Time-temperature effects on Salmonellae and Staphylococci in foods. III. Thermal death time studies.
Thermal death time studies were conducted at 5 F intervals from 130 to 150 F with strains of salmonellae and enterotoxigenic staphylococci. Heat-resistant Salmonella senftenberg strain 775W, Staphylococcus aureus strains 196E and Ms149, and non-heat-resistant Salmonella manhattan were studied in custard, chicken à la king, and ham salad. The F(140) values (minutes of exposure at 140 F required to effect 100% destruction) were as follows: S. senftenberg 775W in custard 78, and chicken à la king 81.5; S. manhattan in custard 19, and chicken à la king 3.1; S. aureus 196E in custard 59, and chicken à la king 47; S. aureus Ms149 in custard 53, and chicken à la king 40. The end points of survival-kill at all the test temperatures for both salmonellae and staphylococci in ham salad were considerably less than for the other foods studied.D(140) values (minutes of exposure at 140 F required to effect a 90% reduction in numbers) were also calculated from the data and presented. Values for z(F) and z(D) (slope of the thermal-death-time and decimal-reduction-time curves) are also presented and discussed in relation to type of food, organism, and temperature. These data indicate that heating perishable foods of the type studied to 150 F and holding every particle of food at this temperature for at least 12 min reduces 10 million or less salmonellae or staphylococci per gram to nondetectable levels. The same degree of destruction is achieved in similarly contaminated foods when held at 140 F for 78 to 83 min. On the basis of the calculation procedures employed, it is estimated that 45-min exposure at 140 F would be necessary to reduce 1,000 organisms per gram to nondetectable levels.


The effect of oral care on intracranial pressure in critically ill adults.
A major goal in the care of patients with neurological problems is to prevent or minimize episodes of increased intracranial pressure (ICP). Elevations in ICP in response to nursing interventions have been acknowledged since the 1960s when ICP monitoring was first introduced in the clinical setting. Until recently, few studies have specifically examined the effect of oral care on ICP, and oral care and other hygiene measures were combined or not specified, prohibiting a direct interpretation of the influence of oral care alone on ICP. The purpose of this study was to describe the relationship between routine oral care interventions and the changes in ICP specifically focusing on the effect of intensity and duration of this intervention. Twenty-three patients with a clinical condition requiring ICP monitoring were enrolled over a 12-month period. Oral care provided by neuroscience intensive care nurses was observed and videotaped. Characteristics of the intervention were documented including products used, patient positioning, and duration of the intervention. A 1-5 subjective scale was used to score intensity of oral care. Wrist actigraphy data were collected from the nurses to provide an objective measure of intensity. Patient physiologic data were collected at 12-second epochs 5 minutes before, during, and 5 minutes after oral care. The mixed-effect repeated measures analysis of variance model indicated that there was a statistically significant increase in ICP in response to oral care (p = .0031). There was, however, no clinically significant effect on ICP. This study provides evidence that oral care is safe to perform in patients in the absence of preexisting elevated ICP.


NICU Couplet Care: metrics to guide an evolving model of care.
The evidence-based science of maternal and neonatal care has been rapidly changing. As a result, clinical practice and the design of physical space have evolved in order to provide neuroprotection for the baby and meet expectations of family presence and participation. The concept and practice of NICU Couplet Care supports positive health well-being/outcomes and early relationship building between the mother-baby-father/partner. Monitoring evidence-based measures and metrics of standardized care, performance competence, neuro-physical and psychosocial outcomes, environmental design, family and staff satisfaction, and sustainability are essential to the evolution of quality, safe, efficient, effective, ethical, and cost-effective care for the mother, baby, and family. Transparency in the dissemination of evidence, practice standards, and outcome data is important to guide parents/families and health professionals in making informed shared decisions regarding the clinical care provided and the environment where care takes place.


[Pelvimetry using magnetic resonance tomography].
20 woman were examined before and after partum by means of conventional and magnetic resonance pelvimetry. Besides the usual data regarding the morphology and diameters of the pelvis, MRI gives information on the soft tissues of the birth canal without the added risk of irradiation to the mother and foetus. As shown by studies done in the USA it is also possible to determine foetal maturity by MRI; this will be done in the future.


Infrared Spectroscopy as a Probe of Electronic Energy Transfer.
We have combined electronic and vibrational spectroscopy in a cryogenic ion trap to produce highly resolved, conformer-selective spectra for the ground and excited states of a peptide containing two chromophores. These spectra permit us to determine the precise three-dimensional structure of the peptide and give insight into the migration of the electronic excitation from phenylalanine to tyrosine because changes in the excited-state infrared spectra are sensitive to localization of the electronic energy in each chromophore. The well-controlled experimental conditions make this result a stringent test for theoretical methods dealing with electronic energy transfer.


Fusarium keratitis: genotyping, in vitro susceptibility and clinical outcomes.
To determine differences in the clinical characteristics and antifungal susceptibility patterns among molecularly characterized ocular Fusarium sp isolates. Fifty-eight isolates of Fusarium sp obtained from 52 eyes of 52 patients were retrieved from the Ocular Microbiology Laboratory of the Bascom Palmer Eye Institute and grown in pure culture. These isolates were characterized based on DNA sequence analysis of the ITS1/2 and ribosomal deoxyribonucleic acid regions. Antifungal susceptibilities were determined for each isolate using broth microdilution methods, and the corresponding medical records were reviewed to determine the clinical outcomes. Fusarium solani isolates had significantly higher values of minimum inhibitory concentration for 90% isolates (MIC90) with voriconazole than F. non-solani organisms (16 and 4 μg/mL, respectively). Isolates of F. solani also exhibited a significantly longer time to cure (65 vs. 40.5 days), a worse follow-up best-corrected visual acuity (20/118 vs. 20/36), and an increased need for urgent surgical management (7 vs. 0 penetrating keratoplasties) when compared with those of F. non-solani. This is the first report to examine the correlation between ocular genotyped Fusarium sp and clinical outcomes. It supports the overall worse prognosis of F. solani versus F. non-solani isolates, including higher voriconazole resistance by the former. The clinical implementation of molecular-based diagnostics and antifungal efficacy testing may yield important prognostic and therapeutic information that could improve the management of fungal ocular infections.


Molecular epidemiology of cystic echinococcosis.
Echinococcus granulosus exhibits substantial genetic diversity that has important implications for the design and development of vaccines, diagnostic reagents and drugs effective against this parasite. DNA approaches that have been used for accurate identification of these genetic variants are presented here as is a description of their application in molecular epidemiological surveys of cystic echinococcosis in different geographical settings and host assemblages. The recent publication of the complete sequences of the mitochondrial (mt) genomes of the horse and sheep strains of E. granulosus and of E. multilocularis, and the availability of mt DNA sequences for a number of other E. granulosus genotypes, has provided additional genetic information that can be used for more in depth strain characterization and taxonomic studies of these parasites. This very rich sequence information has provided a solid molecular basis, along with a range of different biological, epidemiological, biochemical and other molecular-genetic criteria, for revising the taxonomy of the genus Echinococcus. This has been a controversial issue for some time. Furthermore, the accumulating genetic data may allow insight to several other unresolved questions such as confirming the occurrence and precise nature of the E. granulosus G9 genotype and its reservoir in Poland, whether it is present elsewhere, why the camel strain (G6 genotype) appears to affect humans in certain geographical areas but not others, more precise delineation of the host and geographic ranges of the genotypes characterised to date, and whether additional genotypes of E. granulosus remain to be identified.


Which cocoa bean traits persist when eating chocolate? Real-time nosespace analysis by PTR-QiToF-MS.
More consumers have become aware of the existence of different cocoa genotypes and their origins, which resulted in a growing market of premium chocolates with single-origin beans. The question is whether traits of cocoa botanical and geographical origins still persist in the end product, especially when it is consumed. By analysing the concentrations of volatile organic compounds (VOCs) in the nose of subjects over time while they are eating, new insights about aroma release can be gained. In the current study, in vivo release of VOCs during consumption of dark chocolates with different botanical and geographical origins was examined. Proton Transfer Reaction-Quadrupole interface Time of Flight- Mass Spectrometry (PTR-Qi ToF- MS) was applied to analyse nosespace VOC profiles of 10 subjects while they were eating 10 different chocolates manufactured with beans of different botanical origins (Criollo-Forastero-Trinitario) and geographical origins (Africa-South America-Asia). The headspace of the chocolates were also analysed for comparison. Cocoa botanical information appeared to affect the nosespace profiles more than geographical information. The subjects varied considerable in their VOC release, and inter-individual differences were larger than cocoa beans differences. Nevertheless, the botanical origin was consistently reflected in the nosespace profile during eating. It was clearly possible to distinguish Criollo chocolates from the nosespace profiles despite inter-individual differences.


Two-dimensional growth of dendritic islands of NTCDA on Cu(001) studied in real time.
The success of future organic electronic devices distinctively depends on the electronic and geometric properties of thin organic films. Although obviously these properties are strongly influenced by the growth mechanisms, real time growth studies are relatively rare since not many experimental techniques exist that allow in situ studies in ultra high vacuum. In this context, we investigated the prototypical system 1,4,5,8-naphtalene-tetracarboxylic-dianhydride (NTCDA) on Cu(001). We used low-energy electron microscopy (LEEM) for the real-time growth study, and a variety of other techniques for investigating the geometric and electronic structure. While for similar model systems well known and well characterized growth modi occur (e.g., compact, well ordered islands or disordered, gas-like layers), for NTCDA/Cu(001) we observe the growth of dendrite-like, fractal structures. The dendritic structures arise from a strongly preferred one-dimensional growth mode forming a long-range ordered network of thin molecular chains spanning over the entire surface already at small coverages. Later in the growth process, the voids in the network structure are incrementally filled. These results are very unexpected for such a simple adsorbate system consisting of well investigated components, the properties of which were believed to be already well understood. We explain this unexpected behavior by a dendritic growth model that is supported by energetic arguments based on pair-potential calculations. These calculations give reason for the experimentally observed growth of one-dimensional structures, and therefore represent the key to a semi-quantitative understanding of this dendritic growth mode.


