Natural genetic transformation generates a population of merodiploids in Streptococcus pneumoniae.
Partial duplication of genetic material is prevalent in eukaryotes and provides potential for evolution of new traits. Prokaryotes, which are generally haploid in nature, can evolve new genes by partial chromosome duplication, known as merodiploidy. Little is known about merodiploid formation during genetic exchange processes, although merodiploids have been serendipitously observed in early studies of bacterial transformation. Natural bacterial transformation involves internalization of exogenous donor DNA and its subsequent integration into the recipient genome by homology. It contributes to the remarkable plasticity of the human pathogen Streptococcus pneumoniae through intra and interspecies genetic exchange. We report that lethal cassette transformation produced merodiploids possessing both intact and cassette-inactivated copies of the essential target gene, bordered by repeats (R) corresponding to incomplete copies of IS861. We show that merodiploidy is transiently stimulated by transformation, and only requires uptake of a ~3-kb DNA fragment partly repeated in the chromosome. We propose and validate a model for merodiploid formation, providing evidence that tandem-duplication (TD) formation involves unequal crossing-over resulting from alternative pairing and interchromatid integration of R. This unequal crossing-over produces a chromosome dimer, resolution of which generates a chromosome with the TD and an abortive chromosome lacking the duplicated region. We document occurrence of TDs ranging from ~100 to ~900 kb in size at various chromosomal locations, including by self-transformation (transformation with recipient chromosomal DNA). We show that self-transformation produces a population containing many different merodiploid cells. Merodiploidy provides opportunities for evolution of new genetic traits via alteration of duplicated genes, unrestricted by functional selective pressure. Transient stimulation of a varied population of merodiploids by transformation, which can be triggered by stresses such as antibiotic treatment in S. pneumoniae, reinforces the plasticity potential of this bacterium and transformable species generally.


A preclinical model of cutaneous melanoma based on reconstructed human epidermis.
Malignant melanoma is among the tumor entities with the highest increase of incidence worldwide. To elucidate melanoma progression and develop new effective therapies, rodent models are commonly used. While these do not adequately reflect human physiology, two-dimensional cell cultures lack crucial elements of the tumor microenvironment. To address this shortcoming, we have developed a melanoma skin equivalent based on an open-source epidermal model. Melanoma cell lines with different driver mutations were incorporated into these models forming distinguishable tumor aggregates within a stratified epidermis. Although barrier properties of the skin equivalents were not affected by incorporation of melanoma cells, their presence resulted in a higher metabolic activity indicated by an increased glucose consumption. Furthermore, we re-isolated single cells from the models to characterize the proliferation state within the respective model. The applicability of our model for tumor therapeutics was demonstrated by treatment with a commonly used v-raf murine sarcoma viral oncogene homolog B (BRAF) inhibitor vemurafenib. This selective BRAF inhibitor successfully reduced tumor growth in the models harboring BRAF-mutated melanoma cells. Hence, our model is a promising tool to investigate melanoma development and as a preclinical model for drug discovery.


The neuroscience of recovery and rehabilitation: what have we learned from animal research?
To encourage rehabilitation specialists to develop a critical approach to the animal research literature that is relevant to human neurorehabilitation and to encourage clinicians to lend their perspectives to basic research. ATA SOURCES: Scientific publications cited in MEDLINE, PubMed, and PsychInfo, and professional presentations of leading neuroscience researchers. The focus was on current publications to 2001, with historical works included when appropriate. Studies were selected based on their relevance to the objectives. Reviewed study methodology and findings and extracted key principles relevant to rehabilitation. Many themes emerging from neuroscience research are relevant to human rehabilitation, including issues related to timing of intervention and recovery, and characteristics of nervous system plasticity. Although animal research has many limitations, it provides a unique window on nervous system recovery and has generated important directions for future human research. Clinician involvement in basic animal research will improve the extent to which results are relevant to human rehabilitation and recovery.


A descriptive retrospective study of the treatment and outpatient service use in a clinical group of delusional disorder patients.
Gender differences in attendance rates for appointments in delusional disorder have been poorly studied. Furthermore, delusional disorder is traditionally considered a treatment-resistant disorder. We conducted a longitudinal retrospective study with a one-year follow-up, including 78 delusional disorder patients (DSM-IV-TR), consecutively admitted over a period of 10 years. We performed a follow-up for one year in order to describe demographic and clinical variables, and to compare the therapeutic effectiveness. In this study, due to the non-interventional nature of this study, indirect measures were used to measure treatment effectiveness. The sample was divided into three groups according to the antipsychotic received at the first admission. Twenty-three patients received Risperidone Long-Acting Injection (RILD), 30 oral risperidone, and 25 patients received other oral atypical antipsychotics. Delusional disorder women had a later age at onset and needed a longer duration of hospitalization. No statistically significant differences were found between the three treatment groups as regards demographic data, social and personal functioning, and psychopathology at admission. The RILD group showed higher maintenance rates at the end of the follow-up period. Furthermore, the RILD group required treatment with antidepressants and benzodiazepines less often. We concluded that treatment with RLAI may increase maintenance rates at follow-up in DD patients when compared with other oral atypical antipsychotics.


Diluting power of thick limbs of Henle. II. Bumetanide-sensitive 22Na+ influx in medullary vesicles.
We evaluated the effects of osmotic gradients on 22Na+ influx in vesicles prepared from rat outer renal medulla. 22Na+ influx driven in a coflow mode by an inwardly directed 100 mM KCl gradient was measured at 20 and 60 s; 1 mM bumetanide inhibited approximately 30% of 22Na+ influx. The bumetanide-sensitive 22Na+ influx was reduced by approximately 65% when either K+ or Cl- was omitted from the aqueous phases. We found that an osmotic gradient for vesicle shrinkage, that is, 600 mM urea in the extravesicular medium, enhanced the bumetanide-sensitive 22Na+ influx twofold. Conversely, an osmotic gradient for vesicle swelling, that is, with vesicles but not extravesicular media loaded with 600 mM urea, produced a 50% suppression of bumetanide-sensitive 22Na+ influx. Moreover, 600 mM extravesicular urea, an osmotic gradient for vesicle shrinkage, also reduced uptake of the nonspecific marker [14C]mannitol. These effects of osmotic gradients were not due to alterations in ionic driving forces, since bumetanide-sensitive 22Na+ influx driven in a counterflow mode by loading the vesicles with 100 mM NaCl also was activated or suppressed by osmotic gradients for vesicle shrinkage or swelling, respectively. We conclude that osmotic gradients, and/or vesicle volume changes, modulate bumetanide-sensitive Na+:K+:2Cl- activity.


Semiautomated method for microbiological vitamin assays.
A semiautomated method for microbiological vitamin assays is described, which includes separate automated systems for the preparation of the cultures and for the measurement of turbidity. In the dilution and dosage unit based on the continuous-flow principle, vitamin samples were diluted to two different dose levels at a rate of 40 per hr, mixed with the inoculated test broth, and dispensed into culture tubes. After incubation, racks with culture tubes were placed on the sampler of an automatic turbidimeter. This unit, based on the discrete-sample system, measured the turbidity and printed the extinction values at a rate of 300 per hr. Calculations were computerized and the results, including statistical data, are presented in an easily readable form. The automated method is in routine use for the assays of thiamine, riboflavine, pyridoxine, cyanocobalamin, calcium pantothenate, nicotinic acid, pantothenol, and folic acid. Identical vitamin solutions assayed on different days gave variation coefficients for the various vitamin assays of less than 10%.


The Evolutionary Legacy of Diversification Predicts Ecosystem Function.
Theory suggests that the structure of evolutionary history represented in a species community may affect its functioning, but phylogenetic diversity metrics do not allow for the identification of major differences in this structure. Here we propose a new metric, ELDERness (for Evolutionary Legacy of DivERsity) to estimate evolutionary branching patterns within communities by fitting a polynomial function to lineage-through-time (LTT) plots. We illustrate how real and simulated community branching patterns can be more correctly described by ELDERness and can successfully predict ecosystem functioning. In particular, the evolutionary history of branching patterns can be encapsulated by the parameters of third-order polynomial functions and further measured through only two parameters, the "ELDERness surfaces." These parameters captured variation in productivity of a grassland community better than existing phylogenetic diversity or diversification metrics and independent of species richness or presence of nitrogen fixers. Specifically, communities with small ELDERness surfaces (constant accumulation of lineages through time in LTT plots) were more productive, consistent with increased productivity resulting from complementary lineages combined with niche filling within lineages. Overall, while existing phylogenetic diversity metrics remain useful in many contexts, we suggest that our ELDERness approach better enables testing hypotheses that relate complex patterns of macroevolutionary history represented in local communities to ecosystem functioning.


Mixed pyogenic and tuberculous liver abscess: clinical suspicion is what matters.
Tuberculosis (TB) is a rare cause of liver abscess, even in country like India where it is a very common infection. Moreover, tubercular liver abscess (TLA) is the most unusual pattern of hepatic tuberculosis. We report an unusual case of liver abscess in an immune-competent patient presenting only with weight loss. On investigation, initially it appeared pyogenic, but later turn out to be a mixed infection with tuberculosis. He responded well to antibiotic and antitubercular drugs. A mixed pyogenic and TLA is very uncommon. We conclude that, tuberculosis should be suspected in liver abscess, especially in the absence of typical features and failure to respond to antibiotics.


Anesthesia for surgery of pituitary tumors.
Transsphenoidal operations for pituitary tumors have dramatically decreased the morbidity of pituitary surgery in a group of patients with many potential anesthetic problems. Although a good preoperative endocrine workup and anticipation of problems are the key to ensuring a good outcome, it remains a challenge to the "art" of neuroanesthesia to produce a smooth and safe anesthetic.


