Ciliary Entry of the Hedgehog Transcriptional Activator Gli2 Is Mediated by the Nuclear Import Machinery but Differs from Nuclear Transport in Being Imp-α/β1-Independent.
Gli2 is the primary transcriptional activator of Hedgehog signalling in mammals. Upon stimulation of the pathway, Gli2 moves into the cilium before reaching the nucleus. However, the mechanisms underlying its entry into the cilium are not completely understood. Since several similarities have been reported between nuclear and ciliary import, we investigated if the nuclear import machinery participates in Gli2 ciliary entry. Here we show that while two conserved classical nuclear localization signals mediate Gli2 nuclear localization via importin (Imp)-α/β1, these sequences are not required for Gli2 ciliary import. However, blocking Imp-mediated transport through overexpression of GTP-locked Ran reduced the percentage of Gli2 positive cilia, an effect that was not explained by increased CRM1-dependent export of Gli2 from the cilium. We explored the participation of Imp-β2 in Gli2 ciliary traffic and observed that this transporter is involved in moving Gli2 into the cilium, as has been described for other ciliary proteins. In addition, our data indicate that Imp-β2 might also collaborate in Gli2 nuclear entry. How does Imp-β2 determine the final destination of a protein that can localize to two distinct subcellular compartments remains an open question. Therefore, our data shows that the nuclear-cytoplasmic shuttling machinery plays a critical role mediating the subcellular distribution of Gli2 and the activation of the pathway, but distinct importins likely play a differential role mediating its ciliary and nuclear translocation.


Do miRNAs have a deep evolutionary history?
The recent discovery of microRNAs (miRNAs) in unicellular eukaryotes, including miRNAs known previously only from animals or plants, implies that miRNAs have a deep evolutionary history among eukaryotes. This contrasts with the prevailing view that miRNAs evolved convergently in animals and plants. We re-evaluate the evidence and find that none of the 73 plant and animal miRNAs described from protists meet the required criteria for miRNA annotation and, by implication, animals and plants did not acquire any of their respective miRNA genes from the crown ancestor of eukaryotes. Furthermore, of the 159 novel miRNAs previously identified among the seven species of unicellular protists examined, only 28 from the algae Ectocarpus and Chlamydomonas, meet the criteria for miRNA annotation. Therefore, at present only five groups of eukaryotes are known to possess miRNAs, indicating that miRNAs have evolved independently within eukaryotes through exaptation of their shared inherited RNAi machinery.


Choosing dentistry as a career--Part I: A comparison of student motives.
The race/gender composition of first year dental students at the University of the Witwatersrand, Johannesburg has changed over the years. Classes were previously characterised by a predominance of white males, but the race/gender distribution at present reflects a predominance of black students and a significant increase in female students. This may indicate that the motives for choosing dentistry as a career have changed and this possibility was explored in two classes of first year students, those of 1985 and 1990. The black/white and male/female distributions were 29/71 and 85/15 per cent in 1985 and 52.5/47.5 and 57/43 per cent respectively, for the class of 1990. Despite race and gender differences, motives for choosing dentistry appeared to relate to an image of the profession that is perceived as a vehicle for the achievement of personal goals.


Genetic Analysis and Clinical Characteristics of Hereditary Pheochromocytoma and Paraganglioma Syndrome in Korean Population.
Pheochromocytoma and paragangliomas (PPGL) are hereditary in approximately 30% to 40% cases. With the advancement of genetic analysis techniques, including next-generation sequencing (NGS), there were attempts to classify PPGL into molecular clusters. With NGS being applied to clinical settings recently, we aimed to review the results of genetic analysis, including NGS, and investigate the association with clinical characteristics in Korean PPGL patients. We reviewed the medical records of PPGL patients who visited Severance hospital from 2006 to 2019. We documented the clinical phenotype of those who underwent targeted NGS or had known germline mutations of related genes. Among 57 PPGL patients, we found 28 pathogenic germline mutations of susceptibility genes. Before the targeted NGS was implemented, only obvious syndromic feature lead to the Sanger sequencing for the specific genes. Therefore, for the exact prevalence, only patients after the year 2017, when targeted NGS was added, were included (n=43). The positive germline mutations were found in 14 patients; thus, the incidence rate is 32.6%. Patients with germline mutations had a higher likelihood of family history. There were significant differences in the type of PPGLs, percentage of family history, metastasis rate, presence of other tumors, and biochemical profile among three molecular clusters: pseudohypoxic tricarboxylic acid cycle-related, pseudohypoxic von Hippel-Lindau (VHL)/endothelial PAS domain-containing protein 1-related, and kinase-signaling group. Germline mutations were identified in seven PPGL-related genes (SDHB, RET, VHL, NF1, MAX, SDHA, and SDHD). We report the expected prevalence of germline mutations in Korean PPGL patients. NGS is a useful and accessible tool for genetic analysis in patients with PPGLs, and further research on molecular classification is needed for precise management.


Plasma brain natriuretic peptide level after hybrid therapy with pulmonary vein isolation and antiarrhythmic drugs for atrial fibrillation.
The atrial fibrillation (AF) recurrence rate after pulmonary vein isolation (PVI) has been relatively high and in some unsuccessful PVI cases, antiarrhythmic drugs that were ineffective before PVI may become effective (hybrid therapy). The purpose of this study was to investigate the relationship between the brain natriuretic peptide (BNP) level and the effect of the hybrid therapy. In 28 lone AF patients undergoing hybrid therapy, the plasma BNP level was measured before PVI and before and 3 months after administering an antiarrhythmic drug. Fifteen patients were free of AF after the hybrid therapy (effective group), and 13 still had recurrent AF after (noneffective group). At baseline, in all patients the BNP level was elevated, and there were no significant differences in the BNP level between the 2 groups. The BNP level was significantly decreased after PVI in the effective group (56.8 +/- 23.1 versus 37.5 +/- 16.7 pg/mL, P < 0.05) but not in the noneffective group (74.3 +/- 47.8 versus 79.7 +/- 54.4 pg/mL, NS). The elevated BNP level normalized in all effective group patients. The criterion consisting of a net value of < 60.0 pg/mL or a reduction in the BNP level of > 10.0 pg/mL after PVI predicted the effective group patients with a high accuracy. A significant reduction in the BNP level after PVI may be a useful predictor of the responders to antiarrhythmic drug therapy in patients with recurrent AF after PVI. The hybrid therapy may be effective in patients whose PVs serve as a dominant AF substrate.


Health-related quality of life in non-small-cell lung cancer: an update of a systematic review on methodologic issues in randomized controlled trials.
This study is an update of a systematic review of health-related quality-of-life (HRQOL) methodology reporting in non-small-cell lung cancer (NSCLC) randomized controlled trials (RCTs). The objective was to evaluate HRQOL methodology reporting over the last decade and its benefit for clinical decision making. A MEDLINE systematic literature review was performed. Eligible RCTs implemented patient-reported HRQOL assessments and regular oncology treatments for newly diagnosed adult patients with NSCLC. Included studies were published in English from August 2002 to July 2010. Two independent reviewers evaluated all included RCTs. Fifty-three RCTs were assessed. Of the 53 RCTs, 81% reported that there was no significant difference in overall survival (OS). However, 50% of RCTs that were unable to find OS differences reported a significant difference in HRQOL scores. The quality of HRQOL reporting has improved; both reporting of clinically significant differences and statistical testing of HRQOL have improved. A European Organisation for Research and Treatment of Cancer HRQOL questionnaire was used in 57% of the studies. However, reporting of HRQOL hypotheses and rationales for choosing HRQOL instruments were significantly less than before 2002 (P < .05). The number of NSCLC RCTs incorporating HRQOL assessments has considerably increased. HRQOL continues to demonstrate its importance in RCTs, especially in those studies in which no OS difference is found. Despite the improved quality of HRQOL methodology reporting, certain aspects remain underrepresented. Our findings suggest need for an international standardization of HRQOL reporting similar to the CONSORT guidelines for clinical findings.


