Interferon-gamma induction of the human leukocyte antigen-E gene is mediated through binding of a complex containing STAT1alpha to a distinct interferon-gamma-responsive element.
Expression of the human major histocompatibility complex (MHC) class I genes has been shown previously to increase at the transcriptional level following exposure to interferon-gamma (IFN-gamma). In this report we have examined the molecular mechanisms involved in the IFN-gamma-induced transcription of the human MHC class I gene, HLA-E. Functional analysis of CAT reporter gene constructs under the control of the HLA-E promoter transfected into U937 cells revealed the presence of a distinct IFN-gamma-responsive element, termed the interferon response region (IRR), that was necessary and sufficient to mediate the response to IFN-gamma. This cis-acting regulatory sequence contains an imperfect inverted repeat; the 5'-half of the IRR resembles the IFN-gamma activation site (GAS), and the 3'-half of the IRR resembles the interferon-stimulated response element (ISRE). Gel mobility shift assays demonstrated that the IRR bound a single, specific, IFN-gamma-induced complex (IRR-AC), which was formed rapidly following treatment with IFN-gamma and was independent of protein synthesis. Competition experiments with GAS and ISRE sequences from other IFN-inducible genes showed that GAS sequences competed for the IRR-AC, whereas ISRE sequences did not compete. Mutational analysis demonstrated that point mutations in either the 5'-half or 3'-half of the IRR prevented binding of the complex and abrogated or markedly reduced the IFN-gamma responsiveness of reporter gene constructs. Supershift analysis revealed that the IRR-AC contains a factor that was recognized by antibodies specific for the protein STAT1alpha (signal transducer and activator of transcription). Taken together, these findings suggest that the mechanism of IFN-gamma-induced transcription of the HLA-E gene is distinct from that of other MHC class I genes.


Organizational capacity and implementation change: a comparative case study of heart health promotion in Ontario public health agencies.
This paper reports the results of a comparative case study that examines factors influencing changes in implementation of heart health promotion activities in Ontario public health units. The study compared two cases that experienced large changes in implementation from 1994 to 1996, but in opposite directions. Multiple data sources were used, with an emphasis on secondary analyses of quantitative surveys of health units and other community agencies, and in-depth interviews of public health staff, collected as part of the Canadian Heart Health Initiative Ontario Project. Guided by social ecological and organizational theories, changes in implementation were explained by examining changes in (1) organizational predisposition to undertake heart health promotion activities, (2) organizational practices to undertake these activities, (3) other internal organizational factors and (4) external system factors. Findings show that in communities with diverse characteristics, implementation change was most strongly influenced by an interplay of changes in internal features of public health agencies; notably, leadership, structure and staff skills. Findings support a social ecological approach to health promotion by demonstrating the importance of the institutional context in the implementation change process, the interaction of individual (skills) and organizational (structure) levels in explaining implementation change, and community context in shaping the change process. Findings also reinforce the value of strengthening capacity within public health agencies and suggest further research on the implementation change process, especially in different systems and over longer periods of time.


[Basic Clinical Examination of the Shoulder].
Successful treatment of shoulder disease requires an accurate diagnosis. In addition to differentiated history taking, clinical examination is the most important component in the diagnosis of shoulder diseases. The present video explains the common provocation tests and functional tests that are used in the basic clinical examination of the shoulder. In addition to general inspection and palpation, the focus is on different diagnostic tests and clinical signs that improve diagnostic accuracy. The present basic clinical examination methods allow a structured approach to clinical issues and can be a good basis, if supplemented by further specific and individual tests.


Coping with crises: an examination of the impact of traumatic events on religious beliefs.
The impact of traumatic events on empirical and metaphysical assumptions was examined, by comparing assumptions of a group of 25 persons who had recently experienced a major stressor with assumptions of a group of 25 persons who had not had such an experience. Each group was composed of 22 women and 3 men, with a mean age of 20 years. Participants completed written measures assessing levels of adjustment, empirical world assumptions, religious motivation, and religious and spiritual experiences. Naturalistic interviews were conducted with the trauma group. The trauma group obtained significantly higher scores on symptoms of psychological distress but did not differ in evaluations of the empirical world as predictable, safe, or controllable. Interviews suggested that the metaphysical assumptions were not challenged by trauma; rather, they provided a framework for understanding and coping with trauma.


[Effect of limited motor activity on the alanine level of human blood plasma].
The plasma alanine concentration was measured in 28 healthy male test subjects exposed to head-down tilt of various duration (2-hour exposure at-12 degrees, 7-day exposure at-6 degrees, 49-day exposure at-4 degrees, and 120-day exposure at-4 degrees). Head-down tilt led to alanine changes that correlated with exposure time. These results suggest that alanine concentrations in blood reflect to a certain extent the rate of hepatic gluconeogenesis and depend significantly on the hypokinetic time. The findings can be clinically used as a measure of hepatic gluconeogenesis in bed-ridden patients that may require preventive or therapeutic treatment.


Changing patterns of internal migration 1970-1990: a comparative analysis of Jews and whites in the United States.
Independently conducted yet complementary sets of data from the 1970/1971 and 1990 National Jewish Population Surveys and the U.S. censuses of the same years were used to analyze changes in the internal migration of Jews and whites during the periods 1965-1970(1971) and 1985-1990. Interstate lifetime and five-year migration rates among Jews increased to levels significantly surpassing those of whites. Adjusting Jewish migration rates for the educational achievement of their white counterparts did not have much of an effect on lifetime migration or on the recent migration of the 1970/1971 Jewish population; however, it accounted meaningfully for the migration propensities of Jews in the period 1985-1990. These findings suggest that socioeconomic status has begun to play a larger role in promoting different migration patterns than in promoting ethnic group differences. Further, the direction of Jewish migrations followed those of whites (i.e., from the North-east and Midwest to the South and West): and due to their higher migration rates, Jews have considerably narrowed the regional distribution differences between themselves and whites. I interpret these results as evidence of the weakening role of ethnicity in present-day America.


Neuropsychological functioning during the year following severe traumatic brain injury.
The neuropsychological functioning of a group of 65 adults with severe traumatic brain injury was assessed at 6 months and 1 year post-injury. The cognitive domains assessed were pre-morbid intellectual level, current level of general intellectual functioning, simple and complex attention, verbal memory, executive functioning, and perceptual functioning. At least 40%, and up to 74%, of the TBI patients displayed some degree of impairment on tests administered at 6 months. Improvement was found to occur in all areas of cognitive functioning over the first year following injury. Despite this improvement at least 31%, and up to 63%, of TBI patients displayed some degree of impairment on tests administered at 1 year post-injury. The various types of neuropsychological functioning were affected to different degrees, indicating that different aspects of cognition are more susceptible to injury, and that recovery takes place at a differential rate across functions. The implications of these findings for the appropriate planning and allocation of treatment and rehabilitation resources, and the development of effective rehabilitation interventions are outlined.


Interferon at the cellular, individual, and population level in hepatitis C virus infection: Its role in the interferon-free treatment era.
The advent of powerful direct-acting antiviral agents (DAAs) has revolutionized the treatment of hepatitis C. DAAs cure nearly all patients with short duration, oral treatments. Significant efforts are now underway to optimize DAA-based treatments. We discuss the potential role of interferon in this optimization. Clinical studies present compelling evidence that DAAs perform better in treatment-naive individuals than in individuals who previously failed treatment with interferon, a surprising correlation because interferon and DAAs are thought to act independently. Recent mathematical models explore a mechanistic hypothesis underlying this correlation. The hypothesis invokes the action of interferon at the cellular, individual, and population levels. Strong interferon responses prevent the productive infection of cells, reduce viral replication, and impede the development of resistance to DAAs in infected individuals and improve cure rates elicited by DAAs in treated populations. The models develop descriptions of these processes, integrate them into a comprehensive framework, and capture clinical data quantitatively, providing a successful test of the hypothesis. Individuals with strong endogenous interferon responses thus present a promising subpopulation for reducing DAA treatment durations. This review discusses the conceptual advances made by the models, highlights the new insights they unravel, and examines their applicability to optimize DAA-based treatments.


The Sir4 C-terminal coiled coil is required for telomeric and mating type silencing in Saccharomyces cerevisiae.
Saccharomyces cerevisiae Sir4p plays important roles in silent chromatin at telomeric and silent mating type loci. The C terminus of Sir4p (Sir4CT) is critical for its functions in vivo because over-expression or deletion of Sir4CT fragments disrupts normal telomeric structure and abolishes the telomere position effect. The 2.5A resolution X-ray crystal structure of an Sir4CT fragment (Sir4p 1217-1358) reveals a 72 residue homodimeric, parallel coiled coil, burying an extensive 3600A(2) of surface area. The crystal structure is consistent with results of protein cross-linking and analytical ultracentrifugation results demonstrating that Sir4CT exists as a dimer in solution. Disruption of the coiled coil in vivo by point mutagenesis results in total derepression of telomeric and HML silent mating marker genes, suggesting that coiled coil dimerization is essential for Sir4p-mediated silencing. In addition to the coiled coil dimerization interface (Sir4CC interface), a crystallographic interface between pairs of coiled coils is significantly hydrophobic and buries 1228A(2) of surface area (interface II). Remarkably, interface II mutants are deficient in telomeric silencing but not in mating type silencing in vivo. However, point mutants of interface II do not affect the oligomerization state of Sir4CT in solution. These results are consistent with the hypothesis that interface II mimics a protein interface between Sir4p and one of its protein partners that is essential for telomeric silencing but not mating type silencing.