Using the PedsQL™ 3.0 asthma module to obtain scores comparable with those of the PROMIS pediatric asthma impact scale (PAIS).
The National Institutes of Health's Patient-Reported Outcomes Measurement Information System (PROMIS) has developed several scales measuring symptoms and function for use by the clinical research community. One advantage of PROMIS is the ability to link other scales to the PROMIS metric. The objectives of this research are to provide evidence of validity for one of the PROMIS measures, the Pediatric Asthma Impact Scale (PAIS), and to link the PedsQL™ Asthma Symptoms Scale with the metric of the PAIS. Descriptive statistics were computed describing the relationships among scores on the PAIS, the PedsQL™ Asthma Symptoms, Treatment, Worry, and Communication Scales, and the DISABKIDS Asthma Impact and Worry Scales for approximately 300 children ages 8-17. A novel linkage method based on item response theory (IRT), calibrated projection, was used to link scores on the PedsQL™ Asthma Symptoms Scale with the metric of the PAIS. The PAIS exhibited strong convergent validity with the PedsQL™ Asthma Symptoms Scale, and less strong relations with the other five scales. The linkage system uses scores on the PedsQL™ Asthma Symptoms Scale to produce relatively precise score estimates on the metric of the PAIS. Results of this study provide evidence for the validity of the PAIS, and a method to use scores on the PedsQL™ Asthma Symptoms Scale to estimate scores on the metric of the PAIS, in partial fulfillment of the PROMIS goal to provide a lingua franca for health-related quality of life.


Treatment of brain metastases in patients with testicular cancer.
Despite improved cure rates for patients with metastatic testicular cancer with cisplatin-based combination chemotherapy, patients who develop brain metastases are generally considered to possess a poor prognosis. This report summarizes the long-term results in 44 patients with brain metastases from testicular cancer treated between 1978 and 1995 at Hannover University Medical School. Histologically, 42 patients (95%) had a nonseminomatous germ cell cancer and two patients (5%) a seminoma. Thirty-nine patients (89%) had lung metastases and 37 (84%) fulfilled the criteria for advanced disease according to the Indiana University classification even without considering the brain metastases. Eighteen patients (41%) presented with brain metastases at primary diagnosis (group 1), four (9%) developed brain metastases at relapse after a previous favorable response to combination chemotherapy (group 2), and 22 (50%) developed brain metastases during or directly after cisplatin-based chemotherapy. Chemotherapy consisted of cisplatin-based combination treatment and radiotherapy was given as whole-brain irradiation of 30 to 40 Gy and in single cases combined with a boost of 10 Gy to single lesions. Overall, 10 patients achieved long-term survival (23%; 95% confidence interval [CI], 10.1% to 35.4%). The prognosis was significantly better for patients in groups 1 and 2, with six of 18 (33%) and three of four (75%) patients alive, compared with only one of 22 (5%) in group 3 (P < .01). Patients treated with either chemotherapy or radiotherapy alone did not achieve long-term survival, while nine of 28 (32%) who received treatment with both modalities with or without surgery achieved sustained long-term survival. During univariate analysis, patients with the diagnosis of brain metastases at first presentation (P < .01), patients with a single brain lesion (P < .02), and patients who received combined chemotherapy and radiotherapy (P < .03) had a significantly improved outcome. Long-term survival can be achieved in approximately 25% of patients with brain metastases from testicular cancer by combined treatment with brain irradiation and aggressive cisplatin-based chemotherapy. Patients who develop brain metastases during systemic treatment should receive only palliative radiation therapy, since sustained survival will not be reached.


Second-generation Langerhans cells originating from epidermal precursors are essential for CD8+ T cell priming.
In vivo studies questioned the ability of Langerhans cells (LCs) to mediate CD8(+) T cell priming. To address this issue, we used intradermal immunization with plasmid DNA, a system in which activation of CD8(+) T cells depends on delayed kinetics of Ag presentation. We found that dendritic cells (DCs) located in the skin at the time of immunization have limited ability to activate CD8(+) T cells. This activity was mediated by a second generation of DCs that differentiated in the skin several days after immunization, as well as by lymph node-resident DCs. Intriguingly, CD8(+) T cell responses were not affected following treatment with clodronate liposomes, immunization of CCR2(-/-) mice, or local neutralization of CCL20. This suggests that local, rather than blood-derived, DC precursors mediate CD8(+) T cell priming. Analysis of DC differentiation in the immunized skin revealed a gradual increase in the number of CD11c(+) cells, which reached their maximum 2 wk after immunization. A similar differentiation kinetics was observed for LCs, with the majority of differentiating LCs proliferating in situ from epidermal precursors. By using B6/Langerin-diphtheria toxin receptor chimeric mice and LC ablation, we demonstrated that epidermal LCs were crucial for the elicitation of CD8(+) T cell responses in vivo. Furthermore, LCs isolated from lymph nodes 2 wk after immunization contained the immunization plasmid and directly activated Ag-specific CD8(+) T cells ex vivo. Thus, these results indicate that second-generation Ag-expressing LCs differentiating from epidermal precursors directly prime CD8(+) T cells and are essential for optimal cellular immune responses following immunization with plasmid DNA.


Social work in family practice: what are the prospects?
In a study by Frangos and Chase of thirty-nine family practice residents, it was found that the majority had positive attitudes toward having a social worker associated with their future medical practice. Their findings, however, could have been positively skewed since their sample was small and from residents in only one program. This paper reports the findings of 159 board certified family practitioners surveyed in a national sample. This study also yielded positive findings but much less than those in Frangos and Chase's study. These and other findings related to family practitioners' attitudes and practices regarding social work involvement in their practices are reported and discussed.


[Insulin-using woman with type 2 diabetes and weight problems].
A 61-year-old overweight woman had been diagnosed with diabetes mellitus, hypertension and hypothyreosis. Treatment with antidiabetic and antihypertensive medication and thyroxine had been started. Blood sugar had been increasing despite medication and she had started using insulin. In 2003 she used 150 IE insulin per day. She tried hard to adhere to a recommended diet, but gradually became fatter, maximum weight was 120 kg. She started on a low carbohydrate diet on her own and lost 14 kg during 5 months. She had some hypoglycemic episodes and sought advice at Dr. Fedon Lindberg's Clinic. Her low carbohydrate diet was continued, endurance exercise was included, medication with metformin was started and during 8 months she was off insulin and showed much lower blood sugar values than before. She lost 14 kg during this period. She was motivated for loosing more weight and starter on a VLCD (very low caloric diet). She lost another 9 kg on this diet. She than started regular resistance training and her weight stabilized on 80 kg. Her HbA1c value has been reduced from 8.9 to 5.4% and her total/HDL cholesterol ratio has been reduced from 5.4 to 1.7. Her C-peptide value increased in the period when insulin was reduced, but is now reduced to 700 pmol/L. Micro-CRP has been reduced from 9.0 mg/L to 0.4 mg/L. With a low carbohydrate diet and exercise this woman no longer has diabetes or severe overweight. It is our opinion that many patients with type 2 diabetes can manage without medication (especially insulin) by reducing the intake of carbohydrates considerably.


Direct Acylation of C(sp(3))-H Bonds Enabled by Nickel and Photoredox Catalysis.
Using nickel and photoredox catalysis, the direct functionalization of C(sp(3))-H bonds of N-aryl amines by acyl electrophiles is described. The method affords a diverse range of α-amino ketones at room temperature and is amenable to late-stage coupling of complex and biologically relevant groups. C(sp(3))-H activation occurs by photoredox-mediated oxidation to generate α-amino radicals which are intercepted by nickel in catalytic C(sp(3))-C coupling. The merger of these two modes of catalysis leverages nickel's unique properties in alkyl cross-coupling while avoiding limitations commonly associated with transition-metal-mediated C(sp(3))-H activation, including requirements for chelating directing groups and high reaction temperatures.


Global and Regional Tidal Volume Distribution in Spontaneously Breathing Mechanically Ventilated Children.
Allowing the ventilated adult patient to breathe spontaneously may improve tidal volume (VT) distribution toward the dependent lung regions, reduce shunt fraction, and decrease dead space. It has not been studied if these effects under various levels of ventilatory support also occur in children. We sought to explore the effect of level of ventilatory support on VT distribution and end-expiratory lung volume (EELV) in spontaneously breathing ventilated children in the recovery phase of their acute respiratory failure. This is a secondary analysis of data from a prospective clinical trial comparing 2 different ventilator modes during weaning in mechanically ventilated children < 5 y: CPAP + pressure support ventilation (PSV) and pressure control (PC)/intermittent mandatory ventilation (IMV) + PSV with the mandatory breath rate set at 25% of baseline. Using electrical impedance tomography (EIT), we assessed VT distribution by calculating the center of ventilation. Polynomial functions of the second degree were plotted to evaluate regional lung filling characteristics. Changes in end-expiratory impedance were calculated to assess changes in EELV. Baseline measurements were compared with measurements during CPAP/PSV, PC/IMV + PSV, and during a downward titration of the level of pressure support. Thirty-five subjects with a median age 4.5 (2.1-12.9) months and a median ventilation time of 4.9 (3.3-6.9) d were studied. The overall median coefficient of variation was 50.1% and not different between CPAP/PSV or PC/synchronized IMV + PSV. Regional filling characteristics of the lung identified a homogeneous VT distribution under all study conditions. Downtapering of the level of PSV resulted in a significant shift of the coefficient of variation toward the dependent lung regions. Our data showed that allowing ventilated children in the recovery phase of respiratory failure to breathe spontaneously in a continuous spontaneous ventilation mode did not negatively affect VT distribution or EELV.


Adenylate cyclase is inhibited upon depletion of plasma-membrane cholesterol.
A procedure has been developed that allows for the depletion of rat liver plasma membrane cholesterol by incubation with liposomes at 4 degrees C. Upon cholesterol depletion, adenylate cyclase activity was inhibited and the membranes became more rigid, as determined by the flexibility of an incorporated fatty acid spin probe. Decreasing the cholesterol/phospholipid molar ratio elicited a pronounced drop in the net fold-stimulation of adenylate cyclase activity by glucagon. Two lipid phase separations were detected in cholesterol-depleted membranes at around 25 degrees C and 13 degrees C respectively. Breaks at these temperatures were observed in Arrhenius plots of both the mobility of the spin probe and the glucagon-stimulated adenylate cyclase activity for the range 2-40 degrees C, but only the one at the lower temperature for the fluoride-stimulated activity. It is proposed that the lipid phase separation occurring at 25 degrees C is localized in the external half of the bilayer, whereas that at 13 degrees C is due to lipids in the inner half of the bilayer. Similar structural and functional perturbations were manifest if the cholesterol-complexing polyene antibiotic amphotericin B was added to native membranes. The mechanism of adenylate cyclase inhibition achieved by cholesterol depletion and the domain structure of the plasma membrane in relation to cholesterol distribution are discussed. Native cholesterol/phospholipid ratios appear to optimize the functioning of adenylate cyclase in liver plasma membranes.


