Effects of commonly used topical antimicrobial agents on Acinetobacter baumannii: an in vitro study.
Acinetobacter baumannii has recently emerged as an important pathogen among wounded soldiers in Iraq. Because of its ability to develop resistance to antimicrobial agents, wound infections with A. baumannii are difficult to treat and can lead to septicemia and even death. Use of appropriate topical antimicrobial agents in these circumstances could be one of the first steps in the prevention of A. baumannii wound infections. In this study, we present the in vitro effects of seven common topical antimicrobial creams and dressings on A. baumannii. A. baumannii was subjected to sensitivity tests with mupirocin, silver sulfadiazine, mafenide acetate, a double-antibiotic combination of polymyxin and bacitracin, a triple-antibiotic combination of neomycin, bacitracin, and polymyxin, and two silver-containing dressings. Zones of inhibition were measured after 24 hours of incubation. Of the evaluated antimicrobial agents, mafenide acetate was the most efficacious, followed by mupirocin and triple- and double-antibiotic combinations (in decreasing order). The silver-containing dressings yielded smaller zones of inhibition, compared to the previously mentioned agents, and no zone of inhibition was observed with silver sulfadiazine. Further in vivo studies on the effects of antimicrobial agents against A. baumannii are necessary to substantiate these findings and to determine the potential clinical relevance of these therapies.


Quality of life in dementia patients in Athens, Greece: predictive factors and the role of caregiver-related factors.
Quality of life (QOL) is increasingly recognized as the main target of currently available dementia care. Its assessment has grown exponentially in the dementia field, but few studies have examined predictive factors for QOL taking caregiver variables into account. We examined patient and caregiver factors related to the QOL of dementia patients. The study design was cross-sectional. 161 couples of community residing dementia patients and their primary caregivers were interviewed. QOL was measured by the ADRQL, a proxy-rated, dementia-specific QOL instrument. Demographic factors were collected and clinical characteristics assessed using validated scales. In univariate analyses several patient and caregiver characteristics appeared associated with patient QOL. In multivariate analyses, independent predictors of worse patient QOL were behavioral and depressive symptoms of dementia patients, dependency in basic activities of daily living, poorer cognitive function, use of antipsychotic medication, caregiver burden, and caregiver not being an adult child. The adjusted R2 of the final, seven-factor model was 0.598. QOL for a person with dementia is a complex issue that is associated with several patient and caregiver factors. Efforts to improve patients' QOL should be addressed for both patients and caregivers. The measurement of QOL should be included, when possible, as a standard measurement tool, in everyday dementia clinical practice.


Multivariate predictive model for dyslexia diagnosis.
Dyslexia is a specific disorder of language development that mainly affects reading. Etiological researches have led to multiple hypotheses which induced various diagnosis methods and rehabilitation treatments so that many different tests are used by practitioners to identify dyslexia symptoms. Our purpose is to determine a subset of the most efficient ones by integrating them into a multivariate predictive model. A set of screening tasks that are the most commonly used and representative of the different cognitive aspects of dyslexia was proposed to 78 children from elementary school (mean age = 9 years ± 7 months) exempt from identified reading difficulties and to 35 dyslexic children attending a specialized consultation for dyslexia. We proposed a multi-step procedure: within each category, we first selected the most representative tasks using principal component analysis and then we implemented logistic regression models on the preselected variables. Spelling and reading tasks were considered separately. The model with the best predictive performance includes eight variables from four categories of tasks and classifies correctly 94% of the children. The sensitivity (91%) and the specificity (95%) are both high. Forty minutes are necessary to complete the test.


Novel functional roles for PERIANTHIA and SEUSS during floral organ identity specification, floral meristem termination, and gynoecial development.
The gynoecium is the female reproductive structure of angiosperm flowers. In Arabidopsis thaliana the gynoecium is composed of two carpels that are fused into a tube-like structure. As the gynoecial primordium arises from the floral meristem, a specialized meristematic structure, the carpel margin meristem (CMM), develops from portions of the medial gynoecial domain. The CMM is critical for reproductive competence because it gives rise to the ovules, the precursors of the seeds. Here we report a functional role for the transcription factor PERIANTHIA (PAN) in the development of the gynoecial medial domain and the formation of ovule primordia. This function of PAN is revealed in pan aintegumenta (ant) as well as seuss (seu) pan double mutants that form reduced numbers of ovules. Previously, PAN was identified as a regulator of perianth organ number and as a direct activator of AGAMOUS (AG) expression in floral whorl four. However, the seu pan double mutants display enhanced ectopic AG expression in developing sepals and the partial transformation of sepals to petals indicating a novel role for PAN in the repression of AG in floral whorl one. These results indicate that PAN functions as an activator or repressor of AG expression in a whorl-specific fashion. The seu pan double mutants also display enhanced floral indeterminacy, resulting in the formation of "fifth whorl" structures and disruption of WUSCHEL (WUS) expression patterns revealing a novel role for SEU in floral meristem termination.


Lexicase Selection for Multi-Task Evolutionary Robotics.
In Evolutionary Robotics, Lexicase selection has proven effective when a single task is broken down into many individual parameterizations. Evolved individuals have generalized across unique configurations of an overarching task. Here, we investigate the ability of Lexicase selection to generalize across multiple tasks, with each task again broken down into many instances. There are three objectives: to determine the feasibility of introducing additional tasks to the existing platform; to investigate any consequential effects of introducing these additional tasks during evolutionary adaptation; and to explore whether the schedule of presentation of the additional tasks over evolutionary time affects the final outcome. To address these aims we use a quadruped animat controlled by a feed-forward neural network with joint-angle, bearing-to-target, and spontaneous sinusoidal inputs. Weights in this network are trained using evolution with Lexicase-based parent selection. Simultaneous adaptation in a wall crossing task (labelled wall-cross) is explored when one of two different alternative tasks is also present: turn-and-seek or cargo-carry. Each task is parameterized into 100 distinct variants, and these variants are used as environments for evaluation and selection with Lexicase. We use performance in a single-task wall-cross environment as a baseline against which to examine the multi-task configurations. In addition, the objective sampling strategy (the manner in which tasks are presented over evolutionary time) is varied, and so data for treatments implementing uniform sampling, even sampling, or degrees of generational sampling are also presented. The Lexicase mechanism successfully integrates evolution of both turn-and-seek and cargo-carry with wall-cross, though there is a performance penalty compared to single task evolution. The size of the penalty depends on the similarity of the tasks. Complementary tasks (wallcross/turn-and-seek) show better performance than antagonistic tasks (wall-cross/cargo-carry). In complementary tasks performance is not affected by the sampling strategy. Where tasks are antagonistic, uniform and even sampling strategies yield significantly better performance than generational sampling. In all cases the generational sampling requires more evaluations and consequently more computational resources. The results indicate that Lexicase is a viable mechanism for multitask evolution of animat neurocontrollers, though the degree of interference between tasks is a key consideration. The results also support the conclusion that the naive, uniform random sampling strategy is the best choice when considering final task performance, simplicity of implementation, and computational efficiency.


Using Latent Profile Analysis to Examine Cannabis Use Contexts: Associations with Use, Consequences, and Protective Behaviors.
College students represent a large portion of the population, and report high rates of cannabis use and related negative outcomes, including interpersonal problems, risky behaviors, or physical dependency. The contexts in which students use cannabis (e.g., at a party, when feeling down or depressed, after a fight with a loved one) likely affect their risk of experiencing consequences. We aimed to discern profiles of cannabis use contexts and compare profiles on use frequency, consequences, and the use of cannabis protective behavioral strategies (PBS). College students were surveyed regarding their cannabis use contexts, frequency, consequences, and PBS use (n = 265; female = 72.8%). We used Latent Profile Analysis to identify patterns of cannabis use contexts and auxiliary testing to compare profiles on use frequency, consequences, and PBS use. Our examination revealed three latent profiles of cannabis use. The Social Use Profile was associated with use in predominantly social/uplifting contexts. The Physical & Emotional Pain Profile was also associated with use in these contexts but was defined by additional use in response to pain. The All Contexts Profile was associated with frequent use in all contexts, including those that were least endorsed by the other profiles. Profiles differed in cannabis use frequency, PBS use, and the number of consequences experienced, such that profiles were more likely to be associated with more frequent cannabis use, higher risk of experiencing use-consequences, and using fewer PBS as the number of use contexts increased across the profiles. The contexts in which people use cannabis are associated with cannabis risk and protection. Prevention and intervention efforts may benefit from considering contexts of cannabis use.


Hypothermia and neonatal encephalopathy.
Data from large randomized clinical trials indicate that therapeutic hypothermia, using either selective head cooling or systemic cooling, is an effective therapy for neonatal encephalopathy. Infants selected for cooling must meet the criteria outlined in published clinical trials. The implementation of cooling needs to be performed at centers that have the capability to manage medically complex infants. Because the majority of infants who have neonatal encephalopathy are born at community hospitals, centers that perform cooling should work with their referring hospitals to implement education programs focused on increasing the awareness and identification of infants at risk for encephalopathy, and the initial clinical management of affected infants.


Comparison of dominant hand range of motion among throwing types in baseball pitchers.
Previous research on baseball pitchers' wrists, elbows, and should joints contributes to our understanding of pitchers' control over delicate joint motion and ball release. However, limited research on forearm, wrist, and hand joints prevents full comprehension of the throwing mechanism. The present descriptive laboratory study quantifies angular performances of hand and wrist joints while pitching breaking balls, including fastballs, curveballs and sliders, among pitchers with different skill levels. Nineteen baseball pitchers performed required pitching tasks (10 from university and 9 from high school). A three-dimensional motion analysis system collected pitching motion data. The range of joint motion in the wrist and proximal interphalangeal (PIP) and metacarpophalangeal (MP) joints of the index and middle fingers were compared among fastballs, curveballs and sliders. Thirteen reflective markers were placed on selected anatomic landmarks of the wrist, middle and index fingers of the hand. Wrist flexion angle in the pitching acceleration phase was larger in fastballs (20.58±4.07°) and sliders (22.48±5.45°) than in curveballs (9.08±3.03°) (p = .001). The flexion angle of the PIP joint was significantly larger in curveballs (38.5±3.8°) than in fastballs (30.3±4.8°) and sliders (30.2±4.5°) (p=.004) of the middle finger. Abduction angle of MP joint on the middle finger was significantly larger in curveballs (15.4 ±3.6°) than in fastballs (8.9±1.2°) and sliders (6.9±2.9°) (p=.001) of the middle finger, and the abduction angle of index finger was significantly larger in sliders (13.5±15.0°) than in fastballs (7.2 ±2.8°) (p=.007). Hand and wrist motion and grip types affect the relative position between fingers and ball, which produces different types of baseball pitches. A larger extension angle of the wrist joint and the coordination of middle and index fingers are crucial when pitching a fastball. Abduction and flexion movement on the MP joint of the middle finger are important for a curveball. MP joint abduction and flexion movement of the index finger produce sliders. Understanding the control mechanism in a throwing hand can help improve training protocols in either injury prevention or performance improvement for baseball pitchers.