Kearns-Sayre syndrome caused by defective R1/p53R2 assembly.
Mutations in RRM2B encoding ribonucleotide reductase (RNR) p53R2 subunit usually cause paediatric-onset mitochondrial disease associated with mitochondrial DNA (mtDNA) depletion. The importance of RNR dysfunction in adult mitochondrial disease is unclear. To report the RRM2B mutation frequency in adults with multiple mtDNA deletions and examine RNR assembly in a patient with Kearns-Sayre syndrome (KSS) caused by two novel RRM2B mutations. 50 adult patients with multiple mtDNA deletions in skeletal muscle were studied. DNA sequencing of RRM2B was performed in patients without mutations in mtDNA maintenance genes POLG and C10orf2. RNR protein was studied using western blot and Blue-native polyacrylamide gel electrophoresis (BN-PAGE). Four per cent (two unrelated cases) of this adult cohort harboured RRM2B mutations. Patient 1 had KSS and two novel missense mutations: c.122G→A; p.Arg41Gln and c.391G→A; p.Glu131Lys. BN-PAGE demonstrated reduced heterotetrameric R1/p53R2 RNR levels compared with controls, despite normal steady-state p53R2 levels on western blot, suggesting failed assembly of functional RNR as a potential disease mechanism. Patient 2 had late-onset progressive external ophthalmoplegia and fatigue. A heterozygous deletion c.253_255delGAG; p.Glu85del was identified. Muscle histology in both cases showed significant numbers of necrotic muscle fibres, possibly indicating enhanced apoptotic cell death. These data indicate that 4% of adult mitochondrial disease with multiple deletions is caused by RNR dysfunction. KSS has not previously been linked to a nuclear gene defect. Evidence that disease pathogenesis may be caused by defective RNR assembly is given. RRM2B screening should be considered early in the differential diagnosis of adults with multiple mtDNA deletions.


CD10 expression in urothelial bladder carcinomas: a pilot study.
The aims of this study were to investigate the expression of CD10 in normal bladder tissue and urothelial bladder carcinomas and to clarify its association with histopathological variables. A total of 79 urothelial bladder carcinomas were selected from routine archival material. All cases were reevaluated histopathologically and graded according to the World Health Organization (WHO) 1973, WHO/ISUP 1998, and WHO 1999 systems. The TNM system was used for their pathological staging. CD10 immunohistochemical staining was performed in selected slides. Tumoral cases consisted of 74 men (93.7%) and 5 women (6.3%). According to the pathological stage, 25 (31.6%), 33 (41.8%), and 21 (26.6%) cases had pTa, pT1, and pT2-3 carcinomas, respectively. 34 of 79 (43%) urothelial carcinomas and only 1 of 11 (9.1%) nontumoral cases showed positive CD10 immunostaining. It was a cytoplasmic diffuse or granular immunostaining pattern both in nontumoral and tumoral urothelia. There was no statistically significant difference between tumoral and nontumoral cases with respect to CD10 reactivity (p = 0.051), but there was a trend toward significance. In urothelial tumors, there was a significant inverse correlation between pathological stages and CD10 immunoreactivity (p = 0.036, r = -0,237). There was also a statistically significant difference between pTa and pT2-3 urothelial tumors in relation to the CD10 expression (p = 0.034). No association was detected between CD10 expression and grades according to all systems used (p > 0.05). According to our findings, the CD10 expression in noninvasive carcinomas showed a higher level than that in invasive carcinomas, and it is inversely correlated with the pathological stage. CD10 may play an important role in the progression of urothelial bladder carcinomas, and downregulation probably facilitates invasion, especially muscle invasion.


[Expectations of the patient's role].
This study is part of a research on sociological aspects of the doctor-patient relationship. Some previous results have been already published. This paper aims to analyse the influence of some basic variables such as age, sex and socioeconomic level on normative expectations concerning to the patient's role. This role was defined in terms of Parsons' sociological theory, which provides the content of the 20 items of a Likert-type scale constructed by Kassebaum and Baumann. By means of a factorial analysis they discovered four underlying dimensions of the expectations of the patient's role: Dependence, Reciprocity, Role performance and Denial. This scale and a questionnaire built by the author were given to a sample group of 302 subjects, present or potential patients, inhabitants of Buenos Aires city. The results show that expectations of the patient's role are influenced mainly by the subject's socioeconomic level.


Erythroid burst-promoting activity of purified recombinant human GM-CSF and interleukin-3: studies with anti-GM-CSF and anti-IL-3 sera and studies in serum-free cultures.
We studied the erythroid burst-promoting activity (BPA) of recombinant human granulocyte/macrophage colony-stimulating factor (GM-CSF) and Interleukin-3 (IL-3) with two experimental approaches. First we studied the effects of polyclonal antisera prepared against human GM-CSF and gibbon IL-3 on colony formation from 1,000 bone marrow null cells/dish in serum-containing culture. Both GM-CSF and IL-3 independently enhanced erythroid burst formation; however, IL-3 showed more BPA activity than GM-CSF. These data are in agreement with an emerging view that the primary targets of IL-3 are primitive progenitors and that the targets of GM-CSF are intermediate progenitors, including erythroid burst-forming units (BFU-E). The proliferation of one population of BFU-E was independent of GM-CSF or IL-3. To characterize this population of BFU-E further, we developed a serum-free culture assay for the purified progenitors by incorporating insulin-like growth factor-1 (IGF-1) to the serum-free medium. The development of erythroid bursts was supported by IL-3, IGF-1, and erythropoietin (Ep) in a serum-free culture system and to a lesser extent by the combination of GM-CSF, IGF-1, and Ep. Although the burst-promoting ability of GM-CSF and IL-3 was again demonstrated in this system, unlike serum-containing culture Ep alone did not support burst formation. These results indicate that when fetal calf serum (FCS) is present, the culture system contains BPA that is not GM-CSF or IL-3.


Etoposide and cisplatin versus paclitaxel and carboplatin with concurrent thoracic radiotherapy in unresectable stage III non-small cell lung cancer: a multicenter randomized phase III trial.
The optimal chemotherapy regimen administered currently with radiation in patients with stage III non-small cell lung cancer (NSCLC) remains unclear. A multicenter phase III trial was conducted to compare the efficacy of concurrent thoracic radiation therapy with either etoposide/cisplatin (EP) or carboplatin/paclitaxel (PC) in patients with stage III NSCLC. Patients were randomly received 60-66 Gy of thoracic radiation therapy concurrent with either etoposide 50 mg/m2 on days 1-5 and cisplatin 50 mg/m2 on days 1 and 8 every 4 weeks for two cycles (EP arm), or paclitaxel 45 mg/m2 and carboplatin (AUC 2) on day 1 weekly (PC arm). The primary end point was overall survival (OS). The study was designed with 80% power to detect a 17% superiority in 3-year OS with a type I error rate of 0.05. A total of 200 patients were randomized and 191 patients were treated (95 in the EP arm and 96 in the PC arm). With a median follow-up time of 73 months, the 3-year OS was significantly higher in the EP arm than that of the PC arm. The estimated difference was 15.0% (95% CI 2.0%-28.0%) and P value of 0.024. Median survival times were 23.3 months in the EP arm and 20.7 months in the PC arm (log-rank test P = 0.095, HR 0.76, 95%CI 0.55-1.05). The incidence of Grade ≥2 radiation pneumonitis was higher in the PC arm (33.3% versus 18.9%, P = 0.036), while the incidence of Grade ≥3 esophagitis was higher in the EP arm (20.0% versus 6.3%, P = 0.009). EP might be superior to weekly PC in terms of OS in the setting of concurrent chemoradiation for unresectable stage III NSCLC. NCT01494558.


Use of the 'patient journey' model in the internet-based pre-fitting counseling of a person with hearing disability: study protocol for a randomized controlled trial.
Hearing impairment is one of the most frequent chronic conditions. Persons with a hearing impairment (PHI) have various experiences during their 'journey' through hearing loss. In our previous studies we have developed a 'patient journey' model of PHI and their communication partners (CPs). We suggest this model could be useful in internet-based pre-fitting counseling of a person with hearing disability (PHD). A randomized controlled trial (RCT) with waiting list control (WLC) design will be used in this study. One hundred and fifty eight participants with self-reported hearing disability (that is, score > 20 in the Hearing Handicap Questionnaire (HHQ)) will be recruited to participate in this study. They will be assigned to one of two groups (79 participants in each group): (1) Information and counseling provision using the 'patient journey' model; and (2) WLC. They will participate in a 30 day (4 weeks) internet-based counseling program based on the 'patient journey' model. Various outcome measures which focuses on hearing disability, depression and anxiety, readiness to change and acceptance of hearing disability will be administered pre (one week before) and post (one week and six months after) intervention to evaluate the effectiveness of counseling. Internet-based counseling is being introduced as a viable option for audiological rehabilitation. We predict that the 'patient journey' model will have several advantages during counseling of a PHD. Such a program, if proven effective, could yield cost and time-efficient ways of managing hearing disability. ClinicalTrials.gov Protocol Registration System NCT01611129.


Inactivity-induced remodeling of neuromuscular junctions in rat diaphragmatic muscle.
We hypothesized that inactivity-induced remodeling of neuromuscular junctions (NMJs) depends on fiber type and the match between muscle fiber and motoneuron (MN) activities. Two inactivity models were studied in rat diaphragmatic muscle: spinal hemisection at C2 (SH), where both diaphragmatic muscle fibers and phrenic MNs were inactive, and tetrodotoxin (TTX) nerve blockade, where only muscle fibers were inactive. After 2 weeks of inactivity, there was increased number of pre- and postsynaptic branches (fragmentation) of NMJs at type IIx/b fibers in both models. In addition, planar NMJ areas at type IIx/b fibers in the SH model were enlarged. In contrast, NMJs at type I and IIa fibers were unaffected in both SH and TTX models. Functionally, neuromuscular transmission in diaphragmatic muscle fibers improved in the SH model, but worsened in the TTX model, compared to controls. These results suggest that NMJ remodeling depends on the level of MN activity. The relative preservation of NMJs at type I and IIa fibers suggests a potential for recovery from diaphragmatic paralysis in the clinical setting, at least for respiratory behaviors.


Overexpression and Nucleolar Localization of γ-Tubulin Small Complex Proteins GCP2 and GCP3 in Glioblastoma.
The expression, cellular distribution, and subcellular sorting of the microtubule (MT)-nucleating γ-tubulin small complex (γTuSC) proteins, GCP2 and GCP3, were studied in human glioblastoma cell lines and in clinical tissue samples representing all histologic grades of adult diffuse astrocytic gliomas (n = 54). Quantitative real-time polymerase chain reaction revealed a significant increase in the expression of GCP2 and GCP3 transcripts in glioblastoma cells versus normal human astrocytes; these were associated with higher amounts of both γTuSC proteins. GCP2 and GCP3 were concentrated in the centrosomes in interphase glioblastoma cells, but punctate and diffuse localizations were also detected in the cytosol and nuclei/nucleoli. Nucleolar localization was fixation dependent. GCP2 and GCP3 formed complexes with γ-tubulin in the nucleoli as confirmed by reciprocal immunoprecipitation experiments and immunoelectron microscopy. GCP2 and GCP3 depletion caused accumulation of cells in G2/M and mitotic delay but did not affect nucleolar integrity. Overexpression of GCP2 antagonized the inhibitory effect of the CDK5 regulatory subunit-associated tumor suppressor protein 3 (C53) on DNA damage G2/M checkpoint activity. Tumor cell GCP2 and GCP3 immunoreactivity was significantly increased over that in normal brains in glioblastoma samples; it was also associated with microvascular proliferation. These findings suggest that γTuSC protein dysregulation in glioblastomas may be linked to altered transcriptional checkpoint activity or interaction with signaling pathways associated with a malignant phenotype.