A human-derived neurovascular unit in vitro model to study the effects of cellular cross-talk and soluble factors on barrier integrity.
The blood-brain barrier (BBB) restricts paracellular and transcellular diffusion of compounds and is part of a dynamic multicellular structure known as the "neurovascular unit" (NVU), which strictly regulates the brain homeostasis and microenvironment. Several neuropathological conditions (e.g., Parkinson's disease and Alzheimer's disease), are associated with BBB impairment yet the exact underlying pathophysiological mechanisms remain unclear. In total, 90% of drugs that pass animal testing fail human clinical trials, in part due to inter-species discrepancies. Thus, in vitro human-based models of the NVU are essential to better understand BBB mechanisms; connecting its dysfunction to neuropathological conditions for more effective and improved therapeutic treatments. Herein, we developed a biomimetic tri-culture NVU in vitro model consisting of 3 human-derived cell lines: human cerebral micro-vascular endothelial cells (hCMEC/D3), human 1321N1 (astrocyte) cells, and human SH-SY5Y neuroblastoma cells. The cells were grown in Transwell hanging inserts in a variety of configurations and the optimal setup was found to be the comprehensive tri-culture model, where endothelial cells express typical markers of the BBB and contribute to enhancing neural cell viability and neurite outgrowth. The tri-culture configuration was found to exhibit the highest transendothelial electrical resistance (TEER), suggesting that the cross-talk between astrocytes and neurons provides an important contribution to barrier integrity. Lastly, the model was validated upon exposure to several soluble factors [e.g., Lipopolysaccharides (LPS), sodium butyrate (NaB), and retinoic acid (RA)] known to affect BBB permeability and integrity. This in vitro biological model can be considered as a highly biomimetic recapitulation of the human NVU aiming to unravel brain pathophysiology mechanisms as well as improve testing and delivery of therapeutics.


Validation of World Health Organisation HIV/AIDS clinical staging in predicting initiation of antiretroviral therapy and clinical predictors of low CD4 cell count in Uganda.
The WHO clinical guidelines for HIV/AIDS are widely used in resource limited settings to represent the gold standard of CD4 counts for antiviral therapy initiation. The utility of the WHO-defined stage 1 and 2 clinical factors used in WHO HIV/AIDS clinical staging in predicting low CD4 cell count has not been established in Uganda. Although the WHO staging has shown low sensitivity for predicting CD4<200 cells/mm(3), it has not been evaluated at for CD4 cut-offs of <250 cells/mm(3) or <350 cells/mm(3). To validate the World Health Organisation HIV/AIDS clinical staging in predicting initiation of antiretroviral therapy in a low-resource setting and to determine the clinical predictors of low CD4 cell count in Uganda. Data was collected on 395 participants from the Joint Clinical Research Centre, of whom 242 (61.3%) were classified as in stages 1 and 2 and 262 (68%) were females. Participants had a mean age of 36.8 years (SD 8.5). We found a significant inverse correlation between the CD4 lymphocyte count and WHO clinical stages. The sensitivity the WHO clinical staging at CD4 cell count of 250 cells/mm(3) and 350 cells/mm(3) was 53.5% and 49.1% respectively. Angular cheilitis, papular pruritic eruptions and recurrent upper respiratory tract infections were found to be significant predictors of low CD4 cell count among participants in WHO stage 1 and 2. The WHO HIV/AIDS clinical staging guidelines have a low sensitivity and about half of the participants in stages 1 and 2 would be eligible for ART initiation if they had been tested for CD4 count. Angular cheilitis and papular pruritic eruptions and recurrent upper respiratory tract infections may be used, in addition to the WHO staging, to improve sensitivity in the interim, as access to CD4 machines increases in Uganda.


Characterization of the gut microbiota in the golden takin (Budorcas taxicolor bedfordi).
The gut microbiota of mammals is a complex ecosystem, which is essential for maintaining gut homeostasis and the host's health. The high throughput sequencing allowed us to gain a deeper insight into the bacterial structure and diversity. In order to improve the health status of the endangered golden takins, we first characterized the fecal microbiota of healthy golden takins using high throughput sequencing of the 16S rRNA genes V3-V4 hypervariable regions. Our results showed that, Firstly, the gut microbiota community comprised 21 phyla, 40 classes, 62 orders, 96 families, and 216 genera. Firmicutes (67.59%) was the most abundant phylum, followed by Bacteroidetes (23.57%) and Proteobacteria (2.37%). Secondly, the golden takin maintained higher richness in spring than in the winter while community diversity and evenness was not significantly different. Thirdly, four female golden takins demonstrated highly similar microbiota and the five golden takin males had relatively highly similar microbiota. All of our results might indicate that the fecal microbiota of golden takins were influenced by the season and the animal's sex. The findings provided theoretical basis regarding the gut microbiota of golden takins and may offer new insights to protect this endangered species.


Irreducible intertrochanteric fractures of the femur.
A retrospective study during a 3-year interval revealed that four of 112 patients had intertrochanteric fractures that were irreducible by the usual closed manipulation and traction techniques at the time of surgery. Each of these patients' preoperative radiographs showed a fracture line that bisected the lesser trochanter and was relatively uncomminuted. Although longitudinal traction and appropriate closed manipulation provide acceptable reduction for most intertrochanteric fractures, the few with the described fracture pattern may require open reduction with removal of interposed soft tissue to achieve satisfactory alignment.


A review of unusual species of Cotesia (Hymenoptera, Braconidae, Microgastrinae) with the first tergite narrowing at midlength.
The unusual species of Cotesia (Hymenoptera, Braconidae, Microgastrinae) with the first tergite narrowing at midlength are reviewed. One new species, Cotesia trabalae sp. n. is described from India and compared with Cotesia pistrinariae (Wilkinson) from Africa, the only other species sharing the same character of all the described species worldwide. The generic placement of these two species, based on molecular and morphological analyses as well as parasitoid biology is discussed.


Functional neuroanatomy of deductive inference: a language-independent distributed network.
Studies of brain areas supporting deductive reasoning show inconsistent results, possibly because of the variety of tasks and baselines used. In two event-related functional magnetic imaging studies we employed a cognitive load paradigm to isolate the neural correlates of deductive reasoning and address the role (if any) of language in deduction. Healthy participants evaluated the logical status of arguments varying in deductive complexity but matched in linguistic complexity. Arguments also varied in lexical content, involving blocks and pseudo-words in Experiment I and faces and houses in Experiment II. For each experiment, subtraction of simple from complex arguments (collapsing across contents) revealed a network of activations disjoint from regions traditionally associated with linguistic processing and also disjoint from regions recruited by mere reading. We speculate that this network is divided into "core" and "support" regions. The latter include left frontal (BA 6, 47) and parietal (BA 7, 40) cortices, which maintain the formal structure of arguments. Core regions, in the left rostral (BA 10p) and bilateral medial (BA 8) prefrontal cortex, perform deductive operations. Finally, restricting the complex-simple subtraction to each lexical content uncovered additional activations which may reflect the binding of logical variables to lexical items.


Accuracy of gleason grading by practicing pathologists and the impact of education on improving agreement.
The aims of this study were to evaluate the accuracy of Gleason grading for prostatic adenocarcinoma among practicing pathologists in Japan and to determine the influence of education on this accuracy. Using a case-oriented approach, 16 hematoxylin and eosin-;stained glass slides with consensus scores established by 4 urologic pathologists were reviewed by 91 pathologists, divided into 2 groups. In group A, average agreements with consensus scores before and after an educational lecture were 55.7% (n = 17) and 68.4% (n = 25), and average kappa values were 0.43 and 0.67, respectively. Twelve pathologists reviewed slides twice in a different order, with average agreements of 59.5% and 77.6%, and average kappa values of 0.48 and 0.69 before and after the lecture, yielding a statistically significant improvement. In group B, the average agreement before providing an atlas with a tutorial was 61.3% (n = 61), and the kappa value was 0.44. In the second round, the average agreement was 74.5% (n = 39), and the kappa value was 0.68. Among 39 pathologists who reviewed slides twice, the average agreement in the first round was 58.8%, and the kappa value was 0.42. Improvement of both the average agreement and the kappa value were statistically significant. The average improvement in kappa values among participants who reviewed slides twice was 0.22 in group A and 0.27 in group B, a difference that is not statistically significant. Combining both groups, the incidence of concordant scores for 16 cases rose from 58.9% to 75.4%, an average increase of 16.5%. The undergrading of score 5-7 lesions was significantly reduced, from 36.3% to 14.2%. With respect to demographic factors, pathologists signing out more than 5000 cases per year showed a better agreement than those with more than 1000 cases per year (48.9% versus 78.8%; P = 0.031). These results indicate that the general agreement of Gleason scores among practicing pathologists in Japan was comparable with those in the Western countries as reported in the literature. Although this requires further improvement, both the lecture and the printed material had a similar influence on the degree of improvement.


Attenuation of hindlimb vasodilation in heat-stressed baboons during dehydration.
The influence of dehydration on hindlimb vasodilation during environmental heating (EH) was examined in eight unanesthetized chronically instrumented baboons. Mean iliac blood flow (MIBF), arterial blood pressure, and core temperature (Tc) were measured during EH of baboons in euhydrated and dehydrated states. EH consisted of acute exposure to high ambient temperatures (39-44 degrees C) until Tc reached 39.5 degrees C. Dehydration was produced by 68-72 h of fluid deprivation, which caused increases in plasma osmolality [291 +/- 1 (SE) to 338 +/- 6 mosmol/kg H2O] and sodium concentration (143 +/- 2 to 163 +/- 3 meq/l) and a 16% fall in plasma volume. The primary influence of dehydration was attenuation of the progressive rise in MIBF and iliac conductance (IC) during EH. Absolute MIBF and IC levels at Tc = 39.5 degrees C during EH were 44 and 52%, respectively, lower in the dehydrated state. Also, the MIBF-Tc and IC-Tc linear regression coefficients during EH were lower by 33 and 43%, respectively, in the dehydrated state. Since limb skeletal muscle blood flow does not increase during EH, we conclude that dehydration attenuates the heat stress-induced rise in skin blood flow in baboons, an influence that is similar to what occurs in humans.


