Relationship Between Cerebrovascular Reactivity and Cognition Among People With Risk of Cognitive Decline.
Vascular risk factors (e.g., obesity and hypertension) are associated with cerebral small vessel disease, Alzheimer's disease (AD) pathology, and dementia. Reduced perfusion may reflect the impaired ability of blood vessels to regulate blood flow in reaction to varying circumstances such as hypercapnia (increased end-tidal partial pressures of CO2). It has been shown that cerebrovascular reactivity (CVR) measured with blood-oxygen-level-dependent (BOLD) MRI is correlated with cognitive performance and alterations of CVR may be an indicator of vascular disfunction leading to cognitive decline. However, the underlying mechanism of CVR alterations in BOLD signal may not be straight-forward because BOLD signal is affected by multiple physiological parameters, such as cerebral blood flow (CBF), cerebral blood volume, and oxygen metabolism. Arterial spin labeling (ASL) MRI quantitatively measures blood flow in the brain providing images of local CBF. Therefore, in this study, we measured CBF and its changes using a dynamic ASL technique during a hypercapnia challenge and tested if CBF or CVR was related to cognitive performance using the Mini-mental state examination (MMSE) score. Seventy-eight participants underwent cognitive testing and MRI including ASL during a hypercapnia challenge with a RespirAct computer-controlled gas blender, targeting 10 mmHg higher end-tidal CO2 level than the baseline while end-tidal O2 level was maintained. Pseudo-continuous ASL (PCASL) was collected during a 2-min baseline and a 2-min hypercapnic period. CVR was obtained by calculating a percent change of CBF per the end-tidal CO2 elevation in mmHg between the baseline and the hypercapnic challenge. Multivariate regression analyses demonstrated that baseline resting CBF has no significant relationship with MMSE, while lower CVR in the whole brain gray matter (β = 0.689, p = 0.005) and white matter (β = 0.578, p = 0.016) are related to lower MMSE score. In addition, region of interest (ROI) based analysis showed positive relationships between MMSE score and CVR in 26 out of 122 gray matter ROIs.


Health information exchange implementation: lessons learned and critical success factors from a case study.
Much attention has been given to the proposition that the exchange of health information as an act, and health information exchange (HIE), as an entity, are critical components of a framework for health care change, yet little has been studied to understand the value proposition of implementing HIE with a statewide HIE. Such an organization facilitates the exchange of health information across disparate systems, thus following patients as they move across different care settings and encounters, whether or not they share an organizational affiliation. A sociotechnical systems approach and an interorganizational systems framework were used to examine implementation of a health system electronic medical record (EMR) system onto a statewide HIE, under a cooperative agreement with the Office of the National Coordinator for Health Information Technology, and its collaborating organizations. The objective of the study was to focus on the implementation of a health system onto a statewide HIE; provide insight into the technical, organizational, and governance aspects of a large private health system and the Virginia statewide HIE (organizations with the shared goal of exchanging health information); and to understand the organizational motivations and value propositions apparent during HIE implementation. We used a formative evaluation methodology to investigate the first implementation of a health system onto the statewide HIE. Qualitative methods (direct observation, 36 hours), informal information gathering, semistructured interviews (N=12), and document analysis were used to gather data between August 12, 2012 and June 24, 2013. Derived from sociotechnical concepts, a Blended Value Collaboration Enactment Framework guided the data gathering and analysis to understand organizational stakeholders' perspectives across technical, organizational, and governance dimensions. Several challenges, successes, and lessons learned during the implementation of a health system to the statewide HIE were found. The most significant perceived success was accomplishing the implementation, although many interviewees also underscored the value of a project champion with decision-making power. In terms of lessons learned, social reasons were found to be very significant motivators for early implementation, frequently outweighing economic motivations. It was clear that understanding the guides early in the project would have mitigated some of the challenges that emerged, and early communication with the electronic health record vendor so that they have a solid understanding of the undertaking was critical. An HIE implementations evaluation framework was found to be useful for assessing challenges, motivations, value propositions for participating, and success factors to consider for future implementations. This case study illuminates five critical success factors for implementation of a health system onto a statewide HIE. This study also reveals that organizations have varied motivations and value proposition perceptions for engaging in the exchange of health information, few of which, at the early stages, are economically driven.


Exploring the role of delta-V in influencing occupant injury severities - A mediation analysis approach to motor vehicle collisions.
This study investigates the impact that delta-V, the relative change in vehicle velocity pre- and post-crash, has on the severity of motor vehicle collisions (MVCs). We study injury severity using two metrics for each occupant - the number of injuries suffered, and the probability of suffering a serious or worse (MAIS 3+) injury. We use a cross-sectional set of generally-representative MVC data between 2010 and 2015 as a basis for our research. Collision factors that influence the crash environment are combined with the injuries that were suffered in MVCs. The influence of delta-V is captured using a mediation analysis, whereby delta-V acts as the focal point between crash factors and injury outcome. The mediation approach adds to existing research by presenting a detailed view of the relationship between injury severity, delta-V and other collision factors. We find evidence of competitive mediation, wherein a collision factor's positive association with injury severity is offset by a negative association with delta-V. Neglecting to include delta-V in our study would have let the factor's association with injury severity go undiscovered. In addition, certain collision factors are found to be related to injury severity solely because of delta-V, while others are found to have a significant impact regardless of delta-V. Our results support the multitude of policy recommendations that promote seatbelt use and warn against alcohol-impaired driving, and support the proliferation of safety-enabled vehicles whose technology can mitigate the bodily damage associated with detrimental crash types.


Intimate partner violence: development of a brief risk assessment for the emergency department.
Women assaulted by intimate partners are frequently patients in emergency departments (EDs). Many victims and health care providers fail to take into account the potential risks of repeat partner violence. The objective of this study was to use data from a larger study of domestic violence risk assessment methods to develop a brief assessment for acute care settings to identify victims at highest risk for suffering severe injury or potentially lethal assault by an intimate partner or former partner. Victims of intimate partner violence (IPV) were interviewed twice between 2002 and 2004. The baseline interview included the 20 items of Campbell's Danger Assessment (DA; predictor). The follow-up interview, conducted 9 months later on average, assessed abuse inflicted since the baseline interview (outcome). Multiple logistic regression was used to identify questions on the DA most predictive of severe abuse and potentially lethal assaults. Female IPV victims were recruited from New York City family courts, Los Angeles County Sheriff's Department 9-1-1 calls, New York City and Los Angeles shelters, and New York City hospitals; 666 women responded to the DA at baseline, and 60% participated in follow-up interviews. Severe injuries or potentially lethal assaults were experienced by 14.9% of retained study participants between the baseline and follow-up interviews. The best brief prediction instrument has five questions. A positive answer to any three questions has a sensitivity of 83% (95% confidence interval = 70.6% to 91.4%). This instrument can help predict which victims may be at increased risk for severe injury or potentially lethal assault and can aid clinicians in differentiating which patients require comprehensive safety interventions.


The protease of herpes simplex virus type 1 is essential for functional capsid formation and viral growth.
The herpes simplex virus type 1 protease and related proteins are involved in the assembly of viral capsids. The protease encoded by the UL26 gene can process itself and its substrate ICP35, encoded by the UL26.5 gene. To better understand the functions of the protease in infected cells, we have isolated a complementing cell line (BMS-MG22) and constructed and characterized a null UL26 mutant virus, m100. The mutant virus failed to grow on Vero cells and required a complementing cell line for its propagation, confirming that the UL26 gene product is essential for viral growth. Phenotypic analysis of m100 shows that (i) normal amounts of the c and d forms of ICP35 were produced, but they failed to be processed to the cleaved forms, e and f; (ii) viral DNA replication of the mutant proceeded at near wild-type levels, but DNA was not processed to unit length or encapsidated; (iii) capsid structures were observed in thin sections of m100-infected Vero cells by electron microscopy, but assembly of VP5 into hexons of the capsid structure was conformationally altered; and (iv) nuclear localizations of the protease and ICP35 are independent of each other, and the function(s) of Na, at least in part, is to direct the catalytic domain N(o) to the nucleus.


Veterans' reasons for dropping out of prolonged exposure therapy across three delivery modalities: A qualitative examination.
Premature dropout from posttraumatic stress disorder treatment (PTSD) hinders treatment response. Studies have primarily used quantitative methodology to identify factors that contribute to Veterans' premature dropout, which has yielded mixed results. Qualitative methods provide rich data and generate additional hypotheses about why Veterans discontinue PTSD treatment. This study aimed to understand Veterans' reasons for dropping out of prolonged exposure therapy (PE) and to examine if there are differences in reasons for dropout between three delivery modalities: in-home, in-person (IHIP), office-based telehealth (OBT), or home-based telehealth (HBT). Twenty-two Veterans who dropped out of PE from a parent randomized clinical trial participated in individual qualitative interviews about potential contextual and individual factors related to discontinuation. Team-based coding was used to conduct open and focused coding. Themes were generated that described factors that influenced Veterans' dropout from PE and constant comparison was used to explore differences in reasons between the three modalities. Most Veterans had multiple reasons for dropping out and reasons were similar across delivery modalities with few differences. Practical barriers (e.g., scheduling difficulties), attitudes toward mental health providers and therapy (e.g., stigma), psychological and physical health factors (e.g., perceived worsening of symptoms, pain), and the therapeutic context (e.g., disliking aspects of PE) contributed to Veterans' decisions to drop out from PE. Veterans in OBT reported more types of practical barriers than Veterans in HBT and IHIP. These findings can help generate hypotheses about interventions that may promote engagement and future studies should continue to study how to reduce dropout. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


[Efficacy of valve drainage devices in secondary glaucoma in patients with artificial iridolenticular diaphragm].
Results of 5 Akhmed valve implantations in patients with secondary glaucoma various time after artificial iridolenticular diaphragm implantation after concomitant iris and lens trauma were analyzed. Surgery allowed to achieve reliable IOP decrease when severe scar deformation of drainage zone of the anterior chamber prevents traditional hipotensive procedures because of their ineffectiveness. Besides visual acuity stabilization a safe performance of subsequent reconstructive procedures can become possible as well.


