Testing small molecule analogues of the Acanthocheilonema viteae immunomodulator ES-62 against clinically relevant allergens.

ES-62 is a glycoprotein secreted by the filarial nematode Acanthocheilonema viteae that protects against ovalbumin (OVA)-induced airway hyper-responsiveness in mice by virtue of covalently attached anti-inflammatory phosphorylcholine (PC) residues. We have recently generated a library of small molecule analogues (SMAs) of ES-62 based around its active PC moiety as a starting point in novel drug development for asthma and identified two compounds - termed 11a and 12b - that mirror ES-62's protective effects. In this study, we have moved away from OVA, a model allergen, to test the SMAs against two clinically relevant allergens - house dust mite (HDM) and cockroach allergen (CR) extract. We show that both SMAs offer some protection against development of lung allergic responses to CR, in particular reducing eosinophil infiltration, whereas only SMA 12b is effective in protecting against eosinophil-dependent HDM-induced allergy. These data therefore suggest that helminth molecule-induced protection against model allergens may not necessarily translate to clinically relevant allergens. Nevertheless, in this study, we have managed to demonstrate that it is possible to produce synthetic drug-like molecules based on a parasitic worm product that show therapeutic potential with respect to asthma resulting from known triggers in humans.

Genomic comparison of virulent and non-virulent Streptococcus agalactiae in fish.

Streptococcus agalactiae infections in fish are predominantly caused by beta-haemolytic strains of clonal complex (CC) 7, notably its namesake sequence type (ST) 7, or by non-haemolytic strains of CC552, including the globally distributed ST260. In contrast, CC23, including its namesake ST23, has been associated with a wide homeothermic and poikilothermic host range, but never with fish. The aim of this study was to determine whether ST23 is virulent in fish and to identify genomic markers of fish adaptation of S. agalactiae. Intraperitoneal challenge of Nile tilapia, Oreochromis niloticus (Linnaeus), showed that ST260 is lethal at doses down to 10(2) cfu per fish, whereas ST23 does not cause disease at 10(7) cfu per fish. Comparison of the genome sequence of ST260 and ST23 with those of strains derived from fish, cattle and humans revealed the presence of genomic elements that are unique to subpopulations of S. agalactiae that have the ability to infect fish (CC7 and CC552). These loci occurred in clusters exhibiting typical signatures of mobile genetic elements. PCR-based screening of a collection of isolates from multiple host species confirmed the association of selected genes with fish-derived strains. Several fish-associated genes encode proteins that potentially provide fitness in the aquatic environment.

Drug-like analogues of the parasitic worm-derived immunomodulator ES-62 are therapeutic in the MRL/Lpr model of systemic lupus erythematosus.

INTRODUCTION: ES-62, a phosphorylcholine (PC)-containing immunomodulator secreted by the parasitic worm Acanthocheilonema viteae, protects against nephritis in the MRL/Lpr mouse model of systemic lupus erythematosus (SLE). However, ES-62 is not suitable for development as a therapy and thus we have designed drug-like small molecule analogues (SMAs) based around its active PC-moiety. To provide proof of concept that ES-62-based SMAs exhibit therapeutic potential in SLE, we have investigated the capacity of two SMAs to protect against nephritis when administered to MRL/Lpr mice after onset of kidney damage. METHODS: SMAs 11a and 12b were evaluated for their ability to suppress antinuclear antibody (ANA) generation and consequent kidney pathology in MRL/Lpr mice when administered after the onset of proteinuria. RESULTS: SMAs 11a and 12b suppressed development of ANA and proteinuria. Protection reflected downregulation of MyD88 expression by kidney cells and this was associated with reduced production of IL-6, a cytokine that exhibits promise as a therapeutic target for this condition. CONCLUSIONS: SMAs 11a and 12b provide proof of principle that synthetic compounds based on the safe immunomodulatory mechanisms of parasitic worms can exhibit therapeutic potential as a novel class of drugs for SLE, a disease for which current therapies remain inadequate.

Similarity decay of enstrophy in an electron fluid.

A similarity decay law is proposed for enstrophy of a one-signed-vorticity fluid in a circular free-slip domain. It excludes the metastable equilibrium enstrophy which cannot drive turbulence, and approaches Batchelor's t(-2) law for strong turbulence. Measurements of the decay of a turbulent electron fluid agree well with the predictions of the decay law for a variety of initial conditions.

Data and debriefing observations on healthcare simulation to prepare for the COVID-19 pandemic.

