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Aleksandr Kakinen,1,2 Yuhao Jiang,2 Thomas Paul Davis,2 Tambet Teesalu,3,4 Mart Saarma1 1Institute of Biotechnology, HiLIFE, University of Helsinki, Helsinki, Finland; 2Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD, Australia; 3Institute of Biomedicine and Translational Medicine, Faculty of Medicine, University of Tartu, Tartu, Estonia; 4Materials Research Laboratory, University of California Santa Barbara, Santa Barbara, CA, USACorrespondence: Mart Saarma, Institute of Biotechnology, HiLIFE, University of Helsinki, Viikinkaari 5D, Helsinki, 00790, Finland, Tel +358505002726 ; +358294159378, Email mart.saarma@helsinki.fi Aleksandr Kakinen, Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Building 75, Cnr College Road& Cooper Road, St Lucia, QLD, 4067, Australia, Tel +61 7 344 63152, Email a.kakinen@uq.edu.auAbstract: Brain diseases are the most devastating problem among the world’s increasingly aging population, and the number of patients with neurological diseases is expected to increase in the future. Although methods for delivering drugs to the brain have advanced significantly, none of these approaches provide satisfactory results for the treatment of brain diseases. This remains a challenge due to the unique anatomy and physiology of the brain, including tight regulation and limited access of substances across the blood-brain barrier. Nanoparticles are considered an ideal drug delivery system to hard-to-reach organs such as the brain. The development of new drugs and new nanomaterial-based brain treatments has opened various opportunities for scientists to develop brain-specific delivery systems that could improve treatment outcomes for patients with brain disorders such as Alzheimer’s disease, Parkinson’s disease, stroke and brain tumors. In this review, we discuss noteworthy literature that examines recent developments in brain-targeted nanomedicines used in t

Stroke is one of the leading causes of death and disability both within the United States and across the globe. Currently, only one pharmacological treatment (i.e., r-tPA) is FDA approved and has limited effectiveness due to its narrow therapeutic window (4.5 hrs) following stroke onset. Therefore, novel therapeutic paradigms for the treatment of ischemic injury are needed to improve patient outcomes and reduce mortality. Drug delivery at therapeutic concentrations is challenging due to the presence of the blood brain barrier (BBB), limiting passage of small and large molecular drugs. Regulated by numerous cells within the neurovascular unit (NVU), alterations to the BBB during stroke pathogenesis further impair adequate and consistent delivery of neuroprotective compounds. Endogenous transporters within endothelial cells allow for transcellular delivery of pharmacotherapeutics and provide a potential mechanism which can be targeted during ischemic stroke to increase CNS drug delivery. One such group of uptake transporters, the organic anion transporting polypeptides (OATPs in humans; Oatps in rodents), has been shown to transport 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (i.e., statins). Clinical studies provide evidence that statins elicit neuroprotective effects which suggests that statins can be utilized as stroke therapeutic agents. An in depth understanding of statin delivery to the brain and their corresponding effects as a stroke treatment are essential. Additionally, manipulation of CNS drug delivery via uptake transporter expression and function within the BBB is a potential method to increase the translational effectiveness of neuroprotective compounds which are endogenous transporter substrates. Previous work from our lab has shown that Oatp manipulation can be achieved through activation of the TGF-?/ALK1 signaling pathway. We demonstrate regional differences in TGF-?/ALK1 signaling response to bone morphogenic protein (BMP)

Stroke is the 5th most common cause of death and the leading cause of long-term disability in the United States. Drug development for ischemic stroke is challenging as evidenced by the lack of therapeutics that have advanced beyond a phase III clinical trial. There are many reasons for this lack of clinical translation including factors related to the experimental design of preclinical stroke studies. Often overlooked in therapeutic development for ischemic stroke is the requirement of effective drug delivery to the brain, which is critical for neuroprotective efficacy of several small and large molecule drugs. Advancing central nervous system (CNS) drug delivery technologies implies a need for detailed comprehension of the blood-brain barrier (BBB) and neurovascular unit (NVU). Such knowledge will permit the innate biology of the BBB/NVU to be leveraged for improved bench-to-bedside translation of novel stroke therapeutics. At present, FDA-approved drug treatments for ischemic stroke are limited to recombinant-tissue plasminogen activator (r- tPA; Alteplase, Activase®), which is not available to many patients due to its short therapeutic window (i.e., 4.5 h) and clinically significant risk of bleeding complications. Although recanalization in infarcted brain tissue is critical, the adverse events associated with r-tPA treatment are not trivial and can promote neurologic and vascular injury, thus exacerbating post-stroke neurological deficits. Indeed, stroke patients still suffer from debilitating neurocognitive deficits despite the advent of reperfusion therapies. This indicates a need for discovery of stroke therapeutics and/or development of new pharmacological strategies for neuroprotection that are both safe and effective; however, the clinical utility of such compounds is highly dependent upon efficient transport from systemic circulation into ischemic brain tissue. Indeed, the BBB possess several endogenous transporters that can be targeted to promote deliver

Acetaminophen (APAP) is one of the most consumed over-the-counter and prescription medications worldwide due to its effective analgesic and antipyretic properties. Given its routine use in treating everyday ailments, understanding the breadth of its physiological interactions is critical. These interactions become especially pertinent when considering the potential consequences on the blood-brain barrier (BBB), the physical and biochemical barrier that maintains CNS homeostasis and limits brain penetration of potentially toxic xenobiotics. In our study, we have made several novel observations linked to APAP's effects on the BBB. Firstly, we demonstrated that high-dose APAP administration significantly increased BBB permeability by disrupting the BBB tight junctions. This effect resulted in enhanced brain uptake of codeine, a commonly prescribed opioid analgesic drug. This finding alone raises immediate concerns about combined APAP use or misuse with other medications due to the potential for altered CNS bioavailability and/or pharmacological effects. Secondly, our study showed that APAP increases functional expression of the critical BBB efflux transporter breast cancer resistance protein (Bcrp). Since Bcrp is involved in regulating penetrance of exogenous and endogenous molecules across the BBB, the effects of APAP on its transport activity can modulate CNS pharmacokinetics and, consequently, the overall pharmacological efficacy for drugs that are substrates for Bcrp. Taking these findings into the broader context of global health challenges, we cannot ignore the possible ramifications for the ongoing opioid epidemic. If APAP affects the brain uptake of opioids, it might alter their pain-relieving or euphoric potential, which in turn could drive individuals to engage in riskier consumption behaviors in pursuit of desired drug effects. In conclusion, our research uncovers a complex interaction between APAP and the BBB that has potential repercussions for both clinic