Terahertz near-field vectorial imaging of subwavelength apertures and aperture arrays.
We present measurements of the complete terahertz (THz) electric near-field distribution, E(x), E(y) and E(z), in both the time- and frequency-domains, for subwavelength apertures and subsections of subwavelength aperture arrays. Measuring the individual components of the THz near-field with subwavelength spatial resolution, as they emerge from these structures, illustrates how the field interacts with the apertures. We observe the small but measurable y- and z-components of the electric field for both single apertures and arrays. Resonant contributions, attributed to Bloch modes, are detected and we observe the presence of a longitudinal field component, E(z), within the different array apertures, which can be attributed to a diffractive effect. These measurements illustrate in detail the individual THz field components emerging from subwavelength apertures and provide a direct measure of two important mechanisms that contribute to the net transmission of light through arrays.


[Metabolopathies. The importance of a good performance nurse].
The possibility for early detection of metabolic disorders has been a big step forward in medical history due to using a good technique by which diseases can be diagnosed, improving their outcome. The main disorders that these tests are designed to detect are hypothyroidism, congenital adrenal hyperplasia, hyperphenylalaninemia, hemoglobinopathies, and cystic fibrosis, among others. Samples can be collected by two methods: the single draw (taken on the third day of life) or the double draw (the first sample is taken at 48 hours of life and the second after the fourth day). It is important to institute a good technique in order to obtain good results and prevent erroneous samples or false positives.


Effect of Dietary Processed Sulfur Supplementation on Water-holding Capacity, Color, and Lipid Profiles of Pork.
This study was performed to investigate the effect of dietary processed sulfur supplementation on water-holding capacity, color, and lipid profiles of pork according to the level of dietary processed sulfur (0%, CON; 0.3%, S). The pigs were slaughtered at an average final weight of 120 kg, and the longissimus dorsi muscles were collected from the carcasses. As results, pork processed with sulfur had significantly higher moisture and ash contents compared to those of CON but lower crude fat, pH, expressible drip, lower redness and yellowness, and greater lightness. Pork processed with sulfur showed significantly lower total lipid content, triglycerides, and atherosclerosis index but significantly higher high-density lipoprotein cholesterol. Feeding processed sulfur significantly lowered myristic acid, heptadecanoic acid, and stearic acid contents, whereas monounsaturated fatty acids and oleic acids were significantly higher compared to those in the CON. Higher amounts of polyunsaturated fatty acids and n-6 fatty acids were observed in the pork processed with sulfur than that of the CON. Therefore, supplementing pigs with dietary sulfur improved nutrient and meat quality.


Specific and unspecific responses of plants to cold and drought stress.
Different environmental stresses to a plant may result in similar responses at the cellular and molecular level. This is due to the fact that the impacts of the stressors trigger similar strains and downstream signal transduction chains. A good example for an unspecific response is the reaction to stressors which induce water deficiency e.g.drought, salinity and cold, especially frost. The stabilizing effect of liquid water on the membrane bilayer can be supported by compatible solutes and special proteins. At the metabolic level, osmotic adjustment by synthesis of low-molecular osmolytes (carbohydrates, betains, proline) can counteract cellular dehydration and turgor loss. Taking the example of Pinus sylvestris, changes at the level of membrane composition, and concomitantly of photosynthetic capacity during frost hardening is shown. Additionally the effect of photoperiod as measured via the phytochrome system and the effect of subfreezing temperatures on the incidence of frost hardening is discussed. Extremely hydrophilic proteins such as dehydrins are common products protecting not only the biomembranes in ripening seeds (late embryogenesis abundant proteins)but accumulate also in the shoots and roots during cold adaptation, especially in drought tolerant plants. Dehydrins are characterized by conserved amino acid motifs, called the K-,Y-or S-segments. Accumulation of dehydrins can be induced not only by drought, but also by cold,salinity,treatment with abscisic acid and methyl jasmonate. Positive effects of the overexpression of a wild chickpea (Cicer pinnatifidum) dehydrin in tobacco plants on the dehydration tolerance is shown. The presentation discusses the perception of cold and drought,the subsequent signal transduction and expression of genes and their products. Differences and similarities between the plant responses to both stressors are also discussed.


A plant-transformation-competent BIBAC/BAC-based map of rice for functional analysis and genetic engineering of its genomic sequence.
Sequencing of the rice genome has provided a platform for functional genomics research of rice and other cereal species. However, multiple approaches are needed to determine the functions of its genes and sequences and to use the genome sequencing results for genetic improvement of cereal crops. Here, we report a plant-transformation-competent, binary bacterial artificial chromosome (BIBAC) and bacterial artificial chromosome (BAC) based map of rice to facilitate these studies. The map was constructed from 20 835 BIBAC and BAC clones, and consisted of 579 overlapping BIBAC/BAC contigs. To facilitate functional analysis of chromosome 8 genomic sequence and cloning of the genes and QTLs mapped to the chromosome, we anchored the chromosomal contigs to the existing rice genetic maps. The chromosomal map consists of 11 contigs, 59 genetic markers, and 36 sequence tagged sites, spanning a total of ca. 38 Mb in physical length. Comparative analysis between the genetic and physical maps of chromosome 8 showed that there are 3 "hot" and 2 "cold" spots of genetic recombination along the chromosomal arms in addition to the "cold spot" in the centromeric region, suggesting that the sequence component contents of a chromosome may affect its local genetic recombination frequencies. Because of its plant transformability, the BIBAC/BAC map could provide a platform for functional analysis of the rice genome sequence and effective use of the sequencing results for gene and QTL cloning and molecular breeding.


Effect of ribonuclease from Bacillus intermedius on human blood lymphocytes.
By using a rosette formation test the effect of ribonuclease Bacillus intermedius (RNase Bi) on T- and B-lymphocytes in human peripheral blood has been studied in vitro. The RNase effect on T-lymphocytes depends on its concentration: low concentrations (10(-6)-10(-2) microg ml(-1)) stimulate E-rosette formation whereas high concentrations (10 microg ml(-1)) suppress it. The amount of B-lymphocytes decreases under RNase Bi influence in all concentrations tested. RNase Bi like thymus hormones influence immature lymphocytes (0-cells) by inducing the surface expression of E-receptors what leads to rosette formation and, thus, contributes to lymphocyte differentiation. The increase in the amount of active T-cells which represent the mature cell population also confirms the participation of RNase Bi in T-rank lymphocytes differentiation processes. The RNase Bi effect on the human lymphocytes depends on its catalytic activity.


Altered expression of the platelet aggregation-associated protein from Streptococcus sanguis after growth in the presence of collagen.
Certain strains of Streptococcus sanguis adhere selectively to human platelets (Adh+) and, in plasma, induce them to aggregate into in vitro thrombi (Agg+). The induction of aggregation is mediated by the platelet aggregation-associated protein (PAAP) expressed on the cell surface of the streptococcus. In endocarditis, expression of PAAP may be regulated by association with host proteins on damaged heart valves. To begin to test this hypothesis, three strains of S. sanguis were each cultured in the presence or absence of collagens (types I to X), laminin, or PAAP-derived peptide preparations. After harvesting and washing, the platelet-interactive phenotype of strains 133-79 (Adh+ Agg+), L74 (Adh+ Agg-), and 10556 (Adh- Agg-) was unchanged. The cells from each culture were then digested mildly with trypsin to isolate PAAP. PAAP isolated from strain 133-79 (Adh+ Agg+) grown in the absence of added collagen, other proteins, or peptides inhibited platelet aggregation in response to untreated cells of S. sanguis. Platelet aggregation was induced immediately, however, by PAAP from strain 133-79 isolated after growth in the presence of 300 nM type I collagen, while lower concentrations yielded protein fragments that potentiated the response to intact cells. Aggregation-inducing PAAP could be removed by anti-PAAP (PGEQGPK) immunoaffinity chromatography, but only inhibitory activity could be recovered. The agonist effect of PAAP was not associated with collagen itself, since the PAAP preparations did not contain detectable amounts of hydroxyproline. PAAP antigens isolated from cells grown in the presence and absence of collagen had similar apparent molecular weights, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting. When electrophoresis was performed under nondenaturing conditions, however, PAAP isolated from cells grown in type I collagen migrated more slowly. Strain L74 grown with type I collagen yielded tryptic fragments of proteins that inhibited aggregation significantly better than control peptides (no collagen in the medium). Strain 10556 was apparently unaffected by growth in type I collagen. The effect of type I collagen was somewhat unique. Growth in the presence of collagen types II to VI (300 nM) yielded protein fragments that potentiated without inducing platelet aggregation, while other collagens, laminin, and PAAP-derived peptides did not affect platelet aggregation. These results suggest that growth in the presence of type I collagen and, perhaps, collagens II to VI alters the expression and conformation of PAAP in certain strains of S. sanguis.


Targeting GSTP1 as Therapeutic Strategy against Lung Adenocarcinoma Stemness and Resistance to Tyrosine Kinase Inhibitors.
Glutathione S-transferase pi (GSTP1), a phase II detoxification enzyme, is known to be overexpressed and mediates chemotherapeutic resistance in lung cancer. However, whether GSTP1 supports cancer stem cells (CSCs) and the underlying mechanisms in lung adenocarcinoma (LUAD) remain largely unknown. This study unveiled that GSTP1 is upregulated in lung CSCs and supports tumor self-renewal, metastasis, and resistance to targeted tyrosine kinase inhibitors of LUAD both in vitro and in vivo. Mechanistically, CaMK2A (calcium/calmodulin-dependent protein kinase 2 isoform A)/NRF2 (nuclear factor erythroid 2-related factor 2)/GSTP1 is uncovered as a regulatory axis under hypoxia. CaMK2A increased GSTP1 expression through phosphorylating the Sersine558 residue of NRF2 and promoting its nuclear translocation, a novel mechanism for NRF2 activation apart from conventional oxidization-dependent activation. Upregulation of GSTP1 in turn suppressed reactive oxygen species levels and supported CSC phenotypes. Clinically, GSTP1 analyzed by immunohistochemistry is upregulated in a proportion of LUAD and serves as a prognostic marker for survival. Using patient-derived organoids from an ALK-translocated LUAD, the therapeutic potential of a specific GSTP1 inhibitor ezatiostat in combination treatment with the ALK inhibitor crizotinib is demonstrated. This study demonstrates GSTP1 to be a promising therapeutic target for long-term control of LUAD through targeting CSCs.


