of 43.9 kg ha−1 (0.9 lbs/1000 sq. ft.). The dependent variable response over the two growing seasons was highly affected by efficiency enhancement. Following the repeated 16.5-week evaluations, the mean percent of fertilizer N recovered from conventional urea, stabilized urea, and PSCU totaled 57.5, 68.4, and 89.1%, respectively. In the 23 to 51 days from treatment (DFT), recovery of PSCU-N significantly exceeded that from conventional or stabilized urea. [ABSTRACT FROM AUTHOR]

Background: The induction of testicular toxicity by bisphenol A (BPA) may involve oxidative stress. Glutamine (Gln) is a rate limiter for the synthesis of glutathione (GSH), which inhibits oxidative stress. Aim: This study assessed the potential of BPA to preserve testicular structure and function in BPA-treated albino rats. Study Settings and Design: Thirty-two adult male albino rats (210–250 g) were randomly allocated to 4 (A-D) of 8 rats per group with the approval of the Research Ethics Committee. Materials and Methods: Groups B-D were orally treated with Gln (20 mg/kg body weight), BPA (50 mg/kg body weight) and Gln (20 mg/kg body weight) + BPA (50 mg/kg body weight) daily for 65 days, respectively. Group A (control) was orally treated with normal saline (0.2 mL) daily for 65 days. At the termination of treatment, the rats were weighed and anaesthetized blood samples were collected and evaluated for gonadal hormones. The testes were weighed and evaluated for sperm parameters, oxidative stress markers and histology. Statistical Analysis Used: The data were analysed using one-way analysis of variance and Bonferroni post hoc test. Results: BPA caused a significant (P < 0.001) decrease in testis and body weights, sperm count, volume, motility and normal morphology when compared to the control. BPA significantly (P < 0.001) decreased serum testosterone, follicle-stimulating hormone, luteinising hormone, testicular catalase, superoxide dismutase, GSH and GSH peroxidase levels relative to control. Significantly (P < 0.001) increased serum prolactin, estradiol and testicular malondialdehyde levels occurred in BPA-treated rats relative to control. The testes of BPA-treated rats showed sloughing and coalescence of germ cells. However, Gln supplementation prevents BPA-induced testicular toxicity. Gln supplementation restored testis histology. Conclusion: Based on the observation in this study, Gln seems effective against BPA-induced testicular toxicity.

HPF1 controls the ADP-ribosylation activity of PARP1/2 in response to DNA breaks. Here, the authors show that HPF1 regulates the balance between ADP-ribose initiation and elongation through a dynamic interaction that accelerates the initiation rate on serine residues.

Maintaining centromere identity relies upon the persistence of the epigenetic mark provided by the histone H3 variant, centromere protein A (CENP-A), but the molecular mechanisms that underlie its remarkable stability remain unclear. Here, we define the contributions of each of the three candidate CENP-A nucleosome-binding domains (two on CENP-C and one on CENP-N) to CENP-A stability using gene replacement and rapid protein degradation. Surprisingly, the most conserved domain, the CENP-C motif, is dispensable. Instead, the stability is conferred by the unfolded central domain of CENP-C and the folded N-terminal domain of CENP-N that becomes rigidified 1,000-fold upon crossbridging CENP-A and its adjacent nucleosomal DNA. Disrupting the 'arginine anchor' on CENP-C for the nucleosomal acidic patch disrupts the CENP-A nucleosome structural transition and removes CENP-A nucleosomes from centromeres. CENP-A nucleosome retention at centromeres requires a core centromeric nucleosome complex where CENP-C clamps down a stable nucleosome conformation and CENP-N fastens CENP-A to the DNA.

Chromatin assembled with centromere protein A (CENP-A) is the epigenetic mark of centromere identity. Using new reference models, we now identify sites of CENP-A and histone H3.1 binding within the megabase, α-satellite repeat-containing centromeres of 23 human chromosomes. The overwhelming majority (97%) of α-satellite DNA is found to be assembled with histone H3.1-containing nucleosomes with wrapped DNA termini. In both G1 and G2 cell cycle phases, the 2-4% of α-satellite assembled with CENP-A protects DNA lengths centered on 133 bp, consistent with octameric nucleosomes with DNA unwrapping at entry and exit. CENP-A chromatin is shown to contain equimolar amounts of CENP-A and histones H2A, H2B, and H4, with no H3. Solid-state nanopore analyses show it to be nucleosomal in size. Thus, in contrast to models for hemisomes that briefly transition to octameric nucleosomes at specific cell cycle points or heterotypic nucleosomes containing both CENP-A and histone H3, human CENP-A chromatin complexes are octameric nucleosomes with two molecules of CENP-A at all cell cycle phases.