HIV continuum of care in Europe and Central Asia.
The European Centre for Disease Prevention and Control (ECDC) supports countries to monitor progress in their response to the HIV epidemic. In line with these monitoring responsibilities, we assess how, and to what extent, the continuum of care is being measured across countries. The ECDC sent out questionnaires to 55 countries in Europe and Central Asia in 2014. Nominated country representatives were questioned on how they defined and measured six elements of the continuum. We present our results using three previously described frameworks [breakpoints; Joint United Nations Programme on HIV/AIDS (UNAIDS) 90-90-90 targets; diagnosis and treatment quadrant]. Forty countries provided data for at least one element of the continuum. Countries reported most frequently on the number of people diagnosed with HIV infection (37; 93%), and on the number in receipt of antiretroviral therapy (ART) (35; 88%). There was little consensus across countries in their approach to defining linkage to, and retention in, care. The most common breakpoint (>19% reduction between two adjacent elements) related to the estimated number of people living with HIV who were diagnosed (18 of 23; 78%). We present continuum data from multiple countries that provide both a snapshot of care provision and a baseline against which changes over time in care provision across Europe and Central Asia may be measured. To better inform HIV testing and treatment programmes, standard data collection approaches and definitions across the HIV continuum of care are needed. If countries wish to ensure an unbroken HIV continuum of care, people living with HIV need to be diagnosed promptly, and ART needs to be offered to all those diagnosed.


Melatonin signaling affects the timing in the daily rhythm of phagocytic activity by the retinal pigment epithelium.
Earlier studies in Xenopus have indicated a role for melatonin in the regulation of retinal disk shedding, but the role of melatonin in the regulation of daily rhythm in mammalian disk shedding and phagocytosis is still unclear. We recently produced a series of transgenic mice lacking melatonin receptor type 1 (MT1) or type 2 (MT2) in a melatonin-proficient background and have shown that removal of MT1 and MT2 receptors induces significant effects on daily and circadian regulation of the electroretinogram as well as on the viability of photoreceptor cells during aging. In this study we investigated the daily rhythm of phagocytic activity by the retinal pigment epithelium in MT1 and MT2 knock-out mice. Our data indicate that in MT1 and MT2 knock-out mice the peak of phagocytosis is advanced by 3 h with respect to wild-type mice and occurred in dark rather than after the onset of light, albeit the mean phagocytic activity over the 24-h period did not change among the three genotypes. Nevertheless, this small change in the profile of daily phagocytic rhythms may produce a significant effect on retinal health since MT1 and MT2 knock-out mice showed a significant increase in lipofuscin accumulation in the retinal pigment epithelium.


Innervated free flaps for foot reconstruction: a review.
Whether to provide a sensate plantar weight-bearing flap to reconstruct the foot remains an unanswered, but frequently asked, question. It has been more than a decade since Graham and Dellon reviewed this subject. Increasing emphasis on outcome analysis of microsurgical reconstruction has prompted this new review. All published peer-reviewed literature related to reconstruction of the plantar surface of the foot was reviewed to identify whether the flaps were 1) local, regional or distant; 2) innervated or non-innervated and, if innervated, then 3) identified as to the donor and the recipient peripheral nerves. Outcome measures included direct measurement of sensibility, development of ulceration, and activities of daily living. It was concluded that it is still not possible to determine that innervated flap reconstruction of the weight-bearing portion of the foot is critical for either durability or ambulation. It is recommended that the original nerves that innervate the flap be blocked prior to harvest to improve flap design, i.e., that the flap actually contains the intended nerve. It is recommended that appropriate donor nerves are selected to innervate the flaps, e.g., the calcaneal nerve should be used to reinnervate heel reconstructions (rather than the sural nerve), and medial/lateral plantar branches be used to reinnervate the arch and the forefoot. Reinnervating a muscle flap with a sensory nerve will permit reinnervation of the muscle and the overlying skin, but whether this provides a superior result in durability and gait remains to be seen.


C7 reference typing and nomenclature recommendations.
The results of reference typing for C7 are presented and discussed. It appears that the present literature is using consistent nomenclature except that it was not possible to distinguish between the European C7-3 allele and Japanese C7-6. Oriental populations have both a higher frequency of variants and a greater variety of variants than Caucasian populations. Some samples with complex patterns defied classification, and it is speculated that these may be from persons with duplicated C7 genes. It is recommended that no change to the present system of nomenclature be made before a more detailed molecular understanding of the system is achieved.


Effect of tofacitinib on lipid levels and lipid-related parameters in patients with moderate to severe psoriasis.
Psoriasis is a systemic inflammatory disease associated with increased cardiovascular (CV) risk and altered lipid metabolism. Tofacitinib is an oral Janus kinase inhibitor. The aim of the study was to investigate the effects of tofacitinib on traditional and nontraditional lipid parameters and CV risk markers in patients with psoriasis from a phase III study, OPT Pivotal 1. Patients with psoriasis were randomized to tofacitinib 5 or 10 mg twice daily (BID) or placebo BID. Serum samples were collected at baseline, week 4, and week 16. Analyses included serum cholesterol levels, triglycerides, lipoproteins, lipid particles, lipid-related parameters/CV risk markers, and high-density lipoprotein (HDL) function analyses. At week 16, small concurrent increases in mean low-density lipoprotein cholesterol (LDL-C) and HDL cholesterol (HDL-C) levels were observed with tofacitinib; total cholesterol/HDL-C ratio did not change. There was no significant change in the number of small dense LDL particles, which are considered to be more atherogenic than large particles, and oxidized LDL did not increase. Paraoxonase 1 activity, linked to HDL antioxidant capacity, increased, and HDL-associated serum amyloid A, which reduces the anti-atherogenic potential of HDL, decreased. HDL capacity to promote cholesterol efflux from macrophages did not change. Lecithin-cholesterol acyltransferase activity, which is associated with reverse cholesterol transport, increased. Markers of systemic inflammation, serum amyloid A and C-reactive protein, decreased with tofacitinib. While small increases in lipid levels are observed with tofacitinib treatment in patients with psoriasis, effects on selected lipid-related parameters and other circulating CV risk biomarkers are not suggestive of an increased CV risk [NCT01276639].


Existing data sources for clinical epidemiology: Scandinavian Cohort for osteonecrosis of the jaw - work in progress and challenges.
Osteonecrosis of the jaw (ONJ) is a severe side effect associated with antiresorptive treatment. Monitoring of ONJ using routine databases in Scandinavian countries is a challenge owing to lack of valid algorithms and to heterogeneous referral practices. The aim of this paper is to describe the process of establishing a Scandinavian ONJ Cohort enrolling all ONJ cases related to antiresorptive treatment arising in Denmark, Norway, and Sweden between 2011 and 2019. The initial purpose of the cohort is to support an ongoing pharmacovigilance study of denosumab and zoledronic acid in Denmark, Norway, and Sweden. The three countries, with their 199 clinics, departments, and units of oral and maxillofacial surgery, both hospital-based and freestanding, differ somewhat in referral practices of the ONJ patients. By directly contacting all providers of care to ONJ patients in the three countries, we established a network for reporting incident cases to each country's research database directly or through a member of the Scandinavian ONJ task force as a liaison. The task force includes a Scandinavian coordinator and three national coordinators collaborating directly with the clinics. A uniform ONJ registration form has been developed, and the relevant medical community has been informed either directly or through presentations at professional meetings. A website with study information is published in each country, and data entry is ongoing. This large-scale systematic uniform registration of ONJ cases in Denmark, Norway, and Sweden, with an underlying total population of more than 20 million people, merged into the Scandinavian ONJ Cohort, will contribute to better knowledge and understanding of this challenging group of patients, and ultimately, help improve patient care. The Scandinavian ONJ Cohort as a whole and its component national ONJ research databases may offer the potential for large-scale multinational intervention and safety studies in the future.


[Health expenditures in Spain at the dawn of the 3d millennium].
In 1997, per capita health expenditures reached in Spain 805 Euros as compared to 1146 Euros in the entire European Union. In terms of percent GDP, Spanish health expenditures amounted to 7.4 percent, as compared to 7.9 in the European Union. About 80% of Spanish health expenditures are public-funded. Since 1993 onwards, there is a political consensus for keeping at the same pace health expenditures increase and GDP growth. However it is not certain that such a consensus can be maintained, as trade unions are increasingly voicy for a strong growth of salaries in the public sector. There will be also pressure on the demand side, both in the short term (the Spanish people are increasingly expecting to raise their consumption of health services to the level of other European countries) and in the long run (impact of population aging).


Endoscopic transorbital approach to the insular region: cadaveric feasibility study and clinical application (SevEN-005).
The insula is a complex anatomical structure. Accessing tumors in the insula remains a challenge due to its anatomical complexity and the high chance of morbidity. The goal of this study was to evaluate the feasibility of an endoscopic transorbital approach (ETOA) to the insular region based on a cadaveric study. One cadaveric head was used to study the anatomy of the insula and surrounding vessels. Then, anatomical dissection was performed in 4 human cadaveric heads using a dedicated endoscopic system with the aid of neuronavigation guidance. To assess the extent of resection, CT scanning was performed before and after dissection. The insular region was directly exposed by a classic transcranial approach to check the extent of resection from the side with a classic transcranial approach. The entire procedure consisted of two phases: an extradural orbital phase and an intradural sylvian phase. After eyelid incision, the sphenoid bone and orbital roof were extensively drilled out with exposure of the frontal and temporal dural layers. After making a dural window, the anterior ramus of the sylvian fissure was opened and dissected. The M2 segment of the middle cerebral artery (MCA) was identified and traced posterolaterally. A small corticectomy was performed on the posterior orbital gyrus. Through the window between the lateral lenticulostriate arteries and M2, the cortex and medulla of the insula were resected in an anteroposterior direction without violation of the M2 segment of the MCA or its major branches. When confirmed by pterional craniotomy, the sylvian fissure and the MCA were found to be anatomically preserved. After validation of the feasibility and safety based on a cadaveric study, the ETOA was successfully performed in a patient with a high-grade glioma (WHO grade III) in the right insula. The transorbital route can be considered a potential option to access tumors located in the insula. Using an ETOA, the MCA and its major branches were identified and preserved while removal was performed along the long axis of the insula. In particular, lesions in the anterior part of the insula are most benefited by this approach. Because this approach was implemented in only one patient, additional discussion and further verification is required.