INTRODUCTION: It has been recognized that increasing body mass index (BMI) is associated with improved outcome from immune checkpoint inhibitors (ICIs) in patients with various malignancies including non-small cell lung cancer (NSCLC). However, it is unclear whether baseline BMI may influence outcomes from first-line chemoimmunotherapy combinations. METHODS: In this international multicenter study, we evaluated the association between baseline BMI, progression-free survival (PFS) and overall survival (OS) in a cohort of patients with stage IV NSCLC consecutively treated with first-line chemoimmunotherapy combinations. BMI was categorized according to WHO criteria. Results: Among the 853 included patients, 5.3% were underweight; 46.4% were of normal weight; 33.8% were overweight; and 14.5% were obese. Overweight and obese patients were more likely aged ≥70 years (p=0.00085), never smokers (p<0.0001), with better baseline Eastern Cooperative Oncology Group-Performance Status (p=0.0127), and had lower prevalence of central nervous system (p=0.0002) and liver metastases (p=0.0395). Univariable analyses showed a significant difference in the median OS across underweight (15.5 months), normal weight (14.6 months), overweight (20.9 months), and obese (16.8 months) patients (log-rank: p=0.045, log rank test for trend: p=0.131), while no difference was found with respect to the median PFS (log-rank for trend: p=0.510). Neither OS nor PFS was significantly associated with baseline BMI on multivariable analysis. CONCLUSIONS: In contrast to what was observed in the context of chemotherapy-free ICI-based regimens, baseline BMI does not affect clinical outcomes from chemoimmunotherapy combinations in patients with advanced NSCLC.

Background Like most legumes, chickpeas form specialized organs called root nodules. These nodules allow for a symbiotic relationship with rhizobium bacteria. The rhizobia provide fixed atmospheric nitrogen to the plant in a usable form. It is of both basic and practical interest to understand the host plant genetics of legume root nodulation. Chickpea lines PM233 and PM405, which harbor the mutationally identified nodulation genes rn1 and rn4, respectively, both display nodulation-deficient phenotypes. Previous investigators identified the rn1 mutation with the chickpea homolog of Medicago truncatula nodulation gene NSP2, but were unable to define the mutant rn1 allele. We used Illumina and Nanopore sequencing reads to attempt to identify and characterize candidate mutation sites responsible for the PM233 and PM405 phenotypes. Results We aligned Illumina reads to the available desi chickpea reference genome, and did a de novo contig assembly of Nanopore reads. In mutant PM233, the Nanopore contigs allowed us to identify the breakpoints of a ~ 35 kb deleted region containing the CaNSP2 gene, the Medicago truncatula homolog of which is involved in nodulation. In mutant PM405, we performed variant calling in read alignments and identified 10 candidate mutations. Genotyping of a segregating progeny population narrowed that pool down to a single candidate gene which displayed homology to M. truncatula nodulation gene NIN. Conclusions We have characterized the nodulation mutation sites in chickpea mutants PM233 and PM405. In mutant PM233, the rn1 mutation was shown to be due to deletion of the entire CaNSP2 nodulation gene, while in mutant PM405 the rn4 mutation was due to a single base deletion resulting in a frameshift mutation between the predicted RWP-RK and PB1 domains of the NIN nodulation gene. Critical to characterization of the rn1 allele was the generation of Nanopore contigs for mutant PM233 and its wild type parent ICC 640, without which the deletional bounda

Introduction: A significant proportion of patients with cancer who recover from Coronavirus Disease 2019 (COVID-19) may experience COVID-19 sequelae in the early post-infection phase, which negatively affect their continuity of care and oncological outcome. The long-term prevalence and clinical impact of the post-COVID-19 syndrome in patients with cancer are largely unknown. Methods: In this study, we describe the time course of COVID-19 sequelae in patients with non-advanced cancers enrolled in the OnCovid registry. Results: Overall, 186 patients were included, with a median observation period of 9.9 months (95%CI:8,8–11.3) post-COVID-19 resolution. After a median interval of 2.3 months post-COVID-19 (interquartile range: 1.4–3.7), 31 patients (16.6%) reported ≥1 sequelae, including respiratory complications (14, 7.6%), fatigue (13, 7.1%), neuro-cognitive sequelae (7, 3.8%). The vast majority of the patients were not vaccinated prior to COVID-19. COVID-19-related sequelae persisted in 9.8% and 8% of patients 6 and 12 months after COVID-19 resolution. Persistence of sequelae at first oncological follow-up was associated with history of complicated COVID-19 (45.2% vs 24.8%, p = 0.0223), irrespective of oncological features at COVID-19 diagnosis. Conclusion: This study confirms for the first time that, in a largely unvaccinated population, post-COVID-19 syndrome can affect a significant proportion of patients with non-advanced cancer who recovered from the acute illness. COVID-19 sequelae may persist up to 12 months in some patients, highlighting the need for dedicated prevention and supportive strategies.

Hydrogen sulfide (H2S) is an important signalling molecule with potential pharmaceutical applications. In pursuit of a suitable delivery system for H2S, herein we apply an amphiphilic trisulfide to concomitantly alter the mesophase behaviour of dispersed lipid particles and enable triggered H2S release. Amperometric release studies indicate the trisulfide acts as a sustained H2S donor, with inclusion into the mesophase attenuating release vs neat dispersed trisulfide. Taken together the results highlight the potential for including trisulfide-based additives in stimuli-responsive drug delivery vehicles.

