Introduction of handheld computing to a family practice residency program.
Handheld computers are valuable practice tools. It is important for residency programs to introduce their trainees and faculty to this technology. This article describes a formal strategy to introduce handheld computing to a family practice residency program. Objectives were selected for the handheld computer training program that reflected skills physicians would find useful in practice. TRGpro handheld computers preloaded with a suite of medical reference programs, a medical calculator, and a database program were supplied to participants. Training consisted of four 1-hour modules each with a written evaluation quiz. Participants completed a self-assessment questionnaire after the program to determine their ability to meet each objective. Sixty of the 62 participants successfully completed the training program. The mean composite score on quizzes was 36 of 40 (90%), with no significant differences by level of residency training. The mean self-ratings of participants across all objectives was 3.31 of 4.00. Third-year residents had higher mean self-ratings than others (mean of group, 3.62). Participants were very comfortable with practical skills, such as using drug reference software, and less comfortable with theory, such as knowing the different types of handheld computers available. Structured training is a successful strategy for introducing handheld computing to a residency program.


Targeting of BRM Sensitizes BRG1-Mutant Lung Cancer Cell Lines to Radiotherapy.
Targeting of epigenetic regulators as the chromatin remodeler SWI/SNF is proving to be a promising therapeutic strategy for individualized treatment of cancer patients. Here, we tested whether targeting one of the two mutually exclusive subdomains of the SWI/SNF complex BRM/SMARCA2 can sensitize specifically non-small cell lung carcinoma (NSCLC) cells with mutations in the other subunit BRG1/SMARCA4 toward ionizing radiation (IR). Knockdown of BRM with siRNA or shRNA and its consequences for radiation sensitivity as measured by clonogenic survival and plaque-monolayer control was studied in different NSCLC lines with or without BRG1 mutations and in primary fibroblasts. Furthermore, the effect on double-strand break (DSB) repair markers measured by immunofluorescence staining of 53BP1-, γ-H2AX-, and Rad51-foci was investigated. BRG1-mutated cell lines showed an increased surviving fraction compared with BRG1 proficient cells. Depletion of BRM (i) leads to a decreased proliferation rate and plating efficiency specifically in BRG1-mutated cells, (ii) specifically sensitized BRG1-mutant NSCLC cells toward IR as characterized by a survival reducing factor of 0.63 [95% confidence interval (CI), 0.57-0.69] in the dose range between 2 and 6 Gy, and (iii) decreased the tumor control doses after daily fractionation at 4 Gy in BRG1-mutant NSCLC cell lines A549 and H1299 in minimonolayers by 9.9% ± 1.3% and 13.6% ± 1.8%, respectively. In addition, an increase of residual Rad51-foci at 24 hours after irradiation in BRG1-mutant cells was demonstrated. Therefore, targeting of BRM in combination with radiotherapy is supposed to improve the therapeutic outcome of lung cancer patients harboring BRG1 mutations.The present study shows that the moderate radioresponsiveness of NSCLC cells with BRG1 mutations can be increased upon BRM depletion that is associated with a prolonged Rad51-foci prevalence at DNA DSBs.


Do plastids in Dendrobium cv. Lucky Duan petals function similar to autophagosomes and autolysosomes?
In animal cells a double-membrane-bound structure, the autophagosome, encloses a portion of the cytoplasm. The encapsulated material becomes digested after fusion of the autophagosome with a vesicle containing lytic enzymes. The autophagosome is then termed autolysosome. In intact plants, structures similar to animal autophagosomes/autolysosomes have been found only in a few types of cells. Additionally, some early papers indicated that plastids can function similar to autophagosomes/autolysosomes. Here, we report that plastids in Dendrobium cv. Lucky Duan petals produced an endocytosis-like invagination of the two outer membranes. The opening between the invagination space and the cytoplasm was almost isodiametric, less than 0.2 μm in diameter. The volume of the space formed by the invagination had a maximum of about half of the total plastid volume. Staining of the invagination lumen for acid phosphatase, a marker of organelles showing autophagic activity, was positive. Membranes and numerous ribosomes were observed inside the lumen of the invagination. The structure of the material inside the lumen varied from that of the cytoplasm to uniform electron-translucent, indicating that the enclosed cytoplasmic material became completely digested. No support was found for the idea that the material engulfed by the plastid or the whole plastid became transferred to a vacuole. Taken together, the data suggested the hypothesis that plastids in Dendrobium petal mesophyll cells can function in a way similar to both autophagosomes and autolysosomes in animal cells.


[Health promotion and prevention in the Lucerne canton--1995 analysis and prospects].
Considering the legal situation the canton of Lucerne is addressed to make his contribution to health promotion and prevention. His target groups are authorities on cantonal and community levels, specialised organisations and other health professionals, as for example physicians and pharmacists. The objectives of this study are to survey the health promotional and preventive activities, to evaluate the needs of the target groups as well as to focus on the opportunities of improved cooperation. Based on these results recommendations to the cantonal health authorities will be formulated. As far as the allocation of financial and personal resources are concerned health promotion and prevention do not figure as top priorities. Besides, the study results suggest to improve the cooperation between political representatives and health professionals. The cantonal health promotion authorities are well known among the target groups. However, cooperation could be intensified on all levels of action and is regarded as most desirable. A mutual forum focused on information and experience transfer would support these results and could stimulate joint projects. In addition, the needs of the communities have to be given special attention.


Simultaneous profiling of seed-associated bacteria and fungi reveals antagonistic interactions between microorganisms within a shared epiphytic microbiome on Triticum and Brassica seeds.
In order to address the hypothesis that seeds from ecologically and geographically diverse plants harbor characteristic epiphytic microbiota, we characterized the bacterial and fungal microbiota associated with Triticum and Brassica seed surfaces. The total microbial complement was determined by amplification and sequencing of a fragment of chaperonin 60 (cpn60). Specific microorganisms were quantified by qPCR. Bacteria and fungi corresponding to operational taxonomic units (OTU) that were identified in the sequencing study were isolated and their interactions examined. A total of 5477 OTU were observed from seed washes. Neither total epiphytic bacterial load nor community richness/evenness was significantly different between the seed types; 578 OTU were shared among all samples at a variety of abundances. Hierarchical clustering revealed that 203 were significantly different in abundance on Triticum seeds compared with Brassica. Microorganisms isolated from seeds showed 99-100% identity between the cpn60 sequences of the isolates and the OTU sequences from this shared microbiome. Bacterial strains identified as Pantoea agglomerans had antagonistic properties toward one of the fungal isolates (Alternaria sp.), providing a possible explanation for their reciprocal abundances on both Triticum and Brassica seeds. cpn60 enabled the simultaneous profiling of bacterial and fungal microbiota and revealed a core seed-associated microbiota shared between diverse plant genera.


Targeting alzheimer amyloid plaques in vivo.
The only definitive diagnosis for Alzheimer disease (AD) at present is postmortem observation of neuritic plaques and neurofibrillary tangles in brain sections. Radiolabeled amyloid-beta peptide (Abeta), which has been shown to label neuritic plaques in vitro, therefore could provide a diagnostic tool if it also labels neuritic plaques in vivo following intravenous injection. In this study, we show that the permeability of Abeta at the blood-brain barrier can be increased by at least twofold through covalent modification with the naturally occurring polyamine, putrescine. We also show that, following intravenous injection, radiolabeled, putrescine-modified Abeta labels amyloid deposits in vivo in a transgenic mouse model of AD, as well as in vitro in human AD brain sections. This technology, when applied to humans, may be used to detect plaques in vivo, allowing early diagnosis of the disease and therapeutic intervention before cognitive decline occurs.


High-throughput assay to phenotype Salmonella enterica Typhimurium association, invasion, and replication in macrophages.
Salmonella species are zoonotic pathogens and leading causes of food borne illnesses in humans and livestock. Understanding the mechanisms underlying Salmonella-host interactions are important to elucidate the molecular pathogenesis of Salmonella infection. The Gentamicin protection assay to phenotype Salmonella association, invasion and replication in phagocytic cells was adapted to allow high-throughput screening to define the roles of deletion mutants of Salmonella enterica serotype Typhimurium in host interactions using RAW 264.7 murine macrophages. Under this protocol, the variance in measurements is significantly reduced compared to the standard protocol, because wild-type and multiple mutant strains can be tested in the same culture dish and at the same time. The use of multichannel pipettes increases the throughput and enhances precision. Furthermore, concerns related to using less host cells per well in 96-well culture dish were addressed. Here, the protocol of the modified in vitro Salmonella invasion assay using phagocytic cells was successfully employed to phenotype 38 individual Salmonella deletion mutants for association, invasion and intracellular replication. The in vitro phenotypes are presented, some of which were subsequently confirmed to have in vivo phenotypes in an animal model. Thus, the modified, standardized assay to phenotype Salmonella association, invasion and replication in macrophages with high-throughput capacity could be utilized more broadly to study bacterial-host interactions.


Line authority for nurse staffing and costs for acute inpatient care.
There is little empirical evidence evaluating the effects of recent, widespread changes in nurse executive roles and nursing management structures on the costs of patient care. This retrospective cross-sectional study examined the relationship between line authority for nurse staffing and patient care costs (total, nursing, and non-nursing cost) using data from 124 Department of Veterans Affairs (VA) medical centers. After controlling for patient, facility, and market characteristics, nursing line authority was significantly associated with lower nursing cost per admission. Our results provide some evidence that a reduction in nursing line authority may adversely impact nursing costs.