Acupuncture for Climacteric-Like Symptoms in Breast Cancer Improves Sleep, Mental and Emotional Health: A Randomized Trial.
Objective: Breast cancer is the most prevalent malignant neoplasm in women. Optimal treatment frequently includes a hormonal-blockage phase maintained for 5-10 years. Pharmacologic agents used for this blockage induce many climacteric-like symptoms, which often exact a heavy toll on patients' quality of life. Acupuncture has had promising results for treating climacteric-like symptoms induced by hormonal blockage, but there is no evidence of efficacy for controlling hot flashes. Materials and Methods: This trial used acupuncture to treat the climacteric-like symptoms of patients with breast cancer, focusing on the mental, physical, and genitourinary symptoms and sleep disturbances, to determine the influence of acupuncture treatment. The randomized placebo-controlled trial, at a university-based cancer center, with blinded data collectors, compared an Acupuncture group (A), a Sham-Acupuncture group (S), and a Wait-List Control group (C). The patients were receiving tamoxifen. Group A had 10 weekly sessions of manual Acupuncture; Group S had 10 weekly sessions of Sham Acupuncture; and Group C spent 10 weeks on a Wait-List. The main outcome measures were the Beck Depression Inventory-II (BDI-II), the Pittsburgh Sleep Quality Index (PSQI), and the Menopause Rating Scale (MRS). Results: Primary analysis revealed differences among the groups' improvements in scores for the 3 questionnaires (P < 0.001), The A group had significant improvements on the BDI-II (P < 0.001), PSQI (P < 0.002), and MRS (P < 0.004) compared to the S group in a post hoc analysis. Conclusions: Acupuncture improved sleep, and mental and emotional distress symptoms induced by hormonal blockage in patients with breast cancer. Clinical Trial: This trial was registered at CAAE as trial #: 37758414.8.0000.0065.


Use of Dipeptidyl-Peptidase-4 Inhibitors and the Risk of Pneumonia: A Population-Based Cohort Study.
Dipeptidyl-peptidase-4 inhibitors (DPP4Is) are drugs for the treatment of type 2 diabetes mellitus (T2DM). There is increasing evidence that DPP4Is may result in suppression of the immune system and may increase the risk of infections such as pneumonia. Aim of this study was to evaluate the association between the use of DPP4Is and the risk of pneumonia in a population-based study. We conducted a population-based cohort study using data from the world's largest primary care database, the UK Clinical Practice Research Datalink (CPRD). We selected all users of non-insulin antidiabetic drugs (NIADs), including DPP4Is, between 2007 and 2012. To each NIAD user, we matched randomly selected non-users. The NIAD user's first prescription defined the index date, which was then assigned to the matched non-users. Patients were followed from their first prescription until end of data collection or the first event of pneumonia, whichever came first. Cox regression analysis estimated the association between pneumonia and current use of DPP4Is versus 1) current use of other NIADs and 2) non-users. DPP4I use was then stratified to daily and cumulative dose. Analyses were statistically adjusted for age, sex, lifestyle factors and comorbidities and concomitant use of various other drugs. Risk of pneumonia was not increased with current DPP4I use versus use of other NIADs, adjusted Hazard Ratio (HR) 0.70; 95% Confidence Interval (CI) 0.55-0.91. Also higher cumulative doses or daily doses did not further increase risk of pneumonia. We found no increased risk of pneumonia in T2DM patients using DPP4Is compared to T2DM patients using other NIADs. Our finding is in line with direct and indirect evidence from observational studies and RCTs. There is probably no need to avoid prescribing of DPP4Is to elderly patients who are at risk of pneumonia.


Neuromodulatory Burst Therapy for Agent Orange-Induced Peripheral Neuropathy: A Case Report.
The prevalence of neuromuscular diseases and peripheral neuropathies in veterans exposed to Agent Orange (AO) is particularly high. Pharmacologic management has not been effective for these patients. Burst therapy spinal cord stimulation (SCS), recently approved for use in the United States, has had demonstrable success in both Europe and Australia. We present a patient with AO-induced peripheral neuropathy who was successfully treated with burst SCS. To our knowledge, this is the first report describing the use of burst SCS for treatment of peripheral neuropathy related to AO exposure.


Role of calcium in adaptive cytoprotection and cell injury induced by deoxycholate in human gastric cells.
We have developed an in vitro model of adaptive cytoprotection induced by deoxycholate (DC) in human gastric cells and have shown that pretreatment with a low concentration of DC (mild irritant, 50 microM) significantly attenuates injury induced by a damaging concentration of DC (250 microM). This study was undertaken to assess the effect of the mild irritant on changes in intracellular Ca2+ and to determine if these perturbations account for its protective action. Protection conferred by the mild irritant was lost when any of its effects on intracellular Ca2+ were prevented: internal Ca2+ store release via phospholipase C and inositol 1,4, 5-trisphosphate sustained Ca2+ influx through store-operated Ca2+ channels or eventual Ca2+ efflux. We also investigated the relationship between Ca2+ accumulation and cellular injury induced by damaging concentrations of DC. In cells exposed to high concentrations of DC, sustained Ca2+ accumulation as a result of extracellular Ca2+ influx, but not transient changes in intracellular Ca2+ content, appeared to precede and induce cellular injury. We propose that the mild irritant disrupts normal Ca2+ homeostasis and that this perturbation elicits a cellular response (involving active Ca2+ efflux) that subsequently provides a protective action by limiting the magnitude of intracellular Ca2+ accumulation.


Understanding differences between phylogenetic and pedigree-derived mtDNA mutation rate: a model using families from the Azores Islands (Portugal).
We analyzed the control region of the mitochondrial DNA (mtDNA) from maternally related individuals originating from the Azores Islands (Portugal) in order to estimate the mutation rate of mtDNA and to gain insights into the process by which a new mutation arises and segregates into heteroplasmy. Length and/or point heteroplasmies were found at least in one individual of 72% of the studied families. Eleven new point substitutions were found, all of them in heteroplasmy, from which five appear to be somatic mutations and six can be considered germinal, evidencing the high frequency of somatic mutations in mtDNA in healthy young individuals. Different values of the mutation rate according to different assumptions were estimated. When considering all the germinal mutations, the value of the mutation rate obtained is one of the highest reported so far in family studies. However, when corrected for gender (assuming that the mutations present in men have the same evolutionary weight of somatic mutations because they will inevitably be lost) and for the probability of intraindividual fixation, the value for the mutation rate obtained for HVRI and HVRII (0.2415 mutations/site/Myr) was in the upper end of the values provided by phylogenetic estimations. These results indicate that the discrepancy, that has been reported previously, between the human mtDNA mutation rates observed along evolutionary timescales and the estimations obtained using family pedigrees can be minimized when corrections for gender proportions in newborn individuals and for the probability of intraindividual fixation are introduced. The analyses performed support the hypothesis that (1) in a constant, tight bottleneck genetic drift alone can explain different patterns of heteroplasmy segregation and (2) in neutral conditions, the destiny of a new mutation is strictly related to the initial proportion of the new variant. Another important point arising from the data obtained is that, even in the absence of a paternal contribution of mtDNA, recombination may occur between mtDNA molecules present in an individual, which is only observable if it occurs between mtDNA types that differ at two or more positions.


Prospective multi-institutional study of reverse transcriptase polymerase chain reaction for molecular staging of melanoma.
To evaluate the prognostic significance of molecular staging using reverse transcriptase polymerase chain reaction (RT-PCR) in detecting occult melanoma cells in sentinel lymph nodes (SLNs) and circulating bloodstream. In this multicenter study, eligibility criteria included patient age 18 to 71 years, invasive melanoma > or = 1.0 mm Breslow thickness, and no clinical evidence of metastasis. SLN biopsy and wide excision of the primary tumor were performed. SLNs were examined by serial-section histopathology and S-100 immunohistochemistry. A portion of each SLN was frozen for RT-PCR. In addition, RT-PCR was performed on peripheral-blood mononuclear cells (PBMCs). RT-PCR analysis was performed using four markers: tyrosinase, MART1, MAGE3, and GP-100. Disease-free survival (DFS), distant-DFS (DDFS), and overall survival (OS) were analyzed. A total of 1,446 patients with histologically negative SLNs underwent RT-PCR analysis. At a median follow-up of 30 months, there was no difference in DFS, DDFS, or OS between the RT-PCR-positive (n = 620) and RT-PCR-negative (n = 826) patients. Analysis of PBMC from 820 patients revealed significant differences in DFS and DDFS, but not OS, for patients with detection of more than one RT-PCR marker in peripheral blood. In this large, prospective, multi-institutional study, RT-PCR analysis on SLNs and PBMCs provides no additional prognostic information beyond standard histopathologic analysis of SLNs. Detection of more than one marker in PBMC is associated with a worse prognosis. RT-PCR remains investigational and should not be used to direct adjuvant therapy at this time.


Return-to-sports after minimally invasive stabilization of intra-articular calcaneal fractures.
Evaluation of different factors in patient quality of life after minimally invasive stabilization of intra-articular calcaneal fractures, including the return-to-sports rate. Patients with minimally invasive stabilization of intra-articular calcaneal fractures were collected from the database of a Level I trauma center and evaluated in a retrospective and explorative way. The clinical and radiological examination have been done immediately after the operation, after 2 and 6 weeks postoperative and after a minimum follow-up of 2 years. Clinical and radiological examination was performed by applying the American Orthopedic Foot and Ankle Society hindfoot scale score (AOFAS), 36-item Short Form Health Survey (SF-36), the Tegner Activity Scale, the Foot and Ankle Outcome Score (FAOS) and with a questionnaire about pre- and postoperative engagement in sport and recreational activities. Fourty-nine patients with an isolated uni-lateral fracture of the calcaneus who fulfilled all inclusion criteria were assessed. Fourty-two of them were male and 24 were under the age of 50 years. No statistically significant differences were noted between Sanders I/II and Sanders III/IV in terms of SF-36, AOFAS, FAOS or Tegner-scale. A less satisfying result was noticed in Sanders III/IV patients. General health, pain in FAOS, physical functioning and pain in SF-36 were strongly dependent on Tegner score values. Twenty-nine percent of our study population changed sport activities after injury, whereas 22 percent stopped all kinds of sports. Consequently, our overall return-to-sport rate was 78 percent. Clinical results including different scores and quality of life parameters in our study population were satisfying. About 80 percent of patients could return to sports, but there are still many patients that were not able to perform sports and physical activities on the same level as before.