Counselors must make sound ethical decisions but, often, determining the appropriate course to take when confronted with a difficult ethical dilemma can be a challenge. This guide assists counselors in ethical decision making; it is presented as a supplement, and is to be used in conjunction with, the American Counseling Association (ACA) Code of Ethics and Standards of Practice. This practitioner's guide addresses principles that are globally valuable in ethical decision making and provides a model for professionals to utilize in their practice. It opens with a review of the five moral principles that are viewed by many as the cornerstone of ethical guidelines: autonomy, nonmaleficence, beneficence, justice, and fidelity. These principles provide an absolute against which counselors can explore an ethical dilemma, thereby gaining a better understanding of conflicting issues. A sequential, seven-step, ethical, decision-making model is presented next and comprises the following: (1) identify the problem; (2) apply the ACA Code of Ethics; (3) determine the nature and dimensions of the dilemma; (4) generate potential courses of action; (5) consider the potential consequences of all options and then determine a course of action; (6) evaluate the selected course of action; (7) implement this action. (RJM)

To maximize comprehensive school health education, professional preparation would include a focusing component that integrates elements of the comprehensive school health model and develops the leadership and management capabilities of potential school health leaders. The article describes how to integrate the comprehensive school health model and explains program leaders' tasks. (SM)

An important goal of clinical genomics is to be able to estimate the risk of adverse disease outcomes. Between 5% and 10% of individuals with ulcerative colitis (UC) require colectomy within 5 years of diagnosis, but polygenic risk scores (PRSs) utilizing findings from genome-wide association studies (GWASs) are unable to provide meaningful prediction of this adverse status. By contrast, in Crohn disease, gene expression profiling of GWAS-significant genes does provide some stratification of risk of progression to complicated disease in the form of a transcriptional risk score (TRS). Here, we demonstrate that a measured TRS based on bulk rectal gene expression in the PROTECT inception cohort study has a positive predictive value approaching 50% for colectomy. Single-cell profiling demonstrates that the genes are active in multiple diverse cell types from both the epithelial and immune compartments. Expression quantitative trait locus (QTL) analysis identifies genes with differential effects at baseline and week 52 follow-up, but for the most part, differential expression associated with colectomy risk is independent of local genetic regulation. Nevertheless, a predicted polygenic transcriptional risk score (PPTRS) derived by summation of transcriptome-wide association study (TWAS) effects identifies UC-affected individuals at 5-fold elevated risk of colectomy with data from the UK Biobank population cohort studies, independently replicated in an NIDDK-IBDGC dataset. Prediction of gene expression from relatively small transcriptome datasets can thus be used in conjunction with TWASs for stratification of risk of disease complications.

Nanotoxicology and nanomedicine are two sub-disciplines of nanotechnology focusing on the phenomena, mechanisms, and engineering at the nano-bio interface. For the better part of the past three decades, these two disciplines have been largely developing independently of each other. Yet recent breakthroughs in microbiome research and the current COVID-19 pandemic demonstrate that holistic approaches are crucial for solving grand challenges in global health. Here we show the Yin and Yang relationship between the two fields by highlighting their shared goals of making safer nanomaterials, improved cellular and organism models, as well as advanced methodologies. We focus on the transferable knowledge between the two fields as nanotoxicological research is moving from pristine to functional nanomaterials, while inorganic nanomaterials - the main subjects of nanotoxicology - have become an emerging source for the development of nanomedicines. We call for a close partnership between the two fields in the new decade, to harness the full potential of nanotechnology for benefiting human health and environmental safety.

BackgroundDiagnostic options to combat the increasing rates of sexually transmitted infections recorded throughout the world increasingly include multiplex assays. Here we describe the estimated sensitivity and specificity of a triplex molecular assay that simultaneously detects Chlamydia trachomatis (CT), Neisseria gonorrhoeae (or gonococci [GC]), and Trichomonas vaginalis (TV).MethodsParticipants (2547 women and 1159 men) were recruited from 12 clinics in the United States. BD CTGCTV2 for BD MAX System assay (CTGCTV2) results were obtained from vaginal and endocervical swabs, endocervical samples in cytology medium, and female and male urine. Results were compared with infection standards that were sample type and pathogen dependent.ResultsFemale specimen sensitivity estimates ranged from 92.7% to 98.4%, 92.9% to 100%, and 86.6% to 100% for CT, GC and TV, respectively. Male urine sensitivity estimates were 96.7%, 99.2%, and 97.9% for CT, GC, and TV, respectively. Specificity estimates were >98.7% for all sample types.ConclusionsBD CTGCTV2 performed well using a variety of sample types. As a true triplex assay, performed using a benchtop instrument, BD CTGCTV2 may be useful in settings where no testing is currently performed and in settings, such as reference laboratories, where testing turnaround time may be several days. Use of this assay at local laboratories may result in greater access to testing and a shorter time to result, which are important steps for improving our ability to combat sexually transmitted infections.

Amyloid diseases are global epidemics with profound health, social and economic implications and yet remain without a cure. This dire situation calls for research into the origin and pathological manifestations of amyloidosis to stimulate continued development of new therapeutics. In basic science and engineering, the cross-β architecture has been a constant thread underlying the structural characteristics of pathological and functional amyloids, and realizing that amyloid structures can be both pathological and functional in nature has fuelled innovations in artificial amyloids, whose use today ranges from water purification to 3D printing. At the conclusion of a half century since Eanes and Glenner's seminal study of amyloids in humans, this review commemorates the occasion by documenting the major milestones in amyloid research to date, from the perspectives of structural biology, biophysics, medicine, microbiology, engineering and nanotechnology. We also discuss new challenges and opportunities to drive this interdisciplinary field moving forward.