Nutritional profile of Syrian refugee children before resettlement.
The year 2015 marked the highest number of refugees globally and included record numbers of Syrians moving to neighboring countries. Half of the Syrians were children aged ≤18 years. Our study sought to examine undernutrition and overnutrition among a group of Syrian refugee children who underwent medical screening by IOM for resettlement. This is a retrospective review of Syrian refugee children aged 6 to 59 months from January 1, 2015 to December 31, 2016. The World Health Organization (WHO) Stata package computed Z-scores based on available weight and height data. Prevalence estimates of undernutrition (wasting and stunting) and overnutrition (overweight and obesity) were made using WHO standards. Multivariate analysis was used to determine the factors associated with wasting, stunting, and overnutrition, adjusting for age, sex, family size, and country of health assessment. A total of 14,552 Syrian refugee children aged 6 to 59 months underwent health assessments in Jordan (43·1%), Lebanon (38·8%), Turkey (7·0%), Greece (6·7%), Egypt (2·4%), and Iraq (2·1%). Overall, this group of Syrian refugee children had a low prevalence of wasting (< 5%) and stunting (< 10%), and high prevalence of overweight or obese (10.6%). Differences were observed in the prevalence of wasting by country of health assessment. In the multiple regression analysis, the prevalence of stunting and overnutrition decreased with increasing age, and being male was associated with overnutrition but not wasting and stunting. Findings revealed an overall low prevalence of undernutrition among this group of Syrian children assessed, although prevalence varied by age group. This low prevalence may reflect the effectiveness, as well as expose possible gaps, of refugee nutrition programs or interventions in countries of asylum. Further studies are recommended to evaluate other possible contributors to malnutrition in this refugee group.


Vigilance adjustments in relation to long- and short term risk in wild fallow deer (Dama dama).
The risk allocation hypothesis predicts that vigilance should be adjusted to the temporal variation in risk. We test this hypothesis in wild fallow deer exposed to short term (disturbance) and long term (presence of a fawn after parturition) changes in risk. We recorded the proportion, frequency and type of vigilance and size of used area before and after parturition, in GPS-collared wild female fallow deer. Vigilance was divided in two main groups: "non-grazing vigilance" and "grazing vigilance". The latter group was divided into "grazing vigilance while chewing" and a "grazing vigilance when chewing was interrupted". By recording external disturbance in form of passing cars, we were able to investigate if this altered the amount, and type of vigilance. We found that females increased the proportion and frequency of "grazing vigilance stop chewing" after parturition. The "grazing vigilance chewing" was unaffected, but "non-grazing vigilance" decreased. Disturbance increased the proportion "grazing vigilance stop chewing" to the same extent before and after parturition. We found a clear decrease in female home range size after parturition as a possible behavioural adjustment. The increase in "grazing vigilance stop chewing" after parturition is a rarely described but expected cost of reproduction.


Frostbite: a practical approach to hospital management.
Frostbite presentation to hospital is relatively infrequent, and the optimal management of the more severely injured patient requires a multidisciplinary integration of specialist care. Clinicians with an interest in wilderness medicine/freezing cold injury have the awareness of specific potential interventions but may lack the skill or experience to implement the knowledge. The on-call specialist clinician (vascular, general surgery, orthopaedic, plastic surgeon or interventional radiologist), who is likely to receive these patients, may have the skill and knowledge to administer potentially limb-saving intervention but may be unaware of the available treatment options for frostbite. Over the last 10 years, frostbite management has improved with clear guidelines and management protocols available for both the medically trained and winter sports enthusiasts. Many specialist surgeons are unaware that patients with severe frostbite injuries presenting within 24 h of the injury may be good candidates for treatment with either TPA or iloprost. In this review, we aim to give a brief overview of field frostbite care and a practical guide to the hospital management of frostbite with a stepwise approach to thrombolysis and prostacyclin administration for clinicians.


Cultural assemblages show nested structure in humans and chimpanzees but not orangutans.
The evolution of hominin culture is well-documented in the archeological and fossil record, but such a record is largely absent for nonhuman primates. An alternative approach to studying cultural evolution is to examine patterns of modern cultural variation. In this article we measure nestedness across human and great ape "cultural repertoires" to gain insight into the accumulation and maintenance of putative cultural diversity in these species. Cultural assemblages are nested if cultures with a small repertoire of traits tend to comprise a proper subset of those traits present in more complex cultures. This nesting will occur if some traits are sequentially gained or lost, which may be because of the differential dispersal or extinction of traits. Here we apply statistical tools from ecology to examine the degree of nestedness in four datasets documenting the presence or absence of specific cultural traits across indigenous human populations in North America and New Guinea. We then compare the human data to patterns observed for putative cultural traits in chimpanzee and orangutan populations. In both humans and chimpanzees, cultural diversity is highly nonrandom, showing significant nested structure for all of the datasets examined. We find no evidence for nestedness in the orangutan cultural data. These findings are consistent with a sequential "layering" of cultural diversity in humans and chimpanzees, but not orangutans. Such an interpretation implies that the traits required for sequential cultural evolution first appeared in the last common ancestor of chimpanzees and humans.


More than 10 years of unrecognized nosocomial transmission of legionnaires' disease among transplant patients.
To investigate a cluster of cases of legionnaires' disease among patients at a hospital. A university hospital that is a regional transplant center. Retrospective review of microbiology and serology data from the hospital laboratories and prospective surveillance via the radiology department; a case-control study and environmental sampling within the hospital and from nearby cooling towers. Diagnosis of seven cases of legionnaires' disease in the first 9 months of 1996 led to recognition of a nosocomial outbreak that may have begun as early as 1979. Review of charts from 1987 through September 1996 identified 25 culture-confirmed cases of nosocomial or possibly nosocomial legionnaires' disease, including 18 in bone marrow and heart transplant patients. Twelve patients (48%) died. During the first 9 months of 1996, the attack rate was 6% among cardiac and bone marrow transplant patients. For cases that occurred before 1996, intubation was associated with increased risk for disease. High-dose corticosteroid medication was strongly associated with the risk for disease, but other immunosuppressive therapy or cancer chemotherapy was not. Several species and serogroups of Legionella were isolated from numerous sites in the hospital's potable water system. Six of seven available clinical isolates were identical and were indistinguishable from environmental isolates by pulsed-field gel electrophoresis. Initial infection control measures failed to interrupt nosocomial acquisition of infection. After extensive modifications to the water system, closely monitored repeated hyperchlorinations, and reduction of patient exposures to aerosols, transmission was interrupted. No cases have been identified since September 1996. Legionella can colonize hospital potable water systems for long periods of time, resulting in an ongoing risk for patients, especially those who are immunocompromised. In this hospital, nosocomial transmission possibly occurred for more than 17 years and was interrupted in 1996, after a sudden increase in incidence led to its recognition. Hospitals specializing in the care of immunocompromised patients (eg, transplant centers) should prioritize surveillance for cases of legionnaires' disease. Aggressive control measures can interrupt transmission of this disease successfully.


Complementary role of CD4+ T cells and secondary lymphoid tissues for cross-presentation of tumor antigen to CD8+ T cells.
MHC class I-restricted tumor antigens can be presented to CD8+ T cells by two distinct pathways: via direct and indirect presentation. The relative contribution of these two pathways toward the initial activation of tumor antigen-specific CD8+ T cells and their subsequent tumor rejection is still vigorously debated. Using a tumor model able to dissect the relative contributions of direct and indirect presentation, we show unequivocally the inefficiency of direct presentation and the essential requirement of indirect presentation for the priming of naive tumor antigen-specific T cells leading to tumor rejection. Moreover, we characterize the essential environment under which indirect presentation occurs, and find efficient cross-priming of tumor-specific CD8+ T cells in the complete absence of secondary lymphoid tissues. The independence of this process from local lymph nodes is compromised, however, in the absence of CD4+ T cell help. Therefore, our paper demonstrates that effective immune protection against tumors requires the cross-priming of CD8+ T cells under conditions that require either CD4+ T cell help, or draining lymph nodes.


Orthographic effects on phoneme monitoring.
A widely used task in the research on spoken word recognition is phoneme monitoring, in which subjects have to detect phonemes in spoken words. It is generally assumed that this task is performed using phonetic or phonological representations of words only. To test whether an orthographic representation of the words is employed as well, an experiment was conducted in which Dutch subjects monitored for phonemes with either a primary or secondary spelling in phonologically matched spoken words and nonwords. Phoneme monitoring times were slower when the phoneme had a secondary spelling than when it had a primary spelling. The effect was greater after than before the uniqueness point of the word, and monitoring times were faster for words than for nonwords. These findings indicate that an orthographic representation of words is engaged in phoneme monitoring.


[Focal fatty liver lesions: infrequent ultrasonographic presentation].
We present case of focal fatty liver lesions. Echography images revealed multiple hyperechogenic lesions similar to those observed in metastatic disease. Aspiration was negative and diagnosis was based on the results of liver biopsy alone, carried out during laparoscopy. The difficulty of determining the etiology of this type of lesión by echography is discussed.


Course and transmission characteristics of oral low-dose infection of domestic pigs and European wild boar with a Caucasian African swine fever virus isolate.
In 2007, African swine fever virus (ASFV) was introduced into the Transcaucasian countries and Russia. Since then, it has spread alarmingly and reached the European Union. ASFV strains are highly virulent and lead to almost 100% mortality under experimental conditions. However, the possibility of dose-dependent disease courses has been discussed. For this reason, a study was undertaken to assess the risk of chronic disease and the establishment of carriers upon low-dose oronasal infection of domestic pigs and European wild boar. It was demonstrated that very low doses of ASFV are sufficient to infect especially weak or runted animals by the oronasal route. Some of these animals did not show clinical signs indicative of ASF, and they developed almost no fever. However, no changes were observed in individual animal regarding the onset, course and outcome of infection as assessed by diagnostic tests. After amplification of ASFV by these animals, pen- and stablemates became infected and developed acute lethal disease with similar characteristics in all animals. Thus, we found no indication of prolonged or chronic individual courses upon low-dose infection in either species. The scattered onset of clinical signs and pathogen detection within and among groups confirms moderate contagiosity that is strongly linked with blood contact. In conclusion, the prolonged course at the "herd level" together with the exceptionally low dose that proved to be sufficient to infect a runted wild boar could be important for disease dynamics in wild-boar populations and in backyard settings.