Distribution of coronary artery disease in patients with isolate aortic valve disease.
Forty-four adult patients with isolated aortic valve disease underwent coronary arteriography. Seventy precent of patients required aortic valve replacement (AVR). Of 28 men who underwent AVR, 59 percent of those with severe aortic stenosis, 33 percent of those with severe aortic regurgitation and 50 percent of those with mixed aortic stenosis/aortic regurgitation had associated coronary artery disease. Thirty-six percent of men with aortic valve disease not requiring AVR had significant coronary artery disease (CAD). CAD was not found in the 5 female patients studied. In the patients with CAD, the left anterior descending artery was involved 84 percent of the time. Next in frequency were the right coronary artery, the proximal left circumflex artery, and the obtuse marginal artery. Multivessel disease was the rule. All but one patient with significant CAD had angina pectoris, but many patients with angina had normal coronary arteries. The frequent occurence of significant CAD in adult male patients with isolated aortic valve disease argues for the performance of selective coronary arteriography in all such patients in whom AVR is a consideration.


Establishing a children's orthopaedic hospital for Malawi: A review after 10 years.
BEIT CURE International Hospital (BCIH) opened in 2002 providing orthopaedic surgical services to children in Malawi. This study reviews the hospital's progress 10 years after establishment of operational services. In addition we assess the impact of the hospital's Malawi national clubfoot programme (MNCP) and influence on orthopaedic training. All operative paediatric procedures performed by BCIH services in the 10th operative year were included. Data on clubfoot clinic locations and number of patients treated were obtained from the MNCP. BCIH records were reviewed to identify the number of healthcare professionals who have received training at the BCIH. 609 new patients were operated on in the 10th year of hospital service. Patients were treated from all regions; however 60% came from Southern regions compared with the 48% in the 5th year. Clubfoot, burn contracture and angular lower limb deformities were the three most common pathologies treated surgically. In total BCIH managed 9,842 patients surgically over a 10-year period. BCIH helped to establish and co-ordinate the MNCP since 2007. At present the program has a total of 29 clinics, which have treated 5748 patients. Furthermore, BCIH has overseen the full or partial training of 5 orthopaedic surgeons and 82 orthopaedic clinical officers in Malawi. The BCIH has improved the care of paediatric patients in a country that prior to its establishment had no dedicated paediatric orthopaedic service, treating almost 10,000 patients surgically and 6,000 patients in the MNCP. This service has remained consistent over a 10-year period despite times of global austerity. Whilst the type of training placement offered at BCIH has changed in the last 10 years, the priority placed on training has remained paramount. The strategic impact of long-term training commitments are now being realised, in particular by the addition of Orthopaedic surgeons serving the nation.


Phonological Assessment Instrument: evidence of content validity and response processes.
to demonstrate the validity of content and the validity of response processes of an instrument intended for the phonological assessment of children. validation was carried out in two stages by two different groups of judges, a group of specialists and a group of non-specialists. The first group, composed of three expert judges, evaluated the 123 lexical items after creating the instrument, judging the applicability of the figures in the context of child assessment, and suggesting adjustments to compose the content. From the observations, the instrument was adapted and directed to the group of non-specialist judges who, through the application of the instrument, had their responses evaluated according to the ease or difficulty of eliciting the instrument's items. The predictions obtained positive results for content validity and response processes. the study allowed to improve the test items more judiciously, benefiting clinical and scientific use.


High-throughput high-volume nuclear imaging for preclinical in vivo compound screening§.
Preclinical single-photon emission computed tomography (SPECT)/CT imaging studies are hampered by low throughput, hence are found typically within small volume feasibility studies. Here, imaging and image analysis procedures are presented that allow profiling of a large volume of radiolabelled compounds within a reasonably short total study time. Particular emphasis was put on quality control (QC) and on fast and unbiased image analysis. 2-3 His-tagged proteins were simultaneously radiolabelled by 99mTc-tricarbonyl methodology and injected intravenously (20 nmol/kg; 100 MBq; n = 3) into patient-derived xenograft (PDX) mouse models. Whole-body SPECT/CT images of 3 mice simultaneously were acquired 1, 4, and 24 h post-injection, extended to 48 h and/or by 0-2 h dynamic SPECT for pre-selected compounds. Organ uptake was quantified by automated multi-atlas and manual segmentations. Data were plotted automatically, quality controlled and stored on a collaborative image management platform. Ex vivo uptake data were collected semi-automatically and analysis performed as for imaging data. >500 single animal SPECT images were acquired for 25 proteins over 5 weeks, eventually generating >3500 ROI and >1000 items of tissue data. SPECT/CT images clearly visualized uptake in tumour and other tissues even at 48 h post-injection. Intersubject uptake variability was typically 13% (coefficient of variation, COV). Imaging results correlated well with ex vivo data. The large data set of tumour, background and systemic uptake/clearance data from 75 mice for 25 compounds allows identification of compounds of interest. The number of animals required was reduced considerably by longitudinal imaging compared to dissection experiments. All experimental work and analyses were accomplished within 3 months expected to be compatible with drug development programmes. QC along all workflow steps, blinding of the imaging contract research organization to compound properties and automation provide confidence in the data set. Additional ex vivo data were useful as a control but could be omitted from future studies in the same centre. For even larger compound libraries, radiolabelling could be expedited and the number of imaging time points adapted to increase weekly throughput. Multi-atlas segmentation could be expanded via SPECT/MRI; however, this would require an MRI-compatible mouse hotel. Finally, analysis of nuclear images of radiopharmaceuticals in clinical trials may benefit from the automated analysis procedures developed.


Pharmacokinetic and toxicology comparator testing of biosimilar drugs - Assessing need.
A key element in the development of a biosimilar molecule is the comparability of the biological activity/nonclinical similarity to the innovator drug. Although some regulatory guidelines are encouraging little or no in vivo testing, currently a common practice is to perform at least one toxicology and/or one pharmacokinetic (PK) study to assess if any different findings occur for in-life, clinical pathology and histopathological parameters or in exposure. An exercise was performed in which the results of such testing were evaluated. It was found that 10 PK comparison studies in the cynomolgus monkey across 4 monoclonal (Mab) classes showed similar exposure in all cases. In 17 toxicology comparison studies with 5 Mab classes performed in the same species and in 7 toxicology comparison studies with non-Mab biosimilars in the rat, no new/unexpected findings were seen and drug exposure measurement gave comparable values in all cases. Overall, although this work does not rule out possible utility of some in vivo testing (notably in the form of stand-alone PK testing) to confirm similar exposure between the 2 molecules tested, it is unclear what benefit can be gained from toxicology testing, especially if comparability has been demonstrated from physiochemical and in vitro characterisation.


Calcium and EDTA fluxes in dialyzed squid axons.
Ca efflux in dialyzed squid axons was measured with 45Ca as a function of internal ionized Ca in the range 0.005-10 muM. Internal Ca stores were depleted by treatment with CN and dialysis with media free of high energy compounds. The [Ca]iota was stabilized with millimolar concentrations of EDTA, EGTA, or DTPA. Nonspecific leak of chelated Ca was measured with [14C]-EDTA and found to be 0.02 pmol/cm2s/mM EDTA. Correction of the measured Ca efflux for this leak of chelated calcium was made when appropriate. Ca efflux was roughly linear with internal free Ca in the range 0.005-0.1 muM. Above 0.1 muM, efflux was less than proportional to concentration but did not saturate at the highest concentration studied. Ca efflux was reduced about 50% by replacement of external Na with Li at Caiota approximately 1 muM, but was insensitive to such replacement for Ca less than 0.1 muM. Ca efflux was insensitive to internal Mg in the range 0-4 mM, indicating that the Ca pump favors Ca over Mg by a factor of about 10(6). Ca efflux was reduced about 60% by increasing internal Na from 1 to 80 mM. This effect could represent weak interference of a Ca carrier by Na or a loss of driving force because of a reduction in ENa - Em occasioned by an increase in Naiota. A few measurements were made of Ca influx in intact and in dialyzed fibers. In both cases, Ca influx increased when external Na was replaced by Li.


Lactose supports Muller cell protein expression patterns in the absence of the retinal pigment epithelium.
We have previously shown that lactose, but not mannose, promotes the assembly of nascent photoreceptor outer segments in the absence of the retinal pigment epithelium (RPE). The purpose of the present study was to determine if, in addition to the improved outer segment assembly observed in the presence of lactose, biosynthetic changes in Muller cells could also be detected. The RPE was removed from intact isolated Xenopus embryonic eyes that were allowed to complete differentiation in Niu-Twitty medium, Niu-Twitty with mannose, or Niu-Twitty with lactose. Control retinas matured in vitro with an adherent RPE. Retinal morphology was evaluated for organized folding of outer segment membranes and cell loss. In addition, the expression of three Muller cell proteins, glial fibrillary acidic protein (GFAP), cellular retinaldehyde-binding protein (CRALBP), and glutamine synthetase, was examined. In control retinas, GFAP is undetectable, CRALBP heavily immunolabels Muller cells, and radial patterns of glutamine synthetase immunoreactivity are present. In the absence of the RPE, Muller cells upregulate GFAP expression, CRALBP labeling is present at a slightly reduced level, and glutamine synthetase immunolabeling is negligible. Neither mannose nor lactose modify significantly the expression of CRALBP. Similarly, both compounds completely prevent the upregulation of GFAP. However, normal glutamine synthetase expression was observed only in the presence of lactose, but not in the presence of mannose. Statistical analyses of slot blot-based protein quantification confirmed our immunochemical results. In RPE-deprived retinas supplemented with lactose, Muller cells were morphologically normal. The proper photoreceptor outer segment morphogenesis observed under these conditions was uniquely associated with normal levels of glutamine synthetase expression. The exact significance of this finding with respect to photoreceptor outer segment morphogenesis is unknown. We suggest that glutamine synthetase may be a marker of Muller cell metabolic or structural integrity that may reflect the enhanced ability of these cells, in the presence of lactose, to support photoreceptor outer segment morphogenesis.