PurposeRindopepimut is a vaccine targeting the tumor-specific EGF driver mutation, EGFRvIII. The ReACT study investigated whether the addition of rindopepimut to standard bevacizumab improved outcome for patients with relapsed, EGFRvIII-positive glioblastoma.Patients and methodsIn this double-blind, randomized, phase II study (NCT01498328) conducted at 26 hospitals in the United States, bevacizumab-naïve patients with recurrent EGFRvIII-positive glioblastoma were randomized to receive rindopepimut or a control injection of keyhole limpet hemocyanin, each concurrent with bevacizumab. The primary endpoint was 6-month progression-free survival (PFS6) by central review with a one-sided significance of 0.2.ResultsBetween May 2012 and 2014, 73 patients were randomized (36 rindopepimut, 37 control). Rindopepimut toxicity included transient, low-grade local reactions. As primary endpoint, PFS6 was 28% (10/36) for rindopepimut compared with 16% (6/37) for control (P = 0.12, one-sided). Secondary and exploratory endpoints also favored the rindopepimut group including a statistically significant survival advantage [HR, 0.53; 95% confidence interval (CI), 0.32-0.88; two-sided log-rank P = 0.01], a higher ORR [30% (9/30) vs. 18% (6/34; P = 0.38)], median duration of response [7.8 months (95% CI, 3.5-22.2) vs. 5.6 (95% CI, 3.7-7.4)], and ability to discontinue steroids for ≥6 months [33% (6/18) vs. 0% (0/19)]. Eighty percent of rindopepimut-treated patients achieved robust anti-EGFRvIII titers (≥1:12,800), which were associated with prolonged survival (HR = 0.17; 95% CI, 0.07-0.45; P < 0.0001).ConclusionsOur randomized trial supports the potential for targeted immunotherapy among patients with GBM, but the therapeutic benefit requires validation due to the small sample size and potential heterogeneity of bevacizumab response among recurrent patients with GBM.See related commentary by Wick and Wagener, p. 1535.

Cancer is the leading cause of death globally and a major limitation of conventional treatments is toxicity to normal tissues, restricting the dose deliverable to patients. There has been enormous interest in the application of nanotechnology towards therapeutics for cancer, as many of these therapeutics lack specificity and in some cases, efficacy due to limitations in delivering a therapeutic dose. With the hypothesis that delivering increased therapeutic doses specifically to tumour cells will increase the efficacy of treatment, the aim of this thesis is to develop and evaluate targeted therapies for different types of cancer. Initially, the development of targeted magnetic hyperthermia was examined and demonstrated some of the challenges encountered when working with magnetic iron oxide nanoparticles for this application. This highlighted the need for better control of aggregation under clinically relevant conditions. Next, a modular strategy for targeting a clinically-approved liposomal formulation of doxorubicin towards epidermal growth factor receptor on lung cancer cells, prostate specific membrane antigen on prostate cancer cells and disialoganglioside (GD2) on neuroblastoma cells via bispecific antibodies was developed. Targeting resulted in increased internalisation, apoptosis and cytotoxicity in vitro. While initial in vivo experiments were inconclusive, potential reasons for this are discussed. With insights gained regarding nanoparticle properties for targeting, and to broaden the number of payloads that can be effectively targeted towards cancer cells, novel cross-linked poly amino acid micelles were developed and covalently loaded with several payloads including monomethyl auristatin E, a potent anti-microtubule drug. Micelles had a hydrodynamic diameter of 37 nm and were uniform in size with a polydispersity index of 0.1. When applied with bispecific antibodies, the system demonstrated a remarkable increase in internalisation of payload by cancer ce

Funding: D.J.P. is supported by grant funding from the Wellcome Trust Strategic Fund (PS3416) and acknowledges grant support by the Cancer Treatment and Research Trust (CTRT) as well as infrastructural support by the Cancer Research UK Imperial Centre. GG is supported by the AIRC 5 × 1000 Grant, No. 21198, Associazione Italiana per la Ricerca sul Cancro Foundation, Milan, Italy. A.G. is supported by the AIRC IG grant, No. 14230, Associazione Italiana per la Ricerca sul Cancro Foundation, Milan, Italy. A.G., G.G., G.C.A., L.M.C., M.B., P.P.S., M.P. from the University of Piemonte Orientale acknowledge support from the UPO Aging Project., We describe the outcomes in cancer patients during the initial outbreak of the COVID-19 in Europe from the retrospective, multi-center observational OnCovid study. We identified 204 cancer patients from eight centers in the United Kingdom, Italy, and Spain aged > 18 (mean = 69) and diagnosed with COVID-19 between February 26th and April 1st, 2020. A total of 127 (62%) were male, 184 (91%) had a diagnosis of solid malignancy, and 103 (51%) had non-metastatic disease. A total of 161 (79%) had > 1 co-morbidity. A total of 141 (69%) patients had > 1 COVID-19 complication. A total of 36 (19%) were escalated to high-dependency or intensive care. A total of 59 (29%) died, 53 (26%) were discharged, and 92 (45%) were in-hospital survivors. Mortality was higher in patients aged > 65 (36% versus 16%), in those with > 2 co-morbidities (40% versus 18%) and developing > 1 complication from COVID-19 (38% versus 4%, p = 0.004). Multi-variable analyses confirmed age > 65 and > 2 co-morbidities to predict for patient mortality independent of tumor stage, active malignancy, or anticancer therapy. During the early outbreak of SARS-CoV-2 infection in Europe co-morbid burden and advancing age predicted for adverse disease course in cancer patients. The ongoing OnCovid study will allow us to compare risks and outcomes in cancer patients between the initial and later stages of the COVID-19 pandemic.