Myeloma cell growth arrest, apoptosis, and interleukin-6 receptor modulation induced by EB1089, a vitamin D3 derivative, alone or in association with dexamethasone.
We have previously shown that malignant plasma cells expressed the specific receptor for 1,25-dihydroxyvitamin D3 and that this derivative could significantly inhibit the proliferation of such malignant cells. More recently, new vitamin D3 derivatives have been generated with extraordinarily potent inhibitory effects on leukemic cell growth in vitro. These new data prompted us to (re)investigate the capacity of such new vitamin D3 derivatives to inhibit myeloma cell growth in comparison with that of dexamethasone, a potent antitumoral agent in multiple myeloma. In the current study, we show that EB1089, a new vitamin D3 derivative, (1) induces G1 growth arrest of human myeloma cells, which is only partially reversed by interleukin-6 (IL-6); (2) induces apoptosis in synergy with dexamethasone, IL-6, leukemia-inhibitory factor, and Oncostatin M, with an agonistic anti-gp130 monoclonal antibody being unable to prevent this apoptosis; (3) downregulates both the gp80 (ie, the alpha chain of the IL-6 receptor [IL-6Ralpha]) expression on malignant plasma cells and the production of soluble IL-6Ralpha, and finally (4) inhibits the deleterious upregulation of gp80 expression induced by dexamethasone while limiting the dexamethasone-induced upregulation of gp130 expression. Considering that these in vitro effects of EB1089 have been observed at doses obtainable in vivo (without hypercalcemic effects), our present data strongly suggest that EB1089 could have a true interest in the treatment of multiple myeloma, especially in association with dexamethasone.


Alternative poultry litter storage for improved transportation and use as a soil amendment.
Transportation of poultry litter out of nutrient limited watersheds such as the Illinois River basin (eastern Oklahoma) is a logical solution for minimizing phosphorus (P) losses from soils to surface waters. Transportation costs are basedon mass of load and distance transported. This study investigated an alternative litter storage technique designed to promote carbon (C) degradation, thereby concentrating nutrients for the purpose of decreasing transportation costs through decreased mass. Poultry litter was stored in 0.90-Mg conical piles under semipermeable tarps and adjusted to 40% moisture content, tested with and without addition of alum (aluminum sulfate). additional study was conducted using 3.6-Mg piles under the same conditions, except tested with and without use of aeration pipes. Samples were analyzed before and after (8 wk) storage. Litter mass degradation (i.e., loss in mass due to organic matter decomposition) was estimated on the basis of changes in litter total P contents. Additional characterization included pH, total nutrients, moisture content, total C, and degree of humification. Litter storage significantly decreased litter mass (16 to 27%), concentrated nutrients such as P and potassium (K) and increased proportion of fulvic and humic acids. The addition of aeration pipes increased mass degradationrelative to piles without aeration pipes. Nitrogen volatilization losses were minimized with alum additions. Increases in P and K concentrations resulted in greater monetary value per unit mass compared with fresh litter. Such increases translate to increased litter shipping distance and cost savings of $17.2 million over 25 yr for litter movement out of eastern Oklahoma.


Demyelinating neuropathies.
To review recent advances in heritable demyelinating neuropathies and their genotype-phenotype correlations. Two themes dominated the literature on demyelinating neuropathies in the past 2 years: (1) the dissection of the localization and roles of new molecules in the axon-Schwann cell unit, including periaxin and connexin32, which are mutated in specific heritable neuropathies; (2) the recognition of the range of phenotypes associated with individual genetic abnormalities, so that the same defects can even produce both demyelinating and axonal forms of heritable neuropathies. The axonal injury associated with demyelination reflects elaborate interactions between axons and Schwann cells. Demyelinating neuropathies, even those due to abnormalities in intrinsic Schwann cell or myelin proteins, can have clinical manifestations due to distally predominant axonal degeneration.


Reprogramming of lysosomal gene expression by interleukin-4 and Stat6.
Lysosomes play important roles in multiple aspects of physiology, but the problem of how the transcription of lysosomal genes is coordinated remains incompletely understood. The goal of this study was to illuminate the physiological contexts in which lysosomal genes are coordinately regulated and to identify transcription factors involved in this control. As transcription factors and their target genes are often co-regulated, we performed meta-analyses of array-based expression data to identify regulators whose mRNA profiles are highly correlated with those of a core set of lysosomal genes. Among the ~50 transcription factors that rank highest by this measure, 65% are involved in differentiation or development, and 22% have been implicated in interferon signaling. The most strongly correlated candidate was Stat6, a factor commonly activated by interleukin-4 (IL-4) or IL-13. Publicly available chromatin immunoprecipitation (ChIP) data from alternatively activated mouse macrophages show that lysosomal genes are overrepresented among Stat6-bound targets. Quantification of RNA from wild-type and Stat6-deficient cells indicates that Stat6 promotes the expression of over 100 lysosomal genes, including hydrolases, subunits of the vacuolar H⁺ ATPase and trafficking factors. While IL-4 inhibits and activates different sets of lysosomal genes, Stat6 mediates only the activating effects of IL-4, by promoting increased expression and by neutralizing undefined inhibitory signals induced by IL-4. The current data establish Stat6 as a broadly acting regulator of lysosomal gene expression in mouse macrophages. Other regulators whose expression correlates with lysosomal genes suggest that lysosome function is frequently re-programmed during differentiation, development and interferon signaling.


Beneficiation of borax by reverse flotation in boron saturated brine.
Flotation is one of the plausible methods for recovering borax fines discharged as fine waste to the tailings dam in the Kirka borax processing plant. A literature review dealing with the flotation behavior of boron minerals reveals that clay minerals in the boron ores coat boron minerals and thus deteriorate the quality of boron concentrates produced by direct flotation. The main objective of this study is therefore to recover borax fines from the tailings of the concentrator by reverse flotation. A three-level-factor experimental design was used to determine the main and interaction effects of variables selected on the metallurgical performance of reverse flotation. An analysis of variance for experimental results indicates that interaction effects of the variables for concentrate quality and recovery of B2O3 is nonsignificant and the most important variable for grade of concentrate and recovery is the collector dosage. It is shown that a concentrate assaying 11.25% B2O3 with 89.90% B2O3 recovery could be produced by means of single-stage (rougher) reverse flotation. Additionally, in order to produce a sufficient-quality concentrate, a multistage reverse flotation scheme involving rougher, scavenger, and two cleaners was devised. A final concentrate containing 23.47% B2O3 with 81.78% B2O3 recovery was obtained from these tests. The reverse flotation method can be thus considered as an important option for the beneficiation of borax fines.


Are prescription stimulants "smart pills"? The epidemiology and cognitive neuroscience of prescription stimulant use by normal healthy individuals.
Use of prescription stimulants by normal healthy individuals to enhance cognition is said to be on the rise. Who is using these medications for cognitive enhancement, and how prevalent is this practice? Do prescription stimulants in fact enhance cognition for normal healthy people? We review the epidemiological and cognitive neuroscience literatures in search of answers to these questions. Epidemiological issues addressed include the prevalence of nonmedical stimulant use, user demographics, methods by which users obtain prescription stimulants, and motivations for use. Cognitive neuroscience issues addressed include the effects of prescription stimulants on learning and executive function, as well as the task and individual variables associated with these effects. Little is known about the prevalence of prescription stimulant use for cognitive enhancement outside of student populations. Among college students, estimates of use vary widely but, taken together, suggest that the practice is commonplace. The cognitive effects of stimulants on normal healthy people cannot yet be characterized definitively, despite the volume of research that has been carried out on these issues. Published evidence suggests that declarative memory can be improved by stimulants, with some evidence consistent with enhanced consolidation of memories. Effects on the executive functions of working memory and cognitive control are less reliable but have been found for at least some individuals on some tasks. In closing, we enumerate the many outstanding questions that remain to be addressed by future research and also identify obstacles facing this research.


Preferences for Temporal Sequences of Real Outcomes Differ Across Domains but do not Vary by Age.
People's preferences for temporal sequences of events have implications for life-long health and well-being. Prior research suggests that other aspects of intertemporal choice vary by age, but evidence for age differences in sequence-preferences is limited and inconclusive. In response, the present research examined age differences in sequence-preferences for real outcomes administered in a controlled laboratory setting. A pilot study examined sequence-preferences for aversive electrodermal shocks in 30 younger and 30 older adults. The main study examined sequence-preferences for electrodermal shocks, physical effort, and monetary gambles in an adult life-span sample (N = 120). It also examined emotional and physiological responses to sequences as well as underlying mechanisms including time perception and emotion-regulation. There were no significant age differences in sequence-preferences in either of the studies, and there were no age differences in responses to sequences in the main study. Instead, there was a domain effect with participants preferring decreasing sequences for shocks and mixed sequences for effort and money. After considering potential methodological limitations, theoretical contributions and implications for real-life decisions are discussed.