We report on data and debriefing observations in the context of an immersive simulation conducted to (a) train clinicians and (b) test new protocols and kits, developed in table-top exercises without prior clinical experience to fit anticipated clinical encounters in the setting of the rapidly expanding COVID-19 pandemic. We simulated scenarios with particular relevance for anesthesiology, perioperative and critical care, including (1) cardiac arrest, (2) emergency airway management, (3) tele-instruction for remote guidance and supervision, and (4) transporting an intubated patient. Using a grounded theory approach, three authors (MHA, DLR, EHS) developed emergent themes. First alone and then together, we sought consensus in uncovering overarching themes and constructs from the debriefings. We thus performed an informal qualitative thematic analysis based in a critical realist epistemological position - the understanding that our findings, while real, are affected by situational variables and the observer's perspective[1,2]. We compared data from videos and triangulated the data by member checking. All participants and course instructors volunteered to participate in this educational project and contributed as co-authors to this manuscript. During debriefing, we applied crisis resource management concepts including situation awareness, prioritization of tasks, and clear communication practices, conducting the debriefing with emphasis on current TeamStepps 2.0 terminology and concepts. [3,4] In addition, we re-evaluated formerly familiar processes, as shortcomings of protocols, kits, and interdisciplinary cooperation became apparent. The data provide detailed observations on how immersive simulation and debriefing among peers mitigated the unfamiliarity of individual clinicians and the organization at large with the demands of an unprecedented healthcare crisis. We also observed and report on the anxiety caused by resource constraints, risk to clinicians in the face of limited personal equipment, and the overall uncertainty surrounding COVID-19. We began to summarize, interpret, critique, and discuss our data and debriefing observations in a rapid co-publication in the Journal of Clinical Anesthesia. [Healthcare Simulation to Prepare for the COVID-19 Pandemic][5].

Recognition of a DNA operator by the repressor of phage 434: a view at high resolution.

The repressors of temperate bacteriophages such as 434 and lambda control transcription by binding to a set of DNA operator sites. The different affinity of repressor for each of these sites ensures efficient regulation. High-resolution x-ray crystallography was used to study the DNA-binding domain of phage 434 repressor in complex with a synthetic DNA operator. The structure shows recognition of the operator by direct interactions with base pairs in the major groove, combined with the sequence-dependent ability of DNA to adopt the required conformation on binding repressor. In particular, a network of three-centered bifurcated hydrogen bonds among base pairs in the operator helps explain why 434 repressor prefers certain sites over others. These bonds, which stabilize the conformation of the bound DNA, can form only with certain sequences.

Development and evaluation of a one-step real-time reverse transcription polymerase chain reaction assay for the detection of salmonid alphaviruses in serum and tissues.

We designed 4 primer pairs to amplify conserved regions of the E1 or nsP4 genes of salmonid alphavirus (SAV) and evaluated their performance in optimized 1-step SYBR green real-time RT-PCR (RRT-PCR) assays. A single primer pair, amplifying a 227 bp segment of E1 was then chosen for further study. This RRT-PCR was shown to be highly repeatable and reproducible over a wide range of RNA dilutions, with a linear relationship between cycle threshold (Ct) value and RNA concentration over a 10(7) dilution range. The limit of detection was calculated to be < or = 1.5 TCID50 ml(-1). When applied to sera previously screened by virus isolation for SAV viraemia, the RRT-PCR correctly identified all 13 culture-positive samples, as well as finding an additional 28 sera positive. Relative semi-quantitation of sera showed a very highly significant relationship between copy number and TCID50 (p < 0.001, R2 = 0.9563). Following experimental infection of salmon, heart samples were consistently positive until 21 d post infection (dpi), with (weak) positive signals still detectable in 50% of fish 70 dpi.

The complex between phage 434 repressor DNA-binding domain and operator site OR3: structural differences between consensus and non-consensus half-sites.

BACKGROUND: The repressor of phage 434 binds to a set of operator sites as a homodimer. Its relative affinities for these sites determine the switch from lysogenic to lytic growth. The six 434 operator sites (OR1, OR2, OR3, OL1, OL2 and OL3) have a particularly simple organization; all are 14 base pairs long, with a conserved 5'-ACAA sequence symmetrically placed at either end, and a variable central six base pairs. OR3 is unique among naturally-occurring 434 operator sites in that it contains a non-consensus base pair, G.C, at the fourth position of the otherwise invariant 5'-ACAA sequence. Comparisons among structures of the 434 repressor DNA-binding domain, R1-69, bound to various operator sites, allow us to analyze differential specificity in regulatory complexes of this kind. RESULTS: We have determined the structure at 2.5 A resolution of a complex of R1-69 with DNA containing the OR3 site and compared it with previously studied complexes of R1-69 bound to OR1 and OR2. There are surprisingly extensive structural differences between the consensus and non-consensus half-sites of OR3 with respect to their interactions with R1-69, including a shift in the DNA backbone and a small rotation of the entire R1-69 monomer. CONCLUSIONS: Recognition of the base pair difference that is critical for the 434 regulatory switch involves a number of amino acid residues, not just the one or two side chains in direct contact with the G-C base pair. Moreover, the repressor imposes a somewhat altered DNA conformation on the non-consensus half-site.