The receptor for urokinase-type plasminogen activator is deficient on peripheral blood leukocytes in patients with paroxysmal nocturnal hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal defect in bone marrow-derived cells and is clinically associated with intravascular hemolysis, hemoglobinuria, and an increased frequency of venous thrombosis. The common denominator of PNH-affected blood cells appears to be a defect in the membrane attachment of proteins normally anchored by glycosyl-phosphatidylinositol (GPI). We report here that the cellular receptor for urokinase-type plasminogen activator (u-PAR) is deficient on affected peripheral blood monocytes and granulocytes from four individuals with PNH as evidenced by chemical cross-linking analysis as well as by immunofluorescence flow cytometry using a monoclonal anti-u-PAR antibody. In contrast, on normal blood monocytes and granulocytes we find significant amounts of u-PAR, which is attached to the plasma membrane by a GPI-anchor as defined by its sensitivity towards a specific phospholipase treatment. By two-color flow cytometry it was shown that deficiency of u-PAR expression paralleled that of another GPI-anchored protein. As u-PAR is involved in the initiation of pericellular proteolysis, the reduced expression of u-PAR on PNH-affected leukocytes led to an overall reduction in the capacity for plasminogen activation by cell-surface-bound urokinase. Whereas the abnormal susceptibility of PNH-affected erythrocytes to lysis by autologous complement has been related to the low expression of three GPI-anchored complement regulatory proteins on the cell surface, we now propose that lack of u-PAR expression on the surface of peripheral blood leukocytes may be causally related to the high incidence of venous thrombosis observed in PNH patients.


Knowledge in health technology assessment: who, what, how?
Health systems are placing more and more emphasis on designing and delivering services that are focused on the patient, and there is a growing interest in patient aspects of health policy research and health technology assessment (HTA). Only a few HTA agencies use and invest in scientific methods to generate knowledge and evidence about the patient aspects of a given technology. This raises questions about how knowledge is produced in HTA reports and what kind of knowledge is considered relevant. This article uses a Danish HTA on patient education from 2009 as empirical material for a critical examination and discussion of knowledge and knowledge production about the patient aspects of HTA.


[How pediatric residents involve children during medical decision-making?].
To describe how pediatric residents involve children during medical decision-making and evaluate the relationship between practice patterns and residents characteristics. We conducted a prospective multicenter anonymous written survey. Self-reported involvement of children by 45 French pediatric residents in practice pattern was collected and analysed. Most residents reported they informed patients in more than 50% of the cases (82%). Only a minority of the residents asked for consent, respected children refusal or presented other choices to the treatment. The main reasons that explain the lack of partnership are children incompetence and the medical situation. No statistically significant relationship between practice patterns and residents characteristics was found. Partnership with children varies across residents and according to the level of involvement considered. No statistical differences were obtained to explain variations between residents' attitude toward involvement of children. Nevertheless medical education in ethics or decision-making could increase partnership with children.


Determination of Spatially Resolved Tablet Density and Hardness Using Near-Infrared Chemical Imaging (NIR-CI).
Near-infrared chemical imaging (NIR-CI) combines spectroscopy with digital imaging, enabling spatially resolved analysis and characterization of pharmaceutical samples. Hardness and relative density are critical quality attributes (CQA) that affect tablet performance. Intra-sample density or hardness variability can reveal deficiencies in formulation design or the tableting process. This study was designed to develop NIR-CI methods to predict spatially resolved tablet density and hardness. The method was implemented using a two-step procedure. First, NIR-CI was used to develop a relative density/solid fraction (SF) prediction method for pure microcrystalline cellulose (MCC) compacts only. A partial least squares (PLS) model for predicting SF was generated by regressing the spectra of certain representative pixels selected from each image against the compact SF. Pixel selection was accomplished with a threshold based on the Euclidean distance from the median tablet spectrum. Second, micro-indentation was performed on the calibration compacts to obtain hardness values. A univariate model was developed by relating the empirical hardness values to the NIR-CI predicted SF at the micro-indented pixel locations: this model generated spatially resolved hardness predictions for the entire tablet surface.


Data Sharing: HOW to Tell Your Story.
This is the second in a series of articles looking at school health data collection from identification of data points to utilizing data to share your story and submit your data to contribute to the National School Health Data Set: Every Student Counts! This article focuses on using data to share your story. Data storytelling versus data visualization will be discussed as well as HOW schools nurses can utilize easy access programs to support this process. Building on the first article in the series, the school nurse will not only identify the WHY and WHAT related to data collection but also HOW to link school health data to educational data to increase the audience of the story and follow data sharing regulations.


Random selection: a model for poliovirus infection of the central nervous system.
Mixed infections occur in the natural environment, and also result from the use of mixed live vaccines. Some recipients of the trivalent oral poliovirus vaccine develop vaccine-associated paralytic poliomyelitis (VAPP). Numerous serotypes and recombinant genotypes of vaccine-derived polioviruses may be found in stool samples from such cases. To investigate the relationship between the multiplication of various genotypes at the primary replication site in the gut and the infection outcome in the central nervous system (CNS), the viruses excreted on consecutive days by two patients with VAPP were compared with the viruses isolated from the CNS. The genotypes from stools were numerous and varied with time in both cases, suggesting a multiplication of the viruses in multiple foci in the gut. Where the CNS isolated virus clearly corresponded to one of the many viruses detected in stool, this virus was unexpectedly less neurovirulent than others isolated from stool. To assess the mechanism by which viruses with different degrees of neurovirulence are selected in the CNS, transgenic mice sensitive to poliovirus infection were inoculated extraneurally with mixtures of two phenotypically different viruses at different neuropathogenic doses. The virus(es) inducing neurological disease was then isolated from the CNS. At less than 100% input neuropathogenic dose of both inoculated viruses, individual mice were affected stochastically by the virus variants from the mixture. Extrapolated to humans, this selection pattern might explain the occurrence of CNS infections with less neurotropic viruses derived from an extraneural pool containing also highly neurotropic viruses.


Dynamic right ventricular outflow obstruction: A rare cause of hypotension during anestesia induction.
Dynamic obstruction of right ventricle outflow tract (RVOTO) is a rare condition that may acutely cause severe heart failure. It has been reported in some hypertrophic cardiomyopathies, after lung transplantation, and in some cases of hemodynamic instability after cardiopulmonary bypass. We report the case of a 71-year-old man who developed severe hypotension during the induction of general anesthesia for surgical coronary revascularization. Hypotension did not respond to the initial treatment with vasoconstrictors and fluids. RVOTO was suspected during pulmonary artery catheterization because of the difficulty of the catheter tip to move from the right ventricle to the pulmonary artery and, successively, because of the finding of a large gradient between the systolic pressure in the right ventricle and in the pulmonary artery. The diagnosis was confirmed by transesophageal echocardiogram (TEE). Hemodynamics recovered after the infusion of cristalloids, 1L, and the suspension of vasoconstrictors and inotropes. This is the first case in which RVOTO was observed during the induction of general anesthesia. Although this is a rare condition, the diagnostic suspect is of outmost importance because treatment is mainly based on fluid administration, and drugs with positive inotropic properties (like most vasoconstrictors) are contraindicated. RVOTO is an unusual, but possible cause of severe arterial hypotension during general anesthesia induction. TEE is useful for the evaluation of severely hypotensive patients who do not respond to routine treatment with fluids and vasoconstrictors.


Differential timing for programming of glucose homoeostasis, sensitivity to insulin and blood pressure by in utero exposure to dexamethasone in sheep.
Numerous epidemiological studies have related an increased risk of adult-onset cardiovascular and metabolic disease to an adverse intra-uterine environment at critical periods. We have shown that fetal sheep exposed to dexamethasone for only 2 days at 27 days of gestation (term approximately 150 days) became hypertensive adults, whereas those exposed at 64 days of gestation remained normotensive, as did controls. In the same sheep, now nearly 5 years old, we performed glucose tolerance tests and hyperinsulinaemic euglycaemic clamps to study the insulin sensitivity of glucose, amino acid and non-esterified fatty acid metabolism. Glucose tolerance, calculated as the area under the curve, after intravenous administration of bolus glucose and insulin secretion in response to a glucose challenge were not altered in any group. There were no significant differences in the insulin sensitivity of net whole-body glucose or amino acid uptake. However, suppression of lipolysis by insulin, measured as the proportional decrease in the circulating concentration of non-esterified fatty acids during the hyperinsulinaemic clamp, was 69+/-1.2% at steady-state plasma insulin levels ( approximately 1000 m-units/l) in the group exposed to dexamethasone at 27 days of gestation, but only 50.8+/-6.5% in the controls (P<0.05). In the group exposed to dexamethasone at 64 days of gestation, the decrease was 66.4+/-5.1%, which did not reach significance compared with the controls (P=0.10). Thus brief dexamethasone exposure during early gestation programmed hypertension independently of insulin resistance of glucose or amino acid metabolism; however, it did lead to increased insulin sensitivity of the inhibition of lipolysis, which may increase susceptibility to the development of obesity postnatally.