Diagnostic yield of candidate genes in an Australian corneal dystrophy cohort.
Corneal dystrophies describe a clinically and genetically heterogeneous group of inherited disorders. The International Classification of Corneal Dystrophies (IC3D) lists 22 types of corneal dystrophy, 17 of which have been demonstrated to result from pathogenic variants in 19 identified genes. In this study, we investigated the diagnostic yield of genetic testing in a well-characterised cohort of 58 individuals from 44 families with different types of corneal dystrophy. Individuals diagnosed solely with Fuchs endothelial corneal dystrophy were excluded. Clinical details were obtained from the treating ophthalmologist. Participants and their family members were tested using a gene candidate and exome sequencing approach. We identified a likely molecular diagnosis in 70.5% families (31/44). The detection rate was significantly higher among probands with a family history of corneal dystrophy (15/16, 93.8%) than those without (16/28, 57.1%, p = .015), and among those who had undergone corneal graft surgery (9/9, 100.0%) compared to those who had not (22/35, 62.9%, p = .041). We identified eight novel variants in five genes and identified five families with syndromes associated with corneal dystrophies. Our findings highlight the genetic heterogeneity of corneal dystrophies and the clinical utility of genetic testing in reaching an accurate clinical diagnosis.


Calcium Imaging and the Curse of Negativity.
The imaging of neuronal activity using calcium indicators has become a staple of modern neuroscience. However, without ground truths, there is a real risk of missing a significant portion of the real responses. Here, we show that a common assumption, the non-negativity of the neuronal responses as detected by calcium indicators, biases all levels of the frequently used analytical methods for these data. From the extraction of meaningful fluorescence changes to spike inference and the analysis of inferred spikes, each step risks missing real responses because of the assumption of non-negativity. We first show that negative deviations from baseline can exist in calcium imaging of neuronal activity. Then, we use simulated data to test three popular algorithms for image analysis, CaImAn, suite2p, and CellSort, finding that suite2p may be the best suited to large datasets. We also tested the spike inference algorithms included in CaImAn, suite2p, and Cellsort, as well as the dedicated inference algorithms MLspike and CASCADE, and found each to have limitations in dealing with inhibited neurons. Among these spike inference algorithms, FOOPSI, from CaImAn, performed the best on inhibited neurons, but even this algorithm inferred spurious spikes upon the return of the fluorescence signal to baseline. As such, new approaches will be needed before spikes can be sensitively and accurately inferred from calcium data in inhibited neurons. We further suggest avoiding data analysis approaches that, by assuming non-negativity, ignore inhibited responses. Instead, we suggest a first exploratory step, using k-means or PCA for example, to detect whether meaningful negative deviations are present. Taking these steps will ensure that inhibition, as well as excitation, is detected in calcium imaging datasets.


Cell surface expression of MR1B, a splice variant of the MHC class I-related molecule MR1, revealed with antibodies.
The major histocompatibility complex (MHC) class I-related molecule, MR1, is highly conserved in mammals and can present bacteria-derived vitamin B metabolites to mucosal-associated invariant T (MAIT) cells, possibly having important defense function in the microbial infection. MR1B is a splice variant of MR1 and possesses an intriguing domain structure with only two extracellular domains resembling some NKG2D ligand molecules. Thus far, cell surface expression of MR1B could not be analyzed with flow cytometry due to a lack of appropriate antibodies reactive with MR1B. Here we clarified the expression of MR1B recombinant protein on the cell surface of the transfected cells by flow cytometry analyses using the antiserum against MR1. Consistently, MR1B tagged with FLAG peptide at the N-terminus also could be detected with anti-FLAG monoclonal antibodies. Our result showed that MR1B can be recognized on the cell surface by macromolecules such as antibodies, indicating its potential of interaction with certain receptor(s). We discuss possibility of interaction of MR1B and/or the full-length MR1 with some receptor(s) other than αβ T cell receptor (TCR) of MAIT cells based on the highly conserved characteristic residues of the ligand-binding domains of MR1 and its MAIT cells αβTCR footprints.


Revised Korean Cough Guidelines, 2020: Recommendations and Summary Statements.
Cough is the most common respiratory symptom that can have various causes. It is a major clinical problem that can reduce a patient's quality of life. Thus, clinical guidelines for the treatment of cough were established in 2014 by the cough guideline committee under the Korean Academy of Tuberculosis and Respiratory Diseases. From October 2018 to July 2020, cough guidelines were revised by members of the committee based on the first guidelines. The purpose of these guidelines is to help clinicians efficiently diagnose and treat patients with cough. This article highlights the recommendations and summary of the revised Korean cough guidelines. It includes a revised algorithm for the evaluation of acute, subacute, and chronic cough. For a chronic cough, upper airway cough syndrome (UACS), cough variant asthma (CVA), and gastroesophageal reflux disease (GERD) should be considered in differential diagnoses. If UACS is suspected, first-generation antihistamines and nasal decongestants can be used empirically. In cases with CVA, inhaled corticosteroids are recommended to improve cough. In patients with suspected chronic cough due to symptomatic GERD, proton pump inhibitors are recommended. Chronic bronchitis, bronchiectasis, bronchiolitis, lung cancer, aspiration, intake of angiotensin-converting enzyme inhibitor, intake of dipeptidyl peptidase-4 inhibitor, habitual cough, psychogenic cough, interstitial lung disease, environmental and occupational factors, tuberculosis, obstructive sleep apnea, peritoneal dialysis, and unexplained cough can also be considered as causes of a chronic cough. Chronic cough due to laryngeal dysfunction syndrome has been newly added to the guidelines.


Unique lamina propria stromal cells imprint the functional phenotype of mucosal dendritic cells.
Mucosal dendritic cells (DCs) in the intestine acquire the unique capacity to produce retinoic acid (RA), a vitamin A metabolite that induces gut tropism and regulates the functional differentiation of the T cells they prime. Here, we identified a stromal cell (SC) population in the intestinal lamina propria (LP), which is capable of inducing RA production in DCs in a RA- and granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent fashion. Unlike DCs, LP SCs constitutively expressed the enzymatic machinery to produce RA even in the absence of dietary vitamin A, but were not able to do so in germ-free mice implying regulation by microbiota. Interestingly, DCs promoted GM-CSF production by the SCs indicating a two-way cross-talk between both cell types. Furthermore, RA-producing LP SCs and intestinal DCs localized closely in vivo suggesting that the interactions between both cell types might have an important role in the functional education of migratory DCs and therefore in the regulation of immune responses toward oral and commensal antigens.


Use of dried blood spots to define antibody response to the Strongyloides stercoralis recombinant antigen NIE.
An approach to improve the diagnosis of Strongyloides stercoralis infection is the use of serologic assays utilising the NIE antigen from S. stercoralis, with good diagnostic sensitivity and excellent specificity reported. Detection of antibody eluted from dried blood spots (DBS) has shown utility in large-scale seroepidemiological studies for a range of conditions and is appealing for use with children where sample collection is difficult. We adapted an existing NIE-enzyme linked immunosorbent assay (ELISA) for the testing of strongyloides antibody response on DBS, and evaluated it in a population screening and mass drug administration programme (MDA) for strongyloidiasis conducted in an Australian indigenous community. Study participants were treated with 200 μg/kg ivermectin (>15 kg) or 3× 400 mg albendazole (<15kg). The sensitivity of the NIE DBS-ELISA was determined by receiver operator characteristic (ROC) analysis to be 85.7%. A total of 214 DBS were collected from 184 participants across two screening and MDA encounters. A total of 27 of 164 participants (16.5%) tested positive for S. stercoralis NIE-DBS prior to MDA treatment, and 6 of 50 participants (12.0%) tested positive after treatment. These prevalence values are similar to those documented by standard serology in the same community. For 30 participants where a DBS was collected at both MDA 1 and 2, a significant decline in ELISA values was evident post treatment (0.12-0.02, p=0.0012). These results are in agreement with previous studies documenting the high seroprevalence of S. stercoralis in remote Australian Indigenous communities, and suggest that collection of dried blood spots may be a useful approach for field diagnosis of S. stercoralis seroprevalence.


Extracorporeal membrane oxygenation causing asymmetric vasculopathy in neonatal infants.
Extracorporeal membrane oxygenation (ECMO) is a form of cardiopulmonary bypass therapy used in term or near-term infants with severe cardiorespiratory disorders not responsive to conventional intensive care interventions. An ECMO-associated retinal vasculopathy has been described with little reference to the specific condition of the patient. We examined the eyes of 91 infants who underwent ECMO treatment. An assessment was made of the following: (1) when retinal changes occurred, (2) whether there was a particular systemic disorder or ECMO approach associated with these retinal findings, and (3) whether there may be ocular sequelae from this development. Ninety-one neonates were treated with ECMO for meconium aspiration syndrome (MAS), primary persistent pulmonary hypertension of the newborn, sepsis, congenital diaphragmatic hernia (CDH), respiratory distress syndrome (RDS), and blood aspiration. Venoarterial bypass was performed in 73 patients. The remaining 18 patients underwent venovenous bypass. Ophthalmologic examinations were performed during bypass in 6 infants and within 3 weeks of ECMO in the remainder. Asymmetric retinopathy (left eye > right eye) was discovered in six infants with CDH and in one infant with RDS within a 2-week period after bypass, demonstrating venous tortuosity with or without intraretinal hemorrhages. One infant treated for MAS had a left eye intraretinal hemorrhage only. All patients with the noted retinal changes underwent venoarterial cannulation. Six of 9 patients with CDH had retinal findings noted compared with 1 of 10 patients with RDS and 1 of 35 patients with MAS. Because we were able to examine infants while they were receiving ECMO or shortly after termination of bypass, asymmetric vasculopathy was found in a greater percentage of our patients compared with a similar large case series. ECMO-associated retinal vasculopathy appeared to disproportionately occur in those patients with CDH who underwent venoarterial bypass. Further study of retinal vascular changes in patients with CDH should be performed to assess long-term effects.