What is the risk for a permanent stoma after low anterior resection of the rectum for cancer? A six-year follow-up of a multicenter trial.
The aim of this study was to assess the risk for permanent stoma after low anterior resection of the rectum for cancer. In a nationwide multicenter trial 234 patients undergoing low anterior resection of the rectum were randomly assigned to a group with defunctioning stomas (n = 116) or a group with no defunctioning stomas (n = 118). The median age was 68 years, 45% of the patients were women, 79% had preoperative radiotherapy, and 4% had International Union Against Cancer cancer stage IV. The patients were analyzed with regard to the presence of a permanent stoma, the type of stoma, the time point at which the stoma was constructed or considered as permanent, and the reasons for obtaining a permanent stoma. Median follow-up was 72 months (42-108). One patient with a defunctioning stoma who died within 30 days after the rectal resection was excluded from the analysis. During the study period 19% (45/233) of the patients obtained a permanent stoma: 25 received an end sigmoid stoma and 20 received a loop ileostomy. The end sigmoid stomas were constructed at a median of 22 months (1-71) after the low anterior resection of the rectum, and the loop ileostomies were considered as permanent at a median of 12.5 months (1-47) after the initial rectal resection. The reasons for loop ileostomy were metastatic disease (n = 6), unsatisfactory anorectal function (n = 6), deteriorated general medical condition (n = 3), new noncolorectal cancer (n = 2), patient refusal of further surgery (n = 2), and chronic constipation (n = 1). Reasons for end sigmoid stoma were unsatisfactory anorectal function (n = 22) and urgent surgery owing to anastomotic leakage (n = 3). The risk for permanent stomas in patients with symptomatic anastomotic leakage was 56% (25/45) compared with 11% (20/188) in those without symptomatic anastomotic leakage (P < .001). One patient of 5 ended up with a permanent stoma after low anterior resection of the rectum for cancer, and half of the patients with a permanent stoma had previous symptomatic anastomotic leakage.


The Use of Magnetic Resonance Imaging for Non-Invasive Assessment of Venofer® Biodistribution in Rats.
The aim of this study was to determine the potential of magnetic resonance imaging to evaluate the biodistribution of exogenous iron within 24 h after one single injection of Venofer® (iron sucrose). Venofer® was evaluated in vitro for its ability to generate contrast in MR images. Subsequently, iron disposition was assessed in rats with MRI, in vivo up to 3 h and post mortem at 24 h after injection of Venofer®, at doses of 10- and 40 mg/kg body weight (n = 2 × 4), or saline (n = 4). Within 10-20 min after injection of Venofer®, transverse relaxation rates (R2) clearly increased, representative of a local increase in iron concentration, in liver, spleen and kidney, including the kidney medulla and cortex. In liver and spleen R2 values remained elevated up to 3 h post injection, while the initial R2 increase in the kidney was followed by gradual decrease towards baseline levels. Bone marrow and muscle tissue did not show significant increases in R2 values. Whole-body post mortem MRI showed most prominent iron accumulation in the liver and spleen at 24 h post injection, which corroborated the in vivo results. MR imaging is a powerful imaging modality for non-invasive assessment of iron distribution in organs. It is recommended to use this whole-body imaging approach complementary to other techniques that allow quantification of iron disposition at a (sub)cellular level.


Loading of the knee joint during ergometer cycling: telemetric in vivo data.
Within-subject, repeated-measures design. To measure tibiofemoral contact forces during cycling in vivo and to quantify the influences of power, pedaling cadence, and seat height on tibiofemoral contact forces. Cycling is usually classified as a low-demand activity for the knee joint and is therefore recommended for persons with osteoarthritis and rehabilitation programs following knee surgery. However, there are limited data regarding actual joint loading. Instrumented knee implants with telemetric data transmission were used to measure the tibiofemoral contact forces. Data were obtained in 9 subjects, during ergometer cycling and walking, 15 ± 7 months after total knee arthroplasty. Tibiofemoral forces during cycling at power levels between 25 and 120 W, cadences of 40 and 60 rpm, and 2 seat heights were investigated. Within the examined power range, tibiofemoral forces during cycling were smaller than those during walking. At the moderate condition of 60 W and 40 rpm, peak resultant forces of 119% of body weight were measured during the pedal downstroke. Shear forces ranged from 5% to 7% of body weight. Forces increased linearly with cycling power. Higher cadences led to smaller forces. A lower seat height did not increase the resultant force but caused higher posterior shear forces. Due to the relatively small tibiofemoral forces, cycling with moderate power levels is suited for individuals with osteoarthritis and rehabilitation programs following knee surgery, such as cartilage repair or total knee replacement. The lowest forces can be expected while cycling at a low power level, a high cadence, and a high seat height.


Redundancy gain in binocular rivalry.
A behavioural advantage is found across a wide range of stimuli when two targets are presented in opposite hemifields compared with those targets being presented together in one hemifield, or one target being presented alone. This advantage for responses to multiple targets versus a single target is often termed redundancy gain. Here we report on the findings of two experiments investigating redundancy gain in binocular rivalry. Experiment 1 presented a rival pair in one hemifield with an additional image presented to both eyes in the opposite hemifield. There was a weak effect of this stable image on the perceived dominance of the images within the rival pair. Experiment 2 presented a second rival pair in either the same or opposite hemifield and showed that instances of joint predominance were greater when the two pairs were presented in opposite hemifields than within the same hemifield. Therefore, the findings suggest that redundancy gain may be extended to stimuli presented under binocular rivalry conditions.


Dopaminergic involvement in the control of drinking behaviour: a brief review.
This paper briefly reviews the role of the neurotransmitter dopamine (DA) in thirst and drinking behaviour. Electrolytic lesions of the lateral hypothalamus or 6-OHDA lesions of the nigro-striatal DA pathway produce aphaghia and adipsia. Similarly, DA receptor blockade inhibits drinking whereas the administration of DA or DA agonists can facilitate drinking. The involvement of DA in thirst is discussed with respect to the mediation of the motivational and motor responses associated with drinking behaviour.


DO-SRS imaging of metabolic dynamics in aging Drosophila.
Emerging studies have shown that lipid metabolism plays an important role in aging. High resolution in situ imaging of lipid metabolic dynamics inside cells and tissues affords a novel and potent approach for understanding many biological processes such as aging. Here we established a new optical imaging platform that combines D2O-probed stimulated Raman scattering (DO-SRS) imaging microscopy and a Drosophila model to directly visualize metabolic activities in situ during aging. The sub-cellular spatial distribution of de novo lipogenesis in the fat body was quantitatively imaged and examined. We discovered a dramatic decrease in lipid turnover in 35-day-old flies. Decreases in protein turnover occurred earlier than lipids (25-day vs. 35-day), and there are many proteins localized on the cell and lipid droplet membrane. This suggests that protein metabolism may act as a prerequisite for lipid metabolism during aging. This alteration of maintenance of protein turnover indicates disrupted lipid metabolism. We further found a significantly higher lipid turnover rate in large LDs, indicating more active metabolism in large LDs, suggesting that large and small LDs play different roles in metabolism to maintain cellular homeostasis. This is the first study that directly visualizes spatiotemporal alterations of lipid (and protein) metabolism in Drosophila during the aging process. Our study not only demonstrates a new imaging platform for studying lipid metabolism, but also unravels the important interconnections between lipid metabolism and aging.


Characterization, sequence determination, and immunogenicity of a 64-kilodalton protein of Mycobacterium bovis BCG expressed in escherichia coli K-12.
We report the DNA sequence of a previously cloned Mycobacterium bovis BCG gene encoding an immunogenic 64-kilodalton protein. This protein, MbaA, was purified from overproducing Escherichia coli K-12 cells, and the presence of antibodies to MbaA in human sera was determined by an enzyme-linked immunosorbent assay. In about 80% of serum samples from tuberculosis patients and in about 60% of samples from BCG-vaccinated individuals, significant levels of anti-MbaA antibodies were found. Surprisingly, in about 30% of the control serum samples obtained from children, anti-MbaA antibodies were also observed. Guinea pigs sensitized with M. bovis BCG or MbaA showed a delayed-type hypersensitivity reaction after challenge with purified MbaA, supporting the previously observed strong reactivity of human T-cell clones with this, for mycobacteria, common antigen.


Back to the future: forget ultrasound and focus on clinical assessment in rheumatoid arthritis management.
Ultrasound (US) unquestionably improves many aspects of rheumatoid arthritis (RA) diagnosis and management, but no consensus has been reached regarding the optimal US methodology that should be used, and high levels of standardisation have not yet been attained. Current evidence from two randomised controlled trials does not support the US in directing treatment decisions. A return to the endorsement of clinical assessment and the adoption of T2T strategies aiming at ACR/EULAR remission still represent the standard of care for RA and should be pursued through appropriate educational programmes. Thus, for now, the major application of sonography in arthritis should have a focus on diagnostic and especially differential diagnostic aspects.


Evaluation of vincristine-associated myelosuppression in Border Collies.
To determine whether Border Collies (ATP binding cassette subfamily B1 gene [ABCB1] wildtype) were more likely than other breeds to develop vincristine-associated myelosuppression (VAM) and, if so, whether this was caused by a mutation in ABCB1 distinct from ABCB1-1Δ. Phase 1 comprised 36 dogs with the ABCB1 wildtype, including 26 dogs with lymphoma (5 Border Collies and 21 dogs representing 13 other breeds) treated with vincristine in a previous study; phase 2 comprised 10 additional Border Collies, including 3 that developed VAM and 7 with an unknown phenotype. For phase 1, the prevalence of VAM in ABCB1-wildtype Border Collies was compared with that for ABCB1-wildtype dogs of other breeds with data from a previous study. For phase 2, additional Border Collies were included. Hematologic adverse reactions were graded with Veterinary Co-operative Oncology Group criteria. Genomic DNA was used to amplify and sequence all 27 exons of the canine ABCB1. Sequences from affected dogs were compared with those of unaffected dogs and dogs of unknown phenotype. 3 of 5 Border Collies with the ABCB1 wildtype developed VAM; this was significantly higher than the proportion of other dogs that developed VAM (0/21). A causative mutation for VAM in Border Collies was not identified, although 8 single nucleotide polymorphisms in ABCB1 were detected. Breed-associated sensitivity to vincristine unrelated to ABCB1 was detected in Border Collies. Veterinarians should be aware of this breed predisposition to VAM. Causes for this apparent breed-associated sensitivity should be explored.


[Folate food intake and red blood cell folate concentrations in women from Recife, Northeast of Brazil].
Folate food intake and red blood cell folate concentrations were assess in women from Recife, Northeast Brazil. Following a two stages sampling procedure, a cross-sectional study was carried out involving 360 women, between 15-45 y, attending in nine health care public unit in 2007-2008. Folate intake was evaluated by a Quantitative Food Frequency Questionnaire, and compared with the values of the dietary reference intakes-DRI's. Folate status was evaluated by red blood cell folate concentrations. Geometric mean of folate intake was 627.1 [IC 95% 600.4-655.0] microg/day. The frequency of women at risk for folate intake below the recommendation was 16.0% for adolescents (<330 microg/day) and 6.3% for young adult (<320 microg/ day). The prevalence of women whose consumption exceeds the maximum tolerable intake was 48.0% (> 800 microg/day) and 13.7% (> 1000 microg/day) for adolescents and adults, respectively. The mean of red blood cell folate concentrations was 1797.8 +/- 357.1 nmol/L. Folate rich-food intake did not show any correlation with red blood cell folate concentrations (r = 0.058 and p = 0.274). Higher red blood cell folate concentrations were observed in adult young women (p = 0.004) and among those with income up to two minimum wages (p = 0.042). Folate rich-food intake as well as red blood cell folate concentrations among women from Recife were above the international recommendations.