Differences in cytochrome P450-mediated biotransformation of 1,2-dichlorobenzene by rat and man: implications for human risk assessment.
The oxidative biotransformation of 1,2-dichlorobenzene (1,2-DCB) was investigated using hepatic microsomes from male Wistar, Fischer-344 and Sprague-Dawley (SD) rats, phenobarbital (PB)- and isoniazid (ISO) pretreated male Wistar rats and from man. In addition, microsomes from cell lines selectively expressing one cytochrome P450 (P4502E1, 1A1, 1A2, 2B6, 2C9, 2D6, /A6 and 3A4) were used. The rate of conversion was 0.09 nmol/min/mg. protein for both Wistar and Fischer-344 rat microsomes, 0.04 for SD-microsomes and 0.14 for human microsomes. Induction of Wistar rats with isoniazid (ISO, a P4502E1 inducer) or phenobarbital (PB, a P4502B1/2 inducer) resulted in an increased conversion rate of 0.20 and 0.42 nmol/min/mg. protein, respectively. Covalent binding of radioactivity to microsomal protein was similar for Wistar, Fischer and ISO-pretreated rats (16-17% of total metabolites), whereas induction with PB resulted in an increased covalent binding of 23% of total metabolites. Covalent binding was 31% for SD-microsomes and only 4.6% for human microsomes. Ascorbic acid notably reduced the amount of covalently bound metabolites for the SD-microsomes only, indicating that for these microsomes quinones were likely to be involved in this part of the covalent binding. Conjugation of epoxides with glutathione (GSH) inhibited most of the covalent binding for all microsomes. In the absence of GSH, the epoxides were hydrolyzed by epoxide hydrolase, resulting in the formation of dihydrodiols. Inhibition of epoxide hydrolase resulted in a decreased conversion and an increased covalent binding for all microsomes tested, indicating a role of epoxides in the covalent binding. Fischer-344 rat liver microsomes showed a lower epoxide hydrolase activity than microsomes from Wistar and Sprague-Dawley rats, which may explain the higher sensitivity to 1,2-DCB induced hepatotoxicity of Fischer rats in vivo. Conjugation of the epoxides with GSH was predominantly non-enzymatic for the rat, whereas for man, conjugation was nearly exclusively catalyzed by glutathione-S-transferases. This difference may be explained by the formation of a 'non-reactive' 3,4-epoxide by P4502E1 in human microsomes: incubations with microsomes selectively expressing human P4502E1 as well as human liver microsomes, resulted in the formation of similar amounts of 2,3- and 3,4-dichlorophenol (DCP), as well as two GSH-epoxide conjugates in equal amounts. For rat microsomes, one major GSH-epoxide conjugate was found, and a much higher covalent binding, particularly for the PB-microsomes. Therefore, we postulate that rat P4502B1/2 preferentially oxidizes the 4,5-site of 1,2-DCB, resulting in a reactive epoxide. Postulating these epoxides to be involved in the mechanism(s) of toxicity, human risk after exposure to 1,2-DCB will be overestimated when risk assessment is solely based on toxicity studies conducted in rat.


DNA-Conjugated Gold Nanoparticles as High-Mass Probes in Imaging Mass Cytometry.
We employ imaging mass cytometry (IMC) to investigate in vitro uptake and cellular distribution of DNA-functionalized gold nanoparticles (AuNPs). IMC enables the multiparametric imaging of cell components and allows for the detection of AuNPs in cells with >100 times higher sensitivity than conventional confocal fluorescence imaging, as each nanoparticle contains thousands of atoms for signal amplification. Changes in the accumulation of nanoparticles in cells due to oligonucleotide sequence-dependent interactions are exploited to examine a model biomarker for hypoxia-microRNA-210. We find that AuNPs functionalized with microRNA-210-targeting sequence accumulate in hypoxic cells 3 to 4-fold compared to normoxic cells. The work examines the potential use of DNA-AuNP as high-mass probes for the analysis of nonabundant nucleic acids.


Familial dysequilibrium-diplegia with T-lymphocyte deficiency.
A second family is described with a combination of defective thymus-dependent immunity and cerebral palsy. The cerebral palsy comprised nonprogressive dysequilibrium and mild spastic diplegia without limb ataxia. This genetic entity of presumed autosomal recessive inheritance is clearly distinguished from ataxia-telangiectasia. Immunological abnormalities should be sought in other familial or unexplained cerebral palsy syndromes.


Therapeutic Ultrasound for Enhanced Corneal Permeability to Macromolecules.
Topically applied macromolecules have the potential to provide vision-saving treatments for many of the leading causes of blindness in the United States. The aim of this study was to determine if ultrasound can be applied to increase transcorneal drug delivery of macromolecules without dangerously overheating surrounding ocular tissues. Dissected corneas of adult rabbits were placed in a diffusion cell between a donor compartment filled with a solution of macromolecules (40, 70 kDa, or 150 kDa) and a receiver compartment. Each cornea was exposed to the drug solution for 60 minutes, with the experimental group receiving 5 minutes of continuous ultrasound or 10 minutes of pulsed ultrasound at a 50% duty cycle (pulse repetition frequency of 500 ms on, 500 ms off) at the beginning of treatment. Unfocused circular ultrasound transducers were operated at 0.5 to 1 W/cm2 intensity and at 600 kHz frequency. The greatest increase in transcorneal drug delivery seen was 1.2 times (P < .05) with the application of pulsed ultrasound at 0.5 W/cm2 and 600 kHz for 10 minutes with 40 kDa macromolecules. Histological analysis revealed structural damage mostly in the corneal epithelium, with most damage occurring at the epithelial surface. This study suggests that ultrasound may be used for enhancing transcorneal delivery of macromolecules of lower molecular weights. Further research is needed on the long-term effects of ultrasound parameters used in this study on human ocular tissues.


Quantitative expression analyses of candidates for alternative anion conductance in cystic fibrosis mouse models.
Diversity of cystic fibrosis (CF) phenotype in patients with the same CFTR-mutation raised the hypothesis that other factors modulate the phenotype including "alternative" calcium-activated anion currents (CaCC). This study compares the mRNA expression levels of candidate CaCC mediators in CF mouse models with wild type controls. mBEST1, mBEST2, mCLC-3B, mCLC-4, mTTYH3, mTMEM16A, mTMEM16F, mTMEM16K, mCLCA1 to -6 and SLC26A9 mRNA were quantified in CF-relevant tissues in cftr(tm1Cam) and cftr(TgH(neoim)Hgu) mice and controls using real-time RT-qPCR. No consistent differences were observed except for mTTYH3 which was significantly down-regulated throughout the intestinal tract of cftr(tm1Cam) mice. Down-regulation of mTTYH3 may point towards its involvement in the complex CF pathology. However, the markedly reduced expression argues against a direct compensatory action as an alternative anion conductance. If any of the other candidates plays a role as modulator, factors other than transcriptional regulation and mRNA stability may be involved.


Generation and coordination of heartbeat timing oscillation in the medicinal leech. I. Oscillation in isolated ganglia.
1. The interactions among the four pairs of interneurons (HN(1)-HN(4)) of the heartbeat timing oscillator are confined to the third and fourth ganglia (G3 and G4). In isolation, G3 and G4 each produces a rhythm essentially the same as that shown when the two ganglia are linked together. 2. The local circuits in both ganglia have the same general form. In both the oscillation centers on a bilateral pair of HN cells that are linked by reciprocal inhibition (the HN(3) pair in G3 and the HN(4) pair in G4). In addition, there is reciprocal inhibition between an HN(3) or HN(4) cell and the intersegmental processes of the ipsilateral HN(1) and HN(2) cells. 3. These connections account for the phase relationships in an isolated G3 or G4, since cells linked by reciprocal inhibition produce bursts in alternation. 4. In isolated ganglia, reciprocal inhibition not only coordinates the activity of the HN cells but also appears to help generate their bursts. 5. Yet reciprocal inhibition alone cannot account for the activity of the network. An endogenous property of the HN(3) and HN(4) cells appears to create the instability necessary for oscillation.


Enhancement of Naja naja atra antivenin production in horses.
As the conventional hyperimmunization schedule in horses introduced by Tanaka could not produce enough neutralizing antibody against Naja naja atra venom, the mixture of Carboxymethyl cellulose (CMC)-Cobra venom incorporated with adjuvant was used for immunization. The neutralizing antibody produced (30 LD50) seemed to be increased but still not to reach the satisfactory level. By using CMC-Cobratoxin adjuvant mixture as an immunizing agent, highly potent antivenin (220 LD50) was obtained.


Asynchronous origins of ectomycorrhizal clades of Agaricales.
The ectomycorrhizal (ECM) symbiosis is the most widespread biotrophic nutritional mode in mushroom-forming fungi. ECM fungi include, though are not limited to, about 5000 described species of Agaricales from numerous, independently evolved lineages. Two central hypotheses suggest different explanations for the origin of ECM fungal diversity: (i) dual origins, initially with the Pinaceae in the Jurassic and later with angiosperms during the Late Cretaceous, and (ii) a simultaneous and convergent radiation of ECM lineages in response to cooling climate during the Palaeogene and advancing temperate ECM plant communities. Neither of these hypotheses is supported here. While we demonstrate support for asynchronous origins of ECM Agaricales, the timing of such events appears to have occurred more recently than suggested by the first hypothesis, first during the Cretaceous and later during the Palaeogene. We are also unable to reject models of rate constancy, which suggests that the diversity of ECM Agaricales is not a consequence of convergent rapid radiations following evolutionary transitions from saprotrophic to ECM habits. ECM lineages of Agaricales differ not only in age, but also in rates of diversification and rate of substitution at nuclear ribosomal RNA loci. These results question the biological uniformity of the ECM guild.


In vivo biosynthesis of terpene nucleosides provides unique chemical markers of Mycobacterium tuberculosis infection.
Although small molecules shed from pathogens are widely used to diagnose infection, such tests have not been widely implemented for tuberculosis. Here we show that the recently identified compound, 1-tuberculosinyladenosine (1-TbAd), accumulates to comprise >1% of all Mycobacterium tuberculosis lipid. In vitro and in vivo, two isomers of TbAd were detected that might serve as infection markers. Using mass spectrometry and nuclear magnetic resonance, we established the structure of the previously unknown molecule, N(6)-tuberculosinyladenosine (N(6)-TbAd). Its biosynthesis involves enzymatic production of 1-TbAd by Rv3378c followed by conversion to N(6)-TbAd via the Dimroth rearrangement. Intact biosynthetic genes are observed only within M. tuberculosis complex bacteria, and TbAd was not detected among other medically important pathogens, environmental bacteria, and vaccine strains. With no substantially similar known molecules in nature, the discovery and in vivo detection of two abundant terpene nucleosides support their development as specific diagnostic markers of tuberculosis.


Stomach carcinoma incidence patterns in the United States by histologic type and anatomic site.
Using data from the U.S. National Cancer Institute's Surveillance, Epidemiology, and End Results program, we analyzed stomach carcinoma incidence patterns by both histologic type and anatomic site. We calculated age-adjusted (2000 U.S. standard) rates for 1978 to 2005, and for five time periods from 1978-1983 through 2001-2005 according to histologic type and anatomic site, separately and jointly. We also analyzed rates by race, gender, and age group. During 1978 to 2005, more than 54,000 stomach carcinoma cases were diagnosed among residents of the nine Surveillance, Epidemiology, and End Results areas. Total stomach carcinoma rates declined by 34% from the 1978-1983 to the 2001-2005 time periods. By histologic type, intestinal rates decreased consistently, whereas those for diffuse rates increased through 2000 and declined in recent years. By anatomic site, cardia rates increased during earlier years and then decreased, whereas rates for all other sites declined. When considered jointly by histologic type and anatomic site, intestinal carcinoma rates decreased for all sites except the cardia; diffuse rates increased through 2000 and decreased in recent years for all sites except the overlapping/nonspecified sites. Both diffuse and intestinal rates were lowest among whites, intermediate among blacks, and highest among the other, primarily Asian, races, with only modest gender differences for the diffuse type. In contrast, cardia carcinoma rates were highest among whites and were notably higher among males, especially whites among whom the male/female rate ratio was five to one. Stomach carcinoma incidence patterns differ by histologic type, anatomic site, race, gender, and age, suggesting that etiologic heterogeneity should be pursued in future research.