Delayed anastomotic leakage following sphincter-preserving surgery for rectal cancer.
The aim of the present study was to investigate the characteristics of patients who developed delayed anastomotic leakage (DAL) following sphincter-preserving surgery for rectal cancer. We evaluated the following factors: (1) the incidence of DAL, (2) the clinical features of patients who developed DAL, (3) the risk factors for DAL, and (4) treatment outcomes. We reviewed the case histories of 1,838 rectal cancer patients who had undergone curative resection with sphincter preservation and without protective stoma formation between January 2000 and December 2006. DAL was defined as the development of a pelvic abscess or fistula around the anastomosis more than 3 weeks post-surgery in patients without tumor recurrence who had resumed a normal diet and defecation. In 1.3% (24/1,838) of the patients, DAL developed on median postoperative day 99 (range 22-2,069). Pelvic abscess (50%) and anastomotic-vaginal fistula (41.7%) were the most common causes of DAL. Independent risk factors for the development of DAL were: (1) female gender (hazard ratio 3.03; 95% CI 1.06-8.8), (2) low-level anastomosis (< or = 4 cm from the anal verge) (hazard ratio 5.76; 95% CI 1.37-22.39), and (3) preoperative chemoradiation therapy (hazard ratio 4.56; 95% CI 1.4-14.92). Stoma formation was performed in all of the 24 patients. The 3-year stoma-retention rate in patients with DAL was significantly higher than in patients with early anastomotic leakage (72.2% vs 17.5%, P < 0.001). DAL following sphincter-preserving surgery for rectal cancer occurred relatively frequently in our sample and was associated with female gender, a low level of anastomosis, and preoperative radiotherapy. DAL patients required long-term or permanent stomas.


Atonal is a proneural gene for a subset of olfactory sense organs in Drosophila.
The antenna of the adult fruit fly, Drosophila melanogaster, is covered with three morphologically distinct types of olfactory sense organs. In addition, mechano- and hygro-sensitive receptors are also present on its surface. While much has been learnt about the development of peripheral nervous system in Drosophila, the mechanisms underlying the development of olfactory sensilla are just beginning to be unraveled. The antennal sense organs have several properties that make them distinct from other sense organs. While each sensillum type is arranged in a well-defined region of the antenna, the position of an individual sensillum is not fixed. The development of these sense organs appears to combine an initial step of cell recruitment, as in photoreceptors, followed by cell lineage mechanisms, as in the development of other external sense organs. The earliest step in development, the selection of a sensory organ precursor, involves the interaction of proneural and neurogenic genes. The proneural gene for the antennal sense organs has been elusive so far. We show that the basic helix-loop-helix (bHLH) transcription factor encoded by atonal (ato) is a proneural gene for one morphological type of olfactory sensilla on the antenna and for all the olfactory sensilla on the maxillary palp. Loss of function and overexpression experiments together reveal that ato is both necessary and sufficient to specify these sensilla. Immunohistochemical experiments show that Ato expresses in a dynamic pattern in the developing antennal disc. Our results demonstrate that ato acts solely in the specification of antennal sensilla coeloconica. This along with our previous observation that the AS-C genes do not function in antenna allows us to suggest that other proneural genes must operate in the specification of sensilla basiconica and trichoidea. Our experiment involving overexpression of extramacrochaetae, a negative regulator of bHLH encoding genes, results in a significant reduction in the number of all three types of antennal sensilla. This suggests that the unidentified proneural gene(s) possibly encode bHLH factors.


[Endonasal dacryocystorhinostomy (DCR) with protected drill].
To analyze the interest of protected drills to perform endoscopic endonasal DCR. [corrected] Fifty patients suffering from chronic epiphora were operated successively. None of those patients had had prior nasal surgery. The same surgeons (BF and ER) did the surgical procedure for all the cases. The technical steps were: endoscopic visualization of the middle meatus and evaluation of the anatomic variations of the nose; identification of the lacrymal bag with a 1 mm light in the canaliculus; unciformectomy; the maxillary bone was drilled with a 3.2 mm bit (straightshot Xomed), protected and irrigated; the lacrymal bag was opened by cissors and the flap was pushed in the ethmoid and coagulated. The suction-irrigation system was efficient to clean the surgical site, with a good endoscopic control. The protected bit didn't damage the 4-mm optics and the septal wall mucosa, even in narrow noses. The staightshot bit (Xomed) was tough enough to drill the maxillary bone in only 68 % of the surgical cases. In 28 % the drilling was unsatisfactory and in 4 % it was inefficient. The functional results at 6 month were good in 85 %. There was no difference between normal and narrow noses. The concept of protected bits perfectly matches the endoscopic endonasal DCR. The visual comfort and security of the surgical procedure are increased. Short term results are equivalent to external DCR, but are rather inefficient and expensive.


Low incidence of anti-osteoporosis treatment after hip fracture.
Following hip fracture, pharmacologic treatment can reduce the rate of subsequent fragility fractures. The objective of the present study was to assess the proportion of patients who are managed with bisphosphonates or selective estrogen-receptor modulators after hip fracture and to evaluate, among those managed with alendronate, the twelve-month compliance and persistence with treatment. Data were gathered from health insurance companies and were collected by AIM (Agence Intermutualiste) for the Belgian National Social Security Institute (INAMI). We selected all postmenopausal women who had been hospitalized for a hip fracture between April 2001 and June 2004 and had not been previously managed with bisphosphonates. Patients who had received alendronate treatment after the hip fracture were categorized according to their formulation use during the follow-up study (daily, weekly, daily followed by weekly, or weekly followed by weekly). Compliance at twelve months was quantified with use of the medication possession ratio (i.e., the number of days of alendronate supplied during the first year of treatment, divided by 365). Persistence with prescribed treatment was calculated as the number of days from the initial prescription to a lapse of more than five weeks after completion of the previous prescription refill. The cumulative treatment persistence rate was determined with use of Kaplan-Meier survival curves. A total of 23,146 patients who had sustained a hip fracture were identified. Of these patients, 6% received treatment during the study period: 4.6% received alendronate, 0.7% received risedronate, and 0.7% received raloxifene. Bisphosphonate treatment was dispensed to 2.6% and 3.6% of the patients within six months and one year after the occurrence of the hip fracture, respectively. Among women who received alendronate daily (n = 124) or weekly (n = 182) and were followed for at least one year after the hip fracture, the twelve-month mean medication possession ratio was 67% (65.9% in the daily group and 67.7% in the weekly group). The analysis of persistence with treatment included a total of 726 patients (142 in the daily group, 261 in the weekly group, and 323 in the switch group). At twelve months, the rate of persistence was 41% and the median duration of persistence was 40.3 weeks. The vast majority of patients who experience a hip fracture do not take anti-osteoporotic therapy after the fracture. Furthermore, among patients who begin alendronate treatment after the fracture, the adherence to treatment decreases over time and remains suboptimal.


Probabilistic neural-network structure determination for pattern classification.
Network structure determination is an important issue in pattern classification based on a probabilistic neural network. In this study, a supervised network structure determination algorithm is proposed. The proposed algorithm consists of two parts and runs in an iterative way. The first part identifies an appropriate smoothing parameter using a genetic algorithm, while the second part determines suitable pattern layer neurons using a forward regression orthogonal algorithm. The proposed algorithm is capable of offering a fairly small network structure with satisfactory classification accuracy.


In situ characterization of cellular infiltrates in lupus vulgaris indicates lesional T-cell activation.
Skin biopsy specimens from nine patients with lupus vulgaris were examined in situ by means of monoclonal antibodies directed against phenotypes of lymphocyte subsets, Langerhans cells, HLA-DR antigens, and interleukin 2 receptor. The epidermis showed prominent changes, including intense expression of HLA-DR on keratinocytes, increase in epidermal cell layers, moderate to high Langerhans cell hyperplasia, and infiltration by CD3+ pan-T cells as well as CD8+ (cytotoxic/suppressor) and CD4+ (helper/inducer) T cells. The predominant lymphocyte in the dermal granulomas was the activated CD3+ T cell, expressing major histocompatibility complex class II antigens and interleukin 2 receptor. CD4+ and CD8+ cells were randomly distributed among the epithelioid cells, which showed intense staining for major histocompatibility complex class II antigens. In all except two patients, the CD4+ population was greater than that of the CD8+ cells. CD1+ Langerhans cells were scattered in moderate numbers in the dermal granulomas. Acid-fast bacilli were conspicuously absent in the biopsy specimens. These features suggest that T-cell activation and Langerhans cell hyperplasia are prominent features of dermal tuberculosis.


Titin copy number variations associated with dominant inherited phenotypes.
Titinopathies are caused by mutations in the titin gene (TTN). Titin is the largest known human protein; its gene has the longest coding phase with 364 exons. Titinopathies are very complex neuromuscular pathologies due to the variable age of onset of symptoms, the great diversity of pathological and muscular impairment patterns (cardiac, skeletal muscle or mixed) and both autosomal dominant and recessive modes of transmission. Until now, only few CNVs in TTN have been reported without clear genotype-phenotype associations. Our study includes eight families with dominant titinopathies. We performed next-generation sequencing or comparative genomic hybridisation array analyses and found CNVs in the TTN gene. We characterised these CNVs by RNA sequencing (RNAseq) analyses in six patients' muscles and performed genotype-phenotype inheritance association study by combining the clinical and biological data of these eight families. Seven deletion-type CNVs in the TTN gene were identified among these families. Genotype and RNAseq results showed that five deletions do not alter the reading frame and one is out-of-reading frame. The main phenotype identified was distal myopathy associated with contractures. The analysis of morphological, clinical and genetic data and imaging let us draw new genotype-phenotype associations of titinopathies. Identifying TTN CNVs will further increase diagnostic sensitivity in these complex neuromuscular pathologies. Our cohort of patients enabled us to identify new deletion-type CNVs in the TTN gene, with unexpected autosomal dominant transmission. This is valuable in establishing new genotype-phenotype associations of titinopathies, mainly distal myopathy in most of the patients.


Ocular characteristics in a variant microcephalic primordial dwarfism type II.
Microcephalic osteodysplastic primordial dwarfism, type II (MOPD II) is a rare disease that is assumed to be caused by a pericentrin (PCNT) gene mutation. Clinical manifestations have been reported in pediatrics and neurology; however, only a few ocular findings have been documented. We present three unrelated cases of MOPD II with similar facial features and short stature. Unlike the cases described in the literature, all subjects had normal birth weight and height but their growth was retarded thereafter. In addition to delayed milestones, they have a broad forehead, maxillary protrusion, long peaked nose, high nasal bridge, low-set large ears, extreme reromicrogenia, and normal-sized teeth. These three patients had similar ocular manifestations with the short axial length associated with high hyperopia more than + 9 diopters (D) and macular scarring. The oldest subject was a 20 year-old male without neurological symptoms. One female subject had developed alopecia during the previous 2 years. The other female subject had moyamoya disease, but a genetic study revealed a normal PCNT gene. This is the first report of MOPD II focusing on ocular findings, suggesting that macular dystrophy and high hyperopia are the common ocular characteristics of MOPD II. Prompt referral to an ophthalmologist is essential. Although refractive amblyopia can be treated with optical correction, visual prognosis may be poor due to maculopathy.