Rotatable Orifice for Needle-Free Jet Injection.
This research explores a new development in orifice technology for needle-free jet injection. The premise lies in the ability to control the angle at which the drug is delivered into the tissue to increase the lateral dispersion of the drug. Towards this aim, a spherical orifice that can rotate to adjust the injection angle is explored. This work tests the design and feasibility of the spherical orifice, its housing, and the orifice seats. The results show that the most successful way to create a fluid seal within the housing was to use an o-ring to create a fluid seal at the inlet side of the sphere and an extended brass seat on the outlet side of the sphere. This allowed jet speeds up to approximately 123 m/s through a 0.2 mm orifice machined into 9.5 mm diameter brass sphere. Jet speeds large enough to penetrate porcine tissue were reached at jet angles of 0° to 50° relative to the base of the injector. Although the jets successfully penetrated the tissue, the amount of fluid delivered varied depending on the injection angle. With a shallow angle injection, the fluid retention rate (the percentage of the ejected fluid from the injector which the tissue sample retained) was on average 44%. When the spherical orifice was at its maximum angle, the injection achieved an average fluid retention rate of 22%. At its widest angle, lateral dispersion of the drug also increased by approximately 40%, in comparison to conventional needles and traditional perpendicular jet injection. In summary, a spherical orifice needle-free injection system successfully produced high-speed jets and delivered liquid into porcine tissue at injection angles from 0° to 50°, demonstrating the feasibility of this technique that offers unique advantages over typical orifice plates and conventional needles.Clinical Relevance-A rotatable nozzle can be used to control the angle of needle-free drug delivery.


Nucleotide sequence and evolution of ETn elements.
The ETn (for "early transposon") family of long repeated sequences in abundantly transcribed in early mouse embryos from retroviral-like long terminal repeats. Nucleotide sequencing of two elements does not reveal any long open reading frame nor significant homology to retroviral proteins. The genetic polymorphism, monitored by Southern blotting within and across mouse species, reflects a concerted mode of evolution for the ETn sequences.


Road traffic noise model.
The recognition of road traffic noise as one of the main sources of environmental pollution has led to develop models that enable to predict noise level from fundamental variables. Traffic noise prediction models are required as aids for designing roads and highways. In addition, sometimes are used in the assessment of existing or envisaged changes in traffic noise conditions. In this paper a statistical modelling approach has been used for predicting road traffic noise in Iranian road conditions. The study was performed during 2005-2006 in Hamadan city, in the west of Iran. The data set consisted of 282 noise measurements. The entire data set was utilized to develop a new model for Iranian condition using regression analysis. The developed model has twelve explanatory variables in order to achieve a proper fit for measured values of Leq (r2= 0.913). The proposed road traffic noise model can be effectively used as a decision support tools for prediction of traffic noise index of Leq(30min), in Iran's cities.


Pollen dispersal and fruit production in Vaccinium oxycoccos and comparison with its sympatric congener V. uliginosum.
Investigating plant-pollinator interactions and pollen dispersal are particularly relevant for understanding processes ensuring long-term viability of fragmented plant populations. Pollen dispersal patterns may vary strongly, even between similar congeneric species, depending on the mating system, pollinator assemblages and floral traits. We investigated pollen dispersal and fruit production in a population of Vaccinium oxycoccos, an insect-pollinated shrub, and compared the pollen dispersal pattern with a co-flowering, sympatric congener, V. uliginosum. We examined whether they share pollinators (through interspecific fluorescent dye transfers) and may differently attract pollinators, by comparing their floral colour as perceived by insects. Fluorescent dyes were mainly dispersed over short distances (80% within 40.4 m (max. 94.5 m) for V. oxycoccos and 3.0 m (max. 141.3 m) for V. uliginosum). Dye dispersal in V. oxycoccos was not significantly affected by plant area, floral display or the proximity to V. uliginosum plants. Interspecific dye transfers were observed, indicating pollinator sharing. The significantly lower dye deposition on V. oxycoccos stigmas suggests lower visitation rates by pollinators, despite higher flower density and local abundance. The spectral reflectance analysis indicates that bees are unlikely to be able to discriminate between the two species based on floral colour alone. Fruit production increased with increasing floral display, but was not affected by proximity to V. uliginosum plants. Our study highlights that fragmented populations of V. oxycoccos, when sympatric with co-flowering V. uliginosum, might incur increased competition for the shared pollinators in the case of pollination disruption, which might then reduce outcrossed seed set.


Methods used in cross-cultural comparisons of vasomotor symptoms and their determinants.
Methodological differences among studies of vasomotor symptoms limit rigorous comparison or systematic review. Vasomotor symptoms generally include hot flushes and night sweats although other associated symptoms exist. Prevalence rates vary between and within populations, but different studies collect data on frequency, bothersomeness, and/or severity using different outcome measures and scales, making comparisons difficult. We reviewed only cross-cultural studies of menopausal symptoms that explicitly examined symptoms in general populations of women in different countries or different ethnic groups in the same country. This resulted in the inclusion of nine studies: Australian/Japanese Midlife Women's Health Study (AJMWHS), Decisions At Menopause Study (DAMeS), Four Major Ethnic Groups (FMEG), Hilo Women's Health Survey (HWHS), Mid-Aged Health in Women from the Indian Subcontinent (MAHWIS), Penn Ovarian Aging Study (POAS), Study of Women's Health Across the Nation (SWAN), Women's Health in Midlife National Study (WHiMNS), and Women's International Study of Health and Sexuality (WISHeS). These studies highlight the methodological challenges involved in conducting multi-population studies, particularly when languages differ, but also highlight the importance of performing multivariate and factor analyses. Significant cultural differences in one or more vasomotor symptoms were observed in 8 of 9 studies, and symptoms were influenced by the following determinants: menopausal status, hormones (and variance), age (or actually, the square of age, age(2)), BMI, depression, anxiety, poor physical health, perceived stress, lifestyle factors (hormone therapy use, smoking and exposure to passive smoke), and acculturation (in immigrant populations). Recommendations are made to improve methodological rigor and facilitate comparisons in future cross-cultural menopause studies.


ADP-ribosylation factor 1 transiently activates high-affinity adaptor protein complex AP-1 binding sites on Golgi membranes.
Association of the Golgi-specific adaptor protein complex 1 (AP-1) with the membrane is a prerequisite for clathrin coat assembly on the trans-Golgi network (TGN). The AP-1 adaptor is efficiently recruited from cytosol onto the TGN by myristoylated ADP-ribosylation factor 1 (ARF1) in the presence of the poorly hydrolyzable GTP analog guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS). Substituting GTP for GTPgammaS, however, results in only poor AP-1 binding. Here we show that both AP-1 and clathrin can be recruited efficiently onto the TGN in the presence of GTP when cytosol is supplemented with ARF1. Optimal recruitment occurs at 4 microM ARF1 and with 1 mM GTP. The AP-1 recruited by ARF1.GTP is released from the Golgi membrane by treatment with 1 M Tris-HCl (pH 7) or upon reincubation at 37 degreesC, whereas AP-1 recruited with GTPgammaS or by a constitutively active point mutant, ARF1(Q71L), remains membrane bound after either treatment. An incubation performed with added ARF1, GTP, and AlFn, used to block ARF GTPase-activating protein activity, results in membrane-associated AP-1, which is largely insensitive to Tris extraction. Thus, ARF1. GTP hydrolysis results in lower-affinity binding of AP-1 to the TGN. Using two-stage assays in which ARF1.GTP first primes the Golgi membrane at 37 degreesC, followed by AP-1 binding on ice, we find that the high-affinity nucleating sites generated in the priming stage are rapidly lost. In addition, the AP-1 bound to primed Golgi membranes during a second-stage incubation on ice is fully sensitive to Tris extraction, indicating that the priming stage has passed the ARF1.GTP hydrolysis point. Thus, hydrolysis of ARF1.GTP at the priming sites can occur even before AP-1 binding. Our finding that purified clathrin-coated vesicles contain little ARF1 supports the concept that ARF1 functions in the coat assembly process rather than during the vesicle-uncoating step. We conclude that ARF1 is a limiting factor in the GTP-stimulated recruitment of AP-1 in vitro and that it appears to function in a stoichiometric manner to generate high-affinity AP-1 binding sites that have a relatively short half-life.


Myocardial dysfunction is associated with activation of Na+/H+ exchange immediately during reperfusion.
Amiloride analogs block Na+/H+ exchange and thereby protect the heart from myocardial ischemia-reperfusion injury. It is unclear whether drugs must be present before ischemia to be cardioprotective. After 60 min of global ischemia in the coronary-perfused right ventricular wall (RVW), as little as 1 min of exposure to dimethyl amiloride (DMA) immediately at the time of reperfusion protected the RVW. Delaying the drug attenuated the cardioprotection. If DMA was introduced in an ischemic solution near the end of ischemia, the cardioprotective effects were augmented. If the drug was washed out of the RVW vascular space before ischemia, cardioprotection was not observed. In contrast, in whole hearts, preischemic perfusion of the drug was necessary for cardioprotection and the cardioprotection remained even if the drug was washed out before ischemia. We conclude that Na+/H+ exchange is active and contributes to contractile dysfunction during the first seconds of reperfusion. This is difficult to detect in the perfused whole heart, and the washout data suggest that this may be due to a limitation in drug delivery across the vascular wall. The data also suggest that the exchanger is not as active during ischemia itself as it is during reperfusion.


Pattern generation using likelihood inference for cellular automata.
Cellular automata are discrete dynamical systems which evolve on a discrete grid. Recent studies have shown that cellular automata with relatively simple rules can produce highly complex patterns. We develop likelihood-based methods for estimating rules of cellular automata aimed at the re-generation of observed regular patterns. Under noisy data, our approach is equivalent to estimating the local map of a stochastic cellular automaton. Direct computations of the maximum likelihood estimates are possible for regular binary patterns. The likelihood formulation of the problem is congenial with the use of the minimum description length principle as a model selection tool. We illustrate our method with a series of examples using binary images.