On Distribution Dependent Sub-Logarithmic Query Time of Learned Indexing.
A fundamental problem in data management is to find the elements in an array that match a query. Recently, learned indexes are being extensively used to solve this problem, where they learn a model to predict the location of the items in the array. They are empirically shown to outperform non-learned methods (e.g., B-trees or binary search that answer queries in O(logn) time) by orders of magnitude. However, success of learned indexes has not been theoretically justified. Only existing attempt shows the same query time of O(logn), but with a constant factor improvement in space complexity over non-learned methods, under some assumptions on data distribution. In this paper, we significantly strengthen this result, showing that under mild assumptions on data distribution, and the same space complexity as non-learned methods, learned indexes can answer queries in O(loglogn) expected query time. We also show that allowing for slightly larger but still near-linear space overhead, a learned index can achieve O(1) expected query time. Our results theoretically prove learned indexes are orders of magnitude faster than non-learned methods, theoretically grounding their empirical success.


ATP7B variant spectrum in a French pediatric Wilson disease cohort.
The spectrum of ATP7B variants varies significantly according to geographic distribution, and there is insufficient data on the variants observed in the French population. Clinical data of 113 children included in the French WD national registry were gathered from March 01, 1995 to July 01, 2020. Data included epidemiological, clinical, laboratory, genetics. Diagnosis was made at a mean age of 11.0 ± 4.1 years (range 1-18 years). At diagnosis, 91 patients (79.8 %) had hepatic manifestations, 18 (15.8 %) presented neurological manifestations, and 4 patients (3.5 %) were asymptomatic. Only 29 patients (25 %) were homozygous for a variant. We have found a total of 102 different variants including 14 novel variants. Recurrent variant p.His1069Gln was the most prevalent, n = 31 alleles (14,2%), with only seven homozygous; in contrast 55% of variants are identified in only one family. 45% were truncating variants. In respect of mutated exon, the three most prevalent were exon 14 (16.5%), exon 8 (13.8%), and exon 3 (11.5%). When considering patients with two Nonsense / Frameshift variants as a group and those with two Missense variants, we found significantly lower ceruloplasmin for the former: 2.8 ± 0.7 mg/dl vs 8.4 ± 5mg/dl (p<0.05). p.His1069Gln is the most frequent variant (14,2%) and exons 14, 8, and 2 of the ATP7B gene account for 41.7% of total variants. However, there is significant heterogeneity in the French population concerning the other ATP7B variants. Nonsense / Frameshift variants were associated with lower ceruloplasmin levels.


Alternating-script priming in Japanese: Are Katakana and Hiragana characters interchangeable?
Models of written word recognition in languages using the Roman alphabet assume that a word's visual form is quickly mapped onto abstract units. This proposal is consistent with the finding that masked priming effects are of similar magnitude from lowercase, uppercase, and alternating-case primes (e.g., beard-BEARD, BEARD-BEARD, and BeArD-BEARD). We examined whether this claim can be readily generalized to the 2 syllabaries of Japanese Kana (Hiragana and Katakana). The specific rationale was that if the visual form of Kana words is lost early in the lexical access process, alternating-script repetition primes should be as effective as same-script repetition primes at activating a target word. Results showed that alternating-script repetition primes were less effective at activating lexical representations of Katakana words than same-script repetition primes-indeed, they were no more effective than partial primes that contained only the Katakana characters from the alternating-script primes. Thus, the idiosyncrasies of each writing system do appear to shape the pathways to lexical access. (PsycINFO Database Record


An artificial protein cage made from a 12-membered ring.
Artificial protein cages have great potential in diverse fields including as vaccines and drug delivery vehicles. TRAP-cage is an artificial protein cage notable for the way in which the interface between its ring-shaped building blocks can be modified such that the conditions under which cages disassemble can be controlled. To date, TRAP-cages have been constructed from homo-11mer rings, i.e., hendecamers. This is interesting as convex polyhedra with identical regular faces cannot be formed from hendecamers. TRAP-cage overcomes this limitation due to intrinsic flexibility, allowing slight deformation to absorb any error. The resulting TRAP-cage made from 24 TRAP 11mer rings is very close to regular with only very small errors necessary to allow the cage to form. The question arises as to the limits of the error that can be absorbed by a protein structure in this way before the formation of an apparently regular convex polyhedral becomes impossible. Here we use a naturally occurring TRAP variant consisting of twelve identical monomers (i.e., a dodecamer) to probe these limits. We show that it is able to form an apparently regular protein cage consisting of twelve TRAP rings. Comparison of the cryo-EM structure of the new cage with theoretical models and related cages gives insight into the rules of cage formation and allows us to predict other cages that may be formed given TRAP-rings consisting of different numbers of monomers.


Mapping Elastic Properties of Heterogeneous Materials in Liquid with Angstrom-Scale Resolution.
Fast quantitative mapping of mechanical properties with nanoscale spatial resolution represents one of the major goals of force microscopy. This goal becomes more challenging when the characterization needs to be accomplished with subnanometer resolution in a native environment that involves liquid solutions. Here we demonstrate that bimodal atomic force microscopy enables the accurate measurement of the elastic modulus of surfaces in liquid with a spatial resolution of 3 Å. The Young's modulus can be determined with a relative error below 5% over a 5 orders of magnitude range (1 MPa to 100 GPa). This range includes a large variety of materials from proteins to metal-organic frameworks. Numerical simulations validate the accuracy of the method. About 30 s is needed for a Young's modulus map with subnanometer spatial resolution.


Younger age at crisis following parental death in male children and adolescents is associated with higher risk for dementia at old age.
To examine the association of midlife report of crisis following parental death (CFPD) during childhood and adolescence, with dementia at old age. In 1965, 9362 male participants of the Israel Ischemic Heart Disease study were asked whether they have experienced CFPD (paternal or maternal) during the following ages: 0 to 6, 7 to 12, 13 to 18, or >18 years. Dementia was assessed over 3 decades later in 1889 survivors of the original cohort, 1652 of whom were assessed for CFPD in 1965. Controlling for age, the estimated odds ratios for dementia relative to individuals who reported crisis following paternal parental death (CFPD-P) at the age of 18 years and above were 3.06 (95% CI: 1.42-6.61), 2.15 (95% CI: 0.87-5.31), and 2.35 (95% CI: 1.05-5.28) for those who reported CFPD-P at the ages of 0 to 6, 7 to 12, and 13 to 18 years, respectively. Odds ratios for dementia were 0.60 (95% CI: 0.32-1.11) for participants who reported CFPD-P at ages of 18 and above compared with participants who did not report such a crisis. Similar results were obtained for the association of crisis reported following maternal parental death (CFPD-M) at different age groups and dementia. CFPD during childhood is associated with an increased risk for dementia in men who survived until old age.


Prospective study of the pattern of utilization of mental health services of a Nigerian university hospital.
A prospective study of the pattern of utilization of the mental services offered by a Nigerian university teaching hospital was carried out in 1981. Demographic and other characteristics of the 151 patients registered over a 3-month period are described along with their mode of entry into the health care delivery system of the hospital. The significance and implication of the findings are discussed.


Calling Cards enable multiplexed identification of the genomic targets of DNA-binding proteins.
Transcription factors direct gene expression, so there is much interest in mapping their genome-wide binding locations. Current methods do not allow for the multiplexed analysis of TF binding, and this limits their throughput. We describe a novel method for determining the genomic target genes of multiple transcription factors simultaneously. DNA-binding proteins are endowed with the ability to direct transposon insertions into the genome near to where they bind. The transposon becomes a "Calling Card" marking the visit of the DNA-binding protein to that location. A unique sequence "barcode" in the transposon matches it to the DNA-binding protein that directed its insertion. The sequences of the DNA flanking the transposon (which reveal where in the genome the transposon landed) and the barcode within the transposon (which identifies the TF that put it there) are determined by massively parallel DNA sequencing. To demonstrate the method's feasibility, we determined the genomic targets of eight transcription factors in a single experiment. The Calling Card method promises to significantly reduce the cost and labor needed to determine the genomic targets of many transcription factors in different environmental conditions and genetic backgrounds.


Electron transfer between biological molecules by thermally activated tunneling.
A theory of electron transfer between two fixed sites by tunneling is developed. Vibronic coupling in the individual molecules produces an activation energy to transfer at high temperatures, and temperature-independent tunneling (when energetically allowed) at low temperature. The model is compared with known results on electron transfer in Chromatium and in Rhodopseudomonas spheroides. It quantitatively interprets these results, with parameters whose scale is verified by comparison with optical absorption spectra. According to this description, the separation between linking sites for electron transfer is 8-10 A in Chromatium, far smaller than earlier estimates.


Quality of life assessment of Hodgkin's disease survivors: a model for cooperative clinical trials.
With the growing number of patients surviving cancer, there has been an increasing concern with their long-term adaptation. Given the large number of Hodgkin's disease survivors, it was possible to conduct a study of their psychosocial adaptation. Two hundred seventy-three survivors of advanced Hodgkin's disease were interviewed over the telephone concerning the impact of cancer upon their lives, in terms of their psychological, social, and sexual functioning. The level of psychological distress was elevated by one standard deviation above normal on the Brief Symptom Inventory (BSI), for both men and women. Survivors in greater distress reported more problems in other areas of functioning, including sexual, social, vocational, and persistent conditioned nausea.