Free Energy Landscape of Type III Fibronectin Domain with Identified Intermediate State and Hierarchical Symmetry.
The tenth domain of type III fibronectin (FNIII_{10}) mediates cell adhesion to the extracellular matrix. Despite its structural similarity to immunoglobulin domains, FNIII_{10} exhibits unique unfolding behaviors. We employed magnetic tweezers to investigate the unfolding and folding dynamics of FNIII_{10} under physiological forces (4-50 pN). Our results showed that FNIII_{10} follows a consistent transition pathway with an intermediate state characterized by detached A and G β strands. We determined the folding free energies and all force-dependent transition rates of FNIII_{10} and found that both unfolding rates from the native state to the intermediate state and from the intermediate state to the unfolded state deviate from Bell's model. We constructed a quantitative free energy landscape with well-defined traps and barriers that exhibits a hierarchical symmetrical pattern. Our findings provide a comprehensive understanding of FNIII_{10} conformational dynamics and demonstrate how free energy landscape of multistate biomolecules can be precisely mapped, illuminating the relationship between thermal stability, intermediate states, and folding rates in protein folding.


Lignans from Schisandra sphenathera Rehd. et Wils. and semisynthetic schisantherin A analogues: absolute configuration, and their estrogenic and anti-proliferative activity.
A new dibenzocyclooctene-type lignan, named schisandrin A1 (1), together with nine known lignans (2-10), was isolated from the stems of Schisandra sphenathera. The structure of schisandrin A1, which contains a spirocyclic epoxy unit, was established by means of spectroscopic methods. The absolute configurations of schisandrin A1 (1) and schisantherin A (2) were determined by electronic circular dichroism (CD) and TDDFT calculations, with 2 further confirmed by X-ray crystallographic data. Ten new schisantherin A derivatives (11-20) and 6,7-secoschisantherol A (2b) were synthesized. In addition, natural lignans and semisynthetic schisantherin A derivatives showed the antiproliferative activity on four human cancer cell lines and Id1 (an inhibitor of DNA binding protein) and estrogenic potency. Compounds 5, 7, and 8 exhibited very potent estrogenic activity.


Embodying person-centred being and doing: Leading towards person-centred care in nursing homes as narrated by managers.
To explore how managers describe leading towards person-centred care in Swedish nursing homes. Although a growing body of research knowledge exists highlighting the importance of leadership to promote person-centred care, studies focused on nursing home managers' own descriptions of leading their staff towards providing person-centred care is lacking. Descriptive interview study. COREQ guidelines have been applied. The study consisted of semi-structured interviews with 12 nursing home managers within 11 highly person-centred nursing homes purposively selected from a nationwide survey of nursing homes in Sweden. Data collection was performed in April 2017, and the data were analysed using content analysis. Leading towards person-centred care involved a main category; embodying person-centred being and doing, with four related categories: operationalising person-centred objectives; promoting a person-centred atmosphere; maximising person-centred team potential; and optimising person-centred support structures. The findings revealed that leading towards person-centred care was described as having a personal understanding of the PCC concept and how to translate it into practice, and maximising the potential of and providing support to care staff, within a trustful and innovative work place. The findings also describe how managers co-ordinate several aspects of care simultaneously, such as facilitating, evaluating and refining the translation of person-centred philosophy into synchronised care actions. The findings can be used to inspire nursing home leaders' practices and may serve as a framework for implementing person-centred care within facilities. A reasonable implication of these findings is that if organisations are committed to person-centred care provision, care may need to be organised in a way that enables managers to be present on the units, to enact these strategies and lead person-centred care.


A topical antioxidant solution containing vitamins C and E stabilized by ferulic acid provides protection for human skin against damage caused by ultraviolet irradiation.
Skin cancer and photoaging changes result from ultraviolet (UV)-induced oxidative stress. Topical antioxidants may protect skin from these effects. We sought to determine whether a stable topical formulation of 15% L-ascorbic acid, 1% alpha-tocopherol, and 0.5% ferulic acid (CEFer) could protect human skin in vivo from substantial amounts of solar-simulated UV radiation. CEFer and its vehicle were applied to separate patches of normal-appearing human skin for 4 days. Each patch was irradiated with solar-simulated UV, 2 to 10 minimal erythema doses, at 2-minimal erythema dose intervals. One day later, skin was evaluated for erythema and sunburn cells, and immunohistochemically for thymine dimers and p53. UV-induced cytokine formation, including interleukin (IL)-1alpha, IL-6, IL-8, and IL-10, and tumor necrosis factor-alpha, were evaluated by real-time polymerase chain reaction. CEFer provided significant and meaningful photoprotection for skin by all methods of evaluation. The number of patients evaluated was relatively small. CEFer provided substantial UV photoprotection for skin. It is particularly effective for reducing thymine dimer mutations known to be associated with skin cancer. Its mechanism of action is different from sunscreens and would be expected to supplement the sun protection provided by sunscreens.


Preserved endothelial progenitor cell angiogenic activity in African American essential hypertensive patients.
African American (AA) subjects with essential hypertension (EH) have greater inflammation and cardiovascular complications than Caucasian EH. An impaired endogenous cellular repair system may exacerbate vascular injury in hypertension, yet whether these differ between AA EH and Caucasian EH remains unknown. Vascular repair by circulating endothelial progenitor cells (EPCs) is controlled by regulators of EPC mobilization, homing, adhesion and new vessel formation, but can be hindered by various cytokines. We hypothesized that EPC levels and function would be impaired in AA EH compared with Caucasian EH, in association with increased levels of inflammatory mediators and EPC regulators. CD34+/KDR+ EPCs were isolated from inferior vena cava and renal vein blood samples of AA EH and Caucasian EH patients (n = 18 each) and from peripheral veins of 17 healthy volunteers (HVs) and enumerated using fluorescence-activated cell sorting. Angiogenic function of late-outgrowth endothelial cells expanded from these samples for 3 weeks was tested in vitro. Levels of inflammatory mediators, angiogenic factors and EPC regulators were measured by Luminex. EPC levels were decreased in both AA and Caucasian EH compared with HVs, whereas their late-outgrowth endothelial cell angiogenic function was comparable. Levels of several inflammatory mediators were elevated in AA EH compared with Caucasian EH and HVs. Contrarily, vascular endothelial growth factor and its receptor-2 were lower. EPC levels inversely correlated with blood pressure in all hypertensive patients and estimated glomerular filtration rate with inflammatory mediators only in AA EH. Despite lower EPC numbers, decreased vascular endothelial growth factor signaling and inflammation, EPC function is preserved in AA EH compared with Caucasian EH and HVs, suggesting compensatory mechanisms for vascular repair.


A radiographic evaluation of temporomandibular and hand (Metacarpophalangeal) / wrist joints of patients with adult rheumatoid arthritis.
A review of literature revealed that, although the involvement of temporomandibular joint (TMJ) in rheumatoid arthritis (RA) patients is not uncommon, variation in presentation persist. Comparative studies of bony changes in the right and left TMJ with the right and left peripheral hand (Metacarpophalangeal-MCP)/wrist joints have not been done, to the best of our knowledge. In this cross-sectional study, the temporomandibular and hand (MCP) and wrist joints of fifteen rheumatoid arthritis patients were evaluated with questionnaires, clinical and lab assessment and radiographically using conventional radiographs and computed tomography. Students t-test was applied for the statistical analysis of the data obtained and a P value of 0.05 was considered as statistically significant. Comparisons between the right TMJ with right MCP/wrist joint and left TMJ with left MCP/wrist joint did not reveal statistically significant results. Radiographically, flattening and erosions were the common manifestations. MCP joints were more affected than the wrist, but whenever the wrist was involved, it was more likely to be bilaterally affected. Although the TMJ showed osseous changes of a higher grade than the hand (MCP) and wrist joints radiographically, it was observed that patients were more aware of the peripheral joint discomfort. There were no significant differences between TMJ and peripheral joints on both right and left sides.


Specimen stability for DNA-based diagnostic testing.
The use of molecular diagnostic testing is increasing in the clinical setting; therefore, data regarding DNA stability in clinical specimens are essential for correct test performance and interpretation. This study was designed to determine DNA stability in peripheral blood and solid tissue under different storage conditions. DNA quality and yield were assayed by spectrophotometric absorbance, gel electrophoresis, and suitability for Southern hybridization and polymerase chain reaction (PCR), the most widely employed clinical DNA analyses. A second goal of the study was to evaluate DNA stability during storage at 4 degrees C for 1 month to 3 years. The data show that freezing or refrigeration of separated leukocytes is preferable for short- to intermediate-term storage and freezing is preferable for solid tissue. DNA degradation varying from slight to severe is seen inconsistently with such specimens, probably due to sampling of unevenly frozen-tissue areas. Depending on the degree of DNA degradation, analysis may still be possible by PCR and in some cases even by Southern hybridization. Once isolated, DNA was stable at 4 degrees C for at least 3 years. These results suggest a more flexible approach to specimen requirements for molecular pathology, as some samples that would routinely be rejected gave interpretable results.


Cardiac-specific activity of an Nkx2-5 enhancer requires an evolutionarily conserved Smad binding site.
Heart formation in vertebrates and fruit flies requires signaling by bone morphogenetic proteins (BMPs) to cardiogenic mesodermal precursor cells. The vertebrate homeobox gene Nkx2-5 and its Drosophila ortholog, tinman, are the earliest known markers for the cardiac lineage. Transcriptional activation of tinman expression in the cardiac lineage is dependent on a mesoderm-specific enhancer that binds Smad proteins, which activate transcription in response to BMP signaling, and Tinman, which maintains its own expression through an autoregulatory loop. Here, we show that an evolutionarily conserved, cardiac-specific enhancer of the mouse Nkx2-5 gene contains multiple Smad binding sites, as well as a binding site for Nkx2-5. A single Smad site is required for enhancer activity at early and late stages of heart development in vivo, whereas the Nkx2-5 site is not required for enhancer activity. These findings demonstrate that Nkx2-5, like tinman, is a direct target for transcriptional activation by Smad proteins; however, the independence of this Nkx2-5 enhancer of Nkx2-5 binding suggests a fundamental difference in the transcriptional circuitry for activation of Nkx2-5 and tinman expression during cardiogenesis in vertebrates and fruit flies.