Effects of Acupressure on Anxiety: A Systematic Review and Meta-Analysis.
Objectives: The research aim was to perform a systematic review and meta-analysis evaluating the ability of acupressure to reduce anxiety. Design: Randomized controlled trials were obtained through a search of electronic medical databases (four in English and one in Chinese) from inception to October 5, 2020. Two authors searched the databases, evaluated studies' methodological quality, and performed data extraction independently. The final studies for analysis were identified after discussion with the third author. Results: We obtained 27 studies for our systematic review and meta-analysis. Eight studies had a low overall risk of bias, and 13 had some bias concerns with methodological quality. According to the results, acupressure significantly reduced patient anxiety (standardized mean difference = 1.152; 95% confidence interval: 0.847-1.459, p < 0.001), and the study heterogeneity was high (Q = 299.74, p < 0.001, I2 = 91.333%). Two studies reported acupressure-associated adverse events. We also performed a sensitivity analysis by omitting one outlier study, which had the largest effect size; however, high heterogeneity remained (I2 = 87.816%). A subgroup analysis revealed significant differences between participant types (Q = 46.573, p < 0.001), levels of methodological quality (Q = 6.228, p = 0.044), and massage equipment (Q = 4.642, p = 0.031). Conclusions: Our meta-analysis suggests that acupressure can alleviate anxiety. Acupressure was more effective for inpatients and preoperative patients when finger massage was applied. In individuals with anxiety and a stable hemodynamic status, acupressure could be a promising treatment option. However, the substantial heterogeneity across studies means that any inference from the results should be performed cautiously.


The Fundamental Role of Oxime and Oxime Ether Moieties in Improving the Physicochemical and Anticancer Properties of Structurally Diverse Scaffolds.
The present review explores the critical role of oxime and oxime ether moieties in enhancing the physicochemical and anticancer properties of structurally diverse molecular frameworks. Specific examples are carefully selected to illustrate the distinct contributions of these functional groups to general strategies for molecular design, modulation of biological activities, computational modeling, and structure-activity relationship studies. An extensive literature search was conducted across three databases, including PubMed, Google Scholar, and Scifinder, enabling us to create one of the most comprehensive overviews of how oximes and oxime ethers impact antitumor activities within a wide range of structural frameworks. This search focused on various combinations of keywords or their synonyms, related to the anticancer activity of oximes and oxime ethers, structure-activity relationships, mechanism of action, as well as molecular dynamics and docking studies. Each article was evaluated based on its scientific merit and the depth of the study, resulting in 268 cited references and more than 336 illustrative chemical structures carefully selected to support this analysis. As many previous reviews focus on one subclass of this extensive family of compounds, this report represents one of the rare and fully comprehensive assessments of the anticancer potential of this group of molecules across diverse molecular scaffolds.


Promising Antifungal Potential of Engineered Non-ionic Surfactant-Based Vesicles: In Vitro and In Vivo Studies.
Fungal keratitis (FK) is a corneal infection caused by different fungal species. It is treated by the topical application of natamycin (NAT). Nevertheless, this approach faces many limitations like toxic effects, frequent dosing, resistance, and patient discomfort. The present research reports the development of trimethyl chitosan (TMC) coated mucoadhesive cationic niosomes by a modified thin-film hydration method. TMC was synthesized using a one-step carbodiimide method and characterized by 1H-NMR and degree of quaternization (53.74 ± 1.06%). NAT, cholesterol (CHOL), span 60 (Sp60), and dicetyl phosphate (DCP) were used to prepare niosomes which were incubated with TMC to obtain mucoadhesive cationic NAT loaded niosomes (MCNNs). MCNNs showed a spherical shape with 1031.12 ± 14.18 nm size (PDI below 0.3) and 80.23 ± 5.28% entrapment efficiency. In vitro drug release studies showed gradual drug release from TMC coated niosomes as compared to the uncoated niosomes. MIC assay and disk diffusion assay revealed promising in vitro antifungal potential of MCNNs similar to the marketed formulation. For investigating in vivo performance, ocular retention and pharmacokinetics, ocular irritation, and ulcer healing studies were performed using the rabbit model. Mucoadhesive property and prolonged local drug release improved the safety and efficacy of NAT, suggesting that the developed niosomes could be an emerging system for effective treatment of fungal keratitis.


Transcriptome profiling of male and female Ascaris lumbricoides reproductive tissues.
Ascaris lumbricoides causes human ascariasis, the most prevalent helminth disease, infecting approximately 1 billion individuals globally. In 2019 the global disease burden was estimated to be 754,000 DALYs and resulted in 2090 deaths. In the absence of a vaccination strategy, treatment of ascariasis has relied on anthelminthic chemotherapy, but drug resistance is a concern. The propensity for reinfection is also a major challenge to disease control; female worms lay up to 200,000 eggs daily, which contaminate surrounding environments and remain viable for years, resulting in high transmission rates. Understanding the molecular mechanisms of reproductive processes, including control of egg production, spermatogenesis, oogenesis and embryogenesis, will drive the development of new drugs and/or vaccine targets for future ascariasis control. Transcriptome profiles of discrete reproductive and somatic tissue samples were generated from adult male and female worms using Illumina HiSeq with 2 × 150 bp paired-end sequencing. Male tissues included: testis germinal zone, testis part of vas deferens, seminal vesicle and somatic tissue. Female tissues included: ovary germinal zone, ovary part of the oviduct, uterus and somatic tissue. Differentially expressed genes (DEGs) were identified from the fragments per kilobases per million reads (FPKM) profiles. Hierarchical analysis was performed to identify tissue-specific genes. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were employed to identify significant terms and pathways for the DEGs. DEGs involved in protein phosphorylation and adhesion molecules were indicated to play a crucial role in spermatogenesis and fertilization, respectively. Those genes associated with the G-protein-coupled receptor (GPCR) signaling pathway and small GTPase-mediated signal transduction pathway play an essential role in cytoskeleton organization during oogenesis. Additionally, DEGs associated with the SMA genes and TGF-β signaling pathway are crucial in adult female embryogenesis. Some genes associated with particular biological processes and pathways that were identified in this study have been linked to defects in germline development, embryogenesis and reproductive behavior. In the enriched KEGG pathway analysis, Hippo signaling, oxytocin signaling and tight junction pathways were identified to play a role in Ascaris male and female reproductive systems. This study has provided comprehensive transcriptome profiles of discrete A. lumbricoides reproductive tissue samples, revealing the molecular basis of these functionally important tissues. The data generated from this study will provide fundamental knowledge on the reproductive biology of Ascaris and will inform future target identification for anti-ascariasis drugs and/or vaccines.


Anterior optic pathways gliomas. The dilemma of treatment.
Anterior optic pathways gliomas are classifiable histologically and clinically with other pilocytic gliomas of the midline (of hypothalamus and brainstem), especially of childhood. The very slow course, sometimes imperceptible over the years, that characterizes the majority of them should not be taken to mean lack of growth. Tumultuous courses and acute episodic deteriorations are both possible. Hence the need for therapy. Indefinite abstention cannot be upheld, at least in the great majority of cases. In gliomas of a nerve only the object of surgery is usually removal, using the intraorbital, intracanalicular and intracranial approaches, but in gliomas of the chiasm or chiasm-hypothalamus the object is more exploratory and decompressive. Radiotherapy is generally accepted as affective only in the latter cases. No single standard protocol is possible: treatment had to be individualized case by case.


Curated dataset of asphaltene structures.
Asphaltenes, a distinct class of molecules found in crude oil, exhibit insolubility in nonpolar solvents like n-heptane but are soluble in aromatic solvents such as toluene and benzene. Understanding asphaltenes is crucial in the petroleum industry due to their detrimental effects on oil processing, resulting in significant economic losses and production disruptions. While no singular structure defines asphaltenes, two major molecular architectures, namely archipelago and continental models, have gained wide acceptance for their consistency with various experimental investigations and subsequent use in computational studies. The archipelago model comprises two or more polyaromatic hydrocarbon entities interconnected via aliphatic side chains. In contrast, the island or continental model features a unified polyaromatic hydrocarbon moiety with 4 to 10 fused aromatic rings, averaging around 7 rings. To establish a comprehensive collection, we meticulously curated over 250 asphaltene structures derived from previous experimental and computational studies in this field. Our curation process involved an extensive literature survey, conversion of figures from publications into molecular structure files, careful verification of conversion accuracy, and structure editing to ensure alignment with molecular formulas. Our database provides digital structure files and optimized geometries for both predominant structural motifs. The optimization procedure commenced with the PM6 semi-empirical method, followed by further optimization utilizing density functional theory employing the B3LYP functional and the 6-31+G(d,p) basis set. Furthermore, we compiled a range of structural and electronic features for these molecules, serving as a valuable foundation for employing machine learning algorithms to investigate asphaltenes. This work provides a ready to use structural database of asphaltenes and sets the stage for future research endeavours in this domain.