A new model for catalyzing translational science: the early stage investigator mentored research scholar program in HIV vaccines.
Engagement of early stage investigators (ESIs) in the search for a safe and effective vaccine is critical to the success of this highly challenging endeavor. In the wake of disappointing results from a large-scale efficacy trial, the HIV Vaccine Trials Network (HVTN) and Center for HIV/AIDS Vaccine Immunology (CHAVI) developed a novel mentored research program focused on the translation of findings from nonhuman primate studies to human trials of experimental vaccines. From 2008 to 2011, 14 ESI Scholars were selected from 42 complete applications. Post program surveys and tracked outcomes suggest that the combination of flexible funding, transdisciplinary mentorship, and structured training and networking promoted the scientific contributions and career development of promising ESIs. Embedding a multicomponent research program within collaborative clinical trial networks and research consortia is a promising strategy to attract and retain early career investigators and catalyze important translational science.


Should we teach using schemas? Evidence from a randomised trial.
Schema-based instruction may alter knowledge organisation and diagnostic reasoning strategies through the provision of structured knowledge to novice trainees. The effects of schema-based instruction on diagnostic accuracy and knowledge organisation have not been rigorously tested. Year 2 medical students were randomised to learn four cardiac diagnoses using schema-based instruction (n = 26) or traditional instruction (n = 27) on a high-fidelity cardiopulmonary simulator (CPS). Students completed case-based learning in groups of two to five and underwent individual written and practical tests. The written test consisted of questions testing features that linked or distinguished diagnoses (structured knowledge) and questions testing features of individual diagnoses (factual knowledge). A practical test of diagnostic accuracy on the CPS was performed for two diagnoses present in the learning phase (taught lesions) and two untaught lesions. A majority of students (n = 37, 70%) voluntarily returned for follow-up written testing 2-4 weeks later. Learning time and accuracy did not differ between students on schema-based and those on traditional instruction. Students receiving schema-based instruction performed better on structured knowledge questions (p < 0.001) and no differently on factual knowledge questions (p = 0.7). Relative differences between groups remained unchanged on follow-up testing. Diagnostic success was higher in the schema-based instruction group for taught lesions (mean difference = 38%, 95% confidence interval [CI] 20-56; p < 0.001) and untaught lesions (mean difference = 31%, 95% CI 15-48; p < 0.001). Schema-based instruction was associated with improved retention of structured knowledge and diagnostic performance among novices. This study provides important proof-of-concept for a schema-based approach and suggests there is substantial benefit to using this approach with novice trainees.


Diurnal Characteristics of the Orexin System Genes and Its Effects on Pathology at Early Stage in 3xTg-AD Mice.
Orexin and its receptors are closely related to the pathogenesis of Alzheimer's disease (AD). Although the expression of orexin system genes under physiological condition has circadian rhythm, the diurnal characteristics of orexin system genes, and its potential role in the pathogenesis in AD are unknown. In the present study, we hope to elucidate the diurnal characteristics of orexin system genes at the early stage of AD, and to investigate its potential role in the development of AD neuropathology. We firstly detected the mRNA levels of orexin system genes, AD risk genes and core clock genes (CCGs) in hypothalamus and hippocampus in 6-month-old male 3xTg-AD mice and C57BL/6J (wild type, WT) control mice, then analyzed diurnal expression profiles of all genes using JTK_CYCLE algorithm, and did the correlation analysis between expression of orexin system genes and AD risk genes or CCGs. In addition, the expression of β-amyloid protein (Aβ) and phosphorylated tau (p-tau) protein were measured. The results showed that the diurnal mRNA expression profiles of PPO, OX1R, OX2R, Bace2, Bmal1, Per1, Per2 and Cry1 in the hypothalamus, and gene expression of OX1R, OX2R, Bace1, Bmal1, Per1 and Cry2 in the hippocampus in 3xTg-AD mice were different from that in WT mice. Furthermore, there is positive correlation between orexin system genes and AD risk genes or CCGs in the brain in 3xTg-AD mice. In addition, the expression of Aβ and p-tau in hippocampus in 3xTg-AD mice were significantly increased, and the expression of p-tau is higher in night than in day. These results indicate that the abnormal expression profiles of orexin system genes and its interaction with AD risk genes or CCGs might exert important role in the pathogenesis of AD, which will increase the expression of Aβ and p-tau, and accelerate the development of AD.


Comparative Tissue Proteomics of Microdissected Specimens Reveals Novel Candidate Biomarkers of Bladder Cancer.
More than 380,000 new cases of bladder cancer are diagnosed worldwide, accounting for ∼150,200 deaths each year. To discover potential biomarkers of bladder cancer, we employed a strategy combining laser microdissection, isobaric tags for relative and absolute quantitation labeling, and liquid chromatography-tandem MS (LC-MS/MS) analysis to profile proteomic changes in fresh-frozen bladder tumor specimens. Cellular proteins from four pairs of surgically resected primary bladder cancer tumor and adjacent nontumorous tissue were extracted for use in two batches of isobaric tags for relative and absolute quantitation experiments, which identified a total of 3220 proteins. A DAVID (database for annotation, visualization and integrated discovery) analysis of dysregulated proteins revealed that the three top-ranking biological processes were extracellular matrix organization, extracellular structure organization, and oxidation-reduction. Biological processes including response to organic substances, response to metal ions, and response to inorganic substances were highlighted by up-expressed proteins in bladder cancer. Seven differentially expressed proteins were selected as potential bladder cancer biomarkers for further verification. Immunohistochemical analyses showed significantly elevated levels of three proteins-SLC3A2, STMN1, and TAGLN2-in tumor cells compared with noncancerous bladder epithelial cells, and suggested that TAGLN2 could be a useful tumor tissue marker for diagnosis (AUC = 0.999) and evaluating lymph node metastasis in bladder cancer patients. ELISA results revealed significantly increased urinary levels of both STMN1 and TAGLN2 in bladder cancer subgroups compared with control groups. In comparisons with age-matched hernia urine specimens, urinary TAGLN2 in bladder cancer samples showed the largest fold change (7.13-fold), with an area-under-the-curve value of 0.70 (p < 0.001, n = 205). Overall, TAGLN2 showed the most significant overexpression in individual bladder cancer tissues and urine specimens, and thus represents a potential biomarker for noninvasive screening for bladder cancer. Our findings highlight the value of bladder tissue proteome in providing valuable information for future validation studies of potential biomarkers in urothelial carcinoma.


Prostaglandin E2 production in rat IMCD cells. II. Possible role for locally formed dopamine.
Dopamine has been proposed as an intrarenal natriuretic hormone. We reported previously that inner medullary collecting duct (IMCD) cells express a novel DA2-like dopamine receptor (namely, DA2K) that is linked to stimulation of prostaglandin E2 (PGE2) production. In this study we examined whether locally formed dopamine could stimulate PGE2 production in cultured IMCD cells. L-Dopa stimulated PGE2 production dose dependently in cultured IMCD cells (concentration for half-maximal stimulation, 54.3 microM; maximal stimulation, 212.7% of basal), with the maximal stimulation similar to that obtained with dopamine. This effect was blocked by aromatic L-amino acid decarboxylase (AADC) inhibitors and DA2-receptor antagonists. IMCD cells also had measurable AADC activity and produced dopamine from exogenously added L-dopa. AADC inhibitors and DA2 antagonists also lowered basal PGE2 levels, suggesting that dopamine was being formed constitutively in culture. These results suggest that cultured IMCD cells have the capacity to take up and convert L-dopa to dopamine, which then stimulates PGE2 production via DA2K receptors. These results further suggest that locally formed dopamine could act as an autocrine/paracrine hormone in the kidney inner medulla to regulate PGE2 synthesis and water and electrolyte excretion.


Lipase engineering: a window into structure-function relationships.
Utilization of genetic engineering techniques to create novel functional lipases has increased knowledge of structure-function relationships in this important class of enzymes. The examples of engineered lipases presented in this chapter addressed the investigation of domain-specific properties, heparin binding, and subunit orientation. Conclusions reached are credible because the designed lipases retained catalytic activity, implying native, or near-native, conformation. This approach has demonstrated vigor by determining the domain location of several important enzyme functions and by providing the first evidence that LPL subunits are arranged in a head-to-tail orientation. In conjunction with physical techniques, such as crystallography and nuclear magnetic resonance spectroscopy, the engineered lipase approach could reveal new insights into the mechanism by which lipolysis is accomplished. The studies described here represent only the first attempts to explore that subject; more sophisticated lipase engineering will be used in future as a window into structure-function relationships.


Prolonged cutaneous analgesia with transdermal application of amitriptyline and capsaicin.
Capsaicin selectively binds to TRPV1, the vanilloid subtype 1 of the superfamily of transient receptor potential ion channels, which is highly expressed in pain-transmitting C fibers. Recent reports have demonstrated that the coadministration of capsaicin with a local anesthetic (LA) at the rat sciatic nerve elicits a prolonged nociceptive-selective nerve block, suggesting that activation of the TRPV1 receptor may allow LAs to enter the nerve through the TRPV1 pore. In previous studies, we demonstrated that transdermal amitriptyline achieves clinical analgesic effects and is more potent than lidocaine. Here we examine whether the combined application of amitriptyline and capsaicin as a transdermal patch will produce prolonged cutaneous analgesia compared with amitriptyline alone. Male Sprague-Dawley rats (weights 250-300 g) were assigned to five treatment groups (n = 6-8 per group). Transdermal patches containing amitriptyline with different concentrations of capsaicin were applied for 3 hrs to rats' shaved backs: 2.5% amitriptyline alone (control group) and in combination with 0.05%, 0.15%, 1%, and 8% capsaicin. Behavioral testing for cutaneous nociception was conducted before drug application and after patch removal using the cutaneous trunci muscle reflex. In addition, skin appearance was assessed to determine irritation by these formulations. The cutaneous analgesic effect is significantly prolonged when amitriptyline is applied in combination with 8% capsaicin. Amitriptyline alone provided a complete block to pinprick for 4.5 hrs, and the time to full recovery was 96 hrs. Amitriptyline with 8% capsaicin produced a complete block to pinprick for 6 to 9 hrs, and the time to full recovery was 216 hrs (P = 0.002). Amitriptyline alone causes toxic effects in skin, whereas the higher the concentration of capsaicin, the less skin irritation was noted, and the combination of amitriptyline 2.5% with capsaicin 8% caused no adverse skin reactions. This study demonstrates that the combined application of amitriptyline and capsaicin results in prolonged cutaneous analgesia compared with amitriptyline alone, suggesting that the activation of the TRPV1 channel by capsaicin facilitates the passage of amitriptyline into nociceptors. This transdermal patch achieves far longer cutaneous analgesia than currently available patch applications such as EMLA cream. The mechanism that underlies the lesser skin irritation noted when amitriptyline is combined with higher doses of capsaicin compared with amitriptyline alone is unclear and may be related to a counteraction of amitriptyline-induced vasoconstriction.