The Healer's Art: Remembering Our Professional Lineage in Community through the Cultivation of Individual Core Values.
This article was migrated. The article was marked as recommended. Introduction: There is both patient and provider dissatisfaction with the climate of healthcare delivery. Upon review, this is found to be at least in part attributable to the mechanization of health care, which often involves more computer interaction than hands-on care. Despite rising costs, the physical exam is replaced by lab tests and radiologic studies, generating more cost. The time-honored respect for a carefully obtained history from the patient is replaced by a computer check-box template. The humanity of both physician and patient are marginalized, with increased potential for both diagnostic and therapeutic compromise. Though access to medical information about disease is possible with bioinformatics, artificial intelligence cannot substitute for analysis by an informed, attentive, and properly educated physician. The process of healing must begin with the first patient visit - and the presence of an informed, compassionate, and fully attentive physician. Objective: To describe the history of medical educators' grappling with this problem through 3 landmark articles over a 100-year period. To illustrate the challenges of the climate of medical education. To offer some educational strategies (with examples of successful programs) to teach physicians using the Humanities. To illustrate that the art and science of medicine are synergistic, not dichotomous. Methods: Two educational theories ripe for use: Chickering and the Discovery Model, and Osler's recommended bedside reading list, exemplary programs that are being used currently (and over the last 25-plus years) to emphasize the importance of both the science and the practice of medicine in an effort to optimize the medical climate. Conclusion: The problem of physician burnout and patient dissatisfaction is being addressed in the medical literature, by regulatory societies devoted to physician wellness and by medical educators. This is nevertheless a challenge given the current electronic climate (with bioinformatics and artificial intelligence) and revenue-focused agendas of practice management business people. Results: With an awareness of the need for emphasis on the humanities coupled with an historical perspective over the last 100 years, a spirit of hope can be provided to both physician and patient from the lineage of the medical profession, which also is a legacy for our medical students.


The design of a project to assess bilateral versus unilateral hearing aid fitting.
Binaural hearing provides advantages over monaural in several ways, particularly in difficult listening situations. For a person with bilateral hearing loss, the bilateral fitting of hearing aids thus seems like a natural choice. However, surprisingly few studies have been reported in which the additional benefit of bilateral versus unilateral hearing aid use has been investigated based on real-life experiences. Therefore, a project has been designed to address this issue and to find tools to identify people for whom the drawbacks would outweigh the advantages of bilateral fitting. A project following this design is likely to provide reliable evidence concerning differences in benefit between unilateral and bilateral fitting of hearing aids by evaluating correlations between entrance data and outcome measures and final preferences.


Quantification and characterization of virus-like particles by size-exclusion chromatography and nanoparticle tracking analysis.
The rapid quantification of enveloped virus-like particles (VLPs) requires orthogonal methods to obtain reliable results. Three methods-nanoparticle tracking analysis (NTA), size-exclusion HPLC (SE-HPLC) with UV detection, and detection with multi-angle light scattering (MALS)-for quantification of enveloped VLPs have been compared, and the lower and upper limits of detection and quantification have been evaluated. NTA directly counts the enveloped VLPs, and a particle number is obtained with a lower limit of detection (LLOD) of 1.7×107part/mL and lower limit of quantification (LLOQ) of 3.4×108part/mL. SE-HPLC with UV detection was calibrated with standards characterized by NTA, and a LLOD of 6.9×109part/mL and LLOQ of 2.1×1010part/mL were found. SE-HPLC with MALS does not require a pre-calibrated sample because with a spherical model based on the Rayleigh-Gans-Debye approximation, the particle concentration can be directly deduced from the scattered light. A LLOD of 4.8×108part/mL and LLOQ of 2.1×109part/mL were measured and substantially lower compared to the UV method. The absolute particle concentration measured by SE-HPLC-MALS is one order of magnitude lower compared to measurement by NTA, which is explained by the wide size distribution of an enveloped VLP suspension. The model used for evaluation of light scattering data assumes monodisperse, homogeneous, and spherical particles.


Update on avian influenza pandemic threat.
So far, the current risk to people in the United States from the H5N1 bird flu outbreak in Asia is low (CDC, 2005a). The strain of H5N1 virus found in Asia has not been found in the United States; and as of November 2005, there have been no human cases of H5N1 flu reported in the United States. Travel to countries in Asia has not been restricted, but travelers are advised to avoid all direct contact with poultry, poultry farms, animals in live food markets, and any surfaces that appear to be contaminated with feces from poultry or other animals (CDC, 2005a). Meanwhile, medical and public health personnel are watching closely and preparedness plans are under way to respond to the threat of an avian influenza pandemic.


Liver retransplantation of more than two grafts for recurrent failure.
Transplantation of more than two livers for recurring graft failure has not been specifically addressed in the literature. A retrospective analysis was conducted from a total of 2527 overall liver transplants at our institution. Main indications for multiple retransplant included primary nonfunction, chronic rejection, hepatic artery thrombosis, and recurrent disease. We identified 39 patients who received more than two grafts (32 received 3 grafts, 5 received 4 grafts, and 2 received 5 grafts). All patients required interposition arterial grafts from the aorta and hepatojejunostomy for the biliary reconstruction. Seventeen patients are still alive at last follow-up. Perioperative mortality rates after 3rd, 4th, and 5th liver graft were 25%, 14%, and 50%, respectively. Patient and graft survival rates were 72% and 56% at 1 year, respectively. Median length of stay was 27 days and median graft survival was 2.9 years. Selection of patients and a significant use of available resources are some of the important factors that clinicians need to take into account when dealing with multiple retransplantations. With such conditions, however, liver retransplantation of more than two grafts can be a life-saving procedure.


Mangrovitalea sediminis gen. nov., sp. nov., a member of the family Alteromonadaceae isolated from mangrove sediment.
A Gram-stain-negative, facultative anaerobic and oligotrophic, rod-shaped, and motile with single polar flagellum bacterial strain, designed M11-4T was isolated from mangrove sediment in Yunxiao Mangrove National Nature Reserve, China. Growth was observed at temperatures from 10 to 40 °C (optimum 30 °C), at salinities from 0.5 to 6 % (optimum 2-3 %), and at pH from 5 to 8 (optimum 6). Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain M11-4T shared highest sequence similarity with the genus Marinobacter(92.5-95.0 %) and represented a distinct phylogenetic lineage in the family Alteromonadaceae. The G+C content of the genomic DNA was 58.2 mol%. The dominant fatty acids were C16 : 0, C16 : 0 N-alcohol, summed feature 9 (comprising iso-C17 : 1ω9c and/or C16 : 0 10-methyl) and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c). The predominant respiratory quinone was ubiquinone-9 and the major polar lipids were diphosphatidylglycerol; phosphatidylethanolamine; phosphatidylglycerol and an unidentified aminophospholipid. According to its morphology, physiology, fatty acid composition and 16S rRNA gene sequence analysis, the strain M11-4T should be assigned as a novel species of a novel genus for which the name Mangrovitalea sediminis gen. nov., sp. nov. is proposed. The type strain of Mangrovitalea sediminis is M11-4T (=MCCC 1K03312T=JCM 32104T).


Response Characteristics of Hydrogen Sensors Based on PMMA-Membrane-Coated Palladium Nanoparticle Films.
Coating a polymeric membrane for gas separation is a feasible approach to fabricate gas sensors with selectivity. In this study, poly(methyl methacrylate)-(PMMA-)membrane-coated palladium (Pd) nanoparticle (NP) films were fabricated for high-performance hydrogen (H2) gas sensing by carrying out gas-phase cluster deposition and PMMA spin coating. No changes were induced by the PMMA spin coating in the electrical transport and H2-sensing mechanisms of the Pd NP films. Measurements of H2 sensing demonstrated that the devices were capable of detecting H2 gas within the concentration range 0-10% at room temperature and showed high selectivity to H2 due to the filtration effect of the PMMA membrane layer. Despite the presence of the PMMA matrix, the lower detection limit of the sensor is less than 50 ppm. A series of PMMA membrane layers with different thicknesses were spin coated onto the surface of Pd NP films for the selective filtration of H2. It was found that the device sensing kinetics were strongly affected by the thickness of the PMMA layer, with the devices with thicker PMMA membrane layers showing a slower response to H2 gas. Three mechanisms slowing down the sensing kinetics of the devices were demonstrated to be present: diffusion of H2 gas in the PMMA matrix, nucleation and growth of the β phase in the α phase matrix of Pd hydride, and stress relaxation at the interface between Pd NPs and the PMMA matrix. The retardation effect caused by these three mechanisms on the sensing kinetics relied on the phase region of Pd hydride during the sensing reaction. Two simple strategies, minimizing the thickness of the PMMA membrane layer and reducing the size of the Pd NPs, were proposed to compensate for retardation of the sensing response.


Development of the Virtual Kitchen Protocol for Prospective Memory: a virtual reality-based measure of everyday prospective memory abilities.
Prospective memory is the ability to remember to accomplish a task at a specified point in the future. While this cognitive ability has a large impact on daily functioning, it is rarely assessed during neuropsychological evaluations. Furthermore, existing clinical prospective memory measures are few in number and have significant limitations, including applicability to everyday functioning and appropriate norming for older adulthood. There are also many gaps in the literature on prospective memory, such as how environmental factors affect performance across the lifespan. In the current study, we develop and establish a new virtual reality-based measure of prospective memory, the Virtual Kitchen Protocol for Prospective Memory. Young adults (ages 18-29; n = 56), healthy older adults (ages 60-90; n = 94), and clinical older adults (ages 62-90; n = 30) were compared on their performances on both the developed virtual reality prospective memory test and a parallel paper-and-pencil prospective memory test to investigate the impact of test environment across participant group. The Virtual Kitchen Protocol for Prospective Memory was found to adequately differentiate between young adult, healthy older adult, and clinical older adult populations - suggesting baseline ability for prospective memory assessment in clinical settings with older adults and potential for future improvement of neuropsychological evaluations. Additionally, the developed virtual prospective memory task appeared to mimic environmental factors in everyday life but did not replicate the benefit previously measured in healthy older adults' prospective memory ability in naturalistic settings. Overall, the current study built upon extant knowledge of prospective memory in both normal and abnormal aging, suggesting future directions in replicating familiar home environments. Findings provided additional evidence toward future validation of virtual reality assessment tools in clinical neuropsychological evaluations of cognitive abilities, such as prospective memory, with both healthy and clinical older adult populations.