The parasitic worm-derived immunomodulator, ES-62 and its drug-like small molecule analogues exhibit therapeutic potential in a model of chronic asthma.

Chronic asthma is associated with persistent lung inflammation and long-term remodelling of the airways that have proved refractory to conventional treatments such as steroids, despite their efficacy in controlling acute airway contraction and bronchial inflammation. As its recent dramatic increase in industrialised countries has not been mirrored in developing regions, it has been suggested that helminth infection may protect humans against developing asthma. Consistent with this, ES-62, an immunomodulator secreted by the parasitic worm Acanthocheilonema viteae, can prevent pathology associated with chronic asthma (cellular infiltration of the lungs, particularly neutrophils and mast cells, mucus hyper-production and airway thickening) in an experimental mouse model. Importantly, ES-62 can act even after airway remodelling has been established, arresting pathogenesis and ameliorating the inflammatory flares resulting from repeated exposure to allergen that are a debilitating feature of severe chronic asthma. Moreover, two chemical analogues of ES-62, 11a and 12b mimic its therapeutic actions in restoring levels of regulatory B cells and suppressing neutrophil and mast cell responses. These studies therefore provide a platform for developing ES-62-based drugs, with compounds 11a and 12b representing the first step in the development of a novel class of drugs to combat the hitherto intractable disorder of chronic asthma.

Patients' attitudes toward screening for gestational diabetes mellitus in the Illawarra area, Australia.

OBJECTIVE: To examine the attitudes toward screening of a group of women who have had GDM compared with a group who were screened but did not have GDM. RESEARCH DESIGN AND METHODS: We surveyed 90 women with GDM and 100 control women with a postal questionnaire. They were asked to mark on a six-point Likert scale their responses to statements about certain aspects of screening for diabetes in pregnancy. RESULTS: No significant differences were found in mean responses between the two groups. The method of screening was found to be convenient (GDM group, 5.02; control group, 4.77). A strong belief that all women should be screened in pregnancy (GDM group, 5.76; control group, 5.66) and a personal desire for screening in any future pregnancy (GDM group, 5.73; control group, 5.62) were expressed. CONCLUSIONS: Both groups of women surveyed in this study were extremely positive about the convenience of the screening method used, the need for all women to be screened in pregnancy, and their desire for screening in future pregnancies.

Mammography screening and breast cancer tumor size in female members of a managed care organization.

A study of temporal trends in mammography screening and changes in stage of disease at diagnosis was conducted among Hispanic and non-Hispanic white female members of the Lovelace Health Plan, Flexcare Plan, and Lovelace Senior Plan/Senior Options (LHP), a managed care organization. Two-year screening rates for female members ages 50-74 years were calculated for 1989-1996. From 1989-1996, mammography screening rates for non-Hispanic white female members increased from 65.5 to 71.6%, although this was not a statistically significant increase. Screening rates for Hispanic female members also increased from 50.6 to 62.7%, but they were significantly lower than for non-Hispanic white women. All breast cancers occurring among LHP female members ages 40-74 years were also identified for this same time period. A logistic regression model adjusting for age, year of diagnosis, ethnicity, and duration of enrollment prior to diagnosis found that statistically significant predictors of more advanced stage of disease at diagnosis included young age, diagnosis after 1991 for non-Hispanic white women, and diagnosis prior to 1992 for Hispanic women. Longer duration of enrollment prior to diagnosis was predictive of lower stage of disease, but the odds ratio was not statistically significant. For the time period 1992-1996, Hispanic women with breast cancer were more than twice as likely to have advanced stage of breast cancer compared with non-Hispanic white women (odds ratio, 2.12).

Intracoronary thrombus in nontransmural myocardial infarction and in unstable angina pectoris.