EDUCATIONAL SERIES IN CONGENITAL HEART DISEASE: Tetralogy of Fallot: diagnosis to long-term follow-up.
Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart defect, affecting 3 in 10,000 live births. Surgical correction in early childhood is associated with good outcomes, but lifelong follow-up is necessary to identify the long-term sequelae that may occur. This article will cover the diagnosis of TOF in childhood, the objectives of surveillance through adulthood and the value of multi-modality imaging in identifying and guiding timely surgical and percutaneous interventions.


Treatment of muscle mechanoreflex dysfunction in hypertension: effects of L-arginine dialysis in the nucleus tractus solitarii.
What is the central question of this study? Does increasing NO production within the nucleus tractus solitarii (NTS) affect mechanoreflex function in normotensive and hypertensive rats?What is the main finding and its importance? Dialysis of 1 μm l-arginine, an NO precursor, within the NTS significantly attenuated the pressor response to muscle stretch in normotensive and hypertensive rats. In contrast, 10 μm l-arginine had no effect in normotensive animals, while increasing and decreasing the pressor and tachycardic responses to stretch, respectively, in hypertensive rats. This suggests that increasing NO within the NTS using lower doses of l-arginine can partly normalize mechanoreflex overactivity in hypertensive rats, whereas the effects of larger doses are equivocal. The blood pressure response to exercise is exaggerated in hypertension. Recent evidence suggests that an overactive skeletal muscle mechanoreflex contributes significantly to this augmented circulatory responsiveness. Sensory information from the mechanoreflex is processed within the nucleus tractus solitarii (NTS) of the medulla oblongata. Normally, endogenously produced nitric oxide within the NTS attenuates the increase in mean arterial pressure (MAP) induced by mechanoreflex stimulation. Thus, it has been suggested that decreases in NO production in the NTS underlie the generation of mechanoreflex dysfunction in hypertension. Supporting this postulate, it has been shown that blocking NO production within the NTS of normotensive rats reproduces the exaggerated pressor response elicited by mechanoreflex activation in hypertensive animals. What is not known is whether increasing NO production within the NTS of hypertensive rats mitigates mechanoreflex overactivity. In this study, the mechanoreflex was selectively activated by passively stretching hindlimb muscle before and after the dialysis of 1 and 10 μm l-arginine (an NO precursor) within the NTS of decerebrate normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). Stretch induced larger elevations in MAP in SHRs compared with WKY rats. In both groups, dialysis of 1 μm l-arginine significantly attenuated the pressor response to stretch. However, at the 10 μm dose, l-arginine had no effect on the MAP response to stretch in WKY rats, while it enhanced the response in SHRs. The data demonstrate that increasing NO availability within the NTS using lower doses of l-arginine partly normalizes mechanoreflex dysfunction in hypertension, whereas higher doses do not. The findings could prove valuable in the development of treatment options for mechanoreflex overactivity in this disease.


Angiopoietin-1 Knockout Mice as a Genetic Model of Open-Angle Glaucoma.
A leading cause of blindness worldwide, glaucoma is often caused by elevated intraocular pressure (IOP) due to impaired aqueous humor outflow from the anterior chamber through Schlemm's canal (SC) and the trabecular meshwork. Despite the large clinical burden, glaucoma research and drug development are hindered by a limited selection of preclinical models that accurately recapitulate human disease. Here, we propose that Angpt1 conditional knockout mice may provide one such model. Angiopoietin/TEK (ANGPT/TEK) signaling is crucial for SC formation and integrity in mice and humans, and mice lacking TEK or its ligand ANGPT1 develop a hypomorphic SC insufficient for normal aqueous humor outflow. We used a comprehensive histology and physiology approach to characterize the glaucoma phenotype of Angpt1 inducible knockout mice, especially focusing on retina morphology and function. Angpt1 deletion resulted in persistent ocular hypertension beginning in the first month after birth and leading to decreased visual acuity with age due to glaucomatous neuropathy. In the neural retina, we identified marked and specific loss of the retinal ganglion cells, whereas other retinal neurons exhibited largely normal morphology and patterning. Electroretinogram recordings demonstrated reduced scotopic threshold response, further indicating loss of retinal ganglion cell function. These findings highlight the potential of Angpt1 conditional knockout mice as a valuable new glaucoma model. Currently, few reliable, rapid-onset genetic glaucoma models are available, and Angpt1 knockout mice will provide an additional tool for studies of IOP-induced neural damage, mechanisms of disease progression, and novel treatment strategies.


Exploring Women Healthcare Leaders' Perceptions on Barriers to Leadership in Greek Context.
Background: Gender inequalities have been identified as important derailment factors for health workforce and health system sustainability. Literature holds responsible a list of gendered barriers faced by female health workforce. However, there is a gap in the evidence based research on women leaders' own perceptions of barriers to leading positions advancement. This study aims to explore leadership barriers perceived by women healthcare leaders within country's context; research focused on Greece due to country's poor performance on gender equality index and current economic turbulence. Study supplements survey data and provides orientation for further gender sensitive research in health workforce development through country's specificity lens to better inform education and policy makers. Methods: The best-worst object case survey method was used, applying an online questionnaire designed in Qualtrics. The online questionnaire was sent to 30 purposively invited participants. Respondents were asked to tick the most and the least important barriers to women's leadership in provided choice scenarios. Descriptive data analysis was used to understand and interpret the results. Results: Women leaders perceived stereotypes, work/life balance, lack of equal career advancement, lack of confidence, gender gap and gender bias to be the barriers with the greatest relative importance in constraining opportunities for pursuing leading positions in Greek healthcare setting. Twenty more barriers were identified and ranked lower in relative importance. The results are considered exploratory and not to obtain population based outcomes. Conclusion: This exploratory study reports the perceived barriers of women leaders in pursuing leading positions within Greek healthcare context. The findings point mainly to organizational and socio-cultural related barriers potentially aggravated by country's unfortunate current economic turbulence. Further extensive research is required to establish grounded conclusions and better inform education and policy makers in developing gender sensitive strategies to sustainable health workforce development.


Clinical Mimics: An Emergency Medicine-Focused Review of Streptococcal Pharyngitis Mimics.
Pharyngitis is a common disease in the emergency department (ED). Despite a relatively low incidence of complications, there are many dangerous conditions that can mimic this disease and are essential for the emergency physician to consider. This article provides a review of the evaluation and management of group A β-hemolytic Streptococcal (GABHS) pharyngitis, as well as important medical conditions that can mimic this disease. GABHS pharyngitis often presents with fever, sore throat, tonsillar exudates, and anterior cervical lymphadenopathy. History and physical examination are insufficient for the diagnosis. The Centor criteria or McIsaac score can help risk stratify patients for subsequent testing or treatment. Antibiotics may reduce symptom duration and suppurative complications, but the effect is small. Rheumatic fever is uncommon in developed countries, and shared decision making is recommended if antibiotics are used for this indication. Oral analgesics and topical anesthetics are important for symptom management. Physicians should consider alternate diagnoses that may mimic GABHS pharyngitis, which can include epiglottitis, infectious mononucleosis, Kawasaki disease, acute retroviral syndrome, Lemierre's syndrome, Ludwig's angina, peritonsillar abscess, retropharyngeal abscess, and viral pharyngitis. A focused history and physical examination can help differentiate these conditions. GABHS may present similarly to other benign and potentially deadly diseases. Diagnosis and treatment of pharyngitis should be based on clinical evaluation. Consideration of pharyngitis mimics is important in the evaluation and management of ED patients.


Modeling Training Loads and Injuries: The Dangers of Discretization.
To evaluate common modeling strategies in training load and injury risk research when modeling continuous variables and interpreting continuous risk estimates; and present improved modeling strategies. Workload data were pooled from Australian football (n = 2550) and soccer (n = 23,742) populations to create a representative sample of acute:chronic workload ratio observations for team sports. Injuries were simulated in the data using three predefined risk profiles (U-shaped, flat and S-shaped). One-hundred data sets were simulated with sample sizes of 1000 and 5000 observations. Discrete modeling methods were compared with continuous methods (spline regression and fractional polynomials) for their ability to fit the defined risk profiles. Models were evaluated using measures of discrimination (area under receiver operator characteristic [ROC] curve) and calibration (Brier score, logarithmic scoring). Discrete models were inferior to continuous methods for fitting the true injury risk profiles in the data. Discrete methods had higher false discovery rates (16%-21%) than continuous methods (3%-7%). Evaluating models using the area under the ROC curve incorrectly identified discrete models as superior in over 30% of simulations. Brier and logarithmic scoring was more suited to assessing model performance with less than 6% discrete model selection rate. Many studies on the relationship between training loads and injury that have used regression modeling have significant limitations due to improper discretization of continuous variables and risk estimates. Continuous methods are more suited to modeling the relationship between training load and injury. Comparing injury risk models using ROC curves can lead to inferior model selection. Measures of calibration are more informative judging the utility of injury risk models.


A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells.
This paper describes a plasmid vector for cloning cDNAs in Escherichia coli; the same vector also promotes expression of the cDNA segment in mammalian cells. Simian virus 40 (SV40)-derived DNA segments are arrayed in the pcD vector to permit transcription, splicing, and polyadenylation of the cloned cDNA segment. A DNA fragment containing both the SV40 early region promoter and two introns normally used to splice the virus 16S and 19S late mRNAs is placed upstream of the cDNA cloning site to ensure transcription and splicing of the cDNA transcripts. An SV40 late region polyadenylation sequence occurs downstream of the cDNA cloning site, so that the cDNA transcript acquires a polyadenylated 3' end. By using pcD-alpha-globin cDNA as a model, we confirmed that the alpha-globin transcript produced in transfected cells is initiated correctly, spliced at either of the two introns, and polyadenylated either at the site coded in the cDNA segment or at the distal SV40 polyadenylation signal. A cDNA clone library constructed with mRNA from SV40-transformed human fibroblasts and this vector (about 1.4 X 10(6) clones) yielded full-length cDNA clones that express hypoxanthine-guanine phosphoribosyltransferase (Jolly et al., Proc. Natl. Acad. Sci. U.S.A., in press).