Effects of orofacial myofunctional therapy on speech intelligibility in individuals with persistent articulatory impairments.
This study examined the effects of orofacial myofunctional therapy (OMT) on speech intelligibility in adults with persistent articulation impairments. Six adults in the age range of 18-23 years were selected to receive orofacial myofunctional therapy for a period of six weeks. The results showed that five out of six clients made significant progress in oral postures and speech sound production across all three speech production tasks: single words, sentences, and spontaneous speech. Speech intelligibility increased significantly in all clients except the one diagnosed with developmental apraxia of speech. Orofacial myofunctional therapists, speech-language pathologists, and other professionals need to rule out underlying orofacial myofunctional variables when targeting speech sounds for intervention. Speech production tasks may be considered as important measures to understand the efficacy of OMT in clinical settings.


Predominant role of prolactin in stimulating the growth of 7, 12-dimethylbenz(a)anthracene-induced rat mammary tumor.
The effect of prolactin in supporting the growth of 7, 12-dimethylbenz(a)anthracene-induced mammary tumor in adult female Sprague-Dawley rats was investigated when estrogen receptors were blocked by the nonsteroidal antiestrogen, Tamoxifen, ICI 46,474. Following an oophorectomy-induced remission, perphenazine, which stimulates endogenous prolactin release, was able to restore tumor growth whether or not Tamoxifen was added. A second course of perphenazine treatment, instituted after the tumors were allowed to shrink, was again effective in stimulating tumor growth. After a regression in tumor size induced by oophorectomy and daily administration of Tamoxifen, perphenazine was able to restore original tumor size despite continued treatment with Tamoxifen. In intact rats, after regression was obtained by daily administration of Tamoxifen and the prolactin inhibitor, lergotrile mesylate, perphenazine induced tumor growth when the latter was discontinued, even though Tamoxifen was continued for 50 days. Estrogen receptors measured at the time of maximum stimulation by perphenazine were undetectable. On the other hand, estradiol did not stimulate tumor growth when serum prolactin was depressed to undetectable levels by lergotrile. These results indicate that prolactin supports the growth of 7, 12-dimethylbenz(a)anthracene-induced rat mammary tumor and that estrogen receptors are not required under these conditions.


Antibiotic resistance in Burkholderia species.
The genus Burkholderia comprises metabolically diverse and adaptable Gram-negative bacteria, which thrive in often adversarial environments. A few members of the genus are prominent opportunistic pathogens. These include Burkholderia mallei and Burkholderia pseudomallei of the B. pseudomallei complex, which cause glanders and melioidosis, respectively. Burkholderia cenocepacia, Burkholderia multivorans, and Burkholderia vietnamiensis belong to the Burkholderia cepacia complex and affect mostly cystic fibrosis patients. Infections caused by these bacteria are difficult to treat because of significant antibiotic resistance. The first line of defense against antimicrobials in Burkholderia species is the outer membrane penetration barrier. Most Burkholderia contain a modified lipopolysaccharide that causes intrinsic polymyxin resistance. Contributing to reduced drug penetration are restrictive porin proteins. Efflux pumps of the resistance nodulation cell division family are major players in Burkholderia multidrug resistance. Third and fourth generation β-lactam antibiotics are seminal for treatment of Burkholderia infections, but therapeutic efficacy is compromised by expression of several β-lactamases and ceftazidime target mutations. Altered DNA gyrase and dihydrofolate reductase targets cause fluoroquinolone and trimethoprim resistance, respectively. Although antibiotic resistance hampers therapy of Burkholderia infections, the characterization of resistance mechanisms lags behind other non-enteric Gram-negative pathogens, especially ESKAPE bacteria such as Acinetobacter baumannii, Klebsiella pneumoniae and Pseudomonas aeruginosa.


Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction.
A highly sensitive, specific, rapid and simple method to detect Burkholderia pseudomallei in blood samples was developed. Two 22-base oligonucleotide primers, based on sequences from a specific DNA probe, were used for amplification of bacterial DNA by the polymerase chain reaction (PCR). Amplification with these primers yielded a 178-base pair product in 100 clinical isolates of B. pseudomallei. As little as 0.5 fg of B. pseudomallei DNA was detectable by this method. Experiments involving inoculation of the organism into uninfected blood samples showed that the method could be used to detect as few as 1 bacterial cell ml-1 of whole blood. Non-specific amplification of other bacterial DNAs from 18 samples of bacteria was not observed. Blood samples from seven patients proven to have melioidosis by haemoculture were positive using these primers. The total time required for sample processing, amplification and visualization was approximately 3.5 h. The high sensitivity, rapidity and simplicity of this method should make it valuable for diagnosis, monitoring of drug treatment and for epidemiological studies of the melioidosis.


Role of Ion Channels in the Maintenance of Sperm Motility and Swimming Behavior in a Marine Teleost.
In oviparous marine fishes, the hyperosmotic induction of sperm motility in seawater (SW) is well established, however, the potential function of ion channels in the maintenance of post activated spermatozoon swimming performance remains largely unknown. Here, we investigated the influence of ion channels on the spermatozoon swimming parameters using the gilthead seabream (Sparus aurata) as a model for modern marine teleosts. Our data show that the SW-induced activation of seabream sperm motility requires three concomitant processes, the hyperosmotic shock, an ion-flux independent increase of the intracellular concentration of Ca2+ ([Ca2+]i), but not of [K+]i or [Na+]i, and the alkalization of the cytosol. The combination of all three processes is obligatory to trigger flagellar beating. However, the time-course monitoring of sperm motion kinetics and changes in the [Ca2+]i, [K+]i and [Na+]i in SW or in non-ionic activation media, showed that the post activated maintenance of spermatozoa motility is dependent on extracellular Ca2+ and K+. A meta-analysis of a seabream sperm transcriptome uncovered the expression of multiple ion channels, some of which were immunolocalized in the head and/or tail of the spermatozoon. Selective pharmacological inhibition of these ion channel families impaired the long-term motility, progressivity, and velocity of SW-activated spermatozoa. The data further revealed that some antagonists of K+-selective or Ca2+-selective channels, as well as of stretch-activated and mechanosensitive channels, altered the trajectory of spermatozoa, suggesting that these ion channels are likely involved in the control of the swimming pattern of the post activated spermatozoon. These combined findings provide new insight into the signaling pathways regulating spermatozoon activation and swimming performance in marine fishes.


Doping of the hydrogen-passivated Si(100) electronic structure through carborane adsorption studied using density functional theory.
Adsorption of molecular materials with tailored chemical properties represents a new and promising avenue to non-destructively dope silicon. Dithiocarboranes possess large permanent dipoles and readily form stable monolayers on a variety of substrates. Here we use density functional theory to investigate the doping of hydrogen-passivated Si(100) substrates through the adsorption of dithiocarborane molecules. We find that dithiocarboranes can both physisorb and chemisorb on the substrate. Chemisorbed structures arise when a S atom in the molecular linker group replaces a surface H atom. We establish the formation of these Si-molecule bonds and characterize their mechanical and thermal stability. Analysis of the calculated electronic structure of adsorbed interfaces shows that carborane adsorption does not result in interface gap states. The band gap in adsorbed junctions is defined by Si states and its magnitude is almost unchanged with respect to the clean Si slab. The large carborane electrostatic dipole results in the downwards shift of Si spectral features by 0.3 eV, reducing the hole injection barrier by this amount with respect to the pristine Si substrate. Molecular dynamics simulations reveal these structural and electronic features to be stable at room temperature. Our work shows that molecular adsorbates having large electrostatic dipoles are a promising strategy to non-destructively dope semiconductor substrates.


New generation dendritic cell vaccine for immunotherapy of acute myeloid leukemia.
Dendritic cell (DC)-based immunotherapy is a promising strategy for the elimination of minimal residual disease in patients with acute myeloid leukemia (AML). Particularly, patients with a high risk of relapse who are not eligible for hematopoietic stem cell transplantation could benefit from such a therapeutic approach. Here, we review our extensive studies on the development of a protocol for the generation of DCs with improved immunogenicity and optimized for the use in cell-based immunotherapy. This new generation DC vaccine combines the production of DCs in only 3 days with Toll-like receptor-signaling-induced cell maturation. These mature DCs are then loaded with RNA encoding the leukemia-associated antigens Wilm's tumor protein 1 and preferentially expressed antigen in melanoma in order to stimulate an AML-specific T-cell-based immune response. In vitro as well as in vivo studies demonstrated the enhanced capacity of these improved DCs for the induction of tumor-specific immune responses. Finally, a proof-of-concept Phase I/II clinical trial is discussed for post-remission AML patients with high risk for disease relapse.


Lid splinting eyelid retraction technique: a minimised sterile approach for intravitreal injections.
To describe an alternative technique for avoiding contact with the lids and lashes, without the use of a lid speculum, during intravitreal anti-vascular endothelial growth factor injections. Retrospective case series of all patients undergoing intravitreal injections of bevacizumab and ranibizumab, with the lid splinting retraction technique from January 2010 to December 2015. Injections performed by six vitreoretinal specialists were included. The key preinjection ocular surface preparation includes topical anaesthetic, 5% povidone-iodine and a subconjunctival injection of 2% lidocaine with epinephrine. A second instillation of 5% povidone-iodine is given and the intravitreal injection is then performed. No lid speculum is used. A search of the electronic medical records identified patients diagnosed with postinjection endophthalmitis and charts were reviewed to ensure inclusion criteria were met. The main outcome measure was incidence of postinjection endophthalmitis. A total of 78 009 consecutive intravitreal injections were performed, of which 22 207 were bevacizumab and 55 802 were ranibizumab. In this cohort of patients (n=6320), 12 cases of endophthalmitis developed, corresponding to a rate of 0.015%. The technique of eyelid retraction for intravitreal injection has a low rate of endophthalmitis, similar to the reported rates using a metal lid speculum. This is beneficial for both the physician and the patient as it minimises patient discomfort as well as the duration of the procedure. To our knowledge, this is one of the largest studies performed to date evaluating intravitreal injection-related endophthalmitis.