Selective Shielding of Bone Marrow: An Approach to Protecting Humans from External Gamma Radiation.
The current feasibility of protecting emergency responders through bone marrow selective shielding is highlighted in the recent OECD/NEA report on severe accident management. Until recently, there was no effective personal protection from externally penetrating gamma radiation. In Chernobyl, first-responders wore makeshift lead sheeting, whereas in Fukushima protective equipment from gamma radiation was not available. Older protective solutions that use thin layers of shielding over large body surfaces are ineffective for energetic gamma radiation. Acute exposures may result in Acute Radiation Syndrome where the survival-limiting factor up to 10 Gy uniform, homogeneous exposure is irreversible bone marrow damage. Protracted, lower exposures may result in malignancies of which bone marrow is especially susceptible, being compounded by leukemia's short latency time. This highlights the importance of shielding bone marrow for preventing both deterministic and stochastic effects. Due to the extraordinary regenerative potential of hematopoietic stem cells, to effectively prevent the deterministic effects of bone marrow exposure, it is sufficient to protect only a small fraction of this tissue. This biological principle allows for a new class of equipment providing unprecedented attenuation of radiation to select marrow-rich regions, deferring the hematopoietic sub-syndrome of Acute Radiation Syndrome to much higher doses. As approximately half of the body's active bone marrow resides within the pelvis region, shielding this area holds great promise for preventing the deterministic effects of bone marrow exposure and concomitantly reducing stochastic effects. The efficacy of a device that selectively shields this region and other radiosensitive organs in the abdominal area is shown here.


[Is it necessary to vaccinate against varicella?].
Varicella is a frequent viral disease, with a substantial medical and societal impact. For many years, various industrialized countries have adopted an universal mass vaccination against varicella, using a one-dose schedule. In these countries, the global incidence of varicella has decreased by about 90%. A significant reduction in hospitalizations, outpatient visits and medical costs due to varicella has also been observed. Recently, a 2-dose schedule has demonstrated an efficacy of about 98%, as well as herd immunity.


The Ethics of Research in Lower Income Countries: Double Standards Are Not the Problem.
Discussion of the ethics of clinical trials in lower income countries has been dominated by concern over double standards. Most prominently, clinical trials of interventions that are less effective than the worldwide best treatment methods typically are not permitted in higher income countries. Commentators conclude that permitting such trials in lower income countries involves an ethical double standard. Despite significant attention to this concern, and its influence over prominent guidelines for research in lower income countries, there has been little analysis of what constitutes an ethical double standard in clinical research. The present article attempts to address this gap in the literature. This analysis finds that ethical double standards involve a kind of disrespect, and yields a three-step decision procedure for evaluating when trials of less than the worldwide best methods raise this concern. Application of this procedure reveals that permitting these trials in lower income countries rarely involves an ethical double standard. Instead, the real challenge is determining when clinical trials of interventions that are less effective than the worldwide best represent a permissible and effective response to differences in access to healthcare between higher income and lower income countries. To protect research subjects, without blocking clinical trials that have the potential to improve health in lower income countries, research review committees and other stakeholders should focus on this issue, not on ethical double standards.


PD-1 signaling negatively regulates the common cytokine receptor γ chain via MARCH5-mediated ubiquitination and degradation to suppress anti-tumor immunity.
Combination therapy with PD-1 blockade and IL-2 substantially improves anti-tumor efficacy comparing to monotherapy. The underlying mechanisms responsible for the synergistic effects of the combination therapy remain enigmatic. Here we show that PD-1 ligation results in BATF-dependent transcriptional induction of the membrane-associated E3 ubiquitin ligase MARCH5, which mediates K27-linked polyubiquitination and lysosomal degradation of the common cytokine receptor γ chain (γc). PD-1 ligation also activates SHP2, which dephosphorylates γcY357, leading to impairment of γc family cytokine-triggered signaling. Conversely, PD-1 blockade restores γc level and activity, thereby sensitizing CD8+ T cells to IL-2. We also identified Pitavastatin Calcium as an inhibitor of MARCH5, which combined with PD-1 blockade and IL-2 significantly improves the efficacy of anti-tumor immunotherapy in mice. Our findings uncover the mechanisms by which PD-1 signaling antagonizes γc family cytokine-triggered immune activation and demonstrate that the underlying mechanisms can be exploited for increased efficacy of combination immunotherapy of cancer.


Rag defects and thymic stroma: lessons from animal models.
Thymocytes and thymic epithelial cells (TECs) cross-talk is essential to support T cell development and preserve thymic architecture and maturation of TECs and Foxp3(+) natural regulatory T cells. Accordingly, disruption of thymic lymphostromal cross-talk may have major implications on the thymic mechanisms that govern T cell tolerance. Several genetic defects have been described in humans that affect early stages of T cell development [leading to severe combined immune deficiency (SCID)] or late stages in thymocyte maturation (resulting in combined immunodeficiency). Hypomorphic mutations in SCID-causing genes may allow for generation of a limited pool of T lymphocytes with a restricted repertoire. These conditions are often associated with infiltration of peripheral tissues by activated T cells and immune dysregulation, as best exemplified by Omenn syndrome (OS). In this review, we will discuss our recent findings on abnormalities of thymic microenvironment in OS with a special focus of defective maturation of TECs, altered distribution of thymic dendritic cells and impairment of deletional and non-deletional mechanisms of central tolerance. Here, taking advantage of mouse models of OS and atypical SCID, we will discuss how modifications in stromal compartment impact and shape lymphocyte differentiation, and vice versa how inefficient T cell signaling results in defective stromal maturation. These findings are instrumental to understand the extent to which novel therapeutic strategies should act on thymic stroma to achieve full immune reconstitution.


PREVAIL: Predicting Recovery through Estimation and Visualization of Active and Incident Lesions.
The goal of this study was to develop a model that integrates imaging and clinical information observed at lesion incidence for predicting the recovery of white matter lesions in multiple sclerosis (MS) patients. Demographic, clinical, and magnetic resonance imaging (MRI) data were obtained from 60 subjects with MS as part of a natural history study at the National Institute of Neurological Disorders and Stroke. A total of 401 lesions met the inclusion criteria and were used in the study. Imaging features were extracted from the intensity-normalized T1-weighted (T1w) and T2-weighted sequences as well as magnetization transfer ratio (MTR) sequence acquired at lesion incidence. T1w and MTR signatures were also extracted from images acquired one-year post-incidence. Imaging features were integrated with clinical and demographic data observed at lesion incidence to create statistical prediction models for long-term damage within the lesion. The performance of the T1w and MTR predictions was assessed in two ways: first, the predictive accuracy was measured quantitatively using leave-one-lesion-out cross-validated (CV) mean-squared predictive error. Then, to assess the prediction performance from the perspective of expert clinicians, three board-certified MS clinicians were asked to individually score how similar the CV model-predicted one-year appearance was to the true one-year appearance for a random sample of 100 lesions. The cross-validated root-mean-square predictive error was 0.95 for normalized T1w and 0.064 for MTR, compared to the estimated measurement errors of 0.48 and 0.078 respectively. The three expert raters agreed that T1w and MTR predictions closely resembled the true one-year follow-up appearance of the lesions in both degree and pattern of recovery within lesions. This study demonstrates that by using only information from a single visit at incidence, we can predict how a new lesion will recover using relatively simple statistical techniques. The potential to visualize the likely course of recovery has implications for clinical decision-making, as well as trial enrichment.


Acid-base equilibrium dynamics in methanol and dimethyl sulfoxide probed by two-dimensional infrared spectroscopy.
Two-dimensional infrared (2DIR) spectroscopy, which has been proven to be an excellent experimental method for studying thermally-driven chemical processes, was successfully used to investigate the acid dissociation equilibrium of HN3 in methanol (CH3OH) and dimethyl sulfoxide (DMSO) for the first time. Our 2DIR experimental results indicate that the acid-base equilibrium occurs on picosecond timescales in CH3OH but that it occurs on much longer timescales in DMSO. Our results imply that the different timescales of the acid-base equilibrium originate from different proton transfer mechanisms between the acidic (HN3) and basic (N3(-)) species in CH3OH and DMSO. In CH3OH, the acid-base equilibrium is assisted by the surrounding CH3OH molecules which can directly donate H(+) to N3(-) and accept H(+) from HN3 and the proton migrates through the hydrogen-bonded chain of CH3OH. On the other hand, the acid-base equilibrium in DMSO occurs through the mutual diffusion of HN3 and N3(-) or direct proton transfer. Our 2DIR experimental results corroborate different proton transfer mechanisms in the acid-base equilibrium in protic (CH3OH) and aprotic (DMSO) solvents.


[Unified postsynaptic mechanism of plasticity in the striatum, neocortex, hippocampus, and cerebellum].
The unitary postsynaptic mechanism of plasticity in striatum, neocortex, hippocampus and cerebellum involves the LTP/LTD excitation as result of AMPA and NMDA receptor phosphorylation/dephosphorylation, while the LTP/LTD of inhibition is the result of the GABA receptor phosphorylation/dephosphorylation. It follows from this mechanism that when NMDA channels are closed, the determinant role in receptor phosphorylation is played by the PKG. When the NMDA channels are open, the determinant role in receptor phosphorylation is played by the PKC and CaMKII.


Fluid filled oculoplethysmography and carotid artery disease: imperfect but useful.
Fluid filled oculoplethysmography (OPG) is a widely used method of assessing carotid stenosis but it has limitations in the detection of bilateral internal carotid artery lesions and of external carotid artery stenoses. In this study, 157 consecutive patients having carotid angiography and fluid filled OPG were assessed to determine the accuracy of the technique and define the sources of error. Haemodynamically significant stenosis (HDS) was defined as at least 50% stenosis of the internal carotid artery (ICA). Only the most severely stenosed side of the 35 bilateral HDS lesions was detected owing to the poor reliability of ear pulse delays. Eye/eye delays alone detected the most severely stenosed side in 82 of 98 patients with an HDS stenosis of one or both ICAs for a sensitivity of 84% a specificity of 71% (41/59) and accuracy of 79% (81/157). The measurement of ear/ear pulse delays for external carotid artery (ECA) stenosis had a sensitivity of only 15% (5/34). Ear/eye pulse delays detected none of the 35 patients with bilateral HDS ICA stenosis. Bilateral equal HDS ICA stenoses were a significant source of error. Stenotic disease was present in the aortic arch and branches (five patients) or the carotid siphon (eight patients) and in seven cases it resulted in an incorrect localization on OPG. There was no diagnostic relationship between the severity of delay and the presence of total occlusion. Chronic local eye pathology was present in 13 patients and did not affect the results of the OPG. We have ceased to use ear pulse measurements for routine assessment but continue to use the eye/eye delays in conjunction with a carotid doppler imaging system.