Factors of error involved in the diagnosis of juvenile myoclonic epilepsy: A study from South India.
The study was aimed at finding possible factors for delay in the diagnosis of juvenile myoclonic epilepsy (JME) in a developing country. Data was analyzed retrospectively through the medical records and prospectively through a re-evaluation of the history and EEGs of patients with JME registered in a university hospital in south India. Of the 131 patients, 23 (17.5%) patients were seen by neurologists before registration in the clinic. Diagnosis of JME was established in 118 patients at the time of registration and in 13 (10%) patients during follow-up in the clinic. The mean interval between onset of disease and the diagnosis was 6.8 + 6.3 years. In 20 patients the diagnosis was established 10 years after the onset. The mean interval between the first evaluation and diagnosis was 24. 2 months in the 13 patients in whom the diagnosis was established during follow-up in the clinic. Lack of familiarity with the clinical syndrome was probably the factor for delay in the diagnosis in 108 patients seen by practising physicians. The factors for delay in the diagnosis in patients seen by neurologists included failure to ask about myoclonic jerks resulting in misinterpretation of EEGs in 28 patients, misinterpretation of absences and/or unilateral jerks in 4 patients, and failure to ask about myoclonic jerks and misinterpretation of focal EEG abnormalities in 4 patients. This study suggests that the possible factors of error in the diagnosis of JME among the neurologists were similar to the observations reported from the developed countries; whereas the factor for delay in the diagnosis of JME among practising physicians was lack of familiarity with the epileptic syndrome.


Structural analysis of isomeric europium(III) chelates of NB-DOTMA.
Water exchange in lanthanide(III) chelates is a key parameter in developing more effective MRI contrast agents. Our own efforts to optimize water exchange have focused on isolating single coordination geometries of LnDOTA-type chelates (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate.) This isolation may be achieved by appropriately substituting the ligand framework to freeze-out the conformational exchange processes that interconvert coordination geometries. When a single nitrobenzyl substituent is used to "lock" the conformation of the macrocyclic ring, two regioisomeric chelates may be produced; the substituent may be alternatively located on the corner or the side of the ring. Here, we unambiguously demonstrate this regioisomerism by examining the COSY spectra of some conformationally locked Eu(3+) chelates. This exercise also demonstrated that diastereoisomeric chelates arising from racemization of chiral centers during the ligand synthesis, recently discounted as the origin of multiple isomeric chelates, can be produced and isolated. Furthermore, these COSY data revealed several through space NOE correlations that afford a great deal of information about the conformation of the nitrobenzyl substituent. In those isomers in which the substituent is located on the corner of the ring, the nitrobenzyl group is oriented approximately perpendicular to the plane of the macrocycle pointing upward and away from the chelate. In contrast, when the substituent is located on the side of the ring, the nitrobenzyl group is oriented approximately in plane with the macrocycle, pointing along the side of the chelate. Because the main purpose of the nitro group is to facilitate chemical modification and conjugation to biologically relevant molecules, these differences may have important consequences. Specifically, it seems likely that the same chelate may interact very differently with biological systems and molecules depending upon the regioisomer and therefore the orientation of the chelate relative to the biomolecule.


Testicular microlithiasis: ultra sound appearance.
Testicular microlithiasis is a diffuse, benign condition involving both testicles without architectural distortion. Multiple, bright echoes are present on ultrasound examination which rarely cause shadowing. Many associated conditions have been reported in patients with this entity. As this is a benign entity, knowledge of its appearance and associated conditions to prevent unnecessary surgery is important.


Interleukin 17 receptor A modulates monocyte subsets and macrophage generation in vivo.
Interleukin (IL)-17A signaling via Interleukin 17 receptor A (Il17ra) contributes to the inflammatory host response by inducing recruitment of innate immune cells, but also plays a role in homeostatic neutrophilic granulocyte regulation. Monocytes, the other main innate immune cell, have a longer life span and can pursue multiple differentiation pathways towards tissue macrophages. Monocytes are divided into two subpopulations by expression of the Ly6C/Gr1 surface marker in mice. We here investigated the role of Il17ra in monocyte homeostasis and macrophage generation. In Il17ra(-/-) and in mixed bone marrow chimeric wt/Il17ra(-/-) mice, the concentrations of circulating Il17ra(-/-) Gr1(low) monocytes were significantly decreased compared to wt cells. Pulmonary, splenic and resident peritoneal Il17ra(-/-) macrophages were significantly fewer than of wt origin. Bone marrow progenitor and monocyte numbers were equal, but the proportion of Il17ra(-/-) Gr1(low) monocytes was already decreased at bone marrow level. After monocyte depletion, initial Gr1(high) and Gr1(low) monocyte regeneration of Il17ra(-/-) and wt cells was very similar. However, Il17ra(-/-) Gr1(low) counts were not sustained. After labeling with either fluorescent beads or BrdU, Il17ra(-/-) Gr1(high) monocyte transition to Gr1(low) cells was not detectable unlike wt cells. Monocyte recruitment in acute peritonitis, which is known to be largely due to Gr1(high) cell migration, was unaffected in an identical environment. Unilateral ureteral obstruction induces a less acute inflammatory and fibrotic kidney injury. Compared to wt cells in the same environment, Il17ra(-/-) macrophage accumulation in the kidney was decreased. In the absence of Il17ra on all myeloid cells, renal fibrosis was significantly attenuated. Our data show that Il17ra modulates Gr1(low) monocyte counts and suggest defective Gr1(high) to Gr1(low) monocyte transition as an underlying mechanism. Lack of Il17ra altered homeostatic tissue macrophage formation and diminished renal inflammation and fibrosis. Il17ra appears to be a novel modulator of monocyte phenotype and possible therapeutic target in renal fibrosis.


Polyketide intermediate mimics as probes for revealing cryptic stereochemistry of ketoreductase domains.
Among natural product families, polyketides have shown the most promise for combinatorial biosynthesis of natural product-like libraries. Though recent research in the area has provided many mechanistic revelations, a basic-level understanding of kinetic and substrate tolerability is still needed before the full potential of combinatorial biosynthesis can be realized. We have developed a novel set of chemical probes for the study of ketoreductase domains of polyketide synthases. This chemical tool-based approach was validated using the ketoreductase of pikromycin module 2 (PikKR2) as a model system. Triketide substrate mimics 12 and 13 were designed to increase stability (incorporating a nonhydrolyzable thioether linkage) and minimize nonessential functionality (truncating the phosphopantetheinyl arm). PikKR2 reduction product identities as well as steady-state kinetic parameters were determined by a combination of LC-MS/MS analysis of synthetic standards and a NADPH consumption assay. The d-hydroxyl product is consistent with bioinformatic analysis and results from a complementary biochemical and molecular biological approach. When compared to widely employed substrates in previous studies, diketide 63 and trans-decalone 64, substrates 12 and 13 showed 2-10 fold lower K(M) values (2.4 ± 0.8 and 7.8 ± 2.7 mM, respectively), indicating molecular recognition of intermediate-like substrates. Due to an abundance of the nonreducable enol-tautomer, the k(cat) values were attenuated by as much as 15-336 fold relative to known substrates. This study reveals the high stereoselectivity of PikKR2 in the face of gross substrate permutation, highlighting the utility of a chemical probe-based approach in the study of polyketide ketoreductases.


Delivery of affordable and scalable encapsulated allogenic/autologous mesenchymal stem cells in coagulated platelet poor plasma for dental pulp regeneration.
The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, DPSC induced more vessel formation around the dental discs. This study provides evidence of a cost-effective, xenofree scaffold that is compatible with either autologous or allogenic strategy for dental pulp regeneration. This attempt if successfully implemented, could make REPs treatment widely accessible, contributing in improving global health conditions.


Don't worry, be (moderately) happy: Mothers' anxiety and positivity during pregnancy independently predict lower mother-infant synchrony.
Maternal positivity and mother-infant synchrony have been linked, independently, to beneficial infant outcomes; however, research that has examined relations between the two has found that higher positivity is associated with lower synchrony. Methodological issues may inform this counter-intuitive association and clinical theory supports its validity. This study examined the theory that heightened positivity associated with anxiety is a way of avoiding negative emotion and contributes to lower synchrony because it interferes with appropriate responding to infant cues. We examined mothers' (N=75) self-reported anxiety and verbal expression of positivity during pregnancy in relation to mother-infant synchrony at 6 months post-partum. Verbal positivity was assessed using linguistic analysis of interviews about pregnancy experiences. Mother and infant affect and gaze were coded during interaction and synchrony was computed as the correlation between mother and infant behaviors. Higher verbal positivity and anxiety during pregnancy independently predicted lower mother-infant synchrony, suggesting distinct pathways to the same degree of synchrony with potentially different consequences for infant development.


Antiseizure medication discovery: Recent and future paradigm shifts.
Despite the ever-increasing number of available options for the treatment of epilepsies and the remarkable advances on the understanding of their pathophysiology, the proportion of refractory patients has remained approximately unmodified during the last 100 years. How efficient are we translating positive outcomes from basic research to clinical trials and/or the clinical scenario? It is possible that fresh thinking and exploration of new paradigms are required to arrive at truly novel therapeutic solutions, as seemingly proven by recently approved first-in-class antiseizure medications and drug candidates undergoing late clinical trials. Here, the author discusses some approximations in line with the network pharmacology philosophy, which may result in highly innovative (and, hopefully, safer and/or more efficacious) medications for the control of seizures, as embodied with some recent examples in the field, namely tailored multi-target agents and low-affinity ligands.