Memory for faces and objects by deaf and hearing signers and hearing nonsigners.
The memory of 11 deaf and 11 hearing British Sign Language users and 11 hearing nonsigners for pictures of faces of and verbalizable objects was measured using the game Concentration. The three groups performed at the same level for the objects. In contrast the deaf signers were better for faces than the hearing signers, who in turn were superior to the hearing nonsigners, who were the worst. Three hypotheses were made: That there would be no significant difference in terms of the number of attempts between the three groups on the verbalizable object task, that the hearing and deaf signers would demonstrate superior performance to that of the hearing nonsigners on the matching faces task, and that the hearing and deaf signers would exhibit similar performance levels on the matching faces task. The first two hypotheses were supported, but the third was not. Deaf signers were found to be superior for memory for faces to hearing signers and hearing nonsigners. Possible explanations for the findings are discussed, including the possibility that deafness and the long use of sign language have additive effects.


Hand Motor Recovery Following Extensive Frontoparietal Cortical Injury Is Accompanied by Upregulated Corticoreticular Projections in Monkey.
We tested the hypothesis that arm/hand motor recovery after injury of the lateral sensorimotor cortex is associated with upregulation of the corticoreticular projection (CRP) from the supplementary motor cortex (M2) to the gigantocellular reticular nucleus of the medulla (Gi). Three groups of rhesus monkeys of both genders were studied: five controls, four cases with lesions of the arm/hand area of the primary motor cortex (M1) and the lateral premotor cortex (LPMC; F2 lesion group), and five cases with lesions of the arm/hand area of M1, LPMC, S1, and anterior parietal cortex (F2P2 lesion group). CRP strength was assessed using high-resolution anterograde tracers injected into the arm/hand area of M2 and stereology to estimate of the number of synaptic boutons in the Gi. M2 projected bilaterally to the Gi, primarily targeting the medial Gi subsector and, to a lesser extent, lateral, dorsal, and ventral subsectors. Total CRP bouton numbers were similar in controls and F2 lesion cases but F2P2 lesion cases had twice as many boutons as the other two groups (p = 0.0002). Recovery of reaching and fine hand/digit function was strongly correlated with estimated numbers of CRP boutons in the F2P2 lesion cases. Because we previously showed that F2P2 lesion cases experience decreased strength of the M2 corticospinal projection (CSP), whereas F2 lesion monkeys experienced increased strength of the M2 CSP, these results suggest one mechanism underlying arm/hand motor recovery after F2P2 injury is upregulation of the M2 CRP. This M2-CRP response may influence an important reticulospinal tract contribution to upper-limb motor recovery following frontoparietal injury.SIGNIFICANCE STATEMENT We previously showed that after brain injury affecting the lateral motor cortex controlling arm/hand motor function, recovery is variable and closely associated with increased strength of corticospinal projection (CSP) from an uninjured medial cortical motor area. Hand motor recovery also varies after brain injury affecting the lateral sensorimotor cortex, but medial motor cortex CSP strength decreases and cannot account for recovery. Here we observed that motor recovery following sensorimotor cortex injury is closely associated with increased strength of the descending projection from an uninjured medial cortical motor area to a brainstem reticular nucleus involved in control of arm/hand function, suggesting an enhanced corticoreticular projection may compensate for injury to the sensorimotor cortex to enable recovery of arm/hand motor function.


A buffering SERCA pump in models of calcium dynamics.
Many cells use oscillations in calcium concentration to transmit messages. The oscillations largely result from an influx of calcium into the cytosol from the endoplasmic reticulum (ER), followed by an efflux of calcium from the cytosol back into the ER. The sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) pump pumps calcium into the ER. It binds calcium on the cytosolic side and releases it on the ER side and in the delay between binding and release, calcium is buffered by the pump. We developed a model of a buffering SERCA pump and investigated whether including this in a model of calcium oscillations has any significant effects. We found that the oscillations produced when using the SERCA pump, which does not buffer calcium, have a larger amplitude and a slightly smaller period than when using the buffering SERCA pump. We show that the buffering SERCA pump shows adaptation to a stimulus, and we demonstrate that, by using a bidirectional SERCA pump, we are able to eliminate futile cycling of calcium between the cytosol and ER when the cell is at rest.


Opioid side effects, addiction, and anti-inflammatory medications.
Patients in pain often fear medications prescribed or recommend to them by their clinicians. Fear of side effects can contribute greatly to medication non-adherence (noncompliance). Patients often have fears that exceed the potential problems with which their medications are associated. Questions and answers relating to the side effects and the risk of addiction associated with opioids are presented.


Psychological debriefing in cross-cultural contexts: ten implications for practice.
Crisis interventions following exposure to traumatic events have become common in most western and in some non-western countries. The literature regarding early interventions, specifically the use of Psychological Debriefing (PD), is grounded in a western context. Little has been written of its use in different cultural settings. This article focuses on the use of PD in different cultural settings, as well as some of the conceptual issues related to cross-cultural trauma research and practice, which inevitably have implications for the use of early intervention. Ten key implications for practice are suggested.


Induction of colony-stimulating activity in mice by injection of liposomes containing lipophilic muramyl peptide derivatives.
Colony-stimulating factor (CSF) activity induction by lipophilic derivatives of three muramyl peptides, glyceryl dipalmitate-MDP derivatives, was studied in vivo and in vitro and compared to the activity of the same compounds incorporated within freeze-dried liposomes. Two lipophilic derivatives (MDP-GDP and MDPGBe-GDP) were able to induce CSF activity in vivo and in vitro. The incorporation of these compounds within appropriately designed liposomes composed of distearoylphosphatidylcholine and phosphatidylserine (DSPC/PS) increased their ability to induce CSF activity in vivo but completely abrogated their ability to induce CSF activity in vitro. Furthermore, the phospholipid composition of liposomes influenced the efficacy of glycopeptide liposomal incorporation. Thus, the serum CSF-inducing effect of MDP-GDP was considerably enhanced by incorporation of this compound within liposomes composed of DSPC/PS at a molar ratio 7:0.3 but was not modified if the DSPC/PS molar ratio was 7:3. The lipophilic derivative of MDP (D-D), MDP (D-D)-GDP, was unable to induce CSF activity in vivo or in vitro but surprisingly became active in vivo after entrapment within DSPC/PS liposomes (molar ratio 7:0.3). Our results show that appropriate liposomes may be suitable carriers to deliver CSF activity-inducing agents to macrophages in vivo.


Anti-inflammatory treatment and risk for depression after first-time stroke in a cohort of 147 487 Danish patients.
Depression is a common complication after stroke, and inflammation may be a pathophysiological mechanism. This study examines whether anti-inflammatory treatment with acetylsalicylic acid (ASA), nonsteroid anti-inflammatory drugs (NSAIDs) or statins influence the risk of depression after stroke. A register-based cohort including all patients admitted to hospital with a first-time stroke from Jan. 1, 2001, through Dec. 31, 2011, and a nonstroke population with a similar age and sex distribution was followed for depression until Dec. 31, 2014. Depression was defined as having a hospital contact with depression or having filled prescriptions for antidepressant medication. The associations between redeemed prescriptions of ASA, NSAIDs or statins with early- (≤ 1 year after stroke or study entry) and late-onset (> 1 year after stroke or study entry) depression were analyzed using Cox proportional hazard regression. We identified 147 487 patients with first-time stroke and 160 235 individuals without stroke for inclusion in our study. Redeemed prescriptions of ASA, NSAIDs or statins after stroke decreased the risk for early-onset depression, especially in patients with ischemic or severe stroke. Patients who received a combination of anti-inflammatory treatments had the lowest risk for early-onset depression. On the other hand, use of ASA or NSAIDs 1 year after stroke increased the risk for late-onset depression, whereas statin use was associated with a tendency toward a decreased risk. The study used prescription of antidepressant medication as a proxy measure for depression and did not include anti-inflammatory drugs bought over the counter. Anti-inflammatory treatment is associated with a lower risk for depression shortly after stroke but a higher risk for late depression. This suggests that inflammation contributes differently to the development of depression after stroke depending on the time of onset.


Micronutrient Losses during Continuous Renal Replacement Therapy.
Acute kidney injury impacts the micronutrient status by various mechanisms including decreased enteral absorption, changes in redistribution, altered metabolism, and increased consumption. When renal replacement therapy (RRT) is applied, there are additional losses of vitamins, trace elements, and amino acids, and their derivatives due to diffusion or adhesion. Varied data exist regarding the degree of micronutrient losses and plasma concentrations in patients who receive RRT, and these differ by RRT modality, dose, duration, and type of micronutrient. Water-soluble vitamins, selenium, copper, and carnitine are among the most frequently reported depleted nutrients. The role of micronutrient supplementation in critically ill patients undergoing RRT and the optimal dose and mode of administration are yet to be determined.


Two cell-counting factors regulate the aggregate size of the cellular slime mold Dictyostelium discoideum.
Countin, a cell-counting factor in Dictyostelium discoideum, is considered to limit the maximum size of the multicellular structure, because a countin null strain forms a huge fruiting body compared to that of the wild-type. A novel gene, countin2, that is highly homologous to countin (40% identity in amino acid sequence) was identified in the D. discoideum genome. The countin2 null strain formed a 1.7-fold higher number of the aggregates, resulting in smaller fruiting bodies compared with those of wild-type cells. Thus, the Countin2 protein is thought to limit the minimum size of the multicellular structure. The size and number of aggregates formed by a mixture of countin null and countin2 null strains were the same as those of the wild-type. These findings demonstrate that a combination of Countin and Countin2 proteins determines the appropriate size of the multicellular structure of D. discoideum.


Bilateral medial medullary stroke: a challenge in early diagnosis.
Bilateral medial medullary stroke is a very rare type of stroke, with catastrophic consequences. Early diagnosis is crucial. Here, I present a young patient with acute vertigo, progressive generalized weakness, dysarthria, and respiratory failure, who initially was misdiagnosed with acute vestibular syndrome. Initial brain magnetic resonance imaging (MRI) that was done in the acute phase was read as normal. Other possibilities were excluded by lumbar puncture and MRI of cervical spine. MR of C-spine showed lesion at medial medulla; therefore a second MRI of brain was requested, showed characteristic "heart appearance" shape at diffusion weighted (DWI), and confirmed bilateral medial medullary stroke. Retrospectively, a vague-defined hyperintense linear DWI signal at midline was noted in the first brain MRI. Because of the symmetric and midline pattern of this abnormal signal and similarity to an artifact, some radiologists or neurologists may miss this type of stroke. Radiologists and neurologists must recognize clinical and MRI findings of this rare type of stroke, which early treatment could make a difference in patient outcome. The abnormal DWI signal in early stages of this type of stroke may not be a typical "heart appearance" shape, and other variants such as small dot or linear DWI signal at midline must be recognized as early signs of stroke. Also, MRI of cervical spine may be helpful if there is attention to brainstem as well.