The Role of PPARgamma Receptors and Leukotriene B(4) Receptors in Mediating the Effects of LY293111 in Pancreatic Cancer.
Pancreatic cancer is a devastating disease in which current therapies are inadequate. Separate lines of research have identified the 5-lipoxygenase/leukotriene B(4) receptor pathway and the PPARgamma pathway as potential targets for prevention or treatment of this disease. LY293111 was originally designed as a potent leukotriene B(4) receptor antagonist for treatment of inflammatory conditions. LY293111 was also known to have inhibitory effects on 5-lipoxygenase, which is upstream of the production of leukotrienes. LY293111 was shown to have potent anticancer effects in pancreatic cancer and several other solid malignancies, where it caused cell cycle arrest and marked apoptosis. Subsequently, it came to light that LY293111 exhibited PPARgamma agonist activity in addition to its effects on the 5-lipoxygenase pathway. This raises the question of which of the two targets is of greatest importance with regard to the anticancer effects of this agent. The evidence to date is not conclusive, but suggests that the effects of LY293111 may be mediated by both LTB(4) receptors and PPARgamma.


Fostering Interprofessional Geriatric Patient Care Skills for Health Professions Students Through a Nursing Facility-Based Immersion Rotation.
Interprofessional (IP) clinical care is ideally taught in authentic environments; however, training programs often lack authentic opportunities for health professions students to practice IP patient care. Skilled nursing facilities (SNFs) can offer such opportunities, particularly for geriatric patient care, but are underutilized as training sites. We present an IP nursing facility rotation (IP-SNF) in which medical, pharmacy, and physical therapy students provided collaborative geriatric patient care. Our 10-day immersion rotation focused on four geriatric competencies common to all three professions: appropriate/hazardous medications, patient self-care capacity, evaluating and treating falls, and IP collaboration. Activities included conducting medication reviews, quarterly care planning, evaluating functional status/fall risk, and presenting team recommendations at SNF meetings. Facility faculty/staff provided preceptorship and assessed team presentations. Course evaluations included students' pre/post objective-based self-assessment, as well as facility faculty/staff evaluations of interactions with students. Thirty-two students (15 medical, 12 pharmacy, five physical therapy) participated in the first 2 years. Evaluations (n = 31) suggested IP-SNF filled gaps in students' geriatrics and IP education. Pre/post self-assessment showed significant improvement (p < .001) in self-confidence related to course objectives. Faculty/staff indicated students added value to SNF patient care. Challenges included maximizing patient care experiences while allowing adequate team work time. IP-SNF showcases the feasibility of, and potential for, engaging learners in real-world IP geriatric patient care in a SNF. Activities and materials must be carefully designed and implemented to engage all levels/types of IP learners and ensure valuable learning experiences.


AI-IoT Low-Cost Pollution-Monitoring Sensor Network to Assist Citizens with Respiratory Problems.
The proliferation and great variety of low-cost air quality (AQ) sensors, combined with their flexibility and energy efficiency, gives an opportunity to integrate them into Wireless Sensor Networks (WSN). However, with these sensors, AQ monitoring poses a significant challenge, as the data collection and analysis process is complex and prone to errors. Although these sensors do not meet the performance requirements for reference regulatory-equivalent monitoring, they can provide informative measurements and more if we can adjust and add further processing to their raw measurements. Therefore, the integration of these sensors aims to facilitate real-time monitoring and achieve a higher spatial and temporal sampling density, particularly in urban areas, where there is a strong interest in providing AQ surveillance services since there is an increase in respiratory/allergic issues among the population. Leveraging a network of low-cost sensors, supported by 5G communications in combination with Artificial Intelligence (AI) techniques (using Convolutional and Deep Neural Networks (CNN and DNN)) to predict 24-h-ahead readings is the goal of this article in order to be able to provide early warnings to the populations of hazards areas. We have evaluated four different neural network architectures: Multi-Linear prediction (with a dense Multi-Linear Neural Network (NN)), Multi-Dense network prediction, Multi-Convolutional network prediction, and Multi-Long Short-Term Memory (LSTM) network prediction. To perform the training of the prediction of the readings, we have prepared a significant dataset that is analyzed and processed for training and testing, achieving an estimation error for most of the predicted parameters of around 7.2% on average, with the best option being the Multi-LSTM network in the forthcoming 24 h. It is worth mentioning that some pollutants achieved lower estimation errors, such as CO2 with 0.1%, PM10 with 2.4% (as well as PM2.5 and PM1.0), and NO2 with 6.7%.


Antifungal Effect of Magnolol and Honokiol from Magnolia officinalis on Alternaria alternata Causing Tobacco Brown Spot.
In this study, two phenol compounds, magnolol and honokiol, were extracted from Magnolia officinalis and identified by LC-MS, 1H- and 13C-NMR. The magnolol and honokiol were shown to be effective against seven pathogenic fungi, including Alternaria alternata (Fr.) Keissl, Penicillium expansum (Link) Thom, Alternaria dauci f.sp. solani, Fusarium moniliforme J. Sheld, Fusarium oxysporum Schltdl., Valsa mali Miyabe & G. Yamada, and Rhizoctonia solani J.G. Kühn, with growth inhibition of more than 57%. We also investigated the mechanisms underlying the potential antifungal activity of magnolol and honokiol. The results showed that they inhibited the growth of A. alternata in a dose-dependent manner. Moreover, magnolol and honokiol treatment resulted in distorted mycelia and increased the cell membrane permeability of A. alternata, as determined by conductivity measurements. These results suggest that magnolol and honokiol are potential antifungal agents for application against plant fungal diseases.


The specificity of allergic reactions. III. Contact hypersensitivity.
Intradermal injection of a simple hapten (e.g., 1-fluoro-2,4-dinitrobenzene) in water-in-oil emulsion results in contact hypersensitivity to surface application of the homologous hapten and, after appearance of circulating antibody, in Arthus type hypersensitivity to a conjugate of homologous hapten with guinea pig serum. Intradermal administration of this conjugate induces delayed and subsequently Arthus hypersensitivity to the conjugate, but no evidence of a contact reaction to the hapten alone. When a conjugate of hapten plus solubilized guinea pig skin is used as the sensitizing antigen, both contact hypersensitivity to the hapten and delayed and/or Arthus reactions to the conjugate develop. These observations are consistent with the hypothesis that the specificity of contact sensitivity is directed toward some particular protein of the skin which has been modified by combination with hapten.