Distinct and shared roles of β-arrestin-1 and β-arrestin-2 on the regulation of C3a receptor signaling in human mast cells.
The complement component C3a induces degranulation in human mast cells via the activation of cell surface G protein coupled receptors (GPCR; C3aR). For most GPCRs, agonist-induced receptor phosphorylation leads to the recruitment of β-arrestin-1/β-arrestin-2; resulting in receptor desensitization and internalization. Activation of GPCRs also leads to ERK1/2 phosphorylation via two temporally distinct pathways; an early response that reflects G protein activation and a delayed response that is G protein independent but requires β-arrestins. The role of β-arrestins on C3aR activation/regulation in human mast cells, however, remains unknown. We utilized lentivirus short hairpin (sh)RNA to stably knockdown the expression of β-arrestin-1 and β-arrrestin-2 in human mast cell lines, HMC-1 and LAD2 that endogenously expresses C3aR. Silencing β-arrestin-2 attenuated C3aR desensitization, blocked agonist-induced receptor internalization and rendered the cells responsive to C3a for enhanced NF-κB activity as well as chemokine generation. By contrast, silencing β-arrestin-1 had no effect on these responses but resulted in a significant decrease in C3a-induced mast cell degranulation. In shRNA control cells, C3a caused a transient ERK1/2 phosphorylation, which peaked at 5 min but disappeared by 10 min. Knockdown of β-arrestin-1, β-arrestin-2 or both enhanced the early response to C3a and rendered the cells responsive for ERK1/2 phosphorylation at later time points (10-30 min). Treatment of cells with pertussis toxin almost completely blocked both early and delayed C3a-induced ERK1/2 phosphorylation in β-arrestin1/2 knockdown cells. This study demonstrates distinct roles for β-arrestins-1 and β-arrestins-2 on C3aR desensitization, internalization, degranulation, NF-κB activation and chemokine generation in human mast cells. It also shows that both β-arrestin-1 and β-arrestin-2 play a novel and shared role in inhibiting G protein-dependent ERK1/2 phosphorylation. These findings reveal a new level of complexity for C3aR regulation by β-arrestins in human mast cells.


Characterization of the photochromic pigments in red fluorescent proteins purified from the gut juice of the silkworm Bombyx mori L.
Multiple forms of red fluorescent proteins (RFPs) were observed in the gut juice of the silkworm, Bombyx mori L. It is to be noted that only one RFP band is reported in the literature so far. However, we report here three electrophoretically separated RFPs (A, B and C) found to be heterogeneous with respect to their components, namely the protein part and the fluorescent tetrapyrrole pigment moiety. Of the three RFPs, band C was found to be a glycoprotein. The absorption extinction coefficients and fluorescence quantum yields of the three RFPs were estimated. Further, this is the first communication demonstrating the presence of three different chlorophyll derivatives associated with the three different RFPs. The pigments were analyzed by thin layer chromatography followed by their elution to characterize the pigments by spectrophotometric and spectrofluorometric methods. Spectral characteristics have led to the identification of monovinyl chlorophyllide a, divinyl protochlorophyllide a and monovinyl pheophytin a as being associated with RFP bands A, B and C, respectively. These three purified RFPs can serve as the source of the three pigments as the standards.


Daily endocrine profiles in parr and smolt Atlantic salmon.
To elucidate possible mechanisms behind the endocrine control of parr-smolt transformation, the daily plasma profiles in thyroid hormones (TH; free thyroxine (FT(4)), total thyroxine (TT(4)), and total 3,5,3'-triiodothyronine (TT(3))), growth hormone (GH) and cortisol were studied in Atlantic salmon parr and smolts under simulated-natural winter (8 L:16D) and spring (16.5 L:7.5D) photoperiods, respectively. Overall, TT(4), TT(3) and GH levels were higher in smolts than in parr, whereas FT(4) levels fluctuated within the same range in parr and smolts. Significant diurnal changes in plasma TH were present in parr. Both FT(4) and TT(4) levels increased during the photophase and decreased during the scotophase, while TT(3) levels followed an inverse pattern. Growth hormone showed no significant changes in parr. Changes in FT(4), TT(4), GH, and cortisol, but not TT(3), levels, were observed in smolts with peak levels during both the photophase and scotophase for FT(4), TT(4) and GH. Plasma cortisol was not assayed in parr but in smolts the peaks were associated with dusk and dawn. In addition to the general increases in TH, GH and cortisol, the distinct endocrine differences in nighttime levels between parr in the winter and smolts in the spring suggest different interactions between TH, GH, cortisol and melatonin at these different time points. These spring scotophase endocrine profiles may represent synergistic hormone interactions that promote smolt development, similar to the synergistic endocrine interactions shown to accelerate anuran metamorphosis. The variations in these diurnal rhythms between parr and smolts may represent part of the endocrine mechanism for the translation of seasonal information during salmon smoltification.


[Obese woman presenting as vocal cord abductor paralysis and floppy arytenoid associated with early signs of multiple system atrophy].
In multiple system atrophy (MSA), sleep-related breathing disorders are commonly observed, including vocal cord abductor paralysis (VCAP), which can cause sudden death. In its early stage, VCAP occurs only during sleep, but as the disease progresses, it appears when both awake and asleep. We encountered a 59-year-old obese woman who had been under continuous positive airway pressure (CPAP) therapy for obstructive sleep apnea syndrome (OSAS) for approximately one year but later developed acute respiratory failure because of VCAP. VCAP was the predominant finding that led to the diagnosis of MSA in our patient. On laryngoscopic examination, the movement of the patient's larynx was normal during wakefulness, but VCAP, paradoxical movements of the vocal cord and a floppy arytenoid were observed during drug-induced sleep. We suggest that detection of VCAP and laryngopharyngeal abnormalities such as floppy arytenoid in the early stage of MSA is important for determining treatment options.


Hematologic and plasma biochemical reference values for three species of black cockatoos (Calyptorhynchus species).
Black cockatoos in southwest Western Australia face population declines as a result of habitat loss and fragmentation, poaching, competition with other species, vehicle strikes, and shootings. In this study, hematologic and plasma biochemical reference values were determined for 3 endemic Western Australian black cockatoo species: Carnaby's cockatoo (Calyptorhynchus latirostris) (n = 34), Baudin's cockatoo (Calyptorhynchus baudinii) (n = 22), and forest red-tailed black cockatoo (Calyptorhynchus banksii naso) (n = 22). Significant sex-related differences were detected in Carnaby's cockatoos, and differences were also found among the 3 species. The forest red-tailed black cockatoos showed a markedly greater eosinophil count than did the 2 white-tailed species. Blood values from samples obtained by different collection methods differed significantly for Carnaby's cockatoos, which indicates that anesthesia may affect analyte values and that collection methods should remain consistent. The results of this study will be useful for clinically evaluating wild black cockatoos that are undergoing rehabilitation for release and will provide baseline data for future investigations of free-living black cockatoo health.


Heat shock proteins as cellular lifeguards.
Cells have developed complex ways to respond to various stresses. Interestingly, stresses such as heat, ischaemia and radiation can induce different cellular responses depending on their strength. While a mild stress induces a protective heat shock response, a more potent stress stimulus induces apoptosis and an even stronger one leads to necrosis. The heat shock or stress response, ie the synthesis of heat shock proteins (Hsps, stress proteins) in response to a mild stress, allows cells to adapt to gradual changes in their environment and to survive in otherwise lethal conditions. The ability of Hsps to protect cultured cells from both apoptosis and necrosis has been well demonstrated. Novel data suggest an important protective role for them also in vivo as they can protect heart and brain against ischaemia and lungs and liver against sepsis. Moreover, they can render tumours resistant to anticancer therapy. These and other cytoprotective effects of Hsps make them tempting targets for therapeutic interventions in several diseases.


Subcutaneous Semaglutide Use for Weight Management: Practice and Attitudes of Physicians in Israel.
In 2019, 1 mg subcutaneous semaglutide was registered for the treatment of diabetes in Israel. Recognition of its effect on weight has led to its use as a treatment for obesity. To explore physicians' pre-therapy considerations, therapy practices, and attitudes regarding subcutaneous semaglutide for weight loss. A 22-item questionnaire was disseminated to physicians who prescribed semaglutide 1-mg for weight loss using an authorized off-label path. In total, 127 physicians completed the questionnaire. As for pretreatment requirements, in the absence of diabetes, 30% requested a minimal body mass index of 30 kg/m2. Additional requirements were documented lifestyle-change effort (67%) and prior weight loss medication use (13%). Half of the physicians regarded calorie restriction, and 23% considered physical activity as necessary for weight loss while on therapy. As for dose, most physicians (78%) started with a 0.25-mg weekly injection, 57% doubled the dose monthly, and all others recommended doubling when side effects subsided. Regarding weight loss goal, 43% of the physicians set a personal goal with each patient while 26% limited the goal to 10% of initial weight. Fewer than 50% of physicians discussed treatment duration with their patients, and 52% of patients discontinued therapy in the first 3 months. The main reasons for discontinuation were price, lack of effect, and fear of long-term side effects. The diverse approaches regarding off-label use of semaglutide for weight reduction highlight the necessity to guide physicians and standardize treatment regimen.


Gellan gum-based bi-polymeric hydrogel scaffolds loaded with Rosuvastatin calcium: A useful tool for tendon tissue regeneration.
Statins have been long used in tissue engineering, besides their marketed hypolipidemic benefits. The aim of this research was to sustain the release of rosuvastatin calcium from bi-polymeric hydrogel scaffolds. A bi-polymer blend technique was used to enhance the mechanical properties of the fabricated hydrogels. Briefly, hydrogels were prepared via crosslinking gellan gum as the main polymer together with a secondary polymer in the presence of Ca2+. The fabricated hydrogels were assessed in terms of % swelling capacity, hydrolytic degradation and % drug released to determine the most efficient carrier system. The selected hydrogel exhibited a swelling capacity of 131.45±1.49 % following 3 weeks in an aqueous environment with a % weight loss of 15.73±1.86 % after 4 weeks post-equilibrium in aqueous medium. The results ensure a proper window for adequate drug diffusion and nutrient exchange. Sustained release was attained where 94.61±2.77 % of rosuvastatin was released at the 4-week mark. Later, FT-IR and DSC, were carried out and suggested the successful crosslinking and formation of new matrix. SEM images demonstrated the porous surface of the hydrogel while a Young's modulus of 888.558±73.549 kPa indicated the suitability of the hydrogel for soft tissue engineering. In-vivo testing involved implanting the selected hydrogel at precisely surgical cuts in the Achilles tendon of male Wistar Albino rats. Upon visual and microscopic evaluation, enhanced rates of fibrous tissue formation, vascularization and collagen expression were clearly noticed in the treatment group.