Nationwide prevalence and prognostic significance of clinically diagnosable protein-calorie malnutrition in hospitalized inflammatory bowel disease patients.
Inflammatory bowel disease (IBD) patients are at increased risk of protein-calorie malnutrition. We sought to determine the prevalence of clinically diagnosable malnutrition among those hospitalized for IBD throughout the United States and whether this malnutrition influenced health outcomes. We queried the Nationwide Inpatient Sample between 1998 and 2004 to identify admissions for Crohn's disease (CD) or ulcerative colitis (UC) and a representative sample of non-IBD discharges. We assessed the prevalence and predictors of malnutrition and its association with in-hospital mortality and resource utilization. The prevalence of malnutrition was greater in CD and UC patients than in non-IBD patients (6.1% and 7.2% versus 1.8%, P < 0.0001). The adjusted odds ratio for malnutrition among IBD admissions compared with non-IBD admissions was 5.57 [95% confidence interval (CI): 5.29-5.86]. More IBD discharges than non-IBD discharges with malnutrition received parenteral nutrition (26% versus 6%, P < 0.0001). There was increased likelihood of malnutrition among those with fistulizing CD (OR 1.65; 95% CI: 1.50-1.82) and among those who had undergone bowel resection (OR 1.37; 95% CI: 1.27-1.48). Malnutrition was associated with increased in-hospital mortality 3.49 (95% CI: 2.89-4.23), length of stay (11.9 days versus 5.8 days, P < 0.00001), and total charges ($45,188 versus $20,295, P < 0.0001). Clinically apparent malnutrition is more frequent among IBD admissions than among non-IBD admissions. Its association with greater mortality and resource utilization may reflect more severe underlying disease that can lead to both malnutrition and worse outcomes. Nonetheless, diagnosable malnutrition may serve as a clinical marker of poor IBD prognosis in hospitalized patients.


A new method for evaluation of split renal cortical blood flow with contrast echography.
The recent development of contrast echography has made renal enhancement possible through an intravenous injection of microbubble-based contrast. In animal models, tissue perfusion can be quantified using contrast echography by measurement of the rate at which microbubbles replenish tissue after their ultrasound-induced destruction. Our purpose in this study was to evaluate renal blood flow with contrast echography in humans. To increase the sensitivity for microbubbles, we used a combination of power Doppler harmonic and intermittent imaging. The pulsing interval (PI) was changed from 10 cardiac cycles to 1 cardiac cycle during an intravenous infusion of the contrast agent, and alterations in the intensity of the renal cortex were represented as a decline ratio (DR). In 24 patients with various renal diseases, we were able to observe all 48 kidneys with adequate enhancement of the renal cortex. At PI of 10 cardiac cycles, the enhancement was homogeneous and strong, while, obviously, changing PI from 10 to 1 cardiac cycles caused a decline of enhancement. An excellent correlation was found between DR using contrast echography and renal plasma flow determined by clearance and radionuclide measurements. An excellent correlation was found between the DR values determined by contrast echography and the renal plasma flow values determined using clearance and radionuclide measurements. These results suggest that DR may be useful for evaluation of both total and split renal blood flow. Thus the contrast echographic method presented here could succeed in assessing renal cortical blood flow less invasively than conventional methods in humans.


Structural topology and activation of an initial adenylate kinase-substrate complex.
Enzymatic activity is ultimately defined by the structure, chemistry, and dynamics of the Michaelis complex. A large number of experimentally determined structures between enzymes and substrates, substrate analogues, or inhibitors exist. However, transient, short-lived encounter and equilibrium structures also play fundamental roles during enzymatic reaction cycles. Such structures are inherently difficult to study with conventional experimental techniques. The enzyme adenylate kinase undergoes major conformational rearrangements in response to binding of its substrates, ATP and AMP. ATP is sandwiched between two binding surfaces in the closed and active enzyme conformation. Thus, adenylate kinase harbors two spatially distant surfaces in the substrate free open conformation, of which one is responsible for the initial interaction with ATP. Here, we have performed primarily nuclear magnetic resonance experiments on Escherichia coli adenylate kinase (AK(eco)) variants that allowed identification of the site responsible for the initial ATP interaction. This allowed a characterization of the structural topology of an initial equilibrium complex between AK(eco) and ATP. On the basis of the results, we suggest that the ATP binding mechanism for AK(eco) is a mixture between "induced fit" and "conformational selection" models. It is shown that ATP is activated in the initial enzyme-bound complex because it displays an appreciable rate of nonproductive ATP hydrolysis. In summary, our results provide novel structural and functional insights into adenylate kinase catalysis.


Blood Exosomes Have Neuroprotective Effects in a Mouse Model of Parkinson's Disease.
Parkinson's disease (PD) is a common and complex neurodegenerative disease; the pathogenesis of which is still uncertain. Exosomes, nanosized extracellular vesicles, have been suggested to participate in the pathogenesis of PD, but their role is unknown. Here, a metabolomic analysis of serum and brain exosomes showed differentially expressed metabolites between 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride- (MPTP-) induced PD mice and control mice, such as oxidized lipids, vitamins, and cholesterol. These metabolites were enriched in coenzyme, nicotinamide, and amino acid pathways related to PD, and they could be served as preclinical biomarkers. We further found that blood-derived exosomes from healthy volunteers alleviated impaired motor coordination in MPTP-treated mice. Results from immunohistochemistry and western blotting indicated that the loss of dopaminergic neurons in substantia nigra and striatum of PD model mice was rescued by the exosome treatment. The exosome treatment also restored the homeostasis of oxidative stress, neuroinflammation, and cell apoptosis in the model mice. These results suggest that exosomes are important mediators for PD pathogenesis, and exosomes are promising targets for the diagnosis and treatment of PD.


Clinical and Virological Characteristics of Ebola Virus Disease Patients Treated With Favipiravir (T-705)-Sierra Leone, 2014.
During 2014-2015, an outbreak of Ebola virus disease (EVD) swept across parts of West Africa. No approved antiviral drugs are available for Ebola treatment currently. A retrospective clinical case series was performed for EVD patients in Sierra Leone-China Friendship Hospital. Patients with confirmed EVD were sequentially enrolled and treated with either World Health Organization (WHO)-recommended supportive therapy (control group) from 10 to 30 October, or treated with WHO-recommended therapy plus favipiravir (T-705) from 1 to 10 November 2014. Survival and virological characteristics were observed for 85 patients in the control group and 39 in the T-705 treatment group. The overall survival rate in the T-705 treatment group was higher than that of the control group (56.4% [22/39] vs 35.3% [30/85]; P = .027). Among the 35 patients who finished all designed endpoint observations, the survival rate in the T-705 treatment group (64.8% [11/17]) was higher than that of the control group (27.8% [5/18]). Furthermore, the average survival time of the treatment group (46.9 ± 5.6 days) was longer than that of the control group (28.9 ± 4.7 days). Most symptoms of patients in the treatment group improved significantly. Additionally, 52.9% of patients who received T-705 had a >100-fold viral load reduction, compared with only 16.7% of patients in the control group. Treatment of EVD with T-705 was associated with prolonged survival and markedly reduced viral load, which makes a compelling case for further randomized controlled trials of T-705 for treating EVD.


Ice core records of atmospheric N2O covering the last 106,000 years.
Paleoatmospheric records of trace-gas concentrations recovered from ice cores provide important sources of information on many biogeochemical cycles involving carbon, nitrogen, and oxygen. Here, we present a 106,000-year record of atmospheric nitrous oxide (N2O) along with corresponding isotopic records spanning the last 30,000 years, which together suggest minimal changes in the ratio of marine to terrestrial N2O production. During the last glacial termination, both marine and oceanic N2O emissions increased by 40 +/- 8%. We speculate that our records do not support those hypotheses that invoke enhanced export production to explain low carbon dioxide values during glacial periods.


Lung Function in Young Adults and Risk of Cardiovascular Events Over 29 Years: The CARDIA Study.
Background Diminished peak lung function in young adulthood is a risk factor for future chronic obstructive pulmonary disease. The association between lung disease and cardiovascular disease later in life is well documented. Whether peak lung function measured in young adulthood is associated with risk of future cardiovascular events is unknown. Methods and Results CARDIA (The Coronary Artery Risk Development in Young Adults) study is a prospective, multicenter, community-based, longitudinal cohort study including 4761 participants aged 18 to 30 years with lung function testing we investigated the association between lung health in young adulthood and risk of subsequent cardiovascular events. We performed Cox proportional hazards regression to test the association between baseline and years 10 and 20 pulmonary function with incident cardiovascular events. Linear and logistic regression was performed to explore the associations of lung function with development of risk factors for cardiovascular disease as well as carotid intima-media thickness and coronary artery calcified plaque. At baseline, mean age (± SD ) was 24.9±3.6 years. Baseline forced expiratory volume in 1 second (hazard ratio) per -10-unit decrement in percent predicted forced expiratory volume in 1 second (hazard ratio, 1.18; 95% CI, 1.06-1.31 [ P=0.002]) and FVC per -10-unit decrement in percent predicted FVC (hazard ratio, 1.19; 95% CI, 1.06-1.33 [ P=0.003]) were associated with future cardiovascular events independent of traditional cardiovascular risk factors. Baseline lung function was associated with heart failure and cerebrovascular events but not coronary artery disease events. Conclusions Lung function in young adulthood is independently associated with cardiovascular events into middle age. This association appears to be driven by heart failure and cerebrovascular events rather than coronary heart disease. Clinical Trial Registration URL : https://www.clinicaltrials.gov . Unique identifier: NCT 00005130.


