Tetrachlorovancomycin: Total Synthesis of a Designed Glycopeptide Antibiotic of Reduced Synthetic Complexity.

A technically straightforward total synthesis of a new class of vancomycin analogues of reduced synthetic complexity was developed that provided tetrachlorovancomycin (1, LLS = 15 steps, 15% overall yield) and its precursor aglycon 29 (nearly 20% overall yield). The class retains all the intricate vancomycin structural features that contribute to its target binding affinity and selectivity, maintains the antimicrobial activity of vancomycin, and achieves the simplification by an unusual addition, not removal, of benign substituents to the core structure. The modification, accomplished by addition of two aryl chloride substituents to provide 1, permitted a streamlined total synthesis of the new glycopeptide antibiotic class by removing the challenges associated with CD and DE ring system atropisomer stereochemical control. This also enabled their simultaneous and further-activated SNAr macrocyclizations that establish the tricyclic skeleton of 1. Key elements of the approach include catalyst-controlled diastereoselective formation of the AB biaryl axis of chirality (>30:1 dr), an essentially instantaneous macrolactamization of the AB ring system free of competitive epimerization (>30:1 dr), racemization free coupling of the E ring tetrapeptide, room temperature simultaneous CD and DE ring system cyclizations, a highly refined 4-step conversion of the cyclization product to the aglycon, and a protecting-group-free one-pot enzymatic glycosylation for disaccharide introduction. In addition to the antimicrobial evaluation of tetrachlorovancomycin (1), the preparation of key peripherally modified derivatives, which introduce independent and synergistic mechanisms of action, revealed their exceptional antimicrobial potency and provide the foundation for future use of this new class of synthetic glycopeptide analogues.

68Ga-NODAGA-Indole: An Allysine-Reactive Positron Emission Tomography Probe for Molecular Imaging of Pulmonary Fibrogenesis.

Oxidized collagen, wherein lysine residues are converted to the aldehyde allysine, is a universal feature of fibrogenesis, i.e. actively progressive fibrosis. Here we report the small molecule, allysine-binding positron emission tomography probe, 68Ga-NODAGA-indole, that can noninvasively detect and quantify pulmonary fibrogenesis. We demonstrate that the uptake of 68Ga-NODAGA-indole in actively fibrotic lungs is 7-fold higher than in control groups and that uptake is linearly correlated ( R2 = 0.98) with the concentration of lung allysine.

Completing the mechanical energy pathways in turbulent Rayleigh-Bénard convection.

A new, more complete view of the mechanical energy budget for Rayleigh-Bénard convection is developed and examined using three-dimensional numerical simulations at large Rayleigh numbers and Prandtl number of 1. The driving role of available potential energy is highlighted. The relative magnitudes of different energy conversions or pathways change significantly over the range of Rayleigh numbers Ra ~ 10(7)-10(13). At Ra < 10(7) small-scale turbulent motions are energized directly from available potential energy via turbulent buoyancy flux and kinetic energy is dissipated at comparable rates by both the large- and small-scale motions. In contrast, at Ra ≥ 10(10) most of the available potential energy goes into kinetic energy of the large-scale flow, which undergoes shear instabilities that sustain small-scale turbulence. The irreversible mixing is largely confined to the unstable boundary layer, its rate exactly equal to the generation of available potential energy by the boundary fluxes, and mixing efficiency is 50%.

Stability of Free Available Chlorine Levels in Dilute Sodium Hypochlorite Solutions over a 6-Week Period.

Animal care and use programs commonly use chlorine and chlorine-based disinfectants to help prevent facility acquired infections in animals. The Department of Comparative Medicine (DCM) at Oregon Health and Science University (OHSU) follows the Centers for Disease Control and Prevention (CDC) disinfection guidelines for preparing and storing these disinfectants. DCM prepares bottles of dilute solutions of sodium hypochlorite (that is, commercial bleach) daily. In this study, we tested whether dilute bleach solutions, as prepared following the DCM protocol, remained stable under real-world practice conditions for up to 6 wk. We tested 4 groups of spray bottles filled with 0.5% bleach solutions in these experiments. Specifically, we sprayed 2 groups of bottles daily to mimic use while 2 other groups of bottles were not sprayed. We then measured free available chlorine (FAC) using 2 methods, spectrophotometry and colorimetric strips. All 4 test groups showed stable maintenance of FAC concentration for the length of the experiment. Mean FAC loss from baseline levels was not significantly different in the group of bottles not sprayed daily (6% for group 2 at week 5 compared with 7% for Group 4 at week 6). All bottles in Groups 1 and 3 measured by colorimetric strips showed concentrations at or near 5000 mg/L at all weekly time points throughout the experiment. This study shows that 0.5% sodium hypochlorite solutions stored and used in a standard rodent housing room and sprayed daily will maintain acceptable FAC concentrations for at least 5 to 6 wk, perhaps longer. In addition, we report that colorimetric strips may be a useful and accessible quality control tool for testing freshly prepared solutions at regular intervals. We conclude that sodium hypochlorite solutions can be prepared on a weekly, biweekly, or monthly basis with no loss in disinfection effectiveness.