Functional changes in the heart after sacubitril/valsartan use in 5 hemodialysis patients with hypertension. Case report.
The purpose of this report is to describe the efficacy of sacubitril/valsartan in 5 hemodialysis patients with hypertension, including a patient with heart failure with reduced ejection fraction (HFrEF) and a patient with preserved ejection fraction (HFpEF) focused on the functional changes in the heart. We switched from angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor blockers (ARBs) to sacubitril/valsartan and compared blood pressure post dialysis, N-terminal pro-brain natriuretic peptide (NT-pro-BNP) levels and the findings of echocardiography for a period of 6 months. A month after the initiation of sacubitril/valsartan, there was improvement of symptoms and blood pressure post-dialysis, the NT-pro-BNP levels decreased from 23,132.2 ± 16,561.3 pg/mL to 8327 ± 3334.3 pg/mL, and the echocardiography findings showed a decrease in the left atrial dimension from 37.7 ± 5.7 mm to 33 ± 4.9 mm and an increase in the left ventricular ejection fraction from 58.2 ± 16.9% to 66.4 ± 15.0%. These results were sustained for up to 6 months. Also, blood pressure post-dialysis changed from 164 ± 11/77 mmHg to 150 ± 13/72 mmHg over the 6-month period. There were no side effects, such as hyperkalemia and lymphoedema. In conclusion, 5 patients had hypertension, including 2 patients with heart failure. Sacubitril/valsartan improved blood pressure post-dialysis, heart failure symptoms, NT-pro- BNP, the left atrial dimension, the left ventricular ejection fraction, and E/e', E/A found via echocardiography for a 6-months period. Treatment with sacubitril/valsartan was effective in hemodialysis patients in the cardiac function.


[Treatment of urinary stress incontinence using paraurethral injection of autologous fat].
The authors present the late results (greater than 1 year) in the treatment of urinary incontinence in 15 women with stress urinary incontinence and 5 men with post-adenomectomy incontinence using endoscopic injection of autologous fat tissue obtained by microliposuction. Good results were achieved in 23% of the cases with stress urinary incontinence and in 0% of those with post-adenomectomy incontinence. Although the results are disappointing, they are not very different from those obtained with teflon injection. Furthermore, the technique is simple, has no complications and it costs very little to obtain the autologous graft material. A historical review of free fat grafting and injection of liposuction fatty tissue is performed and the hypotheses that have been put forward recently relative to the fate of the fat graft are discussed.


E-ring isoprostane augments cholinergic neurotransmission in bovine trachealis via FP prostanoid receptors.
Isoprostanes are prostaglandin-like molecules that accumulate in oxidative stress and also exert powerful biological effects on a wide variety of tissues. We investigated the effects of several different isoprostanes on contractions evoked by electrical field stimulation (EFS) in bovine trachealis, finding only 15-E2t-IsoP to augment those responses. Many others have shown that isoprostanes act on prostanoid receptors, usually those of the thromboxane-selective prostanoid receptor (TP) subtype, although some describe actions through prostaglandin E2-selective prostanoid receptor (EP) or less frequently through prostaglandin F2alpha-selective prostanoid receptors (FP). We used an extensive panel of highly selective agonists and antagonists of prostanoid receptors to characterize the ones through which 15-E2t-IsoP was acting here. Pretreatment with the FP-selective AL-8810 significantly inhibited the augmentation, whereas TP- and EP-selective blockers did not. On the other hand, the augmentation exerted by 15-E2t-IsoP was mimicked by submicromolar concentrations of the FP-selective agonists PGF2alpha and fluprostenol, as well as by micromolar concentrations of the TP-selective agonist U46619. The concentration-response relationship for exogenously added acetylcholine was not significantly affected by 15-E2t-IsoP, confirming that the effect of the latter on EFS-evoked responses was exerted prejunctionally (i.e., to enhance release of Ach from nerve endings), rather than a direct postjunctional effect via a receptor on the smooth muscle. Finally, we investigated whether the inhibitory (adrenergic) innervation was also modulated by 15-E2t-IsoP, finding EFS-evoked relaxations to be unaffected by the isoprostane. We conclude that 15-E2t-IsoP acts upon an FP receptor on the cholinergic nerve endings, leading to enhanced neurotransmission.


Characterization of the effects of Mg2+ on Ca2+- and Sr2+-activated tension generation of skinned skeletal muscle fibers.
Changes in [Mg2+] in a millimolar range have a significant inverse effect on the Ca2+- (or Sr2+)activated tension generation of skeletal muscle fibers. Single frog (Rana pipiens) semitendinosus muscle fibers were "skinned" (sarcolemma removed) and contracted isometrically in bathing solutions of varying [Ca2+] or [Sr2+] and [Mg2+] but a constant pH, [MgATP2-], [K+], [CP2-], [CPK], and ionic strength. Ca2+- (or Sr2+-)activated steady-state tensions were recorded for three [Mg2+]'s: 5 X 10(-5)M, 1 X 10(-3) M, and 2 X 10(-3) M; and these tensions were expressed as the percentages of maximum tension generation of the fibers for the same [Mg2+]. Maximum tension was not affected by [Mg2+] within Ca2+-activating or Sr2+-activating sets of solutions; however, the submaximum Ca2+-(or Sr2+)activated tension is strongly affected in an inverse fashion by increasing [Mg2+]. Mg2+ behaves as a competitive inhibitor of Ca2+ and also affects the degree of cooperativity in the system. At [Mg2+] = 5 X 10(-5)M the shape of tension versus [Ca2+] (or [Sr2+]) curve showed evidence of cooperativity of Ca2+ (or Sr2+) binding or activation of the contractile system. As [Mg2+] increased, the apparent affinity for Ca2+ or Sr2+ and cooperativity of the contractile system declined. The effect on cooperativity suggests that as [Mg2+] decreases a threshold for Ca2+ activation appears.


A case of spontaneous tumor lysis syndrome in extensive-stage small-cell lung cancer: A rare oncologic emergency.
Tumor lysis syndrome (TLS) is an oncologic emergency. It generally occurs after chemotherapy but sometimes develops spontaneously in hematologic malignancy, such as leukemia. TLS is a rare phenomenon in patients with solid tumors, particularly when it develops spontaneously. Here, we present a case of spontaneous TLS (STLS) in a patient with small-cell lung cancer (SCLC). We report a case of STLS in a 59-year-old male patient who presented with dyspnea and oliguria. Clinicians should suspect TLS in patients with malignancy, who demonstrate the classic electrolyte abnormalities of TLS even if not receiving treatment such as chemotherapy or radiotherapy.


Experimental evidence for a phylogenetic Janzen-Connell effect in a subtropical forest.
Observational evidence increasingly suggests that the Janzen-Connell effect extends beyond the species boundary. However, this has not been confirmed experimentally. Herein, we present both observational and experimental evidence for a phylogenetic Janzen-Connell effect. In a subtropical forest in Guangdong province, China, we observed that co-occurring tree species are less phylogenetically related than expected. The inhibition effects of neighbouring trees on seedling survival decreased with increasing phylogenetic distance between them. In a shade-house experiment, we studied seedling survival of eight species on soil collected close to Castanopsis fissa relative to their survival on soil close to their own adult trees, and found that this relative survival rate increased with phylogenetic distance from C. fissa. This phylogenetic signal disappeared when seedlings were planted in fungicide-treated soil. Our results clearly support negative effects of phylogenetically similar neighbouring trees on seedling survival and suggest that these effects are caused by associated host-specific fungal pathogens.


Growth of nisin-producing lactococci in cooked rice supplemented with soybean extract and its application to inhibition of Bacillus subtilis in rice miso.
Lactic acid fermentation of cooked rice and rice koji by supplementation with soybean extract (SBE) and its application to rice miso fermentation were investigated. By supplementing the cooked rice with SBE, lactic acid bacteria (LAB) grew well without any unfavorable effects on the rice such as off-flavor or coloration. Lactococcus lactis subsp. lactis IFO12007 (Lc. lactis, a producer of the bacteriocin nisin) proliferated at 10(8 to approximately 9) cells/g after 24 h of incubation and produced high activity of nisin. The fermented rice with Lc. lactis strongly inhibited not only Bacillus subtilis ATCC19659 but also the other Bacillus strains. While some strains of LAB markedly inhibited the growth of Asp. oryzae, resulting in failure of koji fermentation, Lc. lactis did not affect the growth of these molds. When Lc. lactis was used for rice miso fermentation as a lactic acid starter culture, Lc. lactis rapidly proliferated and produced high nisin activity of 6,400 IU/g, in the steamed rice, resulting in complete growth inhibition of B. subtilis, which had been inoculated at the beginning of the koji fermentation. The rice miso after 12 weeks of aging had a suitable pH, and favorable taste and color. Furthermore, hyposalting of rice miso could be done without difficulty by lactic acid fermentation of both rice and soybeans.


Familial British dementia: expression and metabolism of BRI.
Vidal et al. (1999. Nature 399: 776-778) discovered that the underlying genetic lesion in familial British dementia (FBD) is a T-A transversion at the termination codon of a membrane protein, termed BRI. The mutation creates an arginine codon; translational read-through generates a novel protein, termed BRI-L, that is extended by 11 amino acids at the carboxyl-terminus. BRI-L is the precursor of the ABri peptide, a component of amyloid deposits in FBD brain. We demonstrate that both BRI and its mutant counterpart are constitutively processed by furin, resulting in the secretion of carboxyl-terminal peptide derivatives that correspond to all, or part of, ABri. Notably, elevated levels of peptides are generated from the mutant BRI precursor, suggesting that subtle conformational alterations at the carboxyl-terminus may influence furin-mediated processing. We have examined BRI/BRI-L processing by other members of the prohormone convertase (PC) family (PACE4, LPC, PC 5/6) and found that these enzymes also process BRI, albeit inefficiently. Moreover, BRI-L processing by the other PC members is severely compromised. Finally, our electron microscopic studies reveal that synthetic ABri peptides assemble into insoluble beta-pleated fibrils. Collectively, our results support the view that enhanced furin-mediated processing of mutant BRI generates amyloidogenic peptides that initiate the pathogenesis of FBD.