Socioeconomic differences in body mass index in Spain: An intersectional multilevel analysis of individual heterogeneity and discriminatory accuracy.
Many studies have demonstrated the existence of simple, unidimensional socioeconomic gradients in body mass index (BMI). However, in the present paper we move beyond such traditional analyses by simultaneously considering multiple demographic and socioeconomic dimensions. Using the Spanish National Health Survey 2011-2012, we apply intersectionality theory and multilevel analysis of individual heterogeneity and discriminatory accuracy (MAIHDA) to analyze 14,190 adults nested within 108 intersectional strata defined by combining categories of gender, age, income, educational achievement and living situation. We develop two multilevel models to obtain information on stratum-specific BMI averages and the degree of clustering of BMI within strata expressed by the intra-class correlation coefficient (ICC). The first model is a simple variance components analysis that provides a detailed mapping of the BMI disparities in the population and measures the accuracy of stratum membership to predict individual BMI. The second model includes the variables used to define the intersectional strata as a way to identify stratum-specific interactions. The first model suggests moderate but meaningful clustering of individual BMI within the intersectional strata (ICC = 12.4%). Compared with the population average (BMI = 26.07 Kg/m2), the stratum of cohabiting 18-35-year-old females with medium income and high education presents the lowest BMI (-3.7 Kg/m2), while cohabiting 36-64-year-old females with low income and low education show the highest BMI (+2.6 Kg/m2). In the second model, the ICC falls to 1.9%, suggesting the existence of only very small stratum specific interaction effects. We confirm the existence of a socioeconomic gradient in BMI. Compared with traditional analyses, the intersectional MAIHDA approach provides a better mapping of socioeconomic and demographic inequalities in BMI. Because of the moderate clustering, public health policies aiming to reduce BMI in Spain should not solely focus on the intersectional strata with the highest BMI, but should also consider whole population polices.


Preservation of the canine liver for 24-48 hours using simple cold storage with UW solution.
The results of a series of 29 orthotopic liver transplants in the dog are described. The livers were preserved in a new cold storage fluid, UW solution, and were successfully transplanted after periods of storage of 24, 30, 36, and 48 hr. All six animals transplanted after 24 hr survived beyond 5 days after transplantation and had excellent graft function. Four of six survived for at least 5 days after 30 hr of cold storage, and five of five after 36 hr. Five of six consecutive dogs that received transplants that had been cold-stored for 48 hr survived for 5 or more days. This solution represents a substantial advance over all existing cold storage solutions for liver preservation.


A purification and some properties of an insecticidal exotoxin from Bacillus thuringiensis Berliner.
An insecticidal exotoxin from Bacillus thuringiensis var. thuringiensis (Berliner) has been purified. The efficiency of each stage of the purification has been ascertained and the yield of toxic material estimated by means of a quantitative bioassay. It is shown that the exotoxin is an adenine derivative substituted at position 9 and having a molecular weight of approximately 825. It can be dephosphorylated enzymically or chemically under conditions that define the exotoxin as a phosphomonoester. This results in loss of toxicity, both to insects and to mice. Spectroscopic and kinetic data are presented which suggest that a beta-ribofuranosyl moiety may be attached to the adenine. Glucose and allomucic acid have been positively identified as hydrolysis fragments from the exotoxin. These results are discussed and compared with the results of others on similar (or possibly identical) compounds.


Catalytic effects of Murchison material: prebiotic synthesis and degradation of RNA precursors.
Mineral components of the Murchison meteorite were investigated in terms of potential catalytic effects on synthetic and hydrolytic reactions related to ribonucleic acid. We found that the mineral surfaces catalyzed condensation reactions of formamide to form carboxylic acids, amino acids, nucleobases and sugar precursors. These results suggest that formamide condensation reactions in the parent bodies of carbonaceous meteorites could give rise to multiple organic compounds thought to be required for the emergence of life. Previous studies have demonstrated similar catalytic effects for mineral assemblies likely to have been present in the early Earth environment. The minerals had little or no effect in promoting hydrolysis of RNA (24mer of polyadenylic acid) at 80°C over a pH range from 4.2 to 9.3. RNA was most stable in the neutral pH range, with a half-life ~5 h, but at higher and lower pH ranges the half-life decreased to ~1 h. These results suggest that if RNA was somehow incorporated into a primitive form of RNA-based thermophilic life, either it must be protected from random hydrolytic events, or the rate of synthesis must exceed the rate of hydrolysis.


Accumulating evidence about what prospective memory costs actually reveal.
Event-based prospective memory (PM) tasks require participants to substitute an atypical PM response for an ongoing task response when presented with PM targets. Responses to ongoing tasks are often slower with the addition of PM demands ("PM costs"). Prominent PM theories attribute costs to capacity-sharing between the ongoing and PM tasks, which reduces the rate of processing of the ongoing task. We modeled PM costs using the Linear Ballistic Accumulator and the Diffusion Decision Model in a lexical-decision task with nonfocal PM targets defined by semantic categories. Previous decision modeling, which attributed costs to changes in caution rather than rate of processing (Heathcote et al., 2015; Horn & Bayen, 2015), could be criticized on the grounds that the PM tasks included did not sufficiently promote capacity-sharing. Our semantic PM task was potentially more dependent on lexical decision resources than previous tasks (Marsh, Hicks, & Cook, 2005), yet costs were again driven by changes in threshold and not by changes in processing speed (drift rate). Costs resulting from a single target focal PM task were also driven by threshold changes. The increased thresholds underlying nonfocal and focal costs were larger for word trials than nonword trials. As PM targets were always words, this suggests that threshold increases are used to extend the time available for retrieval on PM trials. Under nonfocal conditions, but not focal conditions, the nonword threshold also increased. Thus, it seems that only nonfocal instructions cause a global threshold increase because of greater perceived task complexity. (PsycINFO Database Record


Estimating the performance characteristics of quality-control procedures when error persists until detection.
The concepts of the power function for a quality-control rule, the error detection rate, and the false rejection rate were major advances in evaluating the performance characteristics of quality-control procedures. Most early articles published in this area evaluated the performance characteristics of quality-control rules with the assumption that an intermittent error condition occurred only within the current run, as opposed to a persistent error that continued until detection. Difficulties occur when current simulation methods are applied to the persistent error case. Here, I examine these difficulties and propose an alternative method that handles persistent error conditions effectively when evaluating and quantifying the performance characteristics of a quality-control rule.


Dexmedetomidine Dose Dependently Enhances the Local Anesthetic Action of Lidocaine in Inferior Alveolar Nerve Block: A Randomized Double-Blind Study.
Dexmedetomidine (DEX) dose dependently enhances the local anesthetic action of lidocaine in rats. We hypothesized that the effect might also be dose dependent in humans. We evaluated the effect of various concentrations of DEX with a local anesthetic in humans. Eighteen healthy volunteers were randomly assigned by a computer to receive 1.8 mL of 1 of 4 drug combinations: (1) 1% lidocaine with 2.5 ppm (parts per million) (4.5 μg) DEX, (2) lidocaine with 5.0 ppm (9.0 μg) DEX, (3) lidocaine with 7.5 ppm (13.5μg) DEX, or (4) lidocaine with 1:80,000 (22.5 μg) adrenaline (AD), to produce inferior alveolar nerve block. Pulp latency and lower lip numbness (for assessing onset and duration of anesthesia) were tested, and sedation level, blood pressure, and heart rate were recorded every 5 minutes for 20 minutes, and every 10 minutes from 20 to 60 minutes. Pulp latency of each tooth increased compared with baseline, from 5 to 15 minutes until 60 minutes. There were no significant intergroup differences at any time point. Anesthesia onset was not different between groups. Anesthesia duration was different between groups (that with DEX 7.5 ppm was significantly longer than that with DEX 2.5 ppm and AD; there was no difference between DEX 2.5 ppm and AD). Blood pressure decreased from baseline in the 5.0 and 7.5 ppm DEX groups at 30 to 60 minutes, although there was no hypotension; moreover, heart rate did not change in any group. Sedation score did not indicate deep sedation in any of the groups. Dexmedetomidine dose dependently enhances the local anesthetic action of lidocaine in humans. Dexmedetomidine at 2.5 ppm produces similar enhancement of local anesthesia effect as addition of 1:80,000 AD.