Type 2 diabetes (T2D) is a metabolic disease and a global health crisis. Because of the small mass and high dispersity of beta cells in the pancreas, especially among T2D patients, it remains a tremendous challenge to detect and image beta cell mass (BCM) in vitro and in vivo. Herein, a multimodal nanoprobe is constructed by surface functionalization of magnetic iron oxide nanoparticles with a two-photon fluorescent dye (NaP)-labeled polymer. Owing to the nanoparticle surface energy-transfer effect, the nanoprobe enabled pH-triggered fluorescence/magnetic resonance imaging in the acidic beta cell environment. Specifically, confocal one-photon and two-photon modalities revealed prominent fluorescence in BTC-6 pancreatic beta cells among five major cell types, validating the probe as a sensor for BCM quantification. Kinetic assay, transmission electron microscopy, and viability assay further implicated the probe as a potent inhibitor against the aggregation and toxicity of human islet amyloid polypeptide (IAPP), the peptide associated with T2D. This probe presents a first multimodal theranostic system for imaging BCM and inhibition of beta cell degeneration by IAPP amyloidosis. © 2020 American Chemical Society., SCOPUS: ar.j, info:eu-repo/semantics/published

The Rosaceae crop family (including almond, apple, apricot, blackberry, peach, pear, plum, raspberry, rose, strawberry, sweet cherry, and sour cherry) provides vital contributions to human well-being and is economically significant across the U.S. In 2003, industry stakeholder initiatives prioritized the utilization of genomics, genetics, and breeding to develop new cultivars exhibiting both disease resistance and superior horticultural quality. However, rosaceous crop breeders lacked certain knowledge and tools to fully implement DNA-informed breeding—a “chasm” existed between existing genomics and genetic information and the application of this knowledge in breeding. The RosBREED project (“Ros” signifying a Rosaceae genomics, genetics, and breeding community initiative, and “BREED”, indicating the core focus on breeding programs), addressed this challenge through a comprehensive and coordinated 10-year effort funded by the USDA-NIFA Specialty Crop Research Initiative. RosBREED was designed to enable the routine application of modern genomics and genetics technologies in U.S. rosaceous crop breeding programs, thereby enhancing their efficiency and effectiveness in delivering cultivars with producer-required disease resistances and market-essential horticultural quality. This review presents a synopsis of the approach, deliverables, and impacts of RosBREED, highlighting synergistic global collaborations and future needs. Enabling technologies and tools developed are described, including genome-wide scanning platforms and DNA diagnostic tests. Examples of DNA-informed breeding use by project participants are presented for all breeding stages, including pre-breeding for disease resistance, parental and seedling selection, and elite selection advancement. The chasm is now bridged, accelerating rosaceous crop genetic improvement.

Discusses the teaching method of students studying a topic, then explaining the meaning of the concept to each other in their own words using examples from their everyday lives. The instructor acts as a coordinator, mediator, and a resource person. Concludes with the disadvantages of this interactive learning technique on a habitat tour, among which are the need for relatively small lab classes. (AIM)

Increasing evidence recognizes Alzheimer's disease (AD) as a multifactorial and heterogeneous disease with multiple contributors to its pathophysiology, including vascular dysfunction. The recently updated AD Research Framework put forth by the National Institute on Aging-Alzheimer's Association describes a biomarker-based pathologic definition of AD focused on amyloid, tau, and neuronal injury. In response to this article, here we first discussed evidence that vascular dysfunction is an important early event in AD pathophysiology. Next, we examined various imaging sequences that could be easily implemented to evaluate different types of vascular dysfunction associated with, and/or contributing to, AD pathophysiology, including changes in blood-brain barrier integrity and cerebral blood flow. Vascular imaging biomarkers of small vessel disease of the brain, which is responsible for >50% of dementia worldwide, including AD, are already established, well characterized, and easy to recognize. We suggest that these vascular biomarkers should be incorporated into the AD Research Framework to gain a better understanding of AD pathophysiology and aid in treatment efforts.

Human amyloids and plaques uncovered post mortem are highly heterogeneous in structure and composition, yet literature concerning the heteroaggregation of amyloid proteins is extremely scarce. This knowledge deficiency is further exacerbated by the fact that peptide delivery is a major therapeutic strategy for targeting their full-length counterparts associated with the pathologies of a range of human diseases, including dementia and type 2 diabetes (T2D). Accordingly, here we examined the coaggregation of full-length human islet amyloid polypeptide (IAPP), a peptide associated with type 2 diabetes, with its primary and secondary amyloidogenic fragments 19-29 S20G and 8-20. Single-molecular aggregation dynamics was obtained by high-speed atomic force microscopy, augmented by transmission electron microscopy, X-ray diffraction, and super-resolution stimulated emission depletion microscopy. The coaggregation significantly prolonged the pause phase of fibril elongation, increasing its dwell time by 3-fold. Surprisingly, unidirectional elongation of mature fibrils, instead of protofilaments, was observed for the coaggregation, indicating a new form of tertiary protein aggregation unknown to existing theoretical models. Further in vivo zebrafish embryonic assay indicated improved survival and hatching, as well as decreased frequency and severity of developmental abnormalities for embryos treated with the heteroaggregates of IAPP with 19-29 S20G, but not with 8-20, compared to the control, indicating the therapeutic potential of 19-29 S20G against T2D.

Narges Bayat,1,2 Nathan McOrist,1 Nicholas Ariotti,3,4 May Lai,5 Keith CS Sia,1,2 Yuhuan Li,5 James L Grace,5 John F Quinn,5 Michael R Whittaker,5 Maria Kavallaris,2,6,7 Thomas P Davis,5,8,9,* Richard B Lock1,2,* 1Leukemia Biology Program, Children’s Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia; 2School of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, Australia; 3Electron Microscope Unit, Mark Wainwright Analytical Centre, Chemical Sciences Building, University of New South Wales, Sydney, NSW, Australia; 4School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia; 5ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, and Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia; 6Tumor Biology and Targeting Program, Children’s Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia; 7Australian Centre for Nanomedicine, ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney, NSW, Australia; 8Department of Chemistry, University of Warwick, Coventry, UK; 9Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD 4072, Australia*These authors contributed equally to this workCorrespondence: Richard B LockChildren’s Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, PO Box 81, Randwick 2031, NSW, AustraliaTel +61 2 9385 2513Fax +61 2 9662 6583Email rlock@ccia.org.auPurpose: Directing nanoparticles to cancer cells without using antibodies is of great interest. Subtle changes to the surface chemistry of nanoparticles can significantly affect their biological fate, including their propensity to associate with different cell populations.