Protective head-cooling during cardiac arrest and cardiopulmonary resuscitation: the original animal studies.
Prolonged standard cardiopulmonary resuscitation (CPR) does not reliably sustain brain viability during cardiac arrest. Pre-hospital adjuncts to standard CPR are needed in order to improve outcomes. A preliminary dog study demonstrated that surface cooling of the head during arrest and CPR can achieve protective levels of brain hypothermia (30°C) within 10 minutes. We hypothesized that protective head-cooling during cardiac arrest and CPR improves neurological outcomes. Twelve dogs under light ketamine-halothane-nitrous oxide anesthesia were arrested by transthoracic fibrillation. The treated group consisted of six dogs whose shaven heads were moistened with saline and packed in ice immediately after confirmation of ventricular fibrillation. Six control dogs remained at room temperature. All 12 dogs were subjected to four minutes of ventricular fibrillation and 20 minutes of standard CPR. Spontaneous circulation was restored with drugs and countershocks. Intensive care was provided for five hours post-arrest and the animals were observed for 24 hours. In both groups, five of the six dogs had spontaneous circulation restored. After three hours, mean neurological deficit was significantly lower in the treated group (P=0.016, with head-cooled dogs averaging 37% and the normothermic dogs 62%). Two of the six head-cooled dogs survived 24 hours with neurological deficits of 9% and 0%, respectively. None of the control group dogs survived 24 hours. We concluded that head-cooling attenuates brain injury during cardiac arrest with prolonged CPR. We review the literature related to the use of hypothermia following cardiac arrest and discuss some promising approaches for the pre-hospital setting.


Latent nitrate reductase activity is associated with the plasma membrane of corn roots.
Latent nitrate reductase activity (NRA) was detected in corn (Zea mays L., Golden Jubilee) root microsome fractions. Microsome-associated NRA was stimulated up to 20-fold by Triton X-100 (octylphenoxy polyethoxyethanol) whereas soluble NRA was only increased up to 1.2-fold. Microsome-associated NRA represented up to 19% of the total root NRA. Analysis of microsomal fractions by aqueous two-phase partitioning showed that the membrane-associated NRA was localized in the second upper phase (U2). Analysis with marker enzymes indicated that the U2 fraction was plasma membrane (PM). The PM-associated NRA was not removed by washing vesicles with up to 1.0 M NACl but was solubilized from the PM with 0.05% Triton X-100. In contrast, vanadate-sensitive ATPase activity was not solubilized from the PM by treatment with 0.1% Triton X-100. The results show that a protein capable of reducing nitrate is embedded in the hydrophobic region of the PM of corn roots.


Inhibition of Simian virus 40 large T antigen helicase activity by fluoroquinolones.
Fluoroquinolones represent a potent group of antibiotics that inhibit bacterial DNA replication by targeting the essential bacterial enzymes gyrase and topoisomerase IV. Inhibition of gyrase activity by quinolones involves the interaction of these drugs with the helicase component of bacterial gyrase. DNA tumour viruses also encode helicases that are essential for their DNA replication in the host. In this study we have evaluated the effect of fluoroquinolones on viral DNA replication using the DNA tumour virus Simian virus 40 (SV40) as our model. Four different fluoroquinolones, namely, levofloxacin, trovafloxacin, ciprofloxacin and ofloxacin, were tested for their ability to inhibit viral DNA replication. We show here that all four quinolones tested were effective in the inhibition of SV40 plaque formation and DNA replication in CV1-P cells. In addition, we found that each of these quinolones was inhibitory to the helicase activity of SV40 large tumour antigen. Fluoroquinolones and their derivates may therefore be useful in the treatment and/or prevention of infection by SV40-homologous human DNA viruses that encode helicase activity for their survival.


Financing and provision of healthcare for two billion people in low-income nations: Is the cooperative healthcare model a solution?
The international consensus in support of universal health coverage (UHC), though commendable, thus far lacks a clear mechanism to finance and deliver accessible and effective basic healthcare to the two billion rural residents and informal workers of low- and lower-middle-income countries (LLMICs). Importantly, the two preferred financing modes for UHC, general tax revenue and social health insurance, are often infeasible for LLMICs. We identify from historical examples a community-based model that we argue shows promise as a solution to this problem. This model, which we call Cooperative Healthcare (CH), is characterized by community-based risk-pooling and governance and prioritizes primary care. CH leverages communities' existing social capital, such that even those for whom the private benefit of enrolling in a CH scheme is outweighed by the cost may choose to enroll (given sufficient social capital). For CH to be scalable, it needs to demonstrate that it can organize delivery of accessible and reasonable-quality primary healthcare that people value, with management accountable to the communities themselves through structures that people trust, combined with government legitimacy. Once LLMICs with CH programs have industrialized sufficiently to make universal social health insurance feasible, CH schemes can be rolled into such universal programs. We defend cooperative healthcare's suitability for this bridging role and urge LLMIC governments to launch experiments testing it out, with careful adaptation to local conditions.


Mechanistic and structural analyses of the roles of Arg409 and Asp402 in the reaction of the flavoprotein nitroalkane oxidase.
The flavoprotein nitroalkane oxidase (NAO) catalyzes the oxidation of primary and secondary nitroalkanes to the corresponding aldehydes and ketones. The enzyme is a homologue of acyl-CoA dehydrogenase. Asp402 in NAO has been proposed to be the active site base responsible for removing the substrate proton in the first catalytic step; structurally it corresponds to the glutamate which acts as the base in medium chain acyl-CoA dehydrogenase. In the active site of NAO, the carboxylate of Asp402 forms an ionic interaction with the side chain of Arg409. The R409K enzyme has now been characterized kinetically and structurally. The mutation results in a decrease in the rate constant for proton abstraction of 100-fold. Analysis of the three-dimensional structure of the R409K enzyme, determined by X-ray crystallography to a resolution of 2.65 A, shows that the critical structural change is an increase in the distance between the carboxylate of Asp402 and the positively charged nitrogen in the side chain of the residue at position 409. The D402E mutation results in a smaller decrease in the rate constant for proton abstraction of 18-fold. The structure of the D402E enzyme, determined at 2.4 A resolution, shows that there is a smaller increase in the distance between Arg409 and the carboxylate at position 402, and the interaction of this residue with Ser276 is perturbed. These results establish the critical importance of the interaction between Asp402 and Arg409 for proton abstraction by nitroalkane oxidase.


EPICE-HIV: An Epidemiologic Cost-Effectiveness Model for HIV Treatment.
The goal of this research was to establish a new and innovative framework for cost-effectiveness modeling of HIV-1 treatment, simultaneously considering both clinical and epidemiological outcomes. EPICE-HIV is a multi-paradigm model based on a within-host micro-simulation model for the disease progression of HIV-1 infected individuals and an agent-based sexual contact network (SCN) model for the transmission of HIV-1 infection. It includes HIV-1 viral dynamics, CD4+ T cell infection rates, and pharmacokinetics/pharmacodynamics modeling. Disease progression of HIV-1 infected individuals is driven by the interdependent changes in CD4+ T cell count, changes in plasma HIV-1 RNA, accumulation of resistance mutations and adherence to treatment. The two parts of the model are joined through a per-sexual-act and viral load dependent probability of disease transmission in HIV-discordant couples. Internal validity of the disease progression part of the model is assessed and external validity is demonstrated in comparison to the outcomes observed in the STaR randomized controlled clinical trial. We found that overall adherence to treatment and the resulting pattern of treatment interruptions are key drivers of HIV-1 treatment outcomes. Our model, though largely independent of efficacy data from RCT, was accurate in producing 96-week outcomes, qualitatively and quantitatively comparable to the ones observed in the STaR trial. We demonstrate that multi-paradigm micro-simulation modeling is a promising tool to generate evidence about optimal policy strategies in HIV-1 treatment, including treatment efficacy, HIV-1 transmission, and cost-effectiveness analysis.


Prevalence and factors influencing smoking amongst Malay primary school children in Tumpat, Kelantan.
The aims of this study were to determine the prevalence, knowledge and factors that influence smoking in Malay primary school children in Tumpat, Kelantan. A cross-sectional study was conducted in February 2004 among primary school children in Tumpat District. Two hundred-twelve children in standard one to six were randomly selected from three rural schools. An interview that included information on history of ever smoking, knowledge related to smoking and health, and potential factors that could influence smoking was done. Twenty-five children had previously smoked, with a prevalence of 11.8% (95%CI=8.0, 17.0) and 8 were current smokers (3.8%, 95%CI=1.2, 6.4). More than half (64.6%) of the children had a good knowledge of smoking. However, only 105 (49.5%) of them knew that passive smokers have a higher risk of developing diseases. Of those who had ever smoked, 12 (36.6%) were influenced by peers and 17 (51.5%) had a self-desire to smoke. The earliest age to start smoking was at 6 years. Factors found to be significantly associated with smoking on multivariate analysis were increasing age (OR=2.8, 95%CI=1.6, 5.1), being boys (OR=5.8, 95%CI=2.0, 16.8), being at second school level (standard 4, 5, 6)(OR=7.8, 95%CI=1.3, 45.3) and having other family members (excluding father) who smoked (OR=2.8, 95%CI=1.2, 6.5). However, having a father who smoked and a good knowledge were not reported as influencing factors.