Responding to eruptive transitions during the 2020-2021 eruption of La Soufrière volcano, St. Vincent.

A critical challenge during volcanic emergencies is responding to rapid changes in eruptive behaviour. Actionable advice, essential in times of rising uncertainty, demands the rapid synthesis and communication of multiple datasets with prognoses. The 2020-2021 eruption of La Soufrière volcano exemplifies these challenges: a series of explosions from 9-22 April 2021 was preceded by three months of effusive activity, which commenced with a remarkably low level of detected unrest. Here we show how the development of an evolving conceptual model, and the expression of uncertainties via both elicitation and scenarios associated with this model, were key to anticipating this transition. This not only required input from multiple monitoring datasets but contextualisation via state-of-the-art hazard assessments, and evidence-based knowledge of critical decision-making timescales and community needs. In addition, we share strategies employed as a consequence of constraints on recognising and responding to eruptive transitions in a resource-constrained setting, which may guide similarly challenged volcano observatories worldwide.

Causes of ant sting anaphylaxis in Australia: the Australian Ant Venom Allergy Study.

OBJECTIVE: To determine the Australian native ant species associated with ant sting anaphylaxis, geographical distribution of allergic reactions, and feasibility of diagnostic venom-specific IgE (sIgE) testing. DESIGN, SETTING AND PARTICIPANTS: Descriptive clinical, entomological and immunological study of Australians with a history of ant sting anaphylaxis, recruited in 2006-2007 through media exposure and referrals from allergy practices and emergency physicians nationwide. We interviewed participants, collected entomological specimens, prepared reference venom extracts, and conducted serum sIgE testing against ant venom panels relevant to the species found in each geographical region. MAIN OUTCOME MEASURES: Reaction causation attributed using a combination of ant identification and sIgE testing. RESULTS: 376 participants reported 735 systemic reactions. Of 299 participants for whom a cause was determined, 265 (89%; 95% CI, 84%-92%) had reacted clinically to Myrmecia species and 34 (11%; 95% CI, 8%-16%) to green-head ant (Rhytidoponera metallica). Of those with reactions to Myrmecia species, 176 reacted to jack jumper ant (Myrmecia pilosula species complex), 18 to other jumper ants (15 to Myrmecia nigrocincta, three to Myrmecia ludlowi) and 56 to a variety of bulldog ants, with some participants reacting to more than one type of bulldog ant. Variable serological cross-reactivity between bulldog ant species was observed, and sera from patients with bulldog ant allergy were all positive to one or more venoms extracted from Myrmecia forficata, Myrmecia pyriformis and Myrmecia nigriceps. CONCLUSION: Four main groups of Australian ants cause anaphylaxis. Serum sIgE testing enhances the accuracy of diagnosis and is a prerequisite for administering species-specific venom immunotherapy.

The use of an integrated care pathway for evidence-based practice and clinical governance in abortion care.

The aim of the study was to assess the effectiveness of an integrated care pathway (ICP) for delivery of evidence-based practice in abortion care. All women re-admitted after an abortion had their records audited for adherence to national and local guidelines using information in the ICP and general gynaecology case notes. A total of 100 women were re-admitted into the gynaecological wards of hospitals in Hull and East Riding of Yorkshire after an abortion, between January 2000 and December 2006. Out of 8,476 medical or surgical induced abortions undertaken at 14 weeks gestation or under, the overall readmission rate was 1.2%. The ICP showed that 97% of women had chlamydia screening prior to the abortion; all women had a contraceptive discussion and 43% left using a long-acting reversible method of contraception (LARC). However, data outside the care pathway was not documented, and hence the standard of care given on readmission was difficult to locate and variable in quality. The ICP clinical record is demonstrated to be a useful tool for high quality record-keeping and ensuring all patients receive the same standard of pre-assessment care. Although this service has an acceptably low-risk profile in terms of re-admission, we propose the addition of a re-admission episode to the current ICP to further enhance clinical care post-abortion.