Age-related neoplastic risk profiles and penetrance estimations in multiple endocrine neoplasia type 2A caused by germ line RET Cys634Trp (TGC>TGG) mutation.
RET testing in multiple endocrine neoplasia type 2 for molecular diagnosis is the paradigm for the practice of clinical cancer genetics. However, precise data for distinct mutation-based risk profiles are not available. Here, we survey the clinical profile for one specific genotype as a model, TGC to TGG in codon 634 (C634W). By international efforts, we ascertained all available carriers of the RET C634W mutation. Age at diagnosis, penetrance, and clinical complications were analyzed for medullary thyroid carcinoma (MTC), pheochromocytoma, and hyperparathyroidism (HPT), as well as overall survival. Our series comprises 92 carriers from 20 unrelated families worldwide. Sixty-eight subjects had MTC diagnosed at age 3-72 years (mean 29). Lymph node metastases were observed in 16 subjects aged 20-72 and distant metastases in 4 subjects aged 28-69. Forty-one subjects had pheochromocytoma detected at age 18-67 (mean 36). Amongst the 28 subjects with MTC and pheochromocytoma, six developed pheochromocytoma before MTC. Six subjects had HPT diagnosed at age 26-52 (mean 39). Eighteen subjects died; of the 16 with known causes of death, 8 died of pheochromocytoma and 4 of MTC. Penetrance for MTC is 52% by age 30 and 83% by age 50, for pheochromocytoma penetrance is 20% by age 30 and 67% by age 50, and for HPT penetrance is 3% by age 30 and 21% by age 50. These data provide, for the first time, RET C634W-specific neoplastic risk and age-related penetrance profiles. The data may facilitate risk assessment and genetic counseling.


The epidemiology of herpes simplex virus eye disease in Northern California.
To calculate the incidence and prevalence of herpes simplex virus (HSV) eye disease in a large, well-defined population in Northern California, USA, and to determine the recurrence rate following an initial episode of disease in this cohort. A retrospective, observational, cohort study using population-based data and medical record review. The patient database of a large, regional health maintenance organization (Northern California Kaiser Permanente) was searched, and the study population consisted of 1,042,351 people over a 1-year study period from 1 July 1998 through 30 June 1999. Only ocular HSV cases with definitive clinical or laboratory confirmed diagnoses were included. Active and inactive cases were included, however only active cases were used in incidence and prevalence calculations. Bilateral disease was counted as one case. Newly diagnosed cases were followed for recurrence from initial presentation through 31 December 2002. After chart review of 322 possible cases, 71 new cases and 59 previously diagnosed active cases of ocular HSV were confirmed. This resulted in an incidence rate of 6.8 new cases/100,000 person-years (95% confidence interval, CI, 5.3-8.6). Incidence increased with age, and rates were highest in people over 75 years of age (p < 0.001). The recurrence rate in new cases was 18% for the 3-year follow-up time, and was equal to 5% per year (95% CI 3-9%). The incidence and prevalence of ocular herpes simplex in this study was lower than previously reported. Incidence increased with age, and there were significantly higher rates in the older population.


Electrical and secretory manifestations of glucose and amino acid interactions in rat pancreatic islets.
Interactions between glucose and amino acids in rat pancreatic islets were studied by recording the intracellular membrane potential and spike discharges from the isolated perfused pancreas. It was found that L-isoleucine requires the presence of basal glucose (5 mM) in order to increase spike discharge from islet cells and depolarize the cell membrane. Similarly basal glucose is needed for insulin release by L-isoleucine. A physilolgical mixture of twenty amino acids also required the presence of basal glucose in order to increase spike activity and insulin release. In contrast to L-isoleucine the amino acid mixture did not depolarize the beta-cells. Iodoacetate, at concentrations previously shown to block glycolysis completely, did not interfere with any of these permissive actions of glucose, nor did iodoacetate alter the well known electrical manifestations of high levels of glucose itself (i.e. depolarization and increased spike discharge). These data show that glucose plays a pre-eminent role as regulator of islet cell function, governing the efficacy of amino acids as beta-cells stimulants. The results are most easily interpreted if one assumes that glycolysis is not required for glucose to exert its action.


Refining forelimb asymmetry analysis: Correlation with Montoya staircase contralateral function post-stroke.
Forelimb Asymmetry Test is a simple test of motor function, using exploration behavior of a rat in a novel environment and counting the number of times that a rat touches the wall with either forepaw. Our lab has noticed, however, that there appears to be an increased number of fingertip touches to the wall following a stroke in the impaired forelimb. We counted the number of times that the animal either laid its palm flat against the wall of the chamber or touched the wall with only its fingertips, for both the left and right forepaws. We also separated bouts of exploration, so we could clearly determine if fingertip touches normally were associated with a transition from resting state to exploration state. Fishers exact test indicated that there were significant differences in the way that the animals touched the wall pre-stroke compared to post-stroke, with more fingertip touches occurring post-stroke. Counting palm touches as normal and fingertip touches as abnormal increases the sensitivity of the Forelimb Asymmetry analysis and gives a good correlation with the contralateral functional deficits determined by Montoya Staircase post-stroke. If we counted every wall touch as normal (palm touches and fingertip touches), we see a loss of sensitivity and a poor correlation with contralateral function as determined by Montoya Staircase. This refinement of the Forelimb Asymmetry analysis improves correlation with Montoya Staircase contralateral function after stroke.


Advances in Bacterial Oligosaccharyltransferase Structure Elucidation and Potential Application to Glycoconjugate Vaccine Design.
Glycosylation is one of the most common post-translational modifications of proteins across all kingdoms of life. Diverse monosaccharides and polysaccharides can be attached to a range of amino acid residues generating N-glycosylation, O-glycosylation, C-glycosylation, S-glycosylation, as well as P-glycosylation. The functions of the eukaryotic glycosylation system during protein folding in the endoplasmic reticulum (ER) and Golgi are well-studied. Increasing evidence in the recent decade has demonstrated the presence of oligosaccharyltransferases (OSTs) in bacteria and archaea. In particular, the oligosaccharyltransferase (PglB) of Campylobacter jejuni and oligosaccharyltransferase (PglL) enzyme of Neisseria meningitidis are the most characterized OSTs that catalyze bacterial N-linked glycosylation and O-linked glycosylation, respectively. Glycoprotein administered as glycoconjugate vaccines have been shown to be effective prophylactic to protect against numerous pathogenic bacteria. The chemical synthesis of glycoproteins is complex and expensive, which limits its application to the development of glycoconjugate vaccines. However, studies have demonstrated that the biosynthesis of glycoproteins is realizable by transferring PglB, a plasmid encoding a substrate protein, or PglL, a plasmid encoding genes for glycan synthesis to Escherichia coli. This strategy can be applied to the development of glycoconjugate vaccines using engineered host E. coli. This review summarizes the structure and mechanism of action of the bacterial OSTs, PglB and PglL, and discusses their potential application to glycoconjugate vaccine design.


Patterns of primary care among persons with schizophrenia: the role of patients, general practitioners and centre factors.
Patients with schizophrenia and related disorders have more physical problems than the general population. Primary care professionals play an important role in the care of these patients as they are the main entry point into the healthcare system. We aimed to identify patient, general practitioner, and primary care centre factors associated with the number of visits of patients with schizophrenia and related disorders to general practitioners. A descriptive, cross-sectional study was conducted in 13 primary care centres belonging to the Clinical Management Unit of Mental Health of the Regional Hospital of Málaga, Spain. The eligible population was composed of patients with schizophrenia and related disorders attending the primary care centres in the study area, and the general practitioners who attend these patients. Our dependent variable was the total number of general practitioner visits made by patients with schizophrenia and related disorders during the 3.5-year observation period. The independent variables were grouped into three: (a) patient variables (sociodemographic and clinical), (b) general practitioner variables, and (c) primary care centre characteristics. Descriptive, bivariate, and multivariate analyses using the random forest method were performed. A total of 259 patients with schizophrenia and related disorders, 96 general practitioners, and 13 primary care centres were included. The annual mean was 3.9 visits per patient. The results showed that younger general practitioners, patients who were women, patients who were married, some primary care centres to which the patient belonged, taking antipsychotic medication, presenting any cardiovascular risk factor, and more frequency of mental health training sessions at the primary care centre were associated with an increased number of visits to general practitioners. The only general practitioner variable that was associated with the number of visits was age, the older the less contact. There were also patient variables involved in the number of visits. Finally, mental health training for general practitioners was important for these professionals to manage patients with schizophrenia and related disorders.


An interdisciplinary computer-based information tool for palliative severe pain management.
As patient care becomes more collaborative in nature, there is a need for information technology that supports interdisciplinary practices of care. This study developed and performed usability testing of a standalone computer-based information tool to support the interdisciplinary practice of palliative severe pain management (SPM). A grounded theory-participatory design (GT-PD) approach was used with three distinct palliative data sources to obtain and understand user requirements for SPM practice and how a computer-based information tool could be designed to support those requirements. The GT-PD concepts and categories provided a rich perspective of palliative SPM and the process and information support required for different SPM tasks. A conceptual framework consisting of an ontology and a set of three problem-solving methods was developed to reconcile the requirements of different interdisciplinary team members. The conceptual framework was then implemented as a prototype computer-based information tool that has different modes of use to support both day-to-day case management and education of palliative SPM. Usability testing of the computer tool was performed, and the tool tested favorably in a laboratory setting. An interdisciplinary computer-based information tool can be developed to support the different work practices and information needs of interdisciplinary team members, but explicit requirements must be sought from all prospective users of such a tool. Qualitative methods such as the hybrid GT-PD approach used in this research are particularly helpful for articulating computer tool design requirements.


Nutritional devices and hyperalimentation in the elderly.
The feeding devices and delivery systems are only two aspects of the total picture of ideal nutritional support. One needs to (a) perform a complete nutritional assessment to determine the patient's needs (Blackburn et al. 1977); (b) choose appropriately among the currently available formulas for enteral (Bistrian and Wade 1981) and parenteral use; (c) select the necessary feeding device and delivery system; and (d) administer the solution and monitor the patient's progress. There are many options in meeting the nutritional needs of the elderly patient. One needs to consider the patient's ability to ingest, digest, absorb, and metabolize the various foodstuffs. If the gut is functional, it should be used (Bothe et al. 1981). If it is not, the intravenous route is indicated. In light of the vast advances in technology in this area, there exists no excuse to allow our elderly patients to go undernourished.