Centrosomal protein 55 (Cep55) stability is negatively regulated by p53 protein through Polo-like kinase 1 (Plk1).
Centrosomal protein 55 (Cep55), which is localized to the centrosome in interphase cells and recruited to the midbody during cytokinesis, is a regulator required for the completion of cell abscission. Up-regulation of Cep55 and inactivation of p53 occur in the majority of human cancers, raising the possibility of a link between these two genes. In this study we evaluated the role of p53 in Cep55 regulation. We demonstrated that Cep55 expression levels are well correlated with cancer cell growth rate and that p53 is able to negatively regulate Cep55 protein and promoter activity. Down-regulation of expression of Cep55 was accompanied by repression of polo-like kinase 1 (Plk1) levels due to p53 induction. Overexpression of Plk1 and knockdown of p53 expression both enhanced the post-translational protein stability of Cep55. BI 2356, a selective Plk1 inhibitor, however, prevented Cep55 accumulation in p53 knockdown cells while persistently keeping Plk1 levels elevated. Our results, therefore, indicate the existence of a p53-Plk1-Cep55 axis in which p53 negatively regulates expression of Cep55, through Plk1 which, in turn, is a positive regulator of Cep55 protein stability.


Monokine modulation of human astroglial cell production of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. I. Effects of IL-1 alpha and IL-beta.
Granulocyte (G)-CSF and granulocyte-macrophage (GM)-CSF enhance phagocyte survival and function and are produced by fibroblasts and endothelial cells after induction by inflammatory mediators such as IL-1. Our ability to detect G-CSF and GM-CSF activity in the conditioned medium of the human astroglial tumor cell line, U87MG, and molecularly clone the cDNA for G-CSF from a U87MG cDNA library raised the possibility that astroglial cells are capable of G-CSF and GM-CSF production within the central nervous system; if so, the production of these CSF by astroglial cells may be inducible by IL-1. We examined the effects of IL-1 alpha and IL-1 beta on the production of G-CSF and GM-CSF by U87MG and U373MG, another astroglial tumor cell line that does not constitutively produce CSF. We demonstrate that both U87MG and U373MG can be induced to produce G-CSF and GM-CSF by exposure to IL-1 alpha and IL-1 beta. This response, measured by accumulation of increased CSF mRNA, is rapid, sensitive and due to the enhanced stability of CSF message following IL-1 exposure. The implications of these findings to the immunopathogenesis of central nervous system infections are discussed.


Systematic Review of the Management of Local Kidney Cancer Relapse.
Management of locally recurrent renal cancer is complex. In this systematic review we analyse the available literature on the management of local renal cancer recurrence. A systematic search (PubMed, Web of Science, CINAHL, Clinical Trials, and Scopus) of English literature from 2000 to 2017 was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines. The search identified 1838 articles. Of those, 36 were included in the evidence synthesis. The majority of the studies identified were retrospective and not controlled. Local recurrence after thermal ablation (TA) may be managed with repeat TA. Alternatively, salvage nephrectomy is possible. However, a higher rate of complications should be expected than after primary nephrectomy. Salvage nephrectomy and TA represent treatment options for local recurrence after partial nephrectomy. Local retroperitoneal recurrence after radical nephrectomy is ideally treated with surgical resection, for which minimally invasive approaches might be applicable to select patients. For large recurrences, addition of intraoperative radiation may improve local control. Local tumour destruction appears to be more beneficial than systemic therapy alone for local recurrences. Management of local renal cancer relapse varies according to the clinical course and prior treatments. The available data are mainly limited to noncontrolled retrospective series. After nephron-sparing treatment, TA represents an effective treatment with low morbidity. For local recurrence after radical nephrectomy, the low-level evidence available suggests superiority of surgical excision relative to systemic therapy or best supportive care. As a consequence, surgery should be prioritised when feasible and applicable. In renal cell cancer, the occurrence and management of local recurrence depend on the initial treatment. This cancer is a disease with a highly variable clinical course. After initial organ-sparing treatment, thermal ablation offers good cancer control and low rates of complications. For recurrence after radical nephrectomy, surgical excision seems to provide the best long-term cancer control and it is superior to medical therapy alone.


Pharmacist Intervention Program at Different Rent Levels of Geriatric Healthcare.
As a pharmacy service giving pharmaceutical care at different levels of health care for elderly people, we needed a standardization procedure for recording and evaluating pharmacists' interventions. Our objective was to homogenize pharmacist interventions; to know physicians' acceptance of our recommendations, as well as the most prevalent drug related problems (DRP); and the impact of the pharmacists' interventions. To achieve this goal we conducted a one year prospective study at two levels of health care: 176 nursing homes (EAR) (8828 patients) and 2 long-term and subacute care hospitals (HSS) (268 beds). Pharmacists' interventions were recorded using the American Society of Health-System Pharmacists classification as the basis. Frequency of the different DRP and the level of response and acceptance on the part of physicians was determined. The Medication Appropriateness Index (MAI) was used to evaluate the impact of the interventions on the prescription quality. Patients' mean age was 84.2 (EAR) and 80.7 (HSS), and in both cases, polypharmacy ≥ 9 drugs was around 63-69%. There were 4073 interventions done in EAR and 2560 in HSS. Level of response: 44% (EAR), 79% (HSS); degree of acceptance of the recommendations: 84% (EAR), 72% (HSS). Most frequent DRP: inappropriate dose, length of therapy, omissions, and financial impact. Drugs for the nervous system are those with the most DRP. MAI values/medication improved from 4.4 to 2.7 (EAR) and 3.8 to 1.7 (HSS). A normalized way of managing pharmacists' interventions for different health care levels has been established. We are on the way to increasing collaborative work with physicians and we know which DRPs are most prevalent.


Mammal-like striatal functions in Anolis. II. Distribution of dopamine D(1) and D(2) receptors, and a laminar pattern of basal ganglia sub-systems.
We used in situ autoradiographic ligand binding methods to determine the occurrence and distribution of dopamine D(1) and D(2) receptor sub-types in the anole lizard, Anolis carolinensis. Both were present and exhibited pharmacological specificity characteristics similar to those described for mammals. However, unlike in mammals where in the neostriatum [outside the nucleus accumbens/olfactory tubercle complex (NA/OT)] these receptors exhibit only slight dorsolateral (D(2) high, D(1) low) to ventromedial (D(1 )high, D(2) low) gradients that co mingle extensively, in the anole striatum outside the NA/OT there was a striking laminar pattern, with little if any overlap between D(2) (high in a dorsal band) and D(1) (high ventral to the D(2) band) distributions. As D(1) receptors are related to the direct and D(2) to the indirect basal ganglia (BG) subsystems in mammals, we also determined anole striatal distributions of pre-proenkephalin mRNA, a marker for striatal efferents to the indirect BG subsystem in mammals. Here, too, there was a striking laminar pattern, with pre-proenkephalin mRNA in a band similar to that seen for D(2) receptors. The crisp neuroanatomical separation between these classic BG subsystem markers in Anolis striatum make this species attractive for the study of such systems' functions during behavior.


Audit on complicated diverticular disease.
Seventy-three patients were seen between 1970 and 1983 with complicated diverticular disease. There were only six hospital deaths (8%). Two out of 7 patients with faecal peritonitis died, 2 of 27 patients with purulent peritonitis died and there was one death each associated with an inflammatory mass and a peridiverticular abscess. Five of the six hospital deaths were from cardiorespiratory disease and only one was from sepsis. Three of the early deaths were in patients who were receiving steroid therapy. There were three late deaths: one from uncontrolled sepsis, one an anaesthetic death from coronary occlusion during revision of a Hartmann operation and the third was an incidental myocardial infarction. A very conservative surgical policy was adopted, primary resection only being used for an inflammatory mass and selectively for fistula and local purulent disease. Despite our apparent low hospital mortality there was a high incidence of complication; wound sepsis 29%, fistula after colostomy closure 12% and anastomotic dehiscence after primary or secondary reconstruction 12%. These findings indicate the need for a prospective audit which is now in progress.


Neuronal correlates of memory formation in motor cortex after adaptation to force field.
Activity of single neurons in the motor cortex has been shown to change during acquisition of motor skills. We previously reported that the combined activity of cell ensembles in the motor cortex of monkeys (Macaca fascicularis) evolves during adaptation to a novel force field perturbation to encode the direction of compensatory force when reaching to visual targets. We also showed that the population directional signal was altered by the available sensory feedback. Here, we examined whether traces of such activity would linger on to later constitute motor memories of the newly acquired skill and whether memory traces would differ depending on feedback. We found that motor-cortical cell ensembles retained features of their adaptive activity pattern in the absence of perturbation when reaching to both learned and unlearned targets. Moreover, the preferred directions of these cells rotated in the direction of force field while the entire population of cells produced no net rotation of preferred direction when returning to null-field reaches. Whereas the activity pattern and preferred direction rotations were comparable with and without visual feedback, changes in tuning amplitudes differed across feedback conditions. Last, savings in behavioral performance and neuronal activity during later reexposure to force field were apparent. Overall, the findings reflect a novel representation of motor memory by cell ensembles and indicate a putative role of the motor cortex in early acquisition of motor memory.


Genomic variants in members of the Krüppel-like factor gene family are associated with disease severity and hydroxyurea treatment efficacy in β-hemoglobinopathies patients.
Aim: β-Type hemoglobinopathies are characterized by vast phenotypic diversity as far as disease severity is concerned, while differences have also been observed in hydroxyurea (HU) treatment efficacy. These differences are partly attributed to the residual expression of fetal hemoglobin (HbF) in adulthood. The Krüppel-like family of transcription factors (KLFs) are a set of zinc finger DNA-binding proteins which play a major role in HbF regulation. Here, we explored the possible association of variants in KLF gene family members with response to HU treatment efficacy and disease severity in β-hemoglobinopathies patients. Materials & methods: Six tag single nucleotide polymorphisms, located in four KLF genes, namely KLF3, KLF4, KLF9 and KLF10, were analyzed in 110 β-thalassemia major patients (TDT), 18 nontransfusion dependent β-thalassemia patients (NTDT), 82 sickle cell disease/β-thalassemia compound heterozygous patients and 85 healthy individuals as controls. Results: Our findings show that a KLF4 genomic variant (rs2236599) is associated with HU treatment efficacy in sickle cell disease/β-thalassemia compound heterozygous patients and two KLF10 genomic variants (rs980112, rs3191333) are associated with persistent HbF levels in NTDT patients. Conclusion: Our findings provide evidence that genomic variants located in KLF10 gene may be considered as potential prognostic biomarkers of β-thalassemia clinical severity and an additional variant in KLF4 gene as a pharmacogenomic biomarker, predicting response to HU treatment.