Viewing ultrasound scan images prior to termination of pregnancy: choice for women or conflict for ultrasonographers?

Establishing pregnancy location is key to minimising the risks of abortion, and establishing gestational age optimises women's choice of procedure and timing. There is limited information in UK practice about the views of women having an ultrasound before an abortion with regards to whether they would wish to see the scan image of their fetus. We therefore surveyed the views of women having pre-abortion ultrasound scans and the views of ultrasonographers, using self-completed anonymous questionnaires. Responses were available for analysis from 191 women aged 16 to over 45. A total of 25 (13%) women had seen their scan. Of those who did not see their scan, 40 (24%) said they would have liked to see it, to aid their decision-making, as a right, or out of curiosity. Nine out of 17 ultrasonographers completed their questionnaires. Four ultrasonographers thought that women having scans before an abortion should see their scan images if they wished and another four thought they should not because of possible psychological effects. Their response also highlighted possible effects of such scans on those performing them. Given the choice, a larger number of women would like to see their scan images than actually ask. Women request to see their pre-abortion ultrasound images for their own individual reasons, which should be respected. Ultrasonographers may have conflict or ethical dilemmas when performing pre-abortion scans and they should be supported in identifying women for whom looking at their scan images would be of benefit.

Nanopore sequencing of the glucocerebrosidase (GBA) gene in a New Zealand Parkinson's disease cohort.

INTRODUCTION: Bi-allelic mutations in the gene for glucocerebrosidase (GBA) cause Gaucher disease, an autosomal recessive lysosomal storage disorder. Gaucher disease causing GBA mutations in the heterozygous state are also high risk factors for Parkinson's disease (PD). GBA analysis is challenging due to a related pseudogene and structural variations (SVs) that can occur at this locus. We have applied and refined a recently developed nanopore DNA sequencing method to analyze GBA variants in a clinically assessed New Zealand longitudinal cohort of PD. METHOD: We examined amplicons encompassing the coding region of GBA (8.9 kb) from 229 PD cases and 50 healthy controls using the GridION nanopore sequencing platform, and Sanger validation. RESULTS: We detected 23 variants in 21 PD cases (9.2% of patients). We detected modest PD risk variant p.N409S (rs76763715) in one case, p.E365K (rs2230288) in 12 cases, and p.T408 M (rs75548401) in seven cases, one of whom also had p.E365K. We additionally detected the possible risk variants p.R78C (rs146774384) in one case, p.D179H (rs147138516) in one case which occurred on the same haplotype as p.E365K, and one novel variant c.335C > T or p.(L335 = ), that potentially impacts splicing of GBA transcripts. Additionally, we found a higher prevalence of dementia among patients with GBA variants. CONCLUSION: This work confirmed the utility of nanopore sequencing as a high-throughput method to identify known and novel GBA variants, and to assign precise haplotypes. Our observations may contribute to improved understanding of the effects of variants on disease pathogenesis, and to the development of more targeted treatments.

Fibrotic Response to Neoadjuvant Therapy Predicts Survival in Pancreatic Cancer and Is Measurable with Collagen-Targeted Molecular MRI.

PURPOSE: To evaluate the prognostic value of posttreatment fibrosis in human PDAC patients, and to compare a type I collagen targeted MRI probe, CM-101, to the standard contrast agent, Gd-DOTA, for their abilities to identify FOLFIRINOX-induced fibrosis in a murine model of PDAC. EXPERIMENTAL DESIGN: Ninety-three chemoradiation-treated human PDAC samples were stained for fibrosis and outcomes evaluated. For imaging, C57BL/6 and FVB mice were orthotopically implanted with PDAC cells and FOLFIRINOX was administered. Mice were imaged with Gd-DOTA and CM-101. RESULTS: In humans, post-chemoradiation PDAC tumor fibrosis was associated with longer overall survival (OS) and disease-free survival (DFS) on multivariable analysis (OS P = 0.028, DFS P = 0.047). CPA increased the prognostic accuracy of a multivariable logistic regression model comprised of previously established PDAC risk factors [AUC CPA (-) = 0.76, AUC CPA (+) = 0.82]. In multiple murine orthotopic PDAC models, FOLFIRINOX therapy reduced tumor weight (P < 0.05) and increased tumor fibrosis by collagen staining (P < 0.05). CM-101 MR signal was significantly increased in fibrotic tumor regions. CM-101 signal retention was also increased in the more fibrotic FOLFIRINOX-treated tumors compared with untreated controls (P = 0.027), consistent with selective probe binding to collagen. No treatment-related differences were observed with Gd-DOTA imaging. CONCLUSIONS: In humans, post-chemoradiation tumor fibrosis is associated with OS and DFS. In mice, our MR findings indicate that translation of collagen molecular MRI with CM-101 to humans might provide a novel imaging technique to monitor fibrotic response to therapy to assist with prognostication and disease management.