Athlete Monitoring in Rugby Union: Is Heterogeneity in Data Capture Holding Us Back?
In an effort to combat growing demands on players, athlete monitoring has become a central component of professional sport. Despite the introduction of new technologies for athlete monitoring, little is understood about the practices employed in professional rugby clubs. A questionnaire was circulated amongst conditioning staff across the 12 Premiership rugby clubs to capture the methods used, relative importance, perceived effectiveness and barriers to the use of multiple different athlete monitoring measurements. Previous injury, Global Positioning System (GPS) metrics, collision counts and age were deemed the most important risk factors for managing future injury risk. A wide range of GPS metrics are collected across clubs with high-speed running (12/12 clubs), distance in speed zones (12/12 clubs) and total distance (11/12 clubs) the most commonly used. Of the metrics collected, high-speed running was deemed the most important for managing future injury risk (5/12 clubs); however, there was considerable variation between clubs as to the exact definition of high-speed running, with both absolute and relative measures utilised. While the use of such monitoring tools is undertaken to improve athlete welfare by minimising injury risk, this study demonstrates the significant heterogeneity of systems and methods used by clubs for GPS capture. This study therefore questions whether more needs to be done to align practices within the sport to improve athlete welfare.


Sucrose induces expression of the sorbitol-6-phosphate dehydrogenase gene in source leaves of loquat.
Rosaceae fruit trees use sorbitol and sucrose as translocating sugars and the sorbitol-to-sucrose ratio in source leaves determines apple fruit quality. Here, we investigate the effects of sugars on the expression of genes encoding key photosynthetic enzymes, including sorbitol-6-phosphate dehydrogenase (S6PDH, EC 220.127.116.11), sucrose phosphate synthase (SPS, EC 18.104.22.168), and ADP-glucose pyrophosphorylase (ADPGPPase, EC 126.96.36.199) to understand the sugar-signaling mechanism in Rosaceae fruit trees. Mature leaf-petiole cuttings of loquat (Eriobotrya japonica Lindl. cv. Mogi) were supplied with a water, sorbitol or sucrose solution for 2 days at 20°C. The relative levels of the transcripts were analyzed by real-time polymerase chain reaction (PCR). S6PDH transcription was decreased by sorbitol but drastically increased by sucrose. SPS and ADPGPPase large subunit transcription were decreased by sucrose and sorbitol. The simultaneous application of sorbitol and sucrose revealed that S6PDH transcription increased in a dose-dependent manner with sucrose. These results show that both sorbitol and sucrose work as signaling molecules in source organs of Rosaceae fruit trees. These trees have mechanisms to positively keep sorbitol as the dominant translocating sugar, suggesting that sorbitol plays an important role in their survival strategy. Effects of various sugars on S6PDH expression were investigated. Palatinose, a sucrose analog, increased S6PDH transcription much more drastically than sucrose. Mannose and 3-O-methylglucose, glucose analogs, also increased S6PDH transcription; however, glucose did not. Models of sugar signaling in source organs of Rosaceae fruit trees are discussed.


Elevation of HDL-C in response to statin treatment is involved in the regression of carotid atherosclerosis.
Atherosclerosis is strongly associated with an increased mortality in subjects with diabetes. The carotid intima-media thickness (IMT) is commonly measured as a surrogate marker for cardiovascular risk. Statins are well-established protective agents against atherosclerosis and reportedly suppress IMT progression in subjects with diabetes. To clarify the effects of statins on subclinical atherosclerosis, we herein investigated changes in the carotid IMT and lipid profiles in a multi-center, prospective, randomized trial. Hypercholesterolemic subjects with type 2 diabetes were randomly assigned to open-label treatment with either pravastatin or pitavastatin. The primary endpoint of this study was the IMT change after 36 months of statin treatment. A total of 97 subjects (51 pitavastatin; 46 pravastatin) completed this 36-month study. The LDL-C decreased significantly from 163.4 ± 27.9 mg/dl at baseline to 100.4 ± 19.6 mg/dl at 36 months in the pitavastatin group and from 159.7 ± 25.6 mg/dl to 118.5 ± 22.1 mg/dl in the pravastatin group. The mean IMT showed moderate regression in both the pitavastatin (-0.070 ± 0.215 mm, P<0.05) and the pravastatin (-0.067 ± 0.260 mm) group. However, there was no significant difference in the IMT change between the two groups. When the two groups were combined, the 36-month change in the mean IMT was significantly associated with HDL-C change (r=-0.24, P= 0.03). Multiple linear regression analysis revealed the change in HDL-C to be an independent variable showing a positive correlation with the carotid IMT reduction. The administration of statins for 3 years to subjects with type 2 diabetes resulted in a significant regression of the carotid IMT. An elevation of the plasma HDL-C with statin treatment was closely related to a regression of atherosclerosis.


[Intrastromal keratorhexis with the Nd:YAG laser--a possible method of refractive surgery?].
A new method of refractive corneal surgery is presented. With the q-switched Nd:YAG-laser Visulas (Zeiss-Oberkochen) we succeed in altering the optical power of the cornea of rabbits, pigs and humans. The number of applied laser spots and the distribution pattern influence the way and the amount of change of refraction. The method, clinical and histological features, complications and prospects of the Nd: YAG laser keratorhexis are described.


Administration of r-VEGF-A prevents hepatic artery ligation-induced bile duct damage in bile duct ligated rats.
The hepatic artery, through the peribiliary plexus, nourishes the intrahepatic biliary tree. During obstructive cholestasis, the nutritional demands of intrahepatic bile ducts are increased as a consequence of enhanced proliferation; in fact, the peribiliary plexus (PBP) displays adaptive expansion. The effects of hepatic artery ligation (HAL) on cholangiocyte functions during cholestasis are unknown, although ischemic lesions of the biliary tree complicate the course of transplanted livers and are encountered in cholangiopathies. We evaluated the effects of HAL on cholangiocyte functions in experimental cholestasis induced by bile duct ligation (BDL). By using BDL and BDL + HAL rats or BDL + HAL rats treated with recombinant-vascular endothelial growth factor-A (r-VEGF-A) for 1 wk, we evaluated liver morphology, the degree of portal inflammation and periductular fibrosis, microcirculation, cholangiocyte apoptosis, proliferation, and secretion. Microcirculation was evaluated using a scanning electron microscopy vascular corrosion cast technique. HAL induced in BDL rats 1) the disappearance of the PBP, 2) increased apoptosis and impaired cholangiocyte proliferation and secretin-stimulated ductal secretion, and 3) decreased cholangiocyte VEGF secretion. The effects of HAL on the PBP and cholangiocyte functions were prevented by r-VEGF-A, which, by maintaining the integrity of the PBP and cholangiocyte proliferation, prevents damage of bile ducts following ischemic injury.


Cardiovascular consequences of loss of supraspinal control of the sympathetic nervous system after spinal cord injury.
Spinal cord injury (SCI) with resultant quadriplegia or high paraplegia is associated with significant dysfunction of the sympathetic nervous system. This alteration of sympathetic nervous system activity occurs as a consequence of loss of supraspinal control of the sympathetic nervous system and is further complicated by at least three subsequent phenomena that occur below the level of SCI: reduced overall sympathetic activity, morphologic changes in sympathetic preganglionic neurons, and peripheral alpha-adrenoceptor hyperresponsiveness. Reduced sympathetic activity below the level of SCI appears to result in orthostatic hypotension, low resting blood pressure, loss of diurnal fluctuation of blood pressure, reflex bradycardia, and, rarely, cardiac arrest. Peripheral alpha-adrenoceptor hyperresponsiveness likely accounts for some, if not the majority, of the excessive pressor response in autonomic dysreflexia and may also contribute to decreased blood flow in the peripheral microcirculation, potentially increasing susceptibility to pressure sores. What has yet to be established is whether this alpha-adrenoceptor hyperresponsiveness is a consequence of receptor hypersensitivity or a failure of presynaptic reuptake of noradrenaline at the receptor level. Better understanding of the pathophysiology of sympathetic nervous system dysfunction after high-level SCI should allow development of more effective measures to manage clinical complications.


Wellbeing Wednesdays: nurse-led clinic for improving physical health care in a general adolescent inpatient unit.
Young people with mental illness are at high risk of physical health complications. Physical healthcare on a general adolescent inpatient unit is complex. To establish a wellbeing clinic to improve efficiency and quality of the physical healthcare offered and increase health promotion. Plan, Do, Study, Act (PDSA) cycles were used to drive this quality-improvement project. The authors audited 12 records before establishing the clinic and 12 at three further time points (6, 18 and 30 months post-intervention) to guide changes. Results progressively improved over PDSA cycles. Time taken for initial investigations dropped. Compliance with medication monitoring and management of important physical health domains rose from zero in some cases to 100% in all but one area. Establishing a dedicated physical health clinic in this setting is feasible and leads to improved performance against local and national standards. Mental health teams need to ensure physical health is prioritised.


Tension-pCa relations of saponin-skinned rabbit and human heart muscle.
We tested if interaction between the thick and thin filaments, as revealed by the rate of cross-bridge cycling or the pattern of calcium activation, influences the relation between tension and free calcium concentration (pCa) of cardiac muscle. Bundles of rabbit ventricular cells which contained either fast-cycling RV1 myosin or slowly-cycling RV3 myosin and bundles of human atrial cells were chemically skinned by exposure to saponin. Skinned bundles were rapidly activated with calcium using the method of Moisescu and Thieleczek (1978). Our results suggest that the relation between pCa and tension is not different for skinned bundles of rabbit ventricular cells that contain either fast or slowly cycling myosin cross-bridges. The pattern of calcium activation, i.e. maximal to submaximal or submaximal to maximal, was also found not to influence the relation between tension and pCa for either rabbit ventricular or human atrial muscle. The tension-pCa relation of cardiac muscle was not altered by conditions that have been shown to alter the tension-pCa relation of skeletal muscle. These results suggest that the mechanisms responsible for regulating force development at the level of the thick and thin filaments that have been reported for skeletal muscle, are absent in cardiac muscle.