A Comparative Study for Detection of EGFR Mutations in Plasma Cell-Free DNA in Korean Clinical Diagnostic Laboratories.
Liquid biopsies to genotype the epidermal growth factor receptor (EGFR) for targeted therapy have been implemented in clinical decision-making in the field of lung cancer, but harmonization of detection methods is still scarce among clinical laboratories. We performed a pilot external quality assurance (EQA) scheme to harmonize circulating tumor DNA testing among laboratories. For EQA, we created materials containing different levels of spiked cell-free DNA (cfDNA) in normal plasma. The limit of detection (LOD) of the cobas® EGFR Mutation Test v2 (Roche Molecular Systems) was also evaluated. From November 2016 to June 2017, seven clinical diagnostic laboratories participated in the EQA program. The majority (98.94%) of results obtained using the cobas assay and next-generation sequencing (NGS) were acceptable. Quantitative results from the cobas assay were positively correlated with allele frequencies derived from digital droplet PCR measurements and showed good reproducibility among laboratories. The LOD of the cobas assay was 5~27 copies/mL for p.E746_A750del (exon 19 deletion), 35~70 copies/mL for p.L858R, 18~36 copies/mL for p.T790M, and 15~31 copies/mL for p.A767_V769dup (exon 20 insertion). Deep sequencing of materials (>100,000X depth of coverage) resulted in detection of low-level targets present at frequencies of 0.06~0.13%. Our results indicate that the cobas assay is a reliable and rapid method for detecting EGFR mutations in plasma cfDNA. Careful interpretation is particularly important for p.T790M detection in the setting of relapse. Individual laboratories should optimize NGS performance to maximize clinical utility.


Cerebral MR spectroscopy in neurologically asymptomatic HIV-infected patients.
To detect early metabolic changes in the brain of neurologically asymptomatic HIV-infected patients with normal MR imaging and to find the correlation between 1H MR results and immune status. Twenty neurologically asymptomatic HIV seropositive patients underwent MR imaging and single-voxel 1H MR spectroscopy (MRS) using a PRESS sequence. For all patients, the signals from N-acetyl-aspartate (NAA), choline-containing compounds (Cho), creatine-phosphocreatine (Cr) and myoinositol (mI) were compared with 32 healthy volunteers as metabolite ratios and metabolite areas to non-suppressed water area ratios. In HIV patients, the NAA/Cho ratio was significantly lower ( p < 0.01), but there were no changes in NAA/Cr ratio. A statistically significant reduction in NAA/H2O and Cr/H2O (both p < 0.05) was observed. For the immune status there was a statistically significant correlation (r=0.47, p<0.05) between CD4 counts and NAA/H2O ratio. A significant increase in Cho/Cr ( p<0.001) and mI/Cr ( p<0.01) ratios in HIV patients was found, but Cho/H2O and mI/H2O concentrations were non-significantly increased. These results indicate that neuronal loss and gliosis in HIV-infected patients may be associated with impairment of energy metabolism. The spectral changes found suggest that 1H MRS can be used for early detection of brain damage induced by HIV.


Local NO2 concentrations and asthma among over-50s in Ireland: A microdata analysis.
Links between air pollution and asthma are less well established for older adults than some younger groups. Nitrogen dioxide (NO2) concentrations are widely used as an indicator of transport-related air pollution, and some literature suggests NO2 may directly affect asthma. This study used data on 8162 adults >50 years old in the Republic of Ireland to model associations between estimated annual outdoor concentration of NO2 and the probability of having asthma. Individual-level geo-coded survey data from The Irish Longitudinal Study on Ageing (TILDA) were linked to model-based estimates of annual average NO2 at 50 m resolution. Asthma was identified using two methods: self-reported diagnoses and respondents' use of medications related to obstructive airway diseases. Logistic regressions were used to model the relationships. NO2 concentrations were positively associated with the probability of asthma [marginal effect (ME) per 1 ppb of airborne NO2 = 0.24 percentage points asthma self-report, 95% confidence interval (CI) 0.06-0.42, mean asthma prevalence 0.09; for use of relevant medications ME = 0.21 percentage points, 95% CI 0.049-0.37, mean prevalence 0.069]. Results were robust to varying model specification and time period. Respondents in the top fifth percentile of NO2 exposure had a larger effect size but also greater standard error (ME = 2.4 percentage points asthma self-report, 95% CI -0. 49 to 5.3). Associations between local air pollution and asthma among older adults were found at relatively low concentrations. To illustrate this, the marginal effect of an increase in annual average NO2 concentration from sample minimum to median (2.5 ppb) represented about 7-8% of the sample average prevalence of asthma.


A novel immunohistochemical score to predict early mortality in acute myeloid leukemia patients based on indoleamine 2,3 dioxygenase expression.
Indoleamine 2,3 dioxygenase-1 (IDO-1) is an enzyme in the kynurenine pathway which augments tumor-induced immune tolerance. Previous studies in childhood acute myeloid leukemia (AML) have shown a negative correlation of IDO-1 mRNA expression with outcomes. The aim of our study was to develop a practical and objective immunohistochemical technique to quantify IDO-1 expression on diagnostic bone marrow biopsies of AML patients in order to facilitate its use in routine clinical practice. IDO-1 mRNA was extracted from diagnostic bone marrow specimens from 29 AML patients. IDO-1 protein expression was assessed in 40 cases via immunohistochemistry and quantified by a novel 'composite IDO-1 score'. In a univariate analysis, higher age (p = 0.0018), male gender (p = 0.019), high risk cytogenetics (p = 0.002), higher IDO-1 mRNA (p = 0.005), higher composite IDO-1 score (p < 0.0001) and not undergoing allogeneic stem cell transplant (SCT, p = 0.0005) predicted poor overall survival. In a multivariate model that included the aforementioned variables, higher composite IDO-1 score (p = 0.007) and not undergoing allogeneic SCT (p = 0.007) was found to significantly predict poor outcomes. Further, patients who failed induction had higher composite IDO-1 score (p = 0.01). In conclusion, 'composite IDO-1 score' is a prognostic tool that can help identify a certain subset of AML patients with 'early mortality'. This unique subset of patients can potentially benefit from specific IDO-1 inhibitor therapy, currently in clinical trials.


Fidaxomicin for treatment of Clostridium difficile infection in clinical practice: a prospective cohort study in a French University Hospital.
Two randomized controlled trials (RCTs) showed the non-inferiority of fidaxomicin compared with vancomycin for Clostridium difficile infection (CDI) treatment and its superiority regarding recurrence rate. The aim of this study was to evaluate fidaxomicin's efficacy in clinical practice. This single-center prospective cohort study included hospitalized patients treated with fidaxomicin for CDI. Demographic, clinical and biological data were collected. Primary outcome was efficacy of fidaxomicin (clinical cure, recurrence and global cure) at 10 weeks. Secondary outcome was efficacy among different subgroups. Ninety-nine patients were included: 42 severe CDI, 16 complicated CDI and 41 recurrent CDI. Rates of clinical cure, recurrence and global cure were 87, 15 and 59%, respectively. Subgroup analysis showed a higher recurrence rate for patients with recurrent CDI compared with first episode (8 vs. 26%; p = 0.04). Binary toxin was associated with severe/complicated CDI (80 vs. 50%; p < 0.01) and recurrence (32 vs. 7%; p < 0.01). Fidaxomicin was used as a first line for 83% of the patients with recurrence and for only 52% of first episodes even though 86% had recurrence's risk factors. Compared with RCTs, fidaxomicin in real world is used for patients with more severe and recurrent CDI, but clinical cure and recurrence rates were similar. Comparative studies are needed in these specific subgroups. Our data also illustrate clinicians' difficulty to define a "patient at risk for recurrence" among the first episodes. Finally, we showed that binary toxin could be important in the screening for severity and recurrence risks.


Stereo calibration with absolute phase target.
Stereo cameras have been widely used for three-dimensional (3D) photogrammetry, and stereo calibration is a crucial process to estimate the intrinsic and extrinsic parameters. This paper proposes a stereo calibration method with absolute phase target by using horizontal and vertical phase-shifting fringes. The one-to-one mapping from the world points to the image points that can be recovered by referring to the absolute phase and then used to calibrate the stereo cameras. Compared with traditional methods that only use feature points within the overlapping field-of-view (FOV), the proposed method can use all feature points within the overlapping and non-overlapping FOVs. Besides, since phase is more robust against camera defocusing than intensity, the target images can be captured regardless of the depth-of-field (DOF). With the advantages of whole-field capability and defocusing tolerability, the target placement becomes very flexible. Both simulations and experiment results demonstrate the robustness and accuracy of the proposed method.


Particle profiling and classification by a dual-band continuous-wave lidar system.
A dual-band continuous-wave (CW) light detection and ranging (lidar) system has been developed for particle classification. In this lidar system, the range-resolved atmospheric backscattering signal is recorded by an optical imaging system satisfying the Scheimpflug principle instead of the conventional time-of-flight approach. It is thus possible to employ low-cost and compact CW diode lasers, facilitating the development of a robust multiple-wavelength atmospheric lidar system that can attain high accuracy of the retrieved parameters of atmospheric particles. The present work demonstrates a dual-band Scheimpflug lidar system employing two diode lasers at 405 nm (0.5 W) and 808 nm (3.2 W). Exposures are milliseconds apart and interpolated. Measurements of various types of particles and smoke have been performed to verify the feasibility of using the present system for improved particle classification and sizing, for the situation when plumes were dilute and no significant opacity was detected.


Application of 31P-NMR spectroscopy to the study of striated muscle metabolism.
This review presents the principles and limitations of phosphorus nuclear magnetic resonance (31P-NMR) spectroscopy as applied to the study of striated muscle metabolism. Application of the techniques discussed include noninvasive measurement of high-energy phosphate, intracellular pH, intracellular free Mg2+, and metabolite compartmentation. In perfused cat biceps (fast-twitch) muscle, but not in soleus (slow-twitch), NMR spectra indicate a substantially lower (1 mM) free inorganic phosphate level than when measured chemically (6 mM). In addition, saturation and inversion spin-transfer methods that enable direct measurement of the unidirectional fluxes through creatine kinase are described. In perfused cat biceps muscle, results suggest that this enzyme and its substrates are in simple chemical equilibrium.