Peroxidase Sensitive Amplifiable Probe for Molecular Magnetic Resonance Imaging of Pulmonary Inflammation.

An amplifiable magnetic resonance imaging (MRI) probe that combines the stability of the macrocyclic Gd-DOTAGA core with a peroxidase-reactive 5-hydroxytryptamide (5-HT) moiety is reported. The incubation of the complex under enzymatic oxidative conditions led to a 1.7-fold increase in r1 at 1.4 T that was attributed to an oligomerization of the probe upon oxidation. This probe, Gd-5-HT-DOTAGA, provided specific detection of lung inflammation by MRI in bleomycin-injured mice.

Orthotopic and heterotopic murine models of pancreatic cancer and their different responses to FOLFIRINOX chemotherapy.

Syngeneic, immunocompetent allograft tumor models recapitulate important aspects of the tumor microenvironment and have short tumor latency with predictable growth kinetics, making them useful for trialing novel therapeutics. Here, we describe surgical techniques for orthotopic and heterotopic pancreatic ductal adenocarcinoma (PDAC) tumor implantation and characterize phenotypes based on implantation site.Mice (n=8 per group) were implanted with 104 cells in the pancreas or flank. Hy15549 and Han4.13 cell lines were derived from primary murine PDAC (Ptf1-Cre; LSL-KRAS-G12D; Trp53 Lox/+) on C57BL/6 and FVB strains, respectively. Single-cell suspension and solid tumor implants were compared. Tumors were treated with two intravenous doses of FOLFIRINOX and responses evaluated.All mice developed pancreatic tumors within 7 days. Orthotopic tumors grew faster and larger than heterotopic tumors. By 3 weeks, orthotopic mice began losing weight, and showed declines in body condition requiring euthanasia starting at 4 weeks. Single-cell injection into the pancreas had near 100% engraftment, but solid tumor implant engraftment was ∼50% and was associated with growth restriction. Orthotopic tumors were significantly more responsive to intravenous FOLFIRINOX compared with heterotopic tumors, with greater reductions in size and increased apoptosis. Heterotopic tumors were more desmoplastic and hypovascular. However, drug uptake into tumor tissue was equivalent regardless of tumor location or degree of fibrosis, indicating that microenvironment differences between heterotopic and orthotopic tumors influenced response to therapy.Our results show that orthotopic and heterotopic allograft locations confer unique microenvironments that influence growth kinetics, desmoplastic response and angiogenesis. Tumor location influences chemosensitivity to FOLFIRINOX and should inform future preclinical trials.This article has an associated First Person interview with the first author of the paper.

Molecular magnetic resonance imaging accurately measures the antifibrotic effect of EDP-305, a novel farnesoid X receptor agonist.

We examined a novel farnesoid X receptor agonist, EDP-305, for its antifibrotic effect in bile duct ligation (BDL) and choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) models of hepatic injury. We used molecular magnetic resonance imaging with the type 1 collagen-binding probe EP-3533 and the oxidized collagen-specific probe gadolinium hydrazide to noninvasively measure treatment response. BDL rats (n = 8 for each group) were treated with either low or high doses of EDP-305 starting on day 4 after BDL and were imaged on day 18. CDAHFD mice (n = 8 for each group) were treated starting at 6 weeks after the diet and were imaged at 12 weeks. Liver tissue was subjected to pathologic and morphometric scoring of fibrosis, hydroxyproline quantitation, and determination of fibrogenic messenger RNA expression. High-dose EDP-305 (30 mg/kg) reduced liver fibrosis in both the BDL and CDAHFD models as measured by collagen proportional area, hydroxyproline analysis, and fibrogenic gene expression (all P < 0.05). Magnetic resonance signal intensity with both EP-3533 in the BDL model and gadolinium hydrazide in the CDAHFD model was reduced with EDP-305 30 mg/kg treatment (P < 0.01). Histologically, EDP-305 30 mg/kg halted fibrosis progression in the CDAHFD model. Conclusion: EDP-305 reduced fibrosis progression in rat BDL and mouse CDAHFD models. Molecular imaging of collagen and oxidized collagen is sensitive to changes in fibrosis and could be used to noninvasively measure treatment response in clinical trials. (Hepatology Communications 2018;2:821-835).