Refining mimicry: phenotypic variation tracks the local optimum.
Müllerian mimicry between chemically defended preys is a textbook example of natural selection favouring phenotypic convergence onto a shared warning signal. Studies of mimicry have concentrated on deciphering the ecological and genetic underpinnings of dramatic switches in mimicry association, producing a well-known mosaic distribution of mimicry patterns across geography. However, little is known about the accuracy of resemblance between natural comimics when the local phenotypic optimum varies. In this study, using analyses of wing shape, pattern and hue, we quantify multimodal phenotypic similarity between butterfly comimics sharing the so-called postman pattern in different localities with varying species composition. We show that subtle but consistent variation between populations of the localized species, Heliconius timareta thelxinoe, enhance resemblance to the abundant comimics which drive the mimicry in each locality. Those results suggest that rarer comimics track the changes in the phenotypic optimum caused by gradual changes in the composition of the mimicry community, providing insights into the process by which intraspecific diversity of mimetic pattern may arise. Furthermore, our results suggest a multimodal evolution of similarity, with coordinated convergence in different features of the phenotype such as wing outline, pattern and hue. Finally, multilocus genotyping allows estimating local hybridization rates between H. timareta and comimic H. melpomene in different populations, raising the hypothesis that mimicry refinement between closely related comimics may be enhanced by adaptive introgression at loci modifying the accuracy of resemblance.


Development of CD30 CAR-T cells in refractory or relapsed Hodgkin's lymphoma.
After therapy, approximately 15% of individuals with Hodgkin's lymphoma (HL) develop relapsed or drug-resistant Hodgkin's lymphoma (r/rHL). r/rHL has a high fatality rate and poor therapeutic prognosis. CD30 CAR-T-cell therapy has emerged as a new way to treat r/rHL in recent years. However, CD30CAR-T cells are still being explored in clinical trials. To help more patients, this review focuses on current CD30CAR-T-cell advancements as well as clinical breakthroughs in treatment of r/rHL. This research examines the mechanism of action of CD30 CAR-T cells, their function in the real-world therapy of r/rHL, and the influence of different treatment regimens on treatment results. There has been much research into CD30 CAR-T cells as a result of their successful use in treatment of r/rHL. This research has helped us to understand CD30 CAR-T-cell safety as well as the management options available before and after its administration to increase patient survival and reduce side effects.


To Feel in My Flesh: Receptivity, Resonance, Representation, and The Beta Screen.
When we are confronted with the challenge of trying to fully convey or describe something about human life and emotional experience, we find ourselves up against the very limitations of language. This problem becomes especially relevant as we attempt to expand psychoanalytic theory so as to enable us to "approach a mental life unmapped by the theories elaborated for the understanding of neurosis" (Bion1962, p. 37). This paper seeks to aid in that expansion by revisiting Bion's early writings about the beta screen, extending his conclusions about communication from the psychotic part of the mind to the broad area of the unrepresented (the unstructured unconscious), suggesting that there is often a potentially communicative meaning, a mute plea for intersubjective regulatory assistance (alpha function), embedded in the unconscious evocation of emotions in the object and that this cry for help may be encrypted in even the most seemingly destructive, resistant and oppositional patients.


Effects of sodium bicarbonate, magnesium oxide, and a commercial buffer mixture in early lactation cows fed hay crop silage.
Sixteen early lactation Holstein cows fed 70% concentrate: 30% hay crop silage were used to determine effects of .7% sodium bicarbonate, .7% sodium bicarbonate plus .28% magnesium oxide, or 1.8% commercial buffer mixture (total ration dry basis). This mixture contained a variety of buffers, alkalis, and other compounds known to affect milk production or composition in some circumstances. Buffers did not affect dry matter intake, milk yield, or milk composition but decreased efficiency of milk production. Ruminal fluid pH was not affected, but fecal pH and digestibilities of dry matter, organic matter, energy, acid detergent fiber, and cellulose were increased by the mixed buffers compared with sodium bicarbonate alone. Total ruminal volatile fatty acid concentration was reduced by buffers. Compared with sodium bicarbonate alone, mixed buffers increased ruminal ammonia concentration, acetate proportion, and acetate:propionate ratio and decreased proportions of propionate and butyrate. Valerate was reduced by all three buffers. Ruminal volume and liquid dilution rate were unaffected, but buffers increased total fluid outflow from the rumen. Higher amounts of buffers or alkalis may be necessary to offset low rumen pH and affect production with hay crop silage-based diets.


A phylogeny of the family Poritidae (Cnidaria, Scleractinia) based on molecular and morphological analyses.
The family Poritidae formerly included 6 genera: Alveopora, Goniopora, Machadoporites, Porites, Poritipora, and Stylaraea. Morphologically, the genera can be differentiated based on the number of tentacles, the number of septa and their arrangement, the length of the polyp column, and the diameter of the corallites. However, the phylogenetic relationships within and between the genera are unknown or contentious. On the one hand, Alveopora has been transferred to the Acroporidae recently because it was shown to be more closely related to this family than to the Poritidae by previous molecular studies. On the other hand, Goniopora is morphologically similar to 2 recently described genera, Machadoporites and Poritipora, particularly with regard to the number of septa (approximately 24), but they have not yet been investigated at the molecular level. In this study, we analyzed 93 samples from all 5 poritid genera and Alveopora using 2 genetic markers (the barcoding region of the mitochondrial COI and the ITS region of the nuclear rDNA) to investigate their phylogenetic relationships and to revise their taxonomy. The reconstructed molecular trees confirmed that Alveopora is genetically distant from all poritid genera but closely related to the family Acroporidae, whereas the other genera are genetically closely related. The molecular trees also revealed that Machadoporites and Poritipora were indistinguishable from Goniopora. However, Goniopora stutchburyi was genetically isolated from the other congeneric species and formed a sister group to Goniopora together with Porites and Stylaraea, thus suggesting that 24 septa could be an ancestral feature in the Poritidae. Based on these data, we move G. stutchburyi into a new genus, Bernardpora gen. nov., whereas Machadoporites and Poritipora are merged with Goniopora.


Clearance of senescent glial cells prevents tau-dependent pathology and cognitive decline.
Cellular senescence, which is characterized by an irreversible cell-cycle arrest1 accompanied by a distinctive secretory phenotype2, can be induced through various intracellular and extracellular factors. Senescent cells that express the cell cycle inhibitory protein p16INK4A have been found to actively drive naturally occurring age-related tissue deterioration3,4 and contribute to several diseases associated with ageing, including atherosclerosis5 and osteoarthritis6. Various markers of senescence have been observed in patients with neurodegenerative diseases7-9; however, a role for senescent cells in the aetiology of these pathologies is unknown. Here we show a causal link between the accumulation of senescent cells and cognition-associated neuronal loss. We found that the MAPTP301SPS19 mouse model of tau-dependent neurodegenerative disease10 accumulates p16INK4A-positive senescent astrocytes and microglia. Clearance of these cells as they arise using INK-ATTAC transgenic mice prevents gliosis, hyperphosphorylation of both soluble and insoluble tau leading to neurofibrillary tangle deposition, and degeneration of cortical and hippocampal neurons, thus preserving cognitive function. Pharmacological intervention with a first-generation senolytic modulates tau aggregation. Collectively, these results show that senescent cells have a role in the initiation and progression of tau-mediated disease, and suggest that targeting senescent cells may provide a therapeutic avenue for the treatment of these pathologies.


RT qLAMP-Direct Detection of SARS-CoV-2 in Raw Sewage.
A highly efficient, selective, and sensitive method for analysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in raw sewage was developed and tested to illustrate basic characteristics of the procedure. The method uses reverse transcriptase (RT) loop mediated isothermal amplification (LAMP) in a quantitative application, RT qLAMP. The applicability of this procedure to detection of SARS-CoV-2 in clinical samples has been documented in many reports since early 2020. Basic LAMP characteristics depending on the multiple primer design that produce highly selective and sensitive target amplification virtually free of interferences in complex sample media make it ideal for application to target recognition in raw sewage. Three previously described primer sets targeting ORF1a, E- and N-gene regions were selected and tested to define method performance characteristics and performance for SARS-CoV-2 detection in raw sewage samples from a municipal sewage system serving > 600 000, between July and October, 2020. The virus was detected in all samples from each of three independent interceptors near their treatment terminus. Virus quantities varied significantly between samples and between primer targets within samples. Sewage sampling dates corresponded to relatively low COVID-19 incidence rates reported by the local service area health department. The limited number of samples and aggregating downstream sampling locations did not permit resolving concentration differences. The most significant finding was the ability of the RT qLAMP method to detect SARS-CoV-2 in the raw sewage samples directly without preprocessing to isolate or concentrate the virus or to extract and concentrate viral RNA.


High-level expression of the prohormones proenkephalin, pro-neuropeptide Y, proopiomelanocortin, and beta-protachykinin for in vitro prohormone processing.
Prohormone substrates are required for investigation of the proteolytic processing of prohormones and proproteins into active peptide hormones and neurotransmitters. However, the lack of prohormone proteins has been a limiting factor in elucidating proteolytic mechanisms for conversion of prohormones into active peptides. Therefore, in this study, cloned cDNAs encoding the prohormones proenkephalin (PE), pro-neuropeptide Y (pro-NPY), pro-opiomelanocortin (POMC), and beta-protachykinin (beta-PT) were utilized to express recombinant prohormones in Escherichia coli. High-level expression of milligrams of prohormones was achieved with the pET3c expression vector utilizing the T7 promoter for production of PE, pro-NPY, and POMC, as demonstrated by SDS-PAGE gel electrophoresis, Western blots, and 35S-methionine labeling. In addition, beta-PT was expressed at high levels as fusion proteins with the maltose-binding protein and glutathione S-transferase by the pMAL-c and pGEX-2T expression vectors, respectively. Relative rates of processing by the established processing proteases "prohormone thiol protease" (PTP), 70-kDa aspartyl protease, and PC1/ 3 and PC2 (PC, prohormone convertase) were examined with purified PE, pro-NPY, and POMC. Distinct preferences of processing enzymes for different prohormones was demonstrated. PTP preferred PE and pro-NPY substrates, whereas little processing of POMC was detected. In contrast, the 70-kDa aspartyl protease cleaved POMC more readily than pro-NPY or PE. However, PC1/3 and PC2 prefer POMC as substrate. Demonstration of selectivity of processing enzymes for prohormone substrates illustrates the importance of expressing recombinant prohormones for in vitro processing studies.