Electron impact mass spectral fragmentation of 2a,4-disubstituted 2-chloro/2,2-dichloro-2,2a,3,4-tetrahydro-1H-azeto[2,1-d][1,5]benzothia zepin-1-ones.
The mass spectrometric behaviour of nine 2a,4-disubstituted 2-chloro/2,2-dichloro-2,2a,3,4-tetrahydro-1H-azeto[2,1-d][1,5]b enzothiazepin-1-ones has been studied with the aid of mass-analysed ion kinetic energy spectrometry and accurate mass measurements under electron impact ionization. All compounds show a tendency to eliminate a neutral chlorine atom, or a chloroketene, or neutral propene, or styrene or substituted styrene molecule, plus Cl and/or H (or Cl) atom(s), to yield [M-Cl]+ ions, 2,3-dihydro-1,5-benzothiazepine derivative ions, 4,5-dihydro-5H-1,5-benzothiazepin-4-one ions which can further lose CO to give 1,4-benzothiazine ions. Both molecular ions and [M-Cl]+ ions show a tendency to eliminate an ethyl or benzyl/substituted benzyl radical to produce 2,2a-dihydro-1H-azeto[2,1-c][1,4]benzothiazin-1-one ions. The [M-Cl]+ ions could undergo rearrangement to yield 2,2a-dihydro-1H-azeto[2,1-d][1,5]benzothiazepin-1-one ions, 2,2a,3,4-tetrahydro-1H-azeto[1,2-a]quinoline ions or 1,1a,2,3-tetrahydro-azirino[2,1-d][1,5]benzothiazepine ions by loss of an ethane or a benzene/substituted benzene, a SH radical or a CO molecule. The molecular ions could also undergo rearrangement reactions to form other small fragment ions.


Nutrition and hydration concerns of the female football player.
There is little information on the nutritional habits of female football players at any level of the game. There is also a shortage of information on the nutrition and hydration strategies that players should adopt. In general, differences in nutritional needs between males and females are smaller than differences between individuals, so that principles developed for male players also apply to women. There is a need to address energy balance and body composition: prolonged energy deficits cannot be sustained without harm to health and performance. Published reports show mean carbohydrate intakes for female players of about 5 g/kg/day, and this seems to be too low to sustain consistent intensive training. The timing of protein intake may be as important as the amounts consumed, provided that the total intake is adequate. Dehydration adversely affects skill and stamina in women as it does in men, so an individualised hydration strategy should be developed. The prevalence of iron deficiency in women generally is high, but it seems to be alarmingly high in female players. All players should adopt dietary habits that ensure adequate iron intake. Football training seems to increase bone mass in the weight-bearing limbs, with positive implications for bone health in later life, but some players may be at risk from inadequate calcium dietary intake.


Snakes on the Balearic islands: an invasion tale with implications for native biodiversity conservation.
Biological invasions are a major conservation threat for biodiversity worldwide. Islands are particularly vulnerable to invasive species, especially Mediterranean islands which have suffered human pressure since ancient times. In the Balearic archipelago, reptiles represent an outstanding case with more alien than native species. Moreover, in the last decade a new wave of alien snakes landed in the main islands of the archipelago, some of which were originally snake-free. The identification of the origin and colonization pathways of alien species, as well as the prediction of their expansion, is crucial to develop effective conservation strategies. In this study, we used molecular markers to assess the allochthonous status and the putative origin of the four introduced snake species (Hemorrhois hippocrepis, Malpolon monspessulanus, Macroprotodon mauritanicus and Rhinechis scalaris) as well as ecological niche models to infer their patterns of invasion and expansion based on current and future habitat suitability. For most species, DNA sequence data suggested the Iberian Peninsula as the potential origin of the allochthonous populations, although the shallow phylogeographic structure of these species prevented the identification of a restricted source-area. For all of them, the ecological niche models showed a current low habitat suitability in the Balearic, which is however predicted to increase significantly in the next few decades under climate change scenarios. Evidence from direct observations and spatial distribution of the first-occurrence records of alien snakes (but also lizards and worm lizards) suggest the nursery trade, and in particular olive tree importation from Iberian Peninsula, as the main pathway of introduction of alien reptiles in the Balearic islands. This trend has been reported also for recent invasions in NE Spain, thus showing that olive trees transplantation may be an effective vector for bioinvasion across the Mediterranean. The combination of molecular and ecological tools used in this study reveals a promising approach for the understanding of the complex invasion process, hence guiding conservation management actions.


Module-intron correlation and intron sliding in family F/10 xylanase genes.
Xylanases are classified into two families, numbered F/10 and G/11 according to the similarity of amino acid sequences of their catalytic domain (Henrissat, B., Bairoch, A., 1993. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem. J. 293, 781-788). Three-dimensional structure of the catalytic domain of the family F/10 xylanase was reported (White, A., Withers, S.G., Gilkes, N.R., Rose, D.R., 1994. Crystal structure of the catalytic domain of the beta-1,4-glycanase Cex from Cellulomonas fimi. Biochemistry 33, 12546-12552). The domain was decomposed into 22 modules by centripetal profiles (Go, M., Nosaka, M., 1987. Protein architecture and the origin of introns. Cold Spring Harbor Symp. Quant. Biol. 52, 915-924; Noguti, T., Sakakibara, H., Go, M., 1993. Localization of hydrogen-bonds within modules in barnase. Proteins 16, 357-363). A module is a contiguous polypeptide segment of amino acid residues having a compact conformation within a globular domain. Collected 31 intron sites of the family F/10 xylanase genes from fungus were found to be correlated to module boundaries with considerable statistical force (p values <0.001). The relationship between the intron locations and protein structures provides supporting evidence for the ancient origin of introns, because such a relationship cannot be expected by random insertion of introns into eukaryotic genes, but it rather suggests pre-existence of introns in the ancestral genes of prokaryotes and eukaryotes. A phylogenetic tree of the fungal and bacterial xylanase sequences made two clusters; one includes both the bacterial and fungal genes, but the other consists of only fungal genes. The mixed cluster of bacterial genes without introns and the fungal genes with introns further supports the ancient origin of introns. Comparison of the conserved base sequences of introns indicates that sliding of a splice site occurred in Aspergillus kawachii gene by one base from the ancestral position. Substrate-binding sites of xylanase are localized on eight modules, and introns are found at both termini of six out of these functional modules. This result suggests that introns might play a functional role in shuffling the exons encoding the substrate-binding modules.