Mining for diagnostic information in body surface potential maps: a comparison of feature selection techniques.
In body surface potential mapping, increased spatial sampling is used to allow more accurate detection of a cardiac abnormality. Although diagnostically superior to more conventional electrocardiographic techniques, the perceived complexity of the Body Surface Potential Map (BSPM) acquisition process has prohibited its acceptance in clinical practice. For this reason there is an interest in striking a compromise between the minimum number of electrocardiographic recording sites required to sample the maximum electrocardiographic information. In the current study, several techniques widely used in the domains of data mining and knowledge discovery have been employed to mine for diagnostic information in 192 lead BSPMs. In particular, the Single Variable Classifier (SVC) based filter and Sequential Forward Selection (SFS) based wrapper approaches to feature selection have been implemented and evaluated. Using a set of recordings from 116 subjects, the diagnostic ability of subsets of 3, 6, 9, 12, 24 and 32 electrocardiographic recording sites have been evaluated based on their ability to correctly asses the presence or absence of Myocardial Infarction (MI). It was observed that the wrapper approach, using sequential forward selection and a 5 nearest neighbour classifier, was capable of choosing a set of 24 recording sites that could correctly classify 82.8% of BSPMs. Although the filter method performed slightly less favourably, the performance was comparable with a classification accuracy of 79.3%. In addition, experiments were conducted to show how (a) features chosen using the wrapper approach were specific to the classifier used in the selection model, and (b) lead subsets chosen were not necessarily unique. It was concluded that both the filter and wrapper approaches adopted were suitable for guiding the choice of recording sites useful for determining the presence of MI. It should be noted however that in this study recording sites have been suggested on their ability to detect disease and such sites may not be optimal for estimating body surface potential distributions.


Task Programming: Learning Data Efficient Behavior Representations.
Specialized domain knowledge is often necessary to accurately annotate training sets for in-depth analysis, but can be burdensome and time-consuming to acquire from domain experts. This issue arises prominently in automated behavior analysis, in which agent movements or actions of interest are detected from video tracking data. To reduce annotation effort, we present TREBA: a method to learn annotation-sample efficient trajectory embedding for behavior analysis, based on multi-task self-supervised learning. The tasks in our method can be efficiently engineered by domain experts through a process we call "task programming", which uses programs to explicitly encode structured knowledge from domain experts. Total domain expert effort can be reduced by exchanging data annotation time for the construction of a small number of programmed tasks. We evaluate this trade-off using data from behavioral neuroscience, in which specialized domain knowledge is used to identify behaviors. We present experimental results in three datasets across two domains: mice and fruit flies. Using embeddings from TREBA, we reduce annotation burden by up to a factor of 10 without compromising accuracy compared to state-of-the-art features. Our results thus suggest that task programming and self-supervision can be an effective way to reduce annotation effort for domain experts.


Establishing a Telephone Medication Order Policy and Protocol for a Small Private Hospital in Jamaica.
To institutionalize an evidence-based policy/protocol adapted from the Agency for Healthcare Research and Quality (AHRQ) national medication standards for managing telephone medication orders (TMO) and to determine the impact of the policy/protocol on the number of telephone medication errors (TME) on two medical units of a small private hospital in Jamaica. Kotter's Eight-step Change Model was used to facilitate organizational change among nurses and physicians by teaching and implementing the TMO policy/protocol adapted from AHRQ standards and collecting pre-policy and post-policy frequency of TMEs. A convenience sample of 80 nurses and physicians participated in training about the policy/protocol, took post-instructional tests and participated in the implementation of the policy/protocol. Chart audits over six weeks monitored adherence to the policy/protocol. The annual monthly mean of TMEs for the prior year was compared with the number of TMEs just prior to implementation of policy/protocol and at the end of the first six weeks of implementation. One hundred per cent of the convenience sample of 80 nurses and doctors passed the post-instructional test; the workforce adhered fully to the protocol during six weeks of implementation, and there was a 100% reduction in TMEs between the prior year and six weeks after policy/protocol implementation. Kotter's eight-step framework of organizational change was a successful strategy in institutionalizing and sustaining adherence to the TMO policy/protocol, reducing the number of TMEs and positively influencing the organizational culture.


Intracranial placement of a new, compliant guide catheter: technical note.
We describe our initial experience with the use of a novel, compliant guide catheter designed for placement within the cranial vasculature in a series of seven patients who were treated for various intracranial pathologies. Seven patients were deemed to have either tortuous supra-aortic, intracranial, and/or extracranial vasculature or to require additional microcatheter support as the result of lesion location. The patients were treated, in part, with the 6-French Neuron delivery catheter (Penumbra, Inc., San Leandro, CA) at the authors' two institutions. The guide catheter was positioned in various distal locations within the intracranial internal carotid artery or external carotid artery. Three patients were treated for unruptured intracranial aneurysms, 2 patients for intracranial atherosclerosis, 1 patient for an arteriovenous malformation, and 1 patient for a pseudoaneurysm. All lesions were successfully treated through a microcatheter advanced in a coaxial fashion through the guide catheter. There were no complications related to the positioning of the catheter. Distal intra- or extracranial placement of a specially designed, compliant guide catheter can be performed safely and may improve access and microcatheter stability in patients with tortuous vessels or difficult-to-reach lesions.


[Phytochemicals as means to induce sleep].
Phytopharmacons are widely used in Germany. Whereas St. John's wort extracts are prescribed for the treatment of mild forms of depression and kava-kava for unspecified anxiety syndromes, hop, balm, lavender, passiflora and valerian are traditionally administered against nervousness and sleep disturbances. Controlled clinical trials are only available for valerian. However, no sleep inducing potential of valerian was observed, only a certain positive effect on daytime mood. Therefore, the mentioned phytopharmacons cannot be recommended for the treatment of sleep disturbances.


Health literacy in an orthopaedic trauma patient population: improving patient comprehension with informational intervention.
This study aims to evaluate the change in comprehension of diagnoses, treatment plans, and discharge instructions after orthopaedic trauma patients are given an informational document that includes pictorial representations at the time of discharge. It also seeks to determine if the intervention has a greater impact on patients with lower educational backgrounds. Prospective comparative cohort study. Academic level 1 trauma center. From April to December 2011, 529 orthopaedic trauma patients with an operatively fixed isolated fracture were eligible for inclusion. Two hundred ninety-nine eligible questionnaires were collected (56.5% response rate). Patients were administered a questionnaire regarding their treatment and discharge instructions during their first postoperative clinic visit before being seen by a physician. The questionnaire included demographic information and questions regarding: (1) which bone was fractured, (2) type of implanted fixation, (3) weight-bearing status, (4) expected recovery time, and (5) need for deep vein thrombosis prophylaxis. All patients had received verbal instructions outlining this information at postoperative hospital discharge. During the second half of the study, patients were given an additional informational sheet with both text and pictorial representations at discharge. Multivariable log-binomial regression analyses were used to examine the impact of this intervention. One hundred forty-six patients were given only the standard discharge instructions, whereas 153 patients were also administered the additional information document. The mean score for patients who received the intervention was 2.90 (out of 5) compared with the mean score of 2.54 for patients who did not receive the intervention (P = 0.013). Patients who received the intervention were 1.3 times more likely to know which bone was fractured (P = 0.007) and 1.1 times more likely to be able to correctly name the medication(s) they were prescribed for deep vein thrombosis prophylaxis (P = 0.03). Overall performance on comprehension questionnaires in orthopaedic trauma patients was significantly improved via a text and pictorial intervention. The intervention did not preferentially aid patients with lower education backgrounds. Future studies should evaluate long-term postoperative results to determine if improved patient comprehension has an effect on surgical outcomes and patient satisfaction.


Epoxy nanocomposites with two-dimensional transition metal dichalcogenide additives.
Emerging two-dimensional (2D) materials such as transition metal dichalcogenides offer unique and hitherto unavailable opportunities to tailor the mechanical, thermal, electronic, and optical properties of polymer nanocomposites. In this study, we exfoliated bulk molybdenum disulfide (MoS2) into nanoplatelets, which were then dispersed in epoxy polymers at loading fractions of up to 1% by weight. We characterized the tensile and fracture properties of the composite and show that MoS2 nanoplatelets are highly effective at enhancing the mechanical properties of the epoxy at very low nanofiller loading fractions (below 0.2% by weight). Our results show the potential of 2D sheets of transition metal dichalcogenides as reinforcing additives in polymeric composites. Unlike graphene, transition metal dichalcogenides such as MoS2 are high band gap semiconductors and do not impart significant electrical conductivity to the epoxy matrix. For many applications, it is essential to enhance mechanical properties while also maintaining the electrical insulation properties and the high dielectric constant of the polymer material. In such applications, conductive carbon based fillers such as graphene cannot be utilized. This study demonstrates that 2D transition metal dichalcogenide additives offer an elegant solution to such class of problems.


[Nutritional effects following lumbar sympathectomy in arterial occlusive disease (author's transl)].
The effect of lumbar sympathectomy on regional blood-flow and on oxygen uptake of tissue has been investigated in animal experiments and in man. Lumbar sympathectomy did always result in increase of limb blood-flow, but did not significantly improve oxygen consumption of the observed extremity. This finding was manifested by marked narrowing of the arterial-venous oxyhemoglobin saturation difference suggesting that lumbar sympathectomy caused opening of arterial-venous shunts.


[Recurrent familial hyperparathyroidism. A propos of 7 adenomas in 3 members of the same family. Review of the literature].
Over a period of 23 years, 3 members of a family of 5 presented with 7 parathyroid adenomas (4 in the first case, 2 in the second case, 1 in the third case). Excision of each adenoma, with systematic pre-operative assessment of the remaining parathyroid tissue, led to complete clinical and laboratory cure of each episode. The delay in the appearance of recurrence was between 3 and 9 years. After reviewing the literature, the authors stress the importance, in any case of hyperparathyroidism, of routinely investigating the serum calcium levels in members of the patient's family, especially if the patient is young and if he has had several episodes or multiglandular involvement in one episode. The authors discuss the literature concerning recurrent familial hyperparathyroidism with endocrine polyadenomatosis and the "hypercalcaemia-hypocalciuria" syndrome.