Expression of the Na(+)-K(+)-2Cl- cotransporter BSC2 in the nervous system.
We used in situ hybridization and immunocytochemistry with polyclonal antibodies against the mouse bumetanide-sensitive Na(+)-K(+)-2Cl- cotransporter (mBSC2) to determine the location of this cotransporter in rat brain. Northern blots and in situ hybridization showed the presence of cotransporter mRNA in the brain, with an especially high level of expression in the choroid plexus (CP). Affinity-purified anti-BSC2 antibody identified proteins of 145-155 kDa on Western blot analysis and immunoprecipitation of brain and CP membrane protein. Indirect immunofluorescence demonstrated that BSC2 protein is located on the apical surface of the CP and is heterogeneously distributed in cell bodies and dendrites of neurons in the central and peripheral nervous system. The apical localization of BSC2 in the CP was confirmed by 86Rb+ uptakes in primary cultures of CP cells grown on permeable filters and confocal immunofluorescence microscopy. The apical localization of the cotransporter in CP epithelium suggests a role for the cotransporter in cerebrospinal fluid K+ homeostasis. In neurons, the cotransporter may help regulate intracellular Cl- concentration and thereby affect neuronal response to gamma-aminobutyric acid.


A Brownian dynamics tumor progression simulator with application to glioblastoma.
Tumor progression modeling offers the potential to predict tumor-spreading behavior to improve prognostic accuracy and guide therapy development. Common simulation methods include continuous reaction-diffusion (RD) approaches that capture mean spatio-temporal tumor spreading behavior and discrete agent-based (AB) approaches which capture individual cell events such as proliferation or migration. The brain cancer glioblastoma (GBM) is especially appropriate for such proliferation-migration modeling approaches because tumor cells seldom metastasize outside of the central nervous system and cells are both highly proliferative and migratory. In glioblastoma research, current RD estimates of proliferation and migration parameters are derived from computed tomography or magnetic resonance images. However, these estimates of glioblastoma cell migration rates, modeled as a diffusion coefficient, are approximately 1-2 orders of magnitude larger than single-cell measurements in animal models of this disease. To identify possible sources for this discrepancy, we evaluated the fundamental RD simulation assumptions that cells are point-like structures that can overlap. To give cells physical size (~10 μm), we used a Brownian dynamics approach that simulates individual single-cell diffusive migration, growth, and proliferation activity via a gridless, off-lattice, AB method where cells can be prohibited from overlapping each other. We found that for realistic single-cell parameter growth and migration rates, a non-overlapping model gives rise to a jammed configuration in the center of the tumor and a biased outward diffusion of cells in the tumor periphery, creating a quasi-ballistic advancing tumor front. The simulations demonstrate that a fast-progressing tumor can result from minimally diffusive cells, but at a rate that is still dependent on single-cell diffusive migration rates. Thus, modeling with the assumption of physically-grounded volume conservation can account for the apparent discrepancy between estimated and measured diffusion of GBM cells and provide a new theoretical framework that naturally links single-cell growth and migration dynamics to tumor-level progression.


Balloon kyphoplasty for painful C-7 vertebral hemangioma.
The authors report on the efficacy of balloon kyphoplasty applied to obliteration of symptomatic cervical hemangioma. This 49-year-old woman suffered from progressive neck pain, numbness of the upper extremities, and frequent headaches. Hemangioma of the C-7 vertebral body was diagnosed. Kyphoplasty was performed successfully through a right anterolateral approach. The patient became asymptomatic, and follow-up examinations confirmed complete recovery. To the authors' knowledge, this is the first description of cervical hemangioma treated by balloon kyphoplasty.


Evaluation of multi-solvent size exclusion chromatography columns packed with sub-2 μm particles for the characterization of synthetic polymers.
With the recent development of small particle stationary-phases and dedicated instrumentation, the combination of size-exclusion chromatography (SEC) with ultra-high performance liquid chromatography (UHPLC) technology has been realized. It opened up a new polymer analysis technique called UHP-SEC. Although high resolution and fast analysis can be achieved, the multi-solvent suitability for a given column was limited to either organic or aqueous eluents. In this work, the capability of novel SEC columns (AdvanceBio SEC columns) packed with 1.9 μm particles for the characterization of synthetic polymers in organic solvents as well as the multi-solvent compatibility for organic and aqueous eluents have been demonstrated. About six times faster separation for both polystyrene (PS) and polyethylene glycol (PEG) with good peak shape and repeatability were achieved in comparison with standard SEC columns at comparable resolution. Especially for PEG, in contrast to other SEC columns, this column could provide close-to-accurate determination of molecular weights with tetrahydrofuran (THF) as mobile phase. Good reproducibility was obtained after switching several times from water to THF and vice versa with RSD% in retention times less than 0.5 %. Different samples such as polyols, isocyanates and additives can also be analyzed for molecular weight and distribution or composition determination. Volume overload, especially with injection volumes higher than 10 µL needs to be considered. This new column offers a powerful choice for oligomer and polymer analysis with both aqueous and organic mobile phase. Ultimately, hyphenating SEC columns to various detectors can enable more information regarding chemical composition, molecular weight, concentration, and structure.


CCAAT/enhancer-binding protein beta (nuclear factor for interleukin 6) transactivates the human MDR1 gene by interaction with an inverted CCAAT box in human cancer cells.
We investigated the mechanisms of MDR1 gene activation by CCAAT/enhancer binding protein beta (C/EBPbeta, or nuclear factor for interleukin 6) in human cancer cells. Transfection of the breast cancer cell line MCF-7 and its doxorubicin-selected variant MCF-7/ADR by either C/EBPbeta or C/EBPbeta-LIP (a dominant-negative form of C/EBPbeta) confirmed their roles in the activation or repression of the endogenous, chromosomally embedded MDR1 gene. Cotransfection experiments with promoter constructs revealed a C/EBPbeta interaction on the MDR1 promoter via the region within -128 to -75. Deletions within the putative AP-1 box (-123 to -111) increased MDR1 promoter activity when stimulated by C/EBPbeta, suggesting that the AP-1 site negatively regulates MDR1 activation by C/EBPbeta. Mutations within the inverted CCAAT box (Y box) (-82 to -73) abolished the C/EBPbeta-stimulated MDR1 promoter activity, indicating that the Y box is required for MDR1 activation by C/EBPbeta. Chromatin immunoprecipitation (ChIP) revealed that C/EBPbeta precipitates a transcription complex containing C/EBPbeta, the MDR1 promoter sequences (-250 to +54), and the hBrm protein. In conclusion, alteration of expression or function of C/EBPbeta plays an important role in MDR1 gene regulation. C/EBPbeta activates the endogenous MDR1 gene of MCF-7 cells, and this activation was associated with a novel C/EBPbeta interaction region within the proximal MDR1 promoter (-128 to -75). The mechanisms of MDR1 activation by C/EBPbeta include C/EBPbeta binding of the chromatin of the MDR1 gene and interactions of C/EBPbeta with the Y box and Y box-associated proteins.


Effect of black tea intake on blood cholesterol concentrations in individuals with mild hypercholesterolemia: a diet-controlled randomized trial.
Habitual intake of black tea has been associated with relatively lower serum cholesterol concentrations in observational studies. However, clinical trial results evaluating the effects of black tea on serum cholesterol have been inconsistent. Several factors could explain these mixed results, in particular, uncontrolled confounding caused by lifestyle factors (eg, diet). This diet-controlled clinical trial estimates the effect of black tea flavonoid consumption on cholesterol concentrations in 57 borderline hypercholesterolemic individuals (total cholesterol concentrations between 190 and 260 mg/dL [4.9 and 6.7 mmol/L]). A double-blind, randomized crossover trial was conducted in Minneapolis, MN, from April 2002 through April 2004 in which key conditions were tightly controlled to minimize possible confounding. Participants consumed a controlled low-flavonoid diet plus 5 cups per day of black tea or tea-like placebo during two 4-week treatment periods. The flavonoid-free caffeinated placebo matched the tea in color and taste. Differences in cholesterol concentrations at the end of each treatment period were evaluated via linear mixed models. Differences among those treated with tea vs placebo were 3.43 mg/dL (0.09 mmol/L) (95% CI -7.08 to 13.94) for total cholesterol, -1.02 mg/dL (-0.03 mmol/L) (95% CI -11.34 to 9.30) for low-density lipoprotein cholesterol, 0.58 mg/dL (0.02 mmol/L) (95% CI -2.98 to 4.14) for high-density lipoprotein cholesterol, 15.22 mg/dL (0.17 mmol/L) (95% CI -40.91 to 71.35) for triglycerides, and -0.39 mg/dL (-0.01 mmol/L) (95% CI -11.16 to 10.38) for low-density lipoprotein plus high-density lipoprotein cholesterol fraction. The low-density lipoprotein cholesterol to high-density lipoprotein cholesterol ratio decreased by -0.1 units (95% CI -0.41 to 0.21). No results were statistically or clinically significant. The intake of 5 cups of black tea per day did not alter the lipid profile of borderline hypercholesterolemic subjects significantly.


Patient anxiety in the accident and emergency department.
Structured interviews were carried out with a sample of 96 patients to identify the sources of anxiety for patients in the accident and emergency (A & E) department. Only two patients reported that they were not at all anxious about any aspect of being in A & E. The average number of anxieties reported was 6.9. The most frequently reported anxieties were 'not being able to carry on your usual activities', 'not knowing what will happen to you in the department', 'having to undergo an uncomfortable procedure', 'feeling pain' and 'not knowing what is wrong'. The findings indicate that patients were as concerned with psychological and social aspects associated with admission as they were about physical factors. The authors suggest that it is essential for nurses in A & E to assess patients holistically and consider their psychological state and social circumstances as well as their physical condition. There is also a need for closer links to be developed between accident and emergency departments and other community health agencies and for more emphasis to be placed on referral to other bodies.