The compendium of RNA-binding proteins (RBPs) has been greatly expanded by the development of RNA-interactome capture (RIC). However, it remained unknown if the complement of RBPs changes in response to environmental perturbations and whether these rearrangements are important. To answer these questions, we developed “comparative RIC” and applied it to cells challenged with an RNA virus called sindbis (SINV). Over 200 RBPs display differential interaction with RNA upon SINV infection. These alterations are mainly driven by the loss of cellular mRNAs and the emergence of viral RNA. RBPs stimulated by the infection redistribute to viral replication factories and regulate the capacity of the virus to infect. For example, ablation of XRN1 causes cells to be refractory to SINV, while GEMIN5 moonlights as a regulator of SINV gene expression. In summary, RNA availability controls RBP localization and function in SINV-infected cells. Garcia-Moreno, Noerenberg, Ni, and colleagues developed “comparative RNA-interactome capture” to analyze the RNA-bound proteome during virus infection. More than 200 cellular RNA-binding proteins change their binding activity in response to this challenge, mainly driven by transcript availability. Many of these RNA-binding proteins regulate viral replication and can be targeted to influence infection outcome.

BackgroundCentral nervous system (CNS) histoplasmosis is a life-threatening condition and represents a diagnostic and therapeutic challenge. Isolation of Histoplasma capsulatum from cerebrospinal fluid (CSF) or brain tissue is diagnostic; however, culture is insensitive and slow growth may result in significant treatment delay. We performed a retrospective multicenter study to evaluate the sensitivity and specificity of a new anti-Histoplasma antibody enzyme immunoassay (EIA) for the detection of IgG and IgM antibody in the CSF for diagnosis of CNS histoplasmosis, the primary objective of the study. The secondary objective was to determine the effect of improvements in the Histoplasma galactomannan antigen detection EIA on the diagnosis of Histoplasma meningitis.MethodsResidual CSF specimens from patients with Histoplasma meningitis and controls were tested for Histoplasma antigen and anti-Histoplasma immunoglobulin G (IgG) and immunoglobulin M (IgM) antibody using assays developed at MiraVista Diagnostics.ResultsA total of 50 cases and 157 controls were evaluated. Fifty percent of patients with CNS histoplasmosis were immunocompromised, 14% had other medical conditions, and 36% were healthy. Histoplasma antigen was detected in CSF in 78% of cases and the specificity was 97%. Anti-Histoplasma IgG or IgM antibody was detected in 82% of cases and the specificity was 93%. The sensitivity of detection of antibody by currently available serologic testing including immunodiffusion and complement fixation was 51% and the specificity was 96%. Testing for both CSF antigen and antibody by EIA was the most sensitive approach, detecting 98% of cases.ConclusionsTesting CSF for anti-Histoplasma IgG and IgM antibody complements antigen detection and improves the sensitivity for diagnosis of Histoplasma meningitis.

ObjectiveTo evaluate whether the progression of individual motor features was influenced by early deep brain stimulation (DBS), a post hoc analysis of Unified Parkinson's Disease Rating Scale-III (UPDRS-III) score (after a 7-day washout) was conducted from the 2-year DBS in early Parkinson disease (PD) pilot trial dataset. Methods The prospective pilot trial enrolled patients with PD aged 50-75 years, treated with PD medications for 6 months-4 years, and no history of dyskinesia or other motor fluctuations, who were randomized to receive optimal drug therapy (ODT) or DBS plus ODT (DBS + ODT). At baseline and 6, 12, 18, and 24 months, all patients stopped all PD therapy for 1 week (medication and stimulation, if applicable). UPDRS-III "off" item scores were compared between the ODT and DBS + ODT groups (n = 28); items with significant between-group differences were analyzed further. Results UPDRS-III "off" rest tremor score change from baseline to 24 months was worse in patients receiving ODT vs DBS + ODT (p = 0.002). Rest tremor slopes from baseline to 24 months favored DBS + ODT both "off" and "on" therapy (p < 0.001, p = 0.003, respectively). More ODT patients developed new rest tremor in previously unaffected limbs than those receiving DBS + ODT (p = 0.001). Conclusions These results suggest the possibility that DBS in early PD may slow rest tremor progression. Future investigation in a larger cohort is needed, and these findings will be tested in the Food and Drug Administration-approved, phase III, pivotal, multicenter clinical trial evaluating DBS in early PD. Classification of evidence This study provides Class II evidence that for patients with early PD, DBS may slow the progression of rest tremor.

Previous research has demonstrated that BBB structure and function are altered as a result of various neurological disorders including ischemic stroke, traumatic brain injury, epilepsy, and infections of the brain. Additionally, the BBB has also been shown to alter its function in response to nociception. Despite the strong evidence for BBB alterations in both neurological disorders and pain, there is still debate on whether BBB permeability is altered in episodic headache disorders such as migraine. Cortical spreading depression (CSD) in the CNS is suggested as a common mechanism contributing to both headache production and BBB changes. In previous studies examining BBB changes in response to CSD, animals were anesthetized during the study, preventing any behavioral assessments. Additionally, in studies examining CSD induced nociceptive behaviors, BBB permeability was not assessed. Therefore, this work represents the first joint assessment of nociceptive responses and BBB integrity in response to CSD. In these studies, we observed a transient increase in BBB paracellular permeability in the cortex, but not brainstem, in response to KCl induced CSD. Additionally, at corresponding time points, we found that KCl induced CSD reduced periorbital withdrawal thresholds and rearing behavior, indicative of a state of facial mechanical allodynia. Despite strong evidence for CNS involvement in headache disorders, drug development for headache disorders remains focused on peripheral targets. Difficulty in delivering drugs across the BBB may partially account for this disparity. In this work, we demonstrated that KCl induced CSD increased the CNS uptake of radiolabeled sumatriptan in both the cortex and the brainstem. We also found that KCl induced CSD increased the expression of the putative sumatriptan transporter Oatp1a4 in the brainstem, which likely underlies the observed increased brainstem permeability to sumatriptan following CSD induction. Repeated CSD events may be ha