Perceptions, circumstances and motivators that influence implementation of zoonotic control programs on cattle farms.
The implementation of disease control programs on farms requires an act of behavioral change. This study presents a theoretical framework from behavioral science, combined with basic epidemiological principles to investigate and explain the control of zoonotic agents on cattle farms. A pathway to disease control model was adapted from existing models in behavioral science and human medicine. Field data was used to demonstrate the validity of the model to identify and explain motivational factors for implementation of disease control programs among English and Welsh cattle farmers. The field data consisted of interviews conducted with 43 farmers, which were analyzed to investigate the farmers' perception of responsibility for safe cattle produce as well as the intrinsic and extrinsic barriers that inhibited the implementation of a zoonotic control program on their farms. The model was used to illustrate barriers affecting the implementation process and to classify farmers according to their current level of zoonotic control at each stage within the model. Ordinal multivariable logistic regression was used to identify the motivators associated with different levels of implementation. Younger farmers and/or larger herds were more likely to place financial responsibility upon the industry rather than government and all but two farmers accepted a social responsibility for food safety within cattle production. In general, attitudes towards zoonotic control were positive, but approximately half the farmers showed no intent to control and were inhibited by non-supportive social norms and/or a lack of belief in self-efficacy. The remaining farmers showed intent to control, but had not implemented any structured control program due to external barriers including lack of knowledge and both cultural and economic pressure from society and industry. The farmers with no intent to adopt control measures identified their private veterinarian as the preferred motivator, whereas consumer-demand and financial rewards or penalties were significantly associated with farmers who intended to control.


Supported employment for people with severe mental illness: a pilot study of an Italian social enterprise with a special ingredient.
People with mental disorders are far more likely to be unemployed than the general population. Two internationally recognized, evidence-based models of interventions for employment for people with severe mental health problems are Individual Placement Support and the Clubhouse. In Italy, a common model is the 'social enterprise' (SE), which is a programme run by non-profit organisations that help individuals with disabilities to be employed. Despite SEs spread and relevance in Italy, there are no studies about Italian samples. This paper reports on a pilot evaluation of psychosocial and work outcomes of a SE based in Verona, Italy. The study aims to investigate if people with SMI involved in SE job placements may achieve personal recovery and better outcomes over time, and in comparison with a comparable group of users. This is a pilot descriptive study with three components. A longitudinal design that comprised a functioning description of 33 SE members with a psychiatric disability in two time-points (when they joined the SE-on average 5 years before the study recruitment, and at the study recruitment-year 2018); and a repeated collection of job details of the 33 members in three time points: 2 years before the recruitment,-year 2016; 1 year before the recruitment - year 2017; and at the recruitment-year 2018. An assessment at the recruitment time-year 2018, of SE users' satisfaction with the job placement, symptoms, functioning, and quality of life (QoL). A cross-sectional study that compared the 33 SE members at the recruitment time-year 2018, with a matched group of people with the following criteria: living in local supported accommodations, being unemployed and not SE members. The two groups were compared on ratings of psychopathology, functioning, and QoL. Descriptive analyses were done. At the recruitment time - year 2018, all SE participants showed a significant better functioning (p < 0.001) than when they joined the SE-when they had been employed for an average of 5 years. In comparison to the matched group, SE members had significantly better functioning (p = 0.001), psychopathology (p = 0.007), and QoL (p = 0.034). According to their SE membership status, participants comprised trainees (21.2%) and employee members (78.8%). Trainees compared to employees had lower autonomies, functioning, QoL and more severe psychopathology. Over the two years prior to study recruitment, trainees showed stable poor autonomies, while employee members showed a variation from average autonomies in the 2 years before the recruitment time - year 2016, to good ones at the recruitment time - year 2018. Over the two years, all SE members set increasing numbers of objectives in all three domains. All SE participants reported high levels of satisfaction with all aspects of the job placement. SE that provides tailored support to assist people to gain employment skills may be an effective component in helping recovery from SMI.


Neurofibromin-deficient fibroblasts fail to form perineurium in vitro.
To identify cell type(s) that might contribute to nerve sheath tumors (neurofibromas) in patients with neurofibromatosis type 1, we generated cell cultures containing neurons. Schwann cells and fibroblasts from transgenic mouse embryos in which the type 1 neurofibromatosis gene was disrupted by homologous recombination (Brannan et al. (1994) Genes Development, 8,1019-1029). Normal fascicle formation by perineurial cells failed to occur in the absence of neurofibromin. Fascicles were reduced in number and showed abnormal morphology when normal neurons and Schwann cells were cultured up to 37 days with fibroblasts lacking neurofibromin. Proliferation was increased in a majority of fibroblast cell strains analyzed from embryos lacking neurofibromin. These observations suggest that mutations in the neurofibromatosis type I gene affect fibroblast behavior that might contribute to neurofibroma formation in patients with neurofibromatosis type 1.


Single-colour flow cytometric assay to determine NK cell-mediated cytotoxicity and viability against non-adherent human tumor cells.
A flow cytometry-based cytotoxicity (FCC) assay was developed using a single fluorophore, calcein-acetoxymethyl diacetylester (calcein-AM), to measure NK cell-mediated cytotoxicity. Non-adherent human K562 and U937 target cells were individually labelled with calcein-AM and co-incubated with effector NK cells to measure calcein loss, and therefore calculate target cell cytotoxicity. This FCC assay also provided a measure of sample viability. Notably, cell viability measured by traditional calcein/7-amino-actinomycin D (7-AAD) double labelling and Trypan Blue methods were comparable to the viability calculated using calcein-loss FCC. This FCC assay may also be used with various effector and target cell types and as a multi-parameter tool to measure viability and immunophenotype cells for tissue engineering purposes.


Contribution of Gestational Weight Gain on Maternal Glucose Metabolism in Women with GDM and Normal Glucose Tolerance.
Efforts to decrease the risk of developing metabolic complications of pregnancy such as gestational diabetes (GDM) through lifestyle intervention (decreasing excessive gestational weight gain (GWG)) during pregnancy have met with limited success. The purpose of this study was to determine the relationship between the longitudinal changes in weight/body composition and insulin sensitivity and response in women with normal glucose tolerance (NGT) and those who developed GDM. We conducted a secondary analysis of a prospective cohort developed before conception and again at 34 to 36 weeks gestation. A total of 29 NGT and 17 GDM women were evaluated for longitudinal changes in insulin sensitivity/response using the hyperinsulinemic-euglycemic clamp and an IV-glucose tolerance test. Body composition was estimated using hydrodensitometry. Both absolute change (Δ) and relative change (%Δ) between these 2 time points were calculated. We performed simple and multiple linear regression analysis to assess the relationship between GWG and measures of glucose metabolism, ie, insulin sensitivity and response. Based on the primary study design there was no significant difference in clinical characteristics between women with NGT and those developing GDM. Prior to pregnancy, women who developed GDM had lower insulin sensitivity levels (P = 0.01) compared with NGT women. Absolute change and %Δ in insulin sensitivity/insulin response and body weight/body composition were not significantly different between NGT and GDM women. Changes in body weight contributed to only 9% of the Δ in insulin sensitivity both in women developing GDM and NGT women. These data suggest that other factors-such as maternal pre-pregnancy insulin sensitivity and placental derived factors affecting insulin sensitivity-rather than maternal GWG account for the changes in glucose metabolism during human pregnancy.


Ozone exposure and mortality: an empiric bayes metaregression analysis.
Results from time-series epidemiologic studies evaluating the relationship between ambient ozone concentrations and premature mortality vary in their conclusions about the magnitude of this relationship, if any, making it difficult to estimate public health benefits of air pollution control measures. We conducted an empiric Bayes metaregression to estimate the ozone effect on mortality, and to assess whether this effect varies as a function of hypothesized confounders or effect modifiers. We gathered 71 time-series studies relating ozone to all-cause mortality, and we selected 48 estimates from 28 studies for the metaregression. Metaregression covariates included the relationship between ozone concentrations and concentrations of other air pollutants, proxies for personal exposure-ambient concentration relationships, and the statistical methods used in the studies. For our metaregression, we applied a hierarchical linear model with known level-1 variances. We estimated a grand mean of a 0.21% increase (95% confidence interval = 0.16-0.26%) in mortality per 10-microg/m increase of 1-hour maximum ozone (0.41% increase per 10 ppb) without controlling for other air pollutants. In the metaregression, air-conditioning prevalence and lag time were the strongest predictors of between-study variability. Air pollution covariates yielded inconsistent findings in regression models, although correlation analyses indicated a potential influence of summertime PM2.5. These findings, coupled with a greater relative risk of ozone in the summer versus the winter, demonstrate that geographic and seasonal heterogeneity in ozone relative risk should be anticipated, but that the observed relationship between ozone and mortality should be considered for future regulatory impact analyses.


Molecular characterisation of post-bio-electrosprayed human brain astrocytoma cells.
Bio-electrospraying (BES) is a method for directly jetting living cells under conditions that allow their distribution in the x, y, and z axes. Previous work has been focused on achieving jetting in stable cone-jet mode, which is required for precision placement, and these studies have demonstrated that there are no significant effects of bio-electrospraying on cell morphology or viability. In this work, we examine the biological properties of bio-electrosprayed cells using assays of cellular function that range from the molecular level through to integrated cellular systems, and include proteomics, signal transduction, cell growth and proliferation, and the characterisation of apoptotic blebs. From these molecular methods, we have determined that bio-electrospraying, under the electric field conditions used to achieve stable cone-jet mode, causes no alterations to the biological properties and function of the cells being jetted. Bio-electrosprayed and control cells had similar viability, proliferation properties and virtually indistinguishable cell cycle profiles. The biophysical properties of large conducting (BK) potassium channels were unchanged, as were the pharmacological responses of the endogenous muscarinic and exogenous P2Y(11) receptors, both of which are cell surface receptors of the 7TM superfamily. Proteomic analyses revealed that although three proteins had subtle differences in expression level between bio-electrosprayed and control cells, none of these fold differences was above the 1.5-fold cut-off threshold required for further analyses. These findings support the further development of bio-electrosprays as a viable technology for a wide diversity of tissue engineering, regenerative biology, advanced cellular therapeutics and medicinal applications, having significance in the clinic.