A hydrophilic-interaction chromatography tandem mass spectrometry method for quantitation of serum s-adenosylmethionine in patients infected with human immunodeficiency virus.
s-Adenosylmethionine (SAM)1 has been suggested as a diagnostic test and surrogate marker for Pneumocystis jirovecii pneumonia (PCP) in HIV-positive patients. In this study, we report a robust hydrophilic-interaction liquid chromatography tandem mass spectrometry (LC-MS/MS) assay that can be used to quantitate serum SAM in clinical laboratories. Proteins in serum samples were precipitated using trichloroacetic acid. The supernatant was separated after centrifugation. D3d3-SAM was added as the internal standard. SAM and d3-SAM were extracted using a mixed-mode cation exchange column. Extracts were dried under nitrogen and reconstituted in H2O and acetonitrile (1:9, vol:vol). HPLC-tandem mass spectrometry analysis was performed with a silica column and multiple reaction monitoring for SAM and d3-SAM. The limit of quantitation (LOQ) for SAM was 10 ng/mL. The assay was linear between 10 and 500 ng/mL. Intra-assay coefficient of variation (CV) was 8% and inter-assay CV was 17% at the LOQ. Turnaround time for each specimen was approximately 1 h. Using this method, we found that serum SAM concentration was correlated with fasting status, especially methionine intake. We also measured acute and convalescent serum SAM levels of 8 HIV-positive patients with PCP and non-PCP pneumonia. SAM concentrations in convalescent samples were significantly increased compared to acute levels only in patients with PCP. The LC-MS/MS method had sufficient analytical sensitivity for detecting low levels of SAM found in HIV-infected patients and can be used for quantitative measurements in a clinical laboratory. This method facilitates research and possible clinical application of SAM as a marker for PCP.


Teaching little kids big sentences: A randomized controlled trial showing that children with DLD respond to complex syntax intervention embedded within the context of preschool/kindergarten science instruction.
The language of the science curriculum is complex, even in the early grades. To communicate their scientific observations, children must produce complex syntax, particularly complement clauses (e.g., I think it will float; We noticed that it vibrates). Complex syntax is often challenging for children with developmental language disorder (DLD), and thus their learning and communication of science may be compromised. We asked whether recast therapy delivered in the context of a science curriculum led to gains in complement clause use and scientific content knowledge. To understand the efficacy of recast therapy, we compared changes in science and language knowledge in children who received treatment for complement clauses embedded in a first-grade science curriculum to two active control conditions (vocabulary + science, phonological awareness + science). This 2-year single-site three-arm parallel randomized controlled trial was conducted in Delaware, USA. Children with DLD, not yet in first grade and with low accuracy on complement clauses, were eligible. Thirty-three 4-7-year-old children participated in the summers of 2018 and 2019 (2020 was cancelled due to COVID-19). We assigned participants to arms using 1:1:1 pseudo-random allocation (avoiding placing siblings together). The intervention consisted of 39 small-group sessions of recast therapy, robust vocabulary instruction or phonological awareness intervention during eight science units over 4 weeks, followed by two science units (1 week) taught without language intervention. Pre-/post-measures were collected 3 weeks before and after camp by unmasked assessors. Primary outcome measures were accuracy on a 20-item probe of complement clause production and performance on ten 10-item unit tests (eight science + language, two science only). Complete data were available for 31 children (10 grammar, 21 active control); two others were lost to follow-up. Both groups made similar gains on science unit tests for science + language content (pre versus post, d = 2.9, p < 0.0001; group, p = 0.24). The grammar group performed significantly better at post-test than the active control group (d = 2.5, p = 0.049) on complement clause probes and marginally better on science-only unit tests (d = 2.5, p = 0.051). Children with DLD can benefit from language intervention embedded in curricular content and learn both language and science targets taught simultaneously. Tentative findings suggest that treatment for grammar targets may improve academic outcomes. What is already known on the subject We know that recast therapy focused on morphology is effective but very time consuming. Treatment for complex syntax in young children has preliminary efficacy data available. Prior research provides mixed evidence as to children's ability to learn language targets in conjunction with other information. What this study adds This study provides additional data supporting the efficacy of intensive complex syntax recast therapy for children ages 4-7 with Developmental Language Disorder. It also provides data that children can learn language targets and science curricular content simultaneously. What are the clinical implications of this work? As SLPs, we have to talk about something to deliver language therapy; we should consider talking about curricular content. Recast therapy focused on syntactic frames is effective with young children.


Mapping wintering waterfowl distributions using weather surveillance radar.
The current network of weather surveillance radars within the United States readily detects flying birds and has proven to be a useful remote-sensing tool for ornithological study. Radar reflectivity measures serve as an index to bird density and have been used to quantitatively map landbird distributions during migratory stopover by sampling birds aloft at the onset of nocturnal migratory flights. Our objective was to further develop and validate a similar approach for mapping wintering waterfowl distributions using weather surveillance radar observations at the onset of evening flights. We evaluated data from the Sacramento, CA radar (KDAX) during winters 1998-1999 and 1999-2000. We determined an optimal sampling time by evaluating the accuracy and precision of radar observations at different times during the onset of evening flight relative to observed diurnal distributions of radio-marked birds on the ground. The mean time of evening flight initiation occurred 23 min after sunset with the strongest correlations between reflectivity and waterfowl density on the ground occurring almost immediately after flight initiation. Radar measures became more spatially homogeneous as evening flight progressed because birds dispersed from their departure locations. Radars effectively detected birds to a mean maximum range of 83 km during the first 20 min of evening flight. Using a sun elevation angle of -5° (28 min after sunset) as our optimal sampling time, we validated our approach using KDAX data and additional data from the Beale Air Force Base, CA (KBBX) radar during winter 1998-1999. Bias-adjusted radar reflectivity of waterfowl aloft was positively related to the observed diurnal density of radio-marked waterfowl locations on the ground. Thus, weather radars provide accurate measures of relative wintering waterfowl density that can be used to comprehensively map their distributions over large spatial extents.


Intraileal capsaicin inhibits gastrointestinal contractions via a neural reflex in conscious dogs.
The aim of the present study was to determine the effect of intraileal administration of capsaicin on gastrointestinal motility. Mongrel dogs equipped with strain gauge force transducers on the stomach, small intestine, and colon were used. We studied the effects of intraileal capsaicin on gastrointestinal contractions with or without pharmacologic antagonists. The effects of capsaicin administration into the lumen of innervated and extrinsically denervated ileal Thiry loops were also studied. Intraileal capsaicin dose dependently inhibited postprandial contractions at all sites and interdigestive contractions in the upper gastrointestinal tract. Intraileal capsaicin-induced inhibition of gastrointestinal contractions was partially reversed by a nitric oxide (NO) synthase inhibitor, a 5 hydroxytryptamine-3 receptor antagonist (5-HT(3)), and an opiate antagonist. Administration of capsaicin into the innervated ileal Thiry loop had inhibitory effects on gastrointestinal contractions, but gastrointesinal motor activity was not affected by capsaicin administered into the extrinsically denervated Thiry loop. Stimulation of ileal afferent fibers by capsaicin inhibits gastrointestinal contractions via an extrinsic neural reflex.


Photoperiodic control of oestrous cycles in Syrian hamsters: mediation by the mediobasal hypothalamus.
To assess whether the mediobasal hypothalamus (MBH) is necessary for photoperiodic control of oestrous cycles and prolactin secretion, we tested intact female Syrian hamsters (controls) and those that had sustained unilateral or bilateral lesions of the MBH. All hamsters displayed 4-day oestrous cycles postoperatively in the long-day photoperiod (14 h light/day); control females and those with unilateral MBH damage ceased to undergo oestrous cycles approximately 8 weeks after transfer to a short-day photocycle (10 h light/day), whereas 12 of 15 females with bilateral MBH lesions continued to generate 4-day oestrous cycles throughout 22 weeks in short days. Serum prolactin concentrations were either undetectable or low in all hamsters 8 or 14 weeks after the transfer to short-day lengths, but increased above long-day baseline values by week 22. We conclude that melatonin-binding sites in the MBH mediate suppression of oestrous cycles but not prolactin secretion by short-day lengths; recovery of prolactin secretion in females during prolonged exposure to short-day lengths reflects development of refractoriness to melatonin in a substrate distinct from the MBH. These findings suggest that separate neural pathways mediate photoperiodic control of gonadotropin and prolactin secretion in female hamsters.


In Vitro Neurotoxicity of Flumethrin Pyrethroid on SH-SY5Y Neuroblastoma Cells: Apoptosis Associated with Oxidative Stress.
Pyrethroids are neurotoxicants for animals, showing a pattern of toxic action on the nervous system. Flumethrin, a synthetic pyrethroid, is used against ectoparasites in domestic animals, plants, and for public health. This compound has been shown to be highly toxic to bees, while its effects on other animals have been less investigated. However, in vitro studies to evaluate cytotoxicity are scarce, and the mechanisms associated with this effect at the molecular level are still unknown. This study aimed to investigate the oxidative stress and cell death induction in SH-SY5Y neuroblastoma cells in response to flumethrin exposure (1-1000 µM). Flumethrin induced a significant cytotoxic effect, as evaluated by MTT and LDH leakage assays, and produced an increase in the biomarkers of oxidative stress as reactive oxygen species and nitric oxide (ROS and NO) generation, malondialdehyde (MDA) concentration, and caspase-3 activity. In addition, flumethrin significantly increased apoptosis-related gene expressions (Bax, Casp-3, BNIP3, APAF1, and AKT1) and oxidative stress and antioxidative (NFκB and SOD2) mediators. The results demonstrated, by biochemical and gene expression assays, that flumethrin induces oxidative stress and apoptosis, which could cause DNA damage. Detailed knowledge obtained about these molecular changes could provide the basis for elucidating the molecular mechanisms of flumethrin-induced neurotoxicity.


Stable underwater superoleophobic and low adhesive polypyrrole nanowire mesh in highly corrosive environments.
Underwater superoleophobic materials with low adhesion have been widely researched owing to their self-cleaning and anti-corrosive properties. In this study, polypyrrole (PPy) nanowire meshes have been successfully fabricated by in situ electrochemical polymerization on stainless steel mesh substrates in the presence of phosphate buffered saline as both an electrolyte and a dopant. PPy nanowire meshes have high oil contact angles (above 150°) and low sliding angles (less than 10°), and they show underwater superoleophobicity with an excellent self-cleaning performance, not only in pure water, but also in highly corrosive aqueous solutions, including salt solutions, strong acids or basic solutions. PPy nanowire meshes presented here show promise for potential applications in fields such as oil-water separation and marine oil spill clean-up.