Dual macroaneurysms on the same retinal arteriole.

A case of two macroaneurysms on the same retinal arteriole is presented. After observation of these lesions for some weeks, an increased threat to the macula from oedema led to laser photocoagulation. Although treatment did not hasten resolution, the patient has remained asymptomatic. The aetiology and natural history of macroaneurysms are discussed.

A model for developing and pre-testing a multi-media teaching program to enhance the self-care behavior of diabetes insipidus patients.

A patient-education program was designed by members of the Neurology Nursing Service. The program was designed to provide patients with diabetes insipidus with the information needed to help them manage their condition in the hospital and at home upon discharge. The program was approved by the Nursing Department's Patient Education Committee. The nurses worked closely with the hospital's patient-education coordinator and television-production staff to produce a videotape for educating patients about managing diabetes insipidus. A literature review of patient-education resources on diabetes insipidus revealed that no materials were available in either audiovisual or print media. The program utilized the multi-media approach of a videotape that was shown over closed-circuit television and a booklet outlining the salient points of the television program. All patients utilizing the program have been pre-tested and post-tested to assess their achievement of behavioral objectives.

A model for ambulatory care of patients with epilepsy and other neurological disorders.

A model for ambulatory care of patients with epilepsy and other neurological disorders facilitates delivery of high quality nursing care, provides a framework for formulating standard nursing care plans and incorporates patient education components. Education and development of nurses working in this setting is fostered through caregiving activities and development of teaching materials coordinated through the model. The model can be modified to meet the needs of each patient or the requirements of various clinical research protocols and adapted to patients with non-neurological medical or surgical diagnoses.

Professional nursing practice in neurology ambulatory care.

The ambulatory care setting is an important expanding area for professional nursing practice. The demand for ambulatory care nurses is stimulating the definition and development of professional nursing practices in outpatient clinics. Neurology ambulatory care at the National Institutes of Health, Bethesda, Md, is provided in the context of contributing to biomedical research. This article discusses the components of patient selection criteria, experimental therapy, and research data collection that have implications for nursing care. Visit categories are used to discuss administrative and practice issues of the research patient population at all institutes. Management plans are discussed, and a case study is presented along with a care plan.

Collaborative research.

Our experience of collaborative research "the process of working with others in pursuit of scientific discovery" has been a most rewarding experience. The nurses and physicians have developed mutual respect and appreciation of the value of each other's practice, and true collaboration has emerged. The relationships between the nurses and physicians have developed over time and have been supported by our visionary Associate Director for Nursing, Kathryn McKeon. Throughout this process of collaboration, we've grown to appreciate each others strengths and recognize what each discipline has to offer. We have also learned that when we work together, it is the patient who ultimately benefits most. The neurologists' holistic approach to patient care and their respect for nursing practice have greatly contributed to the establishment of truly collaborative research.

Gaucher's disease: a pilot study of the symptomatic responses to enzyme replacement therapy.

Symptomatic responses to enzyme replacement therapy were studied in 12 patients with an inherited lipid storage disorder (Type 1 Gaucher's disease) in order to determine the impact of treatment on perceptions of well being. Before each intravenous infusion of enzyme, patients were asked to comment on the presence or absence of disease-specific symptoms presented in questionnaire format. Symptoms were grouped into five major categories: bleeding abnormalities, chronic fatigue, gastrointestinal complaints, bone pain and psychosocial function. Each reported symptom was discussed in detail with the clinic nurse coordinator and documented in the medical record. After six months of enzyme replacement therapy, each patient's chart was reviewed and changes in the frequency of disease-specific symptoms over time were evaluated. The major subjective changes included a decrease in the frequency and severity of nosebleeds, reduced bruising, increased vigor and energy level, visible reduction in abdominal girth, increased self-esteem and enhanced self-image. Patients frequently described relief of their symptoms well before changes were confirmed by objective laboratory measurements. These observations may prove useful in the comprehensive management of patients with Gaucher's disease as they recover from a chronic, debilitating illness while receiving enzyme replacement therapy. The extent and ease with which these patients can achieve a state of normal, healthy function is unknown at the present time and will require further study.