Neuronal cell surface ATP synthase mediates synthesis of extracellular ATP and regulation of intracellular pH.
The ATP synthase is known to play important roles in ATP generation and proton translocation within mitochondria. Here, we now provide evidence showing the presence of functional ecto-ATP synthase on the neuronal surface. Immunoblotting revealed that the α, β subunits of ATP synthase F1 portion are present in isolated fractions of plasma membrane and biotin-labelled surface protein from primary cultured neurons; the surface distribution of α, β subunits was also confirmed by immunofluorescence staining. Moreover, α and β subunits were also found in fractions of plasma membrane and lipid rafts isolated from rat brain, and flow cytometry analysis showed α subunits on the surface of acutely isolated brain cells. Activity assays showed that the extracellular ATP generation of cultured neurons could be compromised by α, β subunit antibodies and ATP synthase inhibitors. pH(i) (intracellular pH) analysis demonstrated that at low extracellular pH, α or β subunit antibodies decreased pHi of primary cultured neurons. Therefore, ATP synthase on the surface of neurons may be involved in the machineries of extracellular ATP generation and pH(i) homoeostasis.


Polyglutamate-based nanoconjugates for image-guided surgery and post-operative melanoma metastases prevention.
Rationale: Cutaneous melanoma is the most aggressive and deadliest of all skin malignancies. Complete primary tumor removal augmented by advanced imaging tools and effective post-operative treatment is critical in the prevention of tumor recurrence and future metastases formation. Methods: To meet this challenge, we designed novel polymeric imaging and therapeutic systems, implemented in a two-step theranostic approach. Both are composed of the biocompatible and biodegradable poly(α,L-glutamic acid) (PGA) nanocarrier that facilitates extravasation-dependent tumor targeting delivery. The first system is a novel, fluorescent, Turn-ON diagnostic probe evaluated for the precise excision of the primary tumor during image-guided surgery (IGS). The fluorescence activation of the probe occurs via PGA degradation by tumor-overexpressed cathepsins that leads to the separation of closely-packed, quenched FRET pair. This results in the emission of a strong fluorescence signal enabling the delineation of the tumor boundaries. Second, therapeutic step is aimed to prevent metastases formation with minimal side effects and maximal efficacy. To that end, a targeted treatment containing a BRAF (Dabrafenib - mDBF)/MEK (Selumetinib - SLM) inhibitors combined on one polymeric platform (PGA-SLM-mDBF) was evaluated for its anti-metastatic, preventive activity in combination with immune checkpoint inhibitors (ICPi) αPD1 and αCTLA4. Results: IGS in melanoma-bearing mice led to a high tumor-to-background ratio and reduced tumor recurrence in comparison with mice that underwent surgery under white light (23% versus 33%, respectively). Adjuvant therapy with PGA-SLM-mDBF combined with ICPi, was well-tolerated and resulted in prolonged survival and prevention of peritoneal and brain metastases formation in BRAF-mutated melanoma-bearing mice. Conclusions: The results reveal the great clinical potential of our PGA-based nanosystems as a tool for holistic melanoma treatment management.


Is kidney transplantation for everyone? The example of the older dialysis patient.
For the average patient with ESRD, kidney transplantation improves quality of life and prolongs survival compared with patients who are on the transplant waiting list and remain on dialysis. Despite the proven benefits, some patient populations, such as those of older age, may not be referred and accepted for kidney transplantation to the same extent as younger patients. The population with ESRD is aging. Nearly half of all new patients are older than 65 yr. Nephrologists and transplant surgeons will need to make more and more decisions regarding transplantation referral and candidacy in this population. This article reviews the data on survival, quality of life, and cost-effectiveness of kidney transplantation in the older dialysis patient. Clinical guidelines and data on current practice are also reviewed.


Relating surfactant properties to activity and solubilization of the human adenosine a3 receptor.
The effects of various surfactants on the activity and stability of the human adenosine A3 receptor (A3) were investigated. The receptor was expressed using stably transfected HEK293 cells at a concentration of 44 pmol functional receptor per milligram membrane protein and purified using over 50 different nonionic surfactants. A strong correlation was observed between a surfactant's ability to remove A3 from the membrane and the ability of the surfactant to remove A3 selectively relative to other membrane proteins. The activity of A3 once purified also correlates well with the selectivity of the surfactant used. The effects of varying the surfactant were much stronger than those achieved by including A3 ligands in the purification scheme. Notably, all surfactants that gave high efficiency, selectivity and activity fall within a narrow range of hydrophile-lipophile balance values. This effect may reflect the ability of the surfactant to pack effectively at the hydrophobic transmembrane interface. These findings emphasize the importance of identifying appropriate surfactants for a particular membrane protein, and offer promise for the development of rapid, efficient, and systematic methods to facilitate membrane protein purification.


Medical malpractice claims related to cataract surgery complicated by retained lens fragments (an American Ophthalmological Society thesis).
To review malpractice claims associated with retained lens fragments during cataract surgery to identify ways to improve patient outcomes. Retrospective, noncomparative, consecutive case series. Closed claims data related to cataract surgeries complicated by retained lens fragments (1989 through 2009) from an ophthalmic insurance carrier were reviewed. Factors associated with these claims and claims outcomes were analyzed. During the 21-year period, 117 (12.5%) of 937 closed claims associated with cataract surgery were related to retained lens fragments with 108 unique cataract surgeries, 97% against cataract surgeon and 3% against retinal surgeon. Twelve (11%) of 108 claims were resolved by a trial, 30 (28%) were settled, and 66 (61%) were dismissed. The defendant prevailed in 83% of trials. Indemnity payments totaling more than $3,586,000 were made in 32 (30%) of the claims (median payment, $90,000). The difference between the preoperative visual acuity and the final visual acuity was predictive of an indemnity payment (odds ratio [OR], 2.28; P=.001) and going to a trial (OR, 2.93; P=.000). Development of corneal edema was associated with an indemnity payment (OR, 3.50; P=.037). Timing of referral and elevated intraocular pressure (IOP) were statistically significant in univariate analyses but not in multivariate analyses for a trial. Whereas the majority of claims were dismissed, claims associated with greater visual acuity decline, corneal edema, or elevated IOP were more likely to result in a trial or payment. Ways to reduce significant vision loss, including improved management of corneal edema and IOP, and timely referral to a subspecialist should be considered.


Flecainide test in Brugada syndrome: a reproducible but risky tool.
The flecainide test is widely used in Brugada syndrome. However, its reproducibility and safety remain ill-defined. This study included 22 patients (18 men, mean age 34 years). Mutations in the SCN5A gene were found in eight patients. Two patients had aborted sudden cardiac death, 8 had syncope/presyncope, and 12 were asymptomatic. The ECG was diagnostic in 19 patients and suggestive in 3. At baseline, 21 of 22 patients underwent a flecainide test (2 mg/kg IV bolus over 10 minutes). In 21 of 21 patients the test was diagnostic or amplified the typical ECG pattern. At the end of drug infusion, sustained VT lasting 7-10 minutes developed in two patients. A second flecainide test was performed within 2 months in 20 patients. The test was not repeated in the two patients with prior development of VT. The flecainide test was diagnostic in 20 of 20 patients. Sustained VT occurred in one patient and recurrent VF in another. The reproducibility of the flecainide test was 100%. In 4 (18%) of 22 patients major VAs were documented after the end of flecainide infusion. VA occurred in 3 (43%) of 7 patients with, versus 1 (7%) 15 without SCN5A gene mutation (P < 0.05). No diagnostic ECG changes or arrhythmias developed in 25 control patients without structural heart disease who underwent the same study protocol. This study shows a high flecainide reproducibility, supporting its diagnostic value in Brugada syndrome. However, the occurrence of major VA, significantly higher in patients with documented SCN5A gene mutation, including in asymptomatic patients, mandates the performance under appropriate medical supervision. Whether a slower rate of drug infusion can lower the risk of VA induction, while maintaining the sensitivity of the test should be explored.