Clinical outcomes of anterior cruciate ligament reconstruction using LARS artificial graft with an at least 7-year follow-up.
The aim of our study was to assess the medium-term clinical outcomes of anterior cruciate ligament (ACL) reconstruction using the Ligament Advanced Reinforcement System (LARS) artificial ligament.A total of 168 patients who underwent arthroscopic ACL reconstruction with the LARS artificial ligament in our department were enrolled in our research. Only 125 met the inclusion/exclusion criteria, and 91 could ultimately be contacted to participate in our research. The mean follow-up was 92 ± 19 months. Physical examinations and a KT-1000 arthrometer were used to evaluate knee laxity. The International Knee Documentation Committee (IKDC) and Lysholm knee scales were evaluated for knee function. The Tegner score was tested for the condition of return to sport. Range of motion (ROM) and the rates of failure and complications were calculated.Among all patients enrolled in the study, the failure rate was 4.4%, and the overall complication rate was 2.2%. Knee laxity measured by the KT-1000 arthrometer was 1.4 ± 1.5 mm, compared with the preoperative value of 5.1 ± 1.3 mm. The Lysholm score improved from a preoperative value of 54.6 ± 14.3 to a postoperative value of 85.4 ± 12.1. The proportion of return to sport was 86.8% (79/91). The postoperative Tegner score was 4.7 ± 1.3, while its value before injury was 5.5 ± 1.0.In this study, ACL reconstruction using the LARS artificial ligament has a good prognosis with a low failure and complication rate at a mean follow-up of 91 months.


A first insight into the involvement of phytohormones pathways in coffee resistance and susceptibility to Colletotrichum kahawae.
Understanding the molecular mechanisms underlying coffee-pathogen interactions are of key importance to aid disease resistance breeding efforts. In this work the expression of genes involved in salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) pathways were studied in hypocotyls of two coffee varieties challenged with the hemibiotrophic fungus Colletotrichum kahawae, the causal agent of Coffee Berry Disease. Based on a cytological analysis, key time-points of the infection process were selected and qPCR was used to evaluate the expression of phytohormones biosynthesis, reception and responsive-related genes. The resistance to C. kahawae was characterized by restricted fungal growth associated with early accumulation of phenolic compounds in the cell walls and cytoplasmic contents, and deployment of hypersensitive reaction. Similar responses were detected in the susceptible variety, but in a significantly lower percentage of infection sites and with no apparent effect on disease development. Gene expression analysis suggests a more relevant involvement of JA and ET phytohormones than SA in this pathosystem. An earlier and stronger activation of the JA pathway observed in the resistant variety, when compared with the susceptible one, seems to be responsible for the successful activation of defense responses and inhibition of fungal growth. For the ET pathway, the down or non-regulation of ET receptors in the resistant variety, together with a moderate expression of the responsive-related gene ERF1, indicates that this phytohormone may be related with other functions besides the resistance response. However, in the susceptible variety, the stronger activation of ERF1 gene at the beginning of the necrotrophic phase, suggests the involvement of ET in tissue senescence. As far as we know, this is the first attempt to unveil the role of phytohormones in coffee-C. kahawae interactions, thus contributing to deepen our understanding on the complex mechanisms of plant signaling and defense.


Identification of animal movement patterns using tri-axial magnetometry.
Accelerometers are powerful sensors in many bio-logging devices, and are increasingly allowing researchers to investigate the performance, behaviour, energy expenditure and even state, of free-living animals. Another sensor commonly used in animal-attached loggers is the magnetometer, which has been primarily used in dead-reckoning or inertial measurement tags, but little outside that. We examine the potential of magnetometers for helping elucidate the behaviour of animals in a manner analogous to, but very different from, accelerometers. The particular responses of magnetometers to movement means that there are instances when they can resolve behaviours that are not easily perceived using accelerometers. We calibrated the tri-axial magnetometer to rotations in each axis of movement and constructed 3-dimensional plots to inspect these stylised movements. Using the tri-axial data of Daily Diary tags, attached to individuals of number of animal species as they perform different behaviours, we used these 3-d plots to develop a framework with which tri-axial magnetometry data can be examined and introduce metrics that should help quantify movement and behaviour.. Tri-axial magnetometry data reveal patterns in movement at various scales of rotation that are not always evident in acceleration data. Some of these patterns may be obscure until visualised in 3D space as tri-axial spherical plots (m-spheres). A tag-fitted animal that rotates in heading while adopting a constant body attitude produces a ring of data around the pole of the m-sphere that we define as its Normal Operational Plane (NOP). Data that do not lie on this ring are created by postural rotations of the animal as it pitches and/or rolls. Consequently, stereotyped behaviours appear as specific trajectories on the sphere (m-prints), reflecting conserved sequences of postural changes (and/or angular velocities), which result from the precise relationship between body attitude and heading. This novel approach shows promise for helping researchers to identify and quantify behaviours in terms of animal body posture, including heading. Magnetometer-based techniques and metrics can enhance our capacity to identify and examine animal behaviour, either as a technique used alone, or one that is complementary to tri-axial accelerometry.


Heart failure management guidelines: New recommendations and implementation.
The prevalence of heart failure has increased in many developed countries including Japan and the USA, due in large part to the aging of their populations. The lifetime risk of heart failure is now 20-30 % in the USA. Fortunately, there have been important advances in therapy that increase quality and length of life for those with heart failure. This review discusses the important advances in care including treatment and diagnosis and the new recommendations for this care from the recent American College of Cardiology (ACC)/American Heart Association (AHA)/Heart Failure Society of America (HFSA) Guideline. Relevant studies that have been published since the guideline was released are also included. Of the many recommendations in the ACC/AHA/HFSA Guideline, this review focuses on the definition of heart failure, the medical treatments specific to left ventricular ejection fraction, use of devices for treatment and diagnosis, diagnosis and treatment of amyloidosis, treatment of iron deficiency, screening for asymptomatic left ventricular dysfunction, use of patient reported outcomes, and tools for implementation.


The Phosphorus Economy of Mediterranean Oak Saplings Under Global Change.
While a severe decrease in phosphorus (P) availability is already taking place in a large number of ecosystems, drought and nitrogen (N) deposition will likely further decrease the availability of P under global change. Plants have developed physiological strategies to cope with decreasing P resources, but it is unclear how these strategies respond to elevated N deposition and summer droughts. We investigated the influence of N and P availability and soil drought on P uptake (H3 33PO4 feeding experiment) and use efficiencies in young Quercus calliprinos Webb. trees. We hypothesized that (H1) the expected increases in soil N:P ratios will increase the efficiencies of P uptake and use of oak saplings but will decrease the efficiencies of N uptake and use, whereas (H2) drought will affect P uptake efficiency more than N uptake efficiency. In confirmation of (H1) we found that a sharp increase of the soil N:P ratio from 4 to 42 g g-1 significantly increased the instantaneous 33P uptake efficiency (33PUptakeE) by five-fold and long-term P uptake efficiency (PUptakeE) by six-fold, while it decreased N uptake efficiency (NUptakeE) and N use efficiency (NUE). In contradiction to (H1), P use efficiency (PUE) did not respond to the simulated extended gradient of soil N:P ratios but remained relatively constant. (H2) was only partially confirmed as soil drought reduced PUptakeE by up to a fourth at high soil N:P ratios but had no significant effect on NUptakeE. As a consequence, increasing summer droughts may decrease the response of PUptakeE to increasing P limitation, which - in the absence of adjustments of the efficiency of P use - can aggravate growth reductions in this eastern Mediterranean tree species under global change.


Adult ADHD and working memory: neural evidence of impaired encoding.
To investigate neural and behavioural correlates of visual encoding during a working memory (WM) task in young adults with and without Attention-Deficit/Hyperactivity Disorder (ADHD). A sample of 30 college students currently meeting a diagnosis of ADHD and 25 typically developing students, matched on age and gender, performed a delayed match-to-sample task with low and high memory load conditions. Dense-array electroencephalography was recorded. Specifically, the P3, an event related potential (ERP) associated with WM, was examined because of its relation with attentional allocation during WM. Task performance (accuracy, reaction time) as well as performance on other neuropsychological tasks of WM was analyzed. Neural differences were found between the groups. Specifically, the P3 amplitude was smaller in the ADHD group compared to the comparison group for both load conditions at parietal-occipital sites. Lower scores on behavioural working memory tasks were suggestive of impaired behavioural WM performance in the ADHD group. Findings from this study provide the first evidence of neural differences in the encoding stage of WM in young adults with ADHD, suggesting ineffective allocation of attentional resources involved in encoding of information in WM. These findings, reflecting alternate neural functioning of WM, may explain some of the difficulties related to WM functioning that college students with ADHD report in their every day cognitive functioning.


Effects of source of DNA on genotyping success rates and allele percentages in the Preschoolers with Attention-Deficit/Hyperactivity Disorder Treatment Study (PATS).
In children diagnosed with attention-deficit/hyperactivity disorder (ADHD) and their parents, who were participants of the Preschool ADHD Treatment Study (PATS), we assessed the effect of source of DNA (from buccal or blood cells) on the genotyping success rate and allele percentages for the five polymorphisms in three candidate genes (DAT1, DRD4, and SNAP 25) investigated in the PATS pharmacogenetic study of response to stimulant medication. At baseline assessment, 241 individuals (113 probands and 128 parents) consented to participate; 144 individuals (52 probands and 92 parents) provided blood samples from venipuncture, and 97 individuals (61 probands and 36 parents) provided buccal samples from cheek swab as specimens for isolation of DNA. Three types of polymorphisms-variable number of tandem repeat (VNTR) polymorphism, tandem duplication polymorphism (TDP), and single nucleotide polymorphism (SNP)-were evaluated, including the DRD4 gene 48-bp VNTR in exon III, the DAT1 gene 40-bp VNTR in 3'-untranslated region, the DRD4 gene TDP 120-bp duplication in the promoter region, the SNAP-25 gene TC-1069 SNP, and the SNAP-25 gene TG-1065 SNP. Standard procedures were used to genotype individuals for each of these five polymorphisms. Using the methods available in 2004, the genotyping success rate was on the average much greater for DNA from blood cells than buccal cells (e.g., 91% vs. 54% in probands). For some polymorphisms (DRD4-VNTR, DRD4-TDP, and SNAP25-TC SNP), allele proportion also varied by blood versus buccal source of DNA (e.g., 26.5% vs. 18.6% for the 7-repeat allele of the DRD4 gene). The much lower success rate for genotyping based on DNA from buccal than blood cells is likely due to the quality of DNA derived from these two sources. The observed source differences in allele proportion may be due to self-selection related to choice of how specimens were collected (from cheek swab or venipuncture), or to a selective detection of some alleles based on differences in DNA quality.