Protein aggregation into amyloid fibrils is a ubiquitous phenomenon across the spectrum of neurodegenerative disorders and type 2 diabetes. A common strategy against amyloidogenesis is to minimize the populations of toxic oligomers and protofibrils by inhibiting protein aggregation with small molecules or nanoparticles. However, melanin synthesis in nature is realized by accelerated protein fibrillation to circumvent accumulation of toxic intermediates. Accordingly, we designed and demonstrated the use of star-shaped poly(2-hydroxyethyl acrylate) (PHEA) nanostructures for promoting aggregation while ameliorating the toxicity of human islet amyloid polypeptide (IAPP), the peptide involved in glycemic control and the pathology of type 2 diabetes. The binding of PHEA elevated the β-sheet content in IAPP aggregates while rendering a new morphology of "stelliform" amyloids originating from the polymers. Atomistic molecular dynamics simulations revealed that the PHEA arms served as rodlike scaffolds for IAPP binding and subsequently accelerated IAPP aggregation by increased local peptide concentration. The tertiary structure of the star nanoparticles was found to be essential for driving the specific interactions required to impel the accelerated IAPP aggregation. This study sheds new light on the structure-toxicity relationship of IAPP and points to the potential of exploiting star polymers as a new class of therapeutic agents against amyloidogenesis., info:eu-repo/semantics/published

A highly efficient photomediated atom transfer radical polymerization protocol is reported for semi-fluorinated acrylates and methacrylates. Use of the commercially available solvent, 2-trifluoromethyl-2-propanol, optimally balances monomer, polymer, and catalyst solubility while eliminating transesterification as a detrimental side reaction. In the presence of UV irradiation and ppm concentrations of copper(II) bromide and Me6-TREN (TREN = tris(2-aminoethyl amine)), semi-fluorinated monomers with side chains containing between three and 21 fluorine atoms readily polymerize under controlled conditions. The resulting polymers exhibit narrow molar mass distributions (Đ ≈ 1.1) and high end group fidelity, even at conversions greater than 95%. This level of control permits the in situ generation of chain-end functional homopolymers and diblock copolymers, providing facile access to semi-fluorinated macromolecules using a single methodology with unprecedented monomer scope. The results disclosed herein should create opportunities across a variety of fields that exploit fluorine-containing polymers for tailored bulk, interfacial, and solution properties.

Selenium accumulation in vegetation has resulted in toxicity in livestock grazing on phosphate mine soils in Southeastern Idaho. Plant and soil samples were collected from sites located near phosphate mines. Soil physicochemical properties, Se speciation, and Se distribution from a sequential extraction procedure (SEP) were examined in relation to bioavailability in the Se-hyperaccumulator, western aster (Symphyotrichum ascendens Lindl.). Selenium-hyperaccumulators are plants that can absorb over 1000 mg Se kg(-1) (DM) (Dry Matter). Chemical analyses revealed that western aster contained Se exceeding 6000 mg kg(-1) (DM). Soil speciation results indicated that selenite (SeO3 (2-)) was dominant with lower levels of selenate (SeO4 (2-)) present. This was expanded using an SEP that accounted for six fractions. Regression analyses indicated a strong relationship for western aster Se and the water-soluble and phosphate-extractable SEP fractions combined (R (2) = 0.85). Once carbonate, amorphous Fe-oxide, organic, and residual Se fractions were factored into the analysis, the relationship decreased. A strong relationship between selenate and the water-soluble Se fraction was also observed (R-2 = 0.83). Soluble and phosphate-extractable Se were determined to be "bioavailable fractions" for western aster. Thus, simple water extractions can be used for quick assessment of Se bioavailability and provide a means to identify potentially hazardous areas locations.

Selenium accumulation in vegetation has resulted in toxicity in livestock grazing on phosphate mine soils in Southeastern Idaho. Plant and soil samples were collected from sites located near phosphate mines. Soil physicochemical properties, Se speciation, and Se distribution from a sequential extraction procedure (SEP) were examined in relation to bioavailability in the Se-hyperaccumulator, western aster (Symphyotrichum ascendens Lindl.). Selenium-hyperaccumulators are plants that can absorb over 1000 mg Se kg(-1) (DM) (Dry Matter). Chemical analyses revealed that western aster contained Se exceeding 6000 mg kg(-1) (DM). Soil speciation results indicated that selenite (SeO3 (2-)) was dominant with lower levels of selenate (SeO4 (2-)) present. This was expanded using an SEP that accounted for six fractions. Regression analyses indicated a strong relationship for western aster Se and the water-soluble and phosphate-extractable SEP fractions combined (R (2) = 0.85). Once carbonate, amorphous Fe-oxide, organic, and residual Se fractions were factored into the analysis, the relationship decreased. A strong relationship between selenate and the water-soluble Se fraction was also observed (R-2 = 0.83). Soluble and phosphate-extractable Se were determined to be "bioavailable fractions" for western aster. Thus, simple water extractions can be used for quick assessment of Se bioavailability and provide a means to identify potentially hazardous areas locations.