Impact evaluation of free delivery care on maternal health service utilisation and neonatal health in Nepal.
Nepal introduced free delivery services for births in public facilities in 2005 in 25 districts with the intervention initially restricted to women with less than two living children and/or women with obstetric complications. After November 2007, eligibility conditions were relaxed to include all women, and the programme was later expanded to cover an additional 50 districts in December 2008. We exploit the phased expansion of the free birth delivery programme to identify its impact on place of delivery, the presence of skilled birth attendants (SBAs) and neonatal mortality using difference-in-difference methods, on data for 4457 live-births reported between 2001 and 2008 from Nepal Demographic and Health Surveys for 2006 and 2011. Programme impacts were estimated for: (1) initial implementation until the relaxation of eligibility criteria to include all women in November 2007 (early phase); and (2) initial implementation until the programme was expanded nationwide in December 2008 (longer phase). Early implementing districts were treatment districts, while late implementing hill districts were control districts. In the early phase, the likelihood of delivery by SBAs was 5.6 percentage points higher (95%CI 0.002, 0.111) and the likelihood of delivery in a public facility was 5.1 percentage points higher (95%CI -0.003, 0.106) in treatment districts compared with control districts. The programme lowered the likelihood of neonatal mortality by 4.0 (-0.072, -0.009) percentage points for women with less than two living children and by 6.9 percentage points (95%CI -0.104, -0.035) for women from lower castes and indigenous groups in treatment districts compared with women in control districts, during the early phase. Programme effects on use of public facilities for births and deliveries attended by SBAs were not sustained over a longer exposure period. The results on neonatal mortality persisted with longer programme exposure, although the effects were smaller in magnitude.


[Loss of adsorptive ability--the reason for inactivation of the RM2 bacteriophage during storage].
The kinetics of bacteriophage PM2 inactivation at storage was compared with the kinetics of bacteriophage adsorption on Alteromonas espejiana BAL-31 host cells. Adsorption ability and infectivity are lost with the same rate at temperatures 4-28 degrees C suggesting the loss of adsorption ability to result in bacteriophage inactivation. At higher temperatures infectivity is lost more rapidly than the ability of adsorption. The single hit kinetics of adsorption ability loss suggests the simple model of independent inactivation of 12 antireceptors located at the tops of icosaedric capsid to be erroneous. At bacteriophage inactivation the major port of protein I, a fragment of antireceptors, is preserved in the capsid composition.


Diagnosis and classification of autoimmune uveitis.
Uveitis is the most common ophthalmological finding in the practice of rheumatology and clinical immunology. The condition is frequently idiopathic and defined by the inflammatory status of the uvea, the part of the middle eye that includes the iris, ciliary body and choroid. Anterior uveitis involves the iris and ciliary body, while the posterior form is limited to the retina and choroid. Both forms represent indications for an urgent evaluation by an ophthalmologist as untreated cases may cause blindness. Anterior uveitis is associated with the HLA-B27 allele and is a classification criterion for seronegative arthritis forms such as ankylosing spondylitis, psoriatic arthropathy, arthritis associated with Crohn's disease and ulcerative colitis, and reactive arthritis. Posterior uveitis is associated with Behcet's disease and HLA-B51. The clinical suspicion is raised by self-reported symptoms in the case of anterior involvement and floaters for posterior uveitis while the diagnosis, in the absence of established criteria, is made by an experienced ophthalmologist.


Transient increase in intrahepatic pressure mediates successful treatment of the Gunn rat with reduced doses of lentiviral vector.
Lentiviral vectors can stably transduce hepatocytes and are promising tools for gene therapy of hepatic diseases. Although hepatocytes are accessible to blood-borne viral vectors through fenestrations of the hepatic endothelium, improved liver transduction after delivery of vectors to the blood stream is needed. As the normal endothelial fenestration and lentiviral vectors are similar in size (150 nm), we hypothesized that a transient increase in hepatic blood pressure may enhance in vivo gene transfer to hepatocytes. We designed a simple surgical procedure, by which the liver is temporarily excluded from blood flow. Lentiviral vectors were injected in a large volume to increase intrahepatic pressure. We demonstrated that in the Gunn rat, a model of Crigler-Najjar disease, the administration of low vector doses (corresponding to a multiplicity of infection of 0.2) by this procedure resulted in therapeutic correction of hyperbilirubinemia, without toxicity. The correction was sustained for 10 months (end of study). The same vector amounts yielded only partial correction after intraportal delivery. We believe that this new and clinically applicable strategy may broaden the range of genetic liver diseases accessible to gene therapy.


Data-independent Acquisition Improves Quantitative Cross-linking Mass Spectrometry.
Quantitative cross-linking mass spectrometry (QCLMS) reveals structural detail on altered protein states in solution. On its way to becoming a routine technology, QCLMS could benefit from data-independent acquisition (DIA), which generally enables greater reproducibility than data-dependent acquisition (DDA) and increased throughput over targeted methods. Therefore, here we introduce DIA to QCLMS by extending a widely used DIA software, Spectronaut, to also accommodate cross-link data. A mixture of seven proteins cross-linked with bis[sulfosuccinimidyl] suberate (BS3) was used to evaluate this workflow. Out of the 414 identified unique residue pairs, 292 (70%) were quantifiable across triplicates with a coefficient of variation (CV) of 10%, with manual correction of peak selection and boundaries for PSMs in the lower quartile of individual CV values. This compares favorably to DDA where we quantified cross-links across triplicates with a CV of 66%, for a single protein. We found DIA-QCLMS to be capable of detecting changing abundances of cross-linked peptides in complex mixtures, despite the ratio compression encountered when increasing sample complexity through the addition of E. coli cell lysate as matrix. In conclusion, the DIA software Spectronaut can now be used in cross-linking and DIA is indeed able to improve QCLMS.


Reduced health resource use after acupuncture for low-back pain.
Acupuncture is commonly used to treat low-back pain (LBP) and clinical trials have demonstrated its efficacy. However, less is known about how the utilization of acupuncture impacts public health service utilization in the real world. This study investigates the association between acupuncture utilization for LBP and health care utilization by assessing whether patients who undergo acupuncture subsequently use fewer health care resources and whether those patients differ in their health care use from the general population with LBP. This study employed the design of a two-group pre/post secondary data analysis. There were two study populations. To identify patients who received acupuncture for LBP in 2000, patient charts at Alberta registered acupuncture clinics were reviewed. The comparison group was identified from the Alberta physician claims administrative database. Acupuncture group cases were matched with four comparison cases from the general population with LBP based on gender and age. Number of physician visits and physician service cost for LBP-related services for 1 year pre- and postacupuncture treatment period were calculated from the physician claims data for both study groups. For the 201 cases and 804 controls, the mean age was 48 years and 54% were female. The number of physician visits for the 1-year period postacupuncture decreased 49% for the acupuncture group (p<0.01) compared to the 1-year period preacupuncture. For the comparison there was a decrease of 2% in physician visits (p=0.59) for the same time periods. Corresponding to the decrease, physician services cost declined 37% for the case group (p=0.01) and 1% for the comparison (p=0.86). Results suggest that patients with LBP were less likely to visit physicians for LBP after acupuncture treatment. This led to reduced health services spending on LBP.


Partnering to provide simulated learning to address Interprofessional Education Collaborative core competencies.
Learning to effectively communicate and work with other professionals requires skill, yet interprofessional education is often not included in the undergraduate healthcare provider curriculum. Simulation is an effective pedagogy to bring students from multiple professions together for learning. This article describes a pilot study where nursing and social work students learned together in a simulated learning activity, which was evaluated to by the Readiness for Interprofessional Learning Scale (RIPLS). The RIPLS was used before and after the simulated activity to determine if this form of education impacted students' perceptions of readiness to learn together. Students from both professions improved in their RIPLS scores. Students were also asked to identify their interprofessional strengths and challenges before and after the simulation. Changes were identified in qualitative data where reports of strengths and challenges indicated learning and growth had occurred. In conclusion, this pilot study suggests that interprofessional simulation can be an effective method to integrate Interprofessional Education Collaborative core competencies into the curriculum.


ADAM9 enhances CDCP1 by inhibiting miR-1 through EGFR signaling activation in lung cancer metastasis.
MicroRNAs (miRNAs), which are endogenous short noncoding RNAs, can regulate genes involved in important biological and pathological functions. Therefore, dysregulation of miRNAs plays a critical role in cancer progression. However, whether the aberrant expression of miRNAs is regulated by oncogenes remains unclear. We previously demonstrated that a disintegrin and metalloprotease domain 9 (ADAM9) promotes lung metastasis by enhancing the expression of a pro-migratory protein, CUB domain containing protein 1 (CDCP1). In this study, we found that this process occurred via miR-1 down-regulation. miR-1 expression was down-regulated in lung tumors, but increased in ADAM9-knockdown lung cancer cells, and was negatively correlated with CDCP1 expression as well as the migration ability of lung cancer cells. Luciferase-based reporter assays showed that miR-1 directly bound to the 3'-untranslated region of CDCP1 and inhibited its translation. Treatment with a miR-1 inhibitor restored CDCP1 protein levels and enhanced tumor cell mobility. Overexpression of miR-1 decreased tumor metastases and increased the survival rate in mice. ADAM9 knockdown reduced EGFR signaling and increased miR-1 expression. These results revealed that ADAM9 down-regulates miR-1 via activating EGFR signaling pathways, which in turn enhances CDCP1 expression to promote lung cancer progression.


A simple method to determine hematoxylin ripening.
A simple spectroscopic method is proposed to control the "ripening" of Delafield's hematoxylin solution during natural "ripening" or fast oxidation by different agents. The hematoxylin solution has ripened sufficiently for use with tissue sections staining if the absorption is 1.2-1.3 at 560 nm. The hematoxylin solution must first be diluted 15-fold using a 5% ammonium alum solution and the absorption must be measured in 0.5 cm cuvettes.


Impact of Measuring Milk Production by Test Weighing on Breastfeeding Confidence in Mothers of Term Infants.
The duration of exclusive breastfeeding is affected by maternal confidence and perception of milk supply, but objective measurement of milk supply is rarely used. Mothers of preterm infants have found measuring milk supply by in-home test weighing to be helpful to ascertain their infants' breastfeeding intake. This study aimed to determine if this technique affects breastfeeding confidence of mothers of term infants. Participants filled in a survey of their breastfeeding confidence and perceptions of breastfeeding behavior prior to and after a 24-hour period of recording the times of feeds, amounts of each breastfeed or bottle feed (expressed breastmilk or formula), and amounts of breastmilk expressed. The breastfeeding confidence of at least 66% of mothers who were initially confident was maintained, and the breastfeeding confidence of at least 11% of mothers who were initially not confident was improved after objective measurement of milk supply and breastfeeding behavior. Measurement of actual milk supply and breastfeeding behavior provides objective information that can guide clinicians in the management of lactation. It can also help to maintain or improve breastfeeding confidence.