Early assessment of viable myocardium after acute myocardial infarction by low-dose echo-dobutamine.
The aim of the study was to evaluate the usefulness of low-dose dobutamine echocardiographic testing performed within 48 hours from anterior AMI in order to identify the extent of viable myocardium and predict its functional outcome. The early echo-dobutamine test was also compared with a predischarge test in order to evaluate the effects of different timing on the accuracy of the test. Nineteen consecutive patients, aged 54 +/- 11 years, with a first anterior AMI entered the study. All patients underwent a low-dose dobutamine echocardiographic test within 48 hours from hospital admission and at predischarge. In all the patients, a rest follow-up echocardiogram was performed three months after hospital discharge. Eleven patients underwent a revascularization procedure (7 underwent PTCA and 4 CABG). Of the 159 dyssynergic segments, 26% improved spontaneously at predischarge and 51% improved at the three-month follow-up. Of the 145 predischarge dyssynergic segments, 38% improved at three months. Considering the results on a segmental basis, early low-dose dobutamine echocardiography showed a sensitivity of 52%, a specificity of 87%, a positive predictive value of 81%, a negative predictive value of 64% and a diagnostic accuracy of 69% for wall-motion improvement at three months. The predischarge test showed very similar values. A slight enhancement of the sensitivity of both tests was observed considering the akinetic segments only. Finally, considering the amount of segmental reversible dysfunction inside the infarct area in the single patients, early low-dose dobutamine echocardiography showed a sensitivity of 86% and a specificity of 80%. Our results indicate that: 1) recovery of regional wall motion after AMI is slow and progressive, with substantial improvement ensuing within the first days after infarction; 2) considering results on a segmental basis, low-dose dobutamine echocardiography performed within 48 hours of AMI shows a high specificity but a low sensitivity for late recovery of regional function, although it gave information similar to what was obtained performing the test at predischarge; 3) the efficiency of test can be improved by considering the amount of reversible segmental dysfunction inside the infarct area in the single patients.


Characterization of mammalian phosphoribosylglycineamide formyltransferase from transformed cells.
Phosphoribosylglycineamide formyltransferase levels were studied in several mammalian tissues and were found to be elevated 3- to 6-fold in some anaplastic cells, including rat hepatoma H-35 and mouse leukemia L1210, as compared to their corresponding normal tissues. The enzyme was found to reside in the cytosol of chicken liver and L1210 cells. The enzyme was purified to near homogeneity, as judged by a single band on polyacrylamide gels after electrophoresis in the presence of sodium dodecyl sulfate, from two mammalian sources, mouse L1210 and mouse Sarcoma 180. Comparison of the digestion patterns of L1210 enzyme and chicken liver enzyme upon exposure to chymotrypsin showed some similarity between the two, as did cross-reactivity in Western blots of the chicken enzyme with antibodies raised to the L1210 enzyme. Subunit molecular weight of the L1210 and Sarcoma 180 phosphoribosylglycineamide formyltransferases is about 117,000. Steady-state kinetics was performed with the purified murine enzyme in the presence of 5'-phosphoribosyl-N-glycineamide and 10-formyltetrahydrofolate and with 5'-phosphoribosyl-N-glycineamide and 10-formyl-5,8-deazafolate, establishing that these mammalian enzymes utilize 10-formyltetrahydrofolate as the actual cofactor. 11-Formyltetrahydrohomofolate was found to be an inhibitor of the murine and human (HeLa) enzymes, competitive with respect to 10-formyltetrahydrofolate, with KiS of 1 and 3 microM, respectively. 11-Formyldihydrohomofolate was an inhibitor of HeLa cell growth with a 50% effective dose of 8-11 microM.


Stimulation of the neurokinin 3 receptor activates protein kinase C epsilon and protein kinase D in enteric neurons.
Tachykinins, acting through NK(3) receptors (NK(3)R), contribute to excitatory transmission to intrinsic primary afferent neurons (IPANs) of the small intestine. Although this transmission is dependent on protein kinase C (PKC), its maintenance could depend on protein kinase D (PKD), a downstream target of PKC. Here we show that PKD1/2-immunoreactivity occurred exclusively in IPANs of the guinea pig ileum, demonstrated by double staining with the IPAN marker NeuN. PKCepsilon was also colocalized with PKD1/2 in IPANs. PKCepsilon and PKD1/2 trafficking was studied in enteric neurons within whole mounts of the ileal wall. In untreated preparations, PKCepsilon and PKD1/2 were cytosolic and no signal for activated (phosphorylated) PKD was detected. The NK(3)R agonist senktide evoked a transient translocation of PKCepsilon and PKD1/2 from the cytosol to the plasma membrane and induced PKD1/2 phosphorylation at the plasma membrane. PKCepsilon translocation was maximal at 10 s and returned to the cytosol within 2 min. Phosphorylated-PKD1/2 was detected at the plasma membrane within 15 s and translocated to the cytosol by 2 min, where it remained active up to 30 min after NK(3)R stimulation. PKD1/2 activation was reduced by a PKCepsilon inhibitor and prevented by NK(3)R inhibition. NK(3)R-mediated PKCepsilon and PKD activation was confirmed in HEK293 cells transiently expressing NK(3)R and green fluorescent protein-tagged PKCepsilon, PKD1, PKD2, or PKD3. Senktide caused membrane translocation and activation of kinases within 30 s. After 15 min, phosphorylated PKD had returned to the cytosol. PKD activation was confirmed through Western blotting. Thus stimulation of NK(3)R activates PKCepsilon and PKD in sequence, and sequential activation of these kinases may account for rapid and prolonged modulation of IPAN function.


Repeated significance tests for clinical trials with a fixed number of patients and variable follow-up.
Group-sequential tests may be applied to trials in which the number of patients is fixed, a response variable is measured for each patient at successive follow-up visits, and the accumulated responses are compared across treatment groups. The standard theory is inapplicable because the increments in the accumulated responses are no longer independent. An adjustment can be made to allow for the ratio of between-patient to within-patient variance and for possible first-order autocorrelation. The method is illustrated by reference to a dental trial.


[Quality management in sociopsychiatric service: the use of routine data in efficiency control and care planning].
Caring for psychiatric patients, Social Psychiatric Services (SpDi) carry out advising and therapeutic tasks and give medical opinions. They also participate in the management of the community psychiatric care system. The services differ widely in giving priorities to these tasks. In Germany there are no standards concerning quality and quantity of individual help being offered by SpDis. Evaluation of this individual client related help is essential for quality management. Besides rates, which partly describe the utilisation of the service, such parameters are of interest that hint at the psycho-social risk. Furthermore, figures indicating continuity or discontinuity of help are of importance. The statistics of the documentation in 1999 and 2000 of the SpDi Wolfsburg serve as an example for such an evaluation.