Aggregation of the highly amyloidogenic IAPP is endogenously inhibited inside beta-cell granules at millimolar concentrations. Combining in vitro experiments and computer simulations, we demonstrated that the stabilization of IAPP upon the formation of zinc-coordinated ion molecular complex with C-peptide might be important for the endogenous inhibition of IAPP aggregation., info:eu-repo/semantics/published

Human islet amyloid polypeptide (hIAPP or amylin) aggregation is directly associated with pancreatic β-cell death and subsequent insulin deficiency in type 2 diabetes (T2D). Since no cure is currently available for T2D, it is of great benefit to devise new anti-aggregation molecules, which protect β-cells against hIAPP aggregation-induced toxicity. Engineered nanoparticles have been recently exploited as anti-aggregation nanomedicines. In this work, we studied graphene oxide (GO) nanosheets for their potential for hIAPP aggregation inhibition by combining computational modeling, biophysical characterization and cell toxicity measurements. Using discrete molecular dynamics (DMD) simulations and in vitro studies, we showed that GO exhibited an inhibitory effect on hIAPP aggregation. DMD simulations indicated that the strong binding of hIAPP to GO nanosheets was driven by hydrogen bonding and aromatic stacking and that the strong peptide-GO binding efficiently inhibited hIAPP self-association and aggregation on the nanosheet surface. Secondary structural changes of hIAPP upon GO binding derived from DMD simulations were consistent with circular dichroism (CD) spectroscopy measurements. Transmission electron microscopy (TEM) images confirmed the reduction of hIAPP aggregation in the presence of GO. Furthermore, we carried out a cell toxicity assay and found that these nanosheets protected insulin-secreting NIT-1 pancreatic β-cells against hIAPP-induced toxicity. Our multidisciplinary study suggests that GO nanosheets have the potential to be utilized as an anti-aggregation nanomedicine itself in addition to a biosensor or delivery vehicle for the mitigation of T2D progression., info:eu-repo/semantics/published

Aggregation of human islet amyloid polypeptide (hIAPP) into fibrils and plaques is associated with pancreatic β-cell loss in type 2 diabetes (T2D). However, due to the rapidness of hIAPP conversion in aqueous phase, exactly which hIAPP species is responsible for the observed toxicity and through what mechanisms remains ambiguous. In light of the importance of understanding hIAPP toxicity for T2D here we show a biophysical scheme based on the use of a lipophilic Laurdan dye for examining MIN6 cell membranes upon exposure to fresh and oligomeric hIAPP as well as mature amyloid. It has been found that all three hIAPP species, especially fresh hIAPP, enhanced membrane fluidity and caused losses in cell viability. The cell generation of reactive oxygen species (ROS), however, was the most pronounced with mature amyloid hIAPP. The correlation between changes in membrane fluidity and cell viability and their lack of correlation with ROS production suggest hIAPP toxicity is elicited through both physical and biochemical means. This study offers a new insight into β-cell toxicity induced by controlled hIAPP species, as well as new biophysical methodologies that may prove beneficial for the studies of T2D as well as neurological disorders., info:eu-repo/semantics/published

Excavations at the military site of Tell el-Borg, just outside of the eastern Delta has produced two New Kingdom forts that were a part of the East Frontier defense system and the military highway, the Ways of Horus. Among the discoveries at Tell el-Borg were the burnt out remains of a cluster of reed huts. This study will investigate who the occupants of these huts may have been. The pottery and C14 dates suggest that these structures were used in the 2nd Intermediate Period to early New Kingdom. It will be argued provisionally that the occupants of this little community were dessert dwellers, possibly Shasu.

Human islet amyloid polypeptide (hIAPP, or amylin) forms amyloid deposits in the islets of Langerhans, a phenomenon that is associated with type-2 diabetes impacting millions of people worldwide. Accordingly, strategies against hIAPP aggregation are essential for the prevention and eventual treatment of the disease. Here, it is shown that generation-3 OH-terminated poly(amidoamine) dendrimer, a polymeric nanoparticle, can effectively halt the aggregation of hIAPP and shut down hIAPP toxicity in pancreatic MIN6 and NIT-1 cells as well as in mouse islets. This finding is supported by high-throughput dynamic light scattering experiment and thioflavin T assay, where the rapid evolution of hIAPP nucleation and elongation processes is halted by the addition of the dendrimer up to 8 h. Discrete molecular dynamics simulations further reveal that hIAPP residues bound strongly with the dendrimer near the c-terminal portion of the peptide, where the amyloidogenic sequence (residues 22-29) locates. Furthermore, simulations of hIAPP dimerization reveal that binding with the dendrimer significantly reduces formation of interpeptide contacts and hydrogen bonds, thereby prohibiting peptide self-association and amyloidosis. This study points to a promising nanomedicinal strategy for combating type-2 diabetes and may have broader implications for targeting neurological disorders whose distinct hallmark is also amyloid fibrillation., info:eu-repo/semantics/published

Pancreatic cancer (PC) is a lethal disease due to chemoresistance and metastatic spread. We have shown that the microtubule protein βIII-tubulin is upregulated in PC cells, and that inhibition of βIII-tubulin (siRNA or shRNA) increased chemosensitivity in vitro and reduced tumour growth/metastases in vivo (McCarroll et al., Oncotarget, 6, pg.2235-49, 2015). Clinical translation of this work is limited by a lack of pharmacological inhibitors for βIII-tubulin. siRNA-based therapeutics are a potential solution, but require a delivery vehicle (e.g. nanoparticles). Novel star nanoparticles were designed to deliver siRNA to PC cells.The aims of this study were: 1) to design and optimise novel star-shaped nanoparticles that deliver functional siRNA to PC cells in vitro; 2) to examine the biodistribution and gene silencing activity of star-siRNA complexes in vivo using preclinical PC mouse models; and 3) to determine the potential of star-βIII-tubulin siRNA complexes to reduce tumour growth in preclinical PC mouse models. Results of this study demonstrated that star nanoparticles were able to complex and deliver βIII-tubulin siRNA to PC cells and potently silence βIII-tubulin expression. We modified our generation 1 nanoparticles by addition of PEGylation (Star-PEG) which enabled high gene silencing efficiency in PC cells in the presence of serum (an essential property for systemic administration in patients). All the star nanoparticles were non-toxic to PC cells and normal human pancreatic ductal cells. However, star-PEG nanoparticles displayed a superior toxicity profile compared to the non-PEGylated star nanoparticles. Star-PEG 4 (with 12.5% covalently attached PEG and longer cationic side-arms) was non-toxic in vivo and delivered siRNA to pancreatic tumours after systemic administration. Star-PEG 4-βIII-tubulin siRNA potently silenced βIII-tubulin in PC tumours and pilot studies indicated reduced tumour growth in both a subcutaneo