Postmortem molecular diagnosis of sickle beta thalassaemia.
This report describes a case in which the diagnosis of sickle cell disease (SCD) was established after death. The diagnosis of sickle cell syndrome was made in a 68 year old black patient who was found to have sickled red blood cells in many organs at necropsy although the disease had not been diagnosed during her lifetime. DNA was isolated from a peripheral blood smear obtained on the day of the patient's death. The beta globin gene was polymerase chain reaction amplified and sequenced, revealing that the patient had S-beta(+) thalassaemia. This study shows that blood smears are a suitable source for retrospective DNA analysis studies. This case illustrates that relatively "mild" forms of SCD can be overlooked, despite symptomatology suggestive of a sickle syndrome, and demonstrates the feasibility of the postmortem molecular diagnosis of haemoglobinopathies in such cases.


Review: relation between quality-of-care indicators for diabetes and patient outcomes: a systematic literature review.
The authors conducted a systematic literature review to assess whether quality indicators for diabetes care are related to patient outcomes. Twenty-four studies were included that formally tested this relationship. Quality indicators focusing on structure or processes of care were included. Descriptive analyses were conducted on the associations found, differentiating for study quality and level of analysis. Structure indicators were mostly tested in studies with weak designs, showing no associations with surrogate outcomes or mixed results. Process indicators focusing on intensification of drug treatment were significantly associated with better surrogate outcomes in three high-quality studies. Process indicators measuring numbers of tests or visits conducted showed mostly negative results in four high-quality studies on surrogate and hard outcomes. Studies performed on different levels of analysis and studies of lower quality gave similar results. For many widely used quality indicators, there is insufficient evidence that they are predictive of better patient outcomes.


Validation of Anti-Adeno Associated Virus Serotype rh10 (AAVrh.10) Total and Neutralizing Antibody Immunogenicity Assays.
Immunogenicity assessment of Adeno-Associated Virus (AAV) vectors is a critical part of gene therapy drug development. Whether the assays are used for inclusion/exclusion criteria or to monitor the safety and efficacy of the gene therapy, they are critical bioanalytical assessments. While total anti-AAV assays are perceived as easier to develop and implement than neutralizing anti-AAV assays, the gene therapy field is still nascent, and it is not yet clear which of the assays should be implemented at what stage of drug development. Recently AAVrh.10 has gained interest for use in gene therapies targeting cardiac, neurological, and other diseases due to its enhanced transduction efficiency. There is limited information on anti-AAVrh.10 antibodies and their clinical impact; thus, the information presented herein documents the validation of both a total antibody assay (TAb) and a neutralizing antibody (NAb) assay for anti-AAVrh.10 antibodies. In this manuscript, the validation was performed in accordance with the 2019 FDA immunogenicity guidance with additional evaluations to comply with CLIA where applicable. The AAVrh.10 TAb and NAb assays were compared in terms of sensitivity, drug tolerance, and precision, along with a concordance analysis using the same individual serum samples. This comparison gave insight into the utility of each format as a screening assay for inclusion into clinical studies.


Criteria for Clinically Relevant Bleeding in Critically Ill Children: An International Survey.
Bleeding, a feared complication of critical illness, is frequent in critically ill children. However, the concept of clinically relevant bleeding is ill-defined in this population. There are many established diagnostic criteria for bleeding, but only one estimates bleeding in critically ill adults, and none exist for critically ill children. Our objective was to identify the factors that influence pediatric intensivists' perception of clinically relevant bleeding. Self-administered, web-based survey with 9-point Likert scales, to qualify the clinical significance of 103 bleeding characteristics in critically ill children. Online survey. Pediatric critical care physicians and nurse practitioners. None. The response rate was 40%, with 225 respondents from 16 countries. Characteristics most frequently identified as clinically relevant were bleeding in critical locations (e.g., pericardium, pleural space, CNS, and lungs); requiring interventions; leading to physiologic repercussions, including organ failure; and of prolonged duration. Quantifiable bleeding greater than 5 mL/kg/hr for more than 1 hour was frequently considered clinically relevant. Respondents identified the following characteristics as clinically irrelevant: dressings required to be changed no less frequently than every 6 hours, streaks of blood in gastric tubes, streaks of blood in endotracheal tubes or blood in endotracheal tubes only during suctioning, lightly blood-tinged urine, quantifiable bleeding less than 1 mL/kg/hr, and noncoalescing petechiae. Perception of the clinical relevance of bleeding was not associated with the respondent's geographical location of clinical practice or years of experience. This international survey provides a better understanding of the factors that influence the pediatric intensivists' assessment of the clinical relevance of bleeding in critically ill children. It provides the foundation for the development of a validated, diagnostic definition of clinically relevant bleeding in this population.


Experimental fetal endoscopic tracheal occlusion in rhesus and cynomolgus monkeys: nonhuman primate models.
The monkey model is the best model to investigate some physiological response to the fetal transitory tracheal occlusion but it has never been described in Macaca monkeys. The aim of this study was to evaluate the feasibility of fetal endoscopic tracheal occlusion (FETO) in a non-human primate model. Pregnant rhesus monkeys and cynomolgus were tested as a potential experimental model for FETO in the third trimester of pregnancy, by performing fetal tracheoscopies with and without tracheal occlusion. A total of 22 pregnancies were followed in 16 monkeys and underwent fetal surgery. Percutaneous endoscopic access to the uterine cavity was possible in 20 cases (91%). Of these 20 pregnant monkeys, fetal tracheoscopy could be achieved in 15 cases (75%). In rhesus monkeys, the time between the onset of endoscopy and tracheal penetration decreases as operator experience increases. Neither maternal morbidity nor mortality was related to surgery. Two fetal losses were possibly due to the procedure. FETO is feasible in the non-human primate, which closely reflects procedures in humans. The non-human primate model for FETO, specially the rhesus monkeys, may be useful for future studies concerning the mechanisms related to the lung growth after transitory fetal tracheal occlusion.


Antigen-encapsulating host extracellular vesicles derived from Salmonella-infected cells stimulate pathogen-specific Th1-type responses in vivo.
Salmonella Typhimurium is a causative agent of nontyphoidal salmonellosis, for which there is a lack of a clinically approved vaccine in humans. As an intracellular pathogen, Salmonella impacts many cellular pathways. However, the intercellular communication mechanism facilitated by host-derived small extracellular vesicles (EVs), such as exosomes, is an overlooked aspect of the host responses to this infection. We used a comprehensive proteome-based network analysis of exosomes derived from Salmonella-infected macrophages to identify host molecules that are trafficked via these EVs. This analysis predicted that the host-derived small EVs generated during macrophage infection stimulate macrophages and promote activation of T helper 1 (Th1) cells. We identified that exosomes generated during infection contain Salmonella proteins, including unique antigens previously shown to stimulate protective immune responses against Salmonella in murine studies. Furthermore, we showed that host EVs formed upon infection stimulate a mucosal immune response against Salmonella infection when delivered intranasally to BALB/c mice, a route of antigen administration known to initiate mucosal immunity. Specifically, the administration of these vesicles to animals stimulated the production of anti-Salmonella IgG antibodies, such as anti-OmpA antibodies. Exosomes also stimulated antigen-specific cell-mediated immunity. In particular, splenic mononuclear cells isolated from mice administered with exosomes derived from Salmonella-infected antigen-presenting cells increased CD4+ T cells secreting Th1-type cytokines in response to Salmonella antigens. These results demonstrate that small EVs, formed during infection, contribute to Th1 cell bias in the anti-Salmonella responses. Collectively, this study helps to unravel the role of host-derived small EVs as vehicles transmitting antigens to induce Th1-type immunity against Gram-negative bacteria. Understanding the EV-mediated defense mechanisms will allow the development of future approaches to combat bacterial infections.


Distinct mechanisms for the induction and maintenance of allograft tolerance with CTLA4-Fc treatment.
A murine CTLA4/Fc gamma2a heavy chain (mCTLA4-Fc) chimeric fusion molecule was used in B6AF1 recipients of BALB/c pancreatic islet allografts to study the induction and maintenance of tolerance following inhibition of the CD28-B7 pathway for T cell activation. Donor-specific tolerance was achieved by administering 100 microg of mCTLA4-Fc on alternate days for 14 days (8 total doses) or a single 500 microg dose of mCTLA4-Fc on day 2 after transplant. Tolerance was mediated by long-lived peripheral lymphocytes and showed features of organ and alloantigen specificity. Whereas tolerance could not be established in allograft recipients receiving simultaneous mCTLA4-Fc and rIL-2, previously tolerant animals did not reject their grafts when given IL-2, suggesting that the induction and maintenance phases of tolerance were distinct and separate. The maintenance of donor-specific tolerance was an active immunologic process that was CD4+ T cell dependent and could be adoptively transferred to naive lymphocytes, but could not be explained by apoptosis or deletion of alloreactive T cells. Although an IL-2-sensitive mechanism such as anergy may contribute toward the induction of tolerance, its maintenance involves active suppression.


[Somatic hybrids of normal and tumor Djzungarian hamster cells. II. Manifestation of malignancy and karyotype features of hybrid tumors].
The hybrid clones derived from the fusion of tumour and normal cells of Djungarian hamster were tested for their ability to grow progressively in vivo and to form colonies in semisolid medium. In all cases the hybrids were able to produce tumours in animals, but tumorigenicity of different clones varied. Some clones had high take incidence of tumours comparable to that of malignant partner, others had a very low one. The hybrid clones differed in their ability to form colonies in soft agar. No correlation was found between the malignancy of the hybrid clones in vivo and their ability to grow in semisolid medium. Chromosome analysis of 23 hybrid tumours arising from the injections of the hybrid cells showed that in 18 tumours the drastic reduction of chromosomes from tetraploid to near-diploid level, comparable to that of malignant parent, took place. As a rule, morphologically unchanged chromosomes were preferentially lost from the hybrid tumour cells, the markers of the malignant partner being retained. Some hybrid tumours showed insignificant chromosome elimination of all pairs, except chromosomes of the IV and VIII pairs, their number always being reduced.