Large cystic lympangioma of the pancreas: a case reportum.
Lymphangioma of the pancreas is rare and presents as a large cystic mass in the retroperitoneum. The pancreatic origin can be confirmed by careful evaluation of cross sectional imaging. Preoperative differentiation from other pancreatic cystic neoplasms is difficult but possible. Large symptomatic lesions warrant surgery. The diagnosis is confirmed by typical features on histopathology and immunohistochemistry. Presented here is a case report of a pancreatic lymphangioma, discussed in the context of available literature.


Dose uncertainty due to aperture effects in dynamic fields.
Dosimetry of intensity modulated radiation therapy requires accurate modeling of the beamlets that comprise each treatment segment. Planning systems such as Varian Eclipse and Philips Pinnacle recommend measuring dose distributions and output factors for fields as small as possible, generally down to at least 2 x 2 cm2. Conventionally, we perform these measurements for regular fields, defined by the secondary collimators. In practice, it is the multileaf collimation system (MLC) that defines the intensity map and provides dynamic dose modulation in either a moving window or segmented step-and-shoot mode. For this review we have only considered the latter delivery mode. Using this method, we have studied aperture motion effects on the dynamic collimator scatter (S(c)), total scatter (S(c,p)), and phantom scatter (S(p)) factors for various combinations of collimator settings (4 x 4-14 x 40 cm2) and dynamically stepped leaf gaps (0.1 to 1.0 cm) in comparison with those for static field factors. For two different Varian linear accelerators, we found similar results in a systematic dependence of collimator scatter on gap width and collimator setting. As the gap increases from 0.1 to 1.0 cm the dynamic collimator scatter factors converge from a maximum difference of about 30% toward the static field values. At the same time, there is no measurable difference between dynamic field phantom scatter factors and those conventionally obtained for static fields. Second, we evaluated the two planning systems as to how well they account for collimator scatter by attempting to mimic the dynamic apertures used above by planning and measuring dose distributions to several small, cylindrical targets for a similar range of fixed collimator settings. We found that the ratio of measured-to-planned doses as a function of target size were similar to the measured, dynamic S(c) data for the Varian Eclipse planning system, indicating underestimation of dose for targets smaller than 1 cm diameter, but were close to unity for the Philips Pinnacle system, suggestive of the underlying differences in the dose calculation algorithms. We discuss the measurements and results and potential impact on the dosimetry of small clinical targets.


Simple thiazocine-2-acetic acid derivatives via ring-closing metathesis.
A new protocol for synthesis of 2-heterocylylacetic acid derivatives involving conjugate addition of allyl mercaptan to an acrylate containing a tethered olefinic site followed by RCM (ring-closing metathesis) is described. In this series, sulfanyl derivatives were unreactive, while sulfoxide and sulfone analogues provided the corresponding thiazocines in fair to excellent yields. Use of the sulfoxide oxidation state as a protecting group for sulfides inert to RCM is demonstrated also. Thus, oxidation of sulfide 9 [N-allyl-N-[2-(allylthio)-4-(1H-indol-1-yl)-4-oxobutyl]-4-methylbenzenesulfonamide] followed by cyclization yielded the corresponding thiazocine sulfoxide 12. Deprotection (deoxygenation) of 12 was accomplished using Lawesson's reagent, producing 1-[[4-[4-(methylphenyl)sulfonyl]-3,4,5,8-tetrahydro-2H-1,4-thiazocin-2-yl]acetyl]-1H-indole (21) in 67% unoptimized yield.


Trauma-induced expression of astrocytic thrombospondin-1 is regulated by P2 receptors coupled to protein kinase cascades.
Thrombospondin-1 (TSP-1) is an extracellular matrix protein produced by astrocytes, which can promote synaptogenesis. The regulation of astrocytic TSP-1 involves extracellular ATP through the activation of P2Y receptors coupled to various protein kinase signaling pathways. However, not much is known about the mechanisms regulating TSP-1 expression in primary cortical astrocytes after a traumatic brain injury. Using an in-vitro model of central nervous system trauma that stimulates the release of ATP, we found that trauma-induced expression and release of TSP-1 involved purinergic signaling as both expression and release were significantly attenuated by pyridoxalphosphate-6-azophenyl-2'-4'-disulfonic acid, a P2 receptor antagonist. Further antagonist studies with reactive blue 2 point to a role for P2Y4, as reactive blue 2 is a potent antagonist for rat P2Y4 receptors. In addition, the injury-induced expression of TSP-1 was significantly attenuated by the inhibition of extracellular signal-regulated kinase and p38/mitogen-activated protein kinase, whereas injury-induced release of TSP-1 was significantly blocked by the inhibition of extracellular signal-regulated kinase and Akt. Using an in-vivo model of a moderate parasagittal fluid-percussion brain injury, we found that TSP-1 levels were increased when compared with those in sham animals in the cortex, thalamus, and hippocampus. We conclude that TSP-1 expression after injury can be regulated by the activation of P2 receptors coupled with protein kinase signaling pathways and suggest that purinergic signaling, by regulating TSP expression, may play an important role in cell-matrix and cell-cell interactions such as those occurring during central nervous system repair.


Partner Pronoun Use, Communal Coping, and Abstinence during Couple-Focused Intervention for Problematic Alcohol Use.
Communal coping-a process in which romantic partners view a problem as ours rather than yours or mine, and take collaborative action to address it -has emerged as an important predictor of health and treatment outcomes. In a study of partners' pronoun use prior to and during couple-focused alcohol interventions, we examined first-person plural (we-talk) and singular (I-talk) pronouns as linguistic markers of communal coping and behavioral predictors of treatment outcome. Thirty-three couples in which one partner abused alcohol were selected from a randomized control trial (N = 63) of couple-focused Cognitive-Behavioral or Family Systems Therapy if they had unambiguously successful or unsuccessful treatment outcomes (i.e., patient maintained abstinence for 30 days prior to treatment termination or had more than one heavy drinking day in the same period). Pronoun measures for each partner were obtained via computerized text analysis from transcripts of partners' speech, derived from a videotaped pretreatment interaction task and three subsequent therapy sessions. Spouse we-talk during the intervention (accounting for pretreatment we-talk), as an index of communal orientation, uniquely predicted successful treatment outcomes. In contrast, both patient and spouse I-talk during the intervention (accounting for pretreatment I-talk), as a marker of individualistic orientation, uniquely predicted unsuccessful outcomes, especially when distinguishing active and passive (I vs. me/my) pronoun forms. Results strengthen evidence for the prognostic significance of spouse behavior for patient health outcomes and for communal coping (indexed via pronoun use) as a potential mechanism of change in couple-focused interventions for health problems.