Although intracoronary thrombus formation plays a major role in acute transmural myocardial infarction (MI), its occurrence in unstable angina (UA) and nontransmural MI has not clearly been established. To determine whether intracoronary thrombus does occur in these syndromes, coronary arteriography was performed before, during, and after intracoronary nitroglycerin and streptokinase infusion in 17 patients. None of the 8 patients with nontransmural MI and 1 of the 9 patients with UA responded to intracoronary nitroglycerin. Seven of 8 patients with nontransmural MI and 4 of 9 patients with UA responded to streptokinase infusion with opening of an occluded vessel, an increase in stenotic diameter, dissolution of an intracoronary filling defect, or a combination of these. Serial opening and closing of ischemia-related vessels occurred spontaneously and in response to streptokinase in some patients in whom thrombolysis was demonstrated. Evidence of thrombolysis was not seen in any patient studied longer than 1 week from the onset of the rest pain syndrome. The finding of thrombolysis in several patients with nontransmural MI and UA suggests that intracoronary thrombus formation plays a pathogenetic role in some patients with these ischemic syndromes.

Precocious myocardial infarction after radiation treatment for Hodgkin's disease.

A 26-year-old man received extensive cardiac radiation in the course of treatment of mediastinal Hodgkin's disease, and six years later, he experienced an acute myocardial infarction. Angiographic studies demonstrated extensive atherosclerotic abnormalities in the coronary arterial system. It is suggested that radiation-induced injury was a provocation of these precocious arterial abnormalities.

High attack rates and case fatality during a measles outbreak in groups with religious exemption to vaccination.

Beginning in October, 1990, a large measles outbreak involving predominantly unvaccinated preschool age children occurred in Philadelphia. By June, 1991, 938 measles cases had been reported to the Philadelphia Health Department. In addition to these cases, 486 cases and 6 measles-associated deaths occurred between November 4, 1990, and March 24, 1991, among members of 2 Philadelphia church groups that do not accept vaccination. We identified measles cases and collected information on symptoms and potential risk factors for complications. Telephone interviews were conducted to collect demographic and clinical information on measles cases in church member households. We identified 486 measles cases among 892 mostly unvaccinated church members. Age-specific attack rates were highest among children 1 to 4 years of age (94%) and 5 to 14 years of age (91%). Five (83%) of the 6 deaths occurred in females, 3 of whom had underlying illnesses. The overall case-fatality rate was 1.2%. The case-fatality rate was 2% for females, 0.4% for males (P = 0.22), 1.7% for primary cases and 0.7% for secondary household cases (P = 0.67). Only one of the children who died had received medical care. Measles spread rapidly in this group, sparing few susceptible individuals. Lack of medical care and underlying disease appear to have contributed to the high case-fatality rate in the church communities.

Erythrocyte filterability in normal and high-risk pregnancy.

Erythrocyte filterability was studied longitudinally in normal pregnancy and in certain categories of high-risk pregnancy. Study subjects included ten normal controls, 12 insulin-dependent diabetics, eight gestational diabetics, and five essential hypertensives. Our results indicate that erythrocyte filterability remains relatively stable over the course of normal gestation. We noted no differences between controls and essential hypertensives or gestational diabetics, although a favorable effect of insulin therapy was suggested in gestational diabetics. Erythrocyte filterability and mean arterial blood pressure were not related. Insulin-dependent diabetics demonstrated a significantly elevated and widely varying erythrocyte filterability, and individual patient trends correlated well with outcome. Fibrinogen levels in diabetics rose precipitously and were significantly higher than normal throughout gestation. Fibrinogen levels paralleled changes in erythrocyte filterability, with the two parameters positively correlated. Mean glucose control had no influence on filterability. We conclude that in the diabetic pregnancy, varying erythrocyte filterability is related to altered fibrinogen metabolism and may contribute to perinatal morbidity.

Exit-site care--is it time for a change?

Peritoneal catheter exit-site infections (ESI's) continue to impact significantly on morbidity and catheter longevity. The controversy concerning protocols for daily exit-site care continues for frequency, methodology, cleansing agent, and dressing. Routine daily exit-site care prior to January 1991 consisted of daily showers using liquid soap, povidone scrub, rinsing the shower, and drying with a 4 x 4-in. gauze pad. Catheters have always been immobilized, either with tape or an immobilizing device. Hydrogen peroxide was used only when needed to soften crust formation prior to showering. A light dressing, usually 2 x 2 in., was optional. A recent survey revealed that povidone iodine was the antiseptic of choice for catheter care in 75% of the respondents. However, povidone iodine irritates and dries the skin predisposing it to infection. ESI's are prospectively monitored as part of our quality improvement (QI) program. An incidence of 0.76 episodes/patient-year was noted between January 1989 and May 1991. Given the relative high frequency of ESI's, the protocol was modified and introduced during the January-May 1991 time frame. Routine care now consists of daily showers using only CC-500, a gentle antibacterial cleaner (Care-Tech Laboratories, Inc.), rinsing in the shower, and drying with a 4 x 4-in. gauze. Use of hydrogen peroxide and dressings has remained the same. Additionally, a protocol addressing the prophylaxis for traumatized exist sites was initiated. The incidence of ESI's has dropped significantly to 0.12 episodes/patient-year. Although our population size is small (n = 18), this study does point out the utility of prospectively monitoring trends for appropriate indicators within a QI program.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of pH and feeding regime on methylmercury accumulation within aquatic microcosms.