Predicting eruptions at mount st. Helens, june 1980 through december 1982.
Thirteen eruptions of Mount St. Helens between June 1980 and December 1982 were predicted tens of minutes to, more generally, a few hours in advance. The last seven of these eruptions, starting with that of mid-April 1981, were predicted between 3 days and 3 weeks in advance. Precursory seismicity, deformation of the crater floor and the lava dome, and, to a lesser extent, gas emissions provided telltale evidence of forthcoming eruptions. The newly developed capability for prediction reduced risk to life and property and influenced land-use decisions.


Identification of cellular genes targeted by KSHV-encoded microRNAs.
MicroRNAs (miRNAs) are 19 to 23 nucleotide-long RNAs that post-transcriptionally regulate gene expression. Human cells express several hundred miRNAs which regulate important biological pathways such as development, proliferation, and apoptosis. Recently, 12 miRNA genes have been identified within the genome of Kaposi sarcoma-associated herpesvirus; however, their functions are still unknown. To identify host cellular genes that may be targeted by these novel viral regulators, we performed gene expression profiling in cells stably expressing KSHV-encoded miRNAs. Data analysis revealed a set of 81 genes whose expression was significantly changed in the presence of miRNAs. While the majority of changes were below 2-fold, eight genes were down-regulated between 4- and 20-fold. We confirmed miRNA-dependent regulation for three of these genes and found that protein levels of thrombospondin 1 (THBS1) were decreased >10-fold. THBS1 has previously been reported to be down-regulated in Kaposi sarcoma lesions and has known activity as a strong tumor suppressor and anti-angiogenic factor, exerting its anti-angiogenic effect in part by activating the latent form of TGF-beta. We show that reduced THBS1 expression in the presence of viral miRNAs translates into decreased TGF-beta activity. These data suggest that KSHV-encoded miRNAs may contribute directly to pathogenesis by down-regulation of THBS1, a major regulator of cell adhesion, migration, and angiogenesis.


Efficacy of weekly mizoribine pulse therapy in refractory lupus nephritis.
We investigated the efficacy of a high-dose intermittent dosing treatment method (weekly mizoribine pulse therapy) conceived in the hope of achieving better efficacy by increasing the peak blood levels of mizoribine in patients with refractory lupus nephritis. Seventeen patients with lupus nephritis who had been resistant to corticosteroid and immunosuppressant therapy received weekly mizoribine pulse therapy. Mizoribine (350 mg) was administered three times at 12 h intervals over 2 consecutive days (700 mg for day 1 and 350 mg for day 2), followed by a washout period from day 3 to day 7. This therapeutic strategy enabled the peak blood levels of mizoribine to be increased to more than 3 μg/mL in most of the patients. Although SLEDAI, anti-ds-DNA antibody titer, CH-50, and serum albumin level did not significantly improve, urinary protein levels decreased, and it was possible to taper the dose of concomitant steroids. Using our definition of clinical response, 10 of the 17 patients were responders and 4 of them were nonresponders. The average peak serum mizoribine concentration of the responders was as high as 3.5 μg/mL. Elevation of serum liver enzymes was seen in 1 patient, and hyperuricemia occurred in 4 cases, but none of these adverse events were serious. Intermittent administration of mizoribine can increase blood levels and may be effective for refractory lupus nephritis.


Emergency nurse practitioners: a three part study in clinical and cost effectiveness.
To compare the clinical effectiveness and costs of minor injury services provided by nurse practitioners with minor injury care provided by an accident and emergency (A&E) department. A three part prospective study in a city where an A&E department was closing and being replaced by a nurse led minor injury unit (MIU). The first part of the study took a sample of patients attending the A&E department. The second part of the study was a sample of patients from a nurse led MIU that had replaced the A&E department. In each of these samples the clinical effectiveness was judged by comparing the "gold standard" of a research assessment with the clinical assessment. Primary outcome measures were the number of errors in clinical assessment, treatment, and disposal. The third part of the study used routine data whose collection had been prospectively configured to assess the costs and cost consequences of both models of care. The minor injury unit produced a safe service where the total package of care was equal to or in some cases better than the A&E care. Significant process errors were made in 191 of 1447 (13.2%) patients treated by medical staff in the A&E department and 126 of 1313 (9.6%) of patients treated by nurse practitioners in the MIU. Very significant errors were rare (one error). Waiting times were much better at the MIU (mean MIU 19 minutes, A&E department 56.4 minutes). The revenue costs were greater in the MIU (MIU pound 41.1, A&E department pound 40.01) and there was a great difference in the rates of follow up and with the nurses referring 47% of patients for follow up and the A&E department referring only 27%. Thus the costs and cost consequences were greater for MIU care compared with A&E care (MIU pound 12.7 per minor injury case, A&E department pound 9.66 per minor injury case). A nurse practitioner minor injury service can provide a safe and effective service for the treatment of minor injury. However, the costs of such a service are greater and there seems to be an increased use of outpatient services.


Characteristics of pressor response to endothelin in spontaneously hypertensive and Wistar-Kyoto rats.
Endothelin, an endothelium-derived vasoconstrictor peptide, and angiotensin II were intravenously injected into the femoral vein of normotensive Wistar-Kyoto (WKY) rats that had been anesthetized with urethane. Blood pressure and heart rate were recorded from a cannula inserted into the carotid artery. All experiments were carried out after treatment with adrenergic and cholinergic antagonists. Endothelin showed a potent, dose-dependent pressor action. The dose-response relations for the increase in blood pressure of rats receiving endothelin were comparable with those of rats receiving angiotensin II. However, endothelin showed far more long-lasting effects. Endothelin-induced responses consisted of three phases: a rapid and transient depressor phase and then two phases of pressor (transient and long-lasting) response. Nicardipine (0.1 mg/kg), a dihydropyridine Ca2+ channel blocker, markedly attenuated the slow phase of the pressor response but only slightly attenuated the rapid one. The pressor action of endothelin was not inhibited by continuous infusions of saralasin, which almost abolished the angiotensin II-induced pressor response. Endothelin-induced pressor response was also not attenuated by indomethacin, a prostaglandin synthesis inhibitor. These data provide evidence that endothelin produces a unique, potent, and long-lasting pressor response, which appears to be in part related to the activation of Ca2+ channels. In 12-week-old spontaneously hypertensive rats (SHR), the maximal pressor response to endothelin was slightly but significantly greater than that in age-matched WKY rats, but the dose dependency of the response was approximately consistent with that in WKY rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Necrotizing Fasciitis Within 72 hours After Presentation with Skin and Skin Structure Infection.
A small percentage of patients with skin infections later develop necrotizing fasciitis (NF). Diagnostic testing is needed to identify patients with skin infections at low risk of NF who could be discharged from the emergency department (ED) after antibiotic initiation. Elevated lactate has been associated with NF; existing estimates of the frequency of NF are based on retrospective reviews, and cases often lack testing for lactate. We present the incidence of patients with skin infections who developed NF and their baseline lactates. In four phase-3 trials, 2883 adults with complicated or acute bacterial skin and skin structure infections were randomized to dalbavancin or comparator, with early and late follow-up visits through Day 28. We prospectively collected baseline plasma lactates in one trial to assess an association with NF. NF was diagnosed in 3/2883 patients (0.1%); all three survived. In the study with prospectively collected baseline lactates (n = 622), 15/622 (2.4%) had a lactate ≥4 millimoles per liter (mmol/L), including 3/622 (0.5%) with a lactate ≥7 mmol/L. NF was not seen in patients with a lactate <4 mmol/L; NF was seen in 1/15 (6.7%) with a lactate ≥4 mmol/L, including 1/3 (33.3%) with lactate ≥7 mmol/L. NF incidence within 72 hours of antibiotic initiation in patients with complicated or acute bacterial skin and skin structure infections was extremely low (0.1%) and occurred in 6.7% with a lactate ≥4 mmol/L. Lactate <4 mmol/L can be used to identify patients at low risk of NF who could be safely discharged from the ED after antibiotic initiation.


N-tosylhydrazones: versatile reagents for metal-catalyzed and metal-free cross-coupling reactions.
Transition metal-catalyzed cross-coupling reactions have been established as one of the most powerful tools for the construction of C-C and C-X bonds. In this context, the development of novel metal-catalyzed cross-coupling processes that do not require stoichiometric organometallic reagents is particularly attractive. Recently, N-tosylhydrazones have emerged as a new type of versatile coupling partners for transition metal-catalyzed cross-coupling reactions as well as metal-free cross-coupling reactions, and have attracted increasing attention. This tutorial review summarizes recent important developments in this area with N-tosylhydrazones as versatile coupling partners.


Adverse outcomes and predictors of underuse of antithrombotic therapy in medicare beneficiaries with chronic atrial fibrillation.
Antithrombotic therapy can prevent strokes and transient ischemic attacks (TIAs) in carefully selected patients who have chronic nonvalvular atrial fibrillation (NVAF). Our objectives were 3-fold: to document the use of warfarin and aspirin therapy in Missouri Medicare beneficiaries with chronic NVAF; to identify factors associated with warfarin and aspirin underuse; and to determine the association between prescription of warfarin and aspirin at hospital discharge and adverse outcomes in this elderly, frail population. We linked chart reviews from all Missouri hospitals to Medicare claims data from 1993 to 1996. From chart reviews, we documented Medicare beneficiaries' demographic factors, comorbid conditions, and antithrombotic therapy prescribed at the time of hospital discharge. From Medicare claims, we determined the date of outcomes-death from any cause or hospitalization for an ischemic event (a stroke, a TIA, or a myocardial infarction). Only 328 (55%) of the 597 Medicare beneficiaries were prescribed antithrombotic therapy at hospital discharge: 34% received warfarin and 21% received aspirin. Advanced age, female gender, and rural residency predicted underuse of antithrombotic therapy. After controlling for these factors, as well as stroke risk factors and contraindications to anticoagulation, the prescription of warfarin was associated with a 24% relative risk reduction (RRR) in adverse outcomes (P=0.003). Prescription of aspirin was associated with a nonsignificant 5% RRR in these events (P=0.56). The underuse of antithrombotic therapy in Medicare beneficiaries who have NVAF is associated with measurable adverse outcomes. The benefit of warfarin therapy may extend to frail, elderly patients, a group that was excluded from randomized controlled trials. The role of antiplatelet therapy in this population deserves further study because many of these patients have relative contraindications to warfarin.