Mechanical Thrombectomy Outcomes With and Without Intravenous Thrombolysis in Stroke Patients: A Meta-Analysis.
Whether prior intravenous thrombolysis provides any additional benefits to the patients undergoing mechanical thrombectomy for large vessel, acute ischemic stroke remains unclear. We conducted a meta-analysis of 13 studies obtained through PubMed and EMBASE database searches to determine whether functional outcome (modified Rankin Scale) at 90 days, successful recanalization rate, and symptomatic intracerebral hemorrhage rate differed between patients who underwent mechanical thrombectomy with (MT+IVT) and without (MT-IVT) pre-treatment with intravenous thrombolysis. MT+IVT patients compared with MT-IVT patients had better functional outcomes (modified Rankin Scale score, 0-2; summary odds ratio [OR], 1.27 [95% confidence interval (CI), 1.05-1.55]; P=0.02; n=1769/1174), lower mortality (OR, 0.71 [95% CI, 0.55-0.91]; P=0.006; n=1774/1202), and higher rate of successful recanalization (OR, 1.46 [95% CI, 1.09-1.96]; P=0.01; n=1652/1216) without having increased odds of symptomatic intracerebral hemorrhage (OR, 1.11 [95% CI, 0.69-1.77]; P=0.67; n=1471/1143). A greater number of MT+IVT patients required ≤2 passes with a neurothrombectomy device to achieve successful recanalization (OR, 2.06 [95% CI, 1.37-3.10]; P=0.0005; n=316/231). Our results demonstrated that MT+IVT patients had better functional outcomes, lower mortality, higher rate of successful recanalization, requiring lower number of device passes, and equal odds of symptomatic intracerebral hemorrhage compared with MT-IVT patients. The results support the current guidelines of offering intravenous thrombolysis to eligible patients even if they are being considered for mechanical thrombectomy. Because the data are compiled from studies where the 2 groups differed based on eligibility for intravenous thrombolysis, randomized trials are necessary to accurately evaluate the added value of intravenous thrombolysis in patients treated with mechanical thrombectomy.


Management of rib fractures.
Rib fractures are a common reason for hospital admission and are associated with significant morbidity and mortality. This article discusses the management of simple rib fractures and provides practical guidance for junior doctors involved in the care of these patients. Careful assessment to identify patients at high risk of complications is essential and calculation of a rib fracture score can aid management decisions. Pain from rib fractures can be severe and requires multimodal analgesia started promptly and proactively on hospital admission. This may include the use of regional anaesthetic techniques such as thoracic epidurals and erector spinae blocks for patients with significant chest trauma or those at high risk of pulmonary complications.


Two new species of the genus Leptomyxa (Goodey, 1915) - Leptomyxa botanica n. sp. and Leptomyxa monrepos n. sp. from the soil habitats of Northwestern Russia.
Amoebae of the genus Leptomyxa have variable morphologies and can only be reliably identified using molecular data. However, species distinction based on the 18S rRNA gene sequence is difficult due to the very low level of sequence divergence among morphologically different species. The database for other genes is much smaller, and genomic data on Leptomyxa is almost absent. In this study, we describe two new terrestrial species of the genus Leptomyxa isolated from Northwestern Russia, Leptomyxa botanica n. sp. and Leptomyxa monrepos n. sp. Both species easily adopt an expanded fan-shaped form and have a complex structure of the nucleolar material. Phylogenetic analyses show a derived status of these two species. They form a clade with Leptomyxa valladaresi. Our tree confirms that the 18S rRNA gene sequences of Leptomyxa species are split into two large clades. The morphological synapomorphies of these clades are not obvious. This analysis is complicated by the lack of reliable morphological data on many sequenced strains and probable misidentification of some isolates.


Reversal of desipramine toxicity in rats using drug-specific antibody Fab' fragment: effects on hypotension and interaction with sodium bicarbonate.
The effect of drug-specific antibody Fab' fragment on desipramine (DMI) toxicity was studied in anesthetized rats to determine 1) whether DMI-induced hypotension can be reversed, and 2) whether the effect of this Fab' fragment can be enhanced by the concurrent administration of hypertonic NaHCO3. DMI (60 mg/kg) was administered i.p. to produce marked hypotension. Antitricyclic antidepressant (TCA) Fab' (molar Fab'/DMI ratio = 0.11) or control Fab' was administered 15 min later as a 10 min i.v. infusion. The mean arterial pressure was higher at the end of anti-TCA Fab' infusion than after control Fab' (58 +/- 8 vs. 17 +/- 7 mm Hg, P less than .001). In a second protocol, DMI (30 mg/kg) was administered to prolong QRS duration. Anti-TCA Fab' alone (molar Fab'/DMI ratio = 0.09) and NaHCO3 alone both reduced QRS prolongation compared to control treatment, and combined therapy was more effective than either one alone. In both protocols, anti-TCA Fab' markedly increased the total DMI concentration and the bound fraction of DMI in serum, but did not alter the unbound DMI concentration. In the low DMI dose protocol, anti-TCA Fab' also reduced the cardiac DMI concentration. Concurrent treatment with anti-TCA Fab' and NaHCO3 substantially increased urinary DMI and anti-TCA Fab' excretion compared to treatment with anti-TCA Fab' alone.(ABSTRACT TRUNCATED AT 250 WORDS)


Impact of gut microbiota on gut-distal autoimmunity: a focus on T cells.
The immune system is essential for maintaining a delicate balance between eliminating pathogens and maintaining tolerance to self-tissues to avoid autoimmunity. An enormous and complex community of gut microbiota provides essential health benefits to the host, particularly by regulating immune homeostasis. Many of the metabolites derived from commensals can impact host health by directly regulating the immune system. Many autoimmune diseases arise from an imbalance between pathogenic effector T cells and regulatory T (Treg) cells. Recent interest has emerged in understanding how cross-talk between gut microbiota and the host immune system promotes autoimmune development by controlling the differentiation and plasticity of T helper and Treg cells. At the molecular level, our recent study, along with others, demonstrates that asymptomatic colonization by commensal bacteria in the gut is capable of triggering autoimmune disease by molecular mimicking self-antigen and skewing the expression of dual T-cell receptors on T cells. Dysbiosis, an imbalance of the gut microbiota, is involved in autoimmune development in both mice and humans. Although it is well known that dysbiosis can impact diseases occurring within the gut, growing literature suggests that dysbiosis also causes the development of gut-distal/non-gut autoimmunity. In this review, we discuss recent advances in understanding the potential molecular mechanisms whereby gut microbiota induces autoimmunity, and the evidence that the gut microbiota triggers gut-distal autoimmune diseases.


Concentration of dentin sialoprotein at the initial stage of orthodontic treatment using self-ligating and conventional preadjusted brackets: A pilot study.
This study evaluated differences in concentration of dentin sialoprotein (DSP) in gingival crevicular fluid (GCF) relating to orthodontically induced inflammatory root resorption (OIIRR) at the initial stage of orthodontic treatment using self-ligating and conventional preadjusted brackets. Eighteen patients were assigned to three groups of equal size. Two experimental groups received non-extraction orthodontic treatment using passive self-ligating or conventional preadjusted bracket. The control group included patients without orthodontic treatment. GCF was collected from five proximal sites of maxillary anterior teeth at subsequent intervals: immediately prior to orthodontic treatment (T0), and at three and 12 weeks after initiation of treatment (T1 and T2). DSP concentration was evaluated by enzyme-linked immunoabsorbent assay and the differences in DSP levels were analyzed between and within groups. There were no significant differences in DSP levels within both experimental groups and the control group during T0-T1-T2 (P ≥ 0.05). A significant difference of DSP concentration was found between the conventional preadjusted bracket and the control group at T2 (P = 0.038). However, it was thought to be clinically insignificant. The study showed no significant difference in DSP concentration at the initial stage of orthodontic treatment with either self