The effect of pH (pH 5, 6 or 7) on accumulation of radiolabelled methylmercury was examined using a laboratory microcosm system. Accumulation of labelled mercury at three trophic levels, primary consumers (Daphnia magna), secondary consumers (rainbow trout, Salmo gairdneri) and tertiary consumers (walleye, Stizostedion vitreum) was not significantly affected by pH. Our results are in direct contrast with field observations of elevated methylmercury concentrations in fish from low pH water and indicate that the elevated mercury residues observed in the field result from factors other than the direct effects of pH on accumulation of methylmercury by aquatic organisms. Both water and diet were important as sources of the labelled mercury which was accumulated by walleye. Walleye which were fed rainbow trout, that had accumulated labelled mercury from within the experimental microcosms, accumulated almost twice as much labelled mercury as walleye fed non-labelled prey, or walleye which were not fed. Walleye fed non-labelled rainbow trout accumulated slightly more labelled mercury than walleye which were not fed, presumably as a result of the higher metabolic rate of the fish which were fed.

Cross-cultural studies in Alzheimer's disease.

The search for risk factors for Alzheimer's disease would be greatly enhanced by identification of populations with significantly different prevalence rates, particularly if these populations consisted of ethnic groups now living in different environments and cultures. Evidence is presented that two such groups are worthy of further study: subjects of African origin living in Africa and in the West and Native Americans living on and off reserves.

Cognitive therapy for multiple sclerosis: a preliminary study.

BACKGROUND: Drug treatments for multiple sclerosis are expensive, may cause side effects, and do not have demonstrated efficacy for cognitive deficits associated with this disease. OBJECTIVE: To test the effectiveness of a multimodal cognitive therapy on cognitive and physical measures known to be affected in multiple sclerosis. DESIGN: Quasi-experimental wait-list control. SETTING: Alternative medicine clinic. PATIENTS: 27 persons with clinically definite multiple sclerosis. INTERVENTION: Multimodal cognitive therapy. MAIN OUTCOME MEASURES: Neuropsychological measures of verbal learning and memory, abstraction, vocabulary, and information processing speed; Beck Depression Inventory; tactile sensitivity of the hands; grip strength; and visual acuity. MAIN RESULTS: 12 of 14 patients in the therapy group and 10 of 13 patients in the control group completed 24 weeks of treatment and all assessments. Patients who received therapy showed significantly greater improvement in verbal learning, verbal abstraction, depression, and some measures of grip strength and tactile sensitivity than did patients in the untreated control group. The groups did not differ in the magnitude of change on vocabulary, information processing speed, or visual acuity. CONCLUSION: Cognitive therapy appears to be a promising treatment for ameliorating some symptoms of multiple sclerosis. A larger study with a randomized design and additional outcome measures is warranted.

Knowledge and care of chronic illness in three ethnic minority groups.

BACKGROUND AND OBJECTIVES: Despite advances in medical approaches to the management of chronic illnesses, relatively little is known about how older members of ethnic minority groups view their chronic illnesses or how they manage them in daily life. METHODS: We recruited 35 African-Americans, 61 Latinos, and 55 Filipino-Americans, all over age 50. Criteria for entry into the study was the presence of one or more chronic illnesses. Findings are based on structured and semi-structured questions in one in-depth interview. Qualitative data on transcribed interviews with 151 respondents was analyzed. RESULTS: Comparison of the three groups revealed social and cultural differences and similarities that affected the management of chronic illness. The extent to which respondents demonstrated an understanding of their illnesses as chronic varied considerably, with discernible differences among groups about knowledge of illness and self-care practices. CONCLUSIONS: Our findings showed that although major chronic illnesses were, for the most part, the same for all three groups, each group differed in its response to and management of its illnesses. These findings have implications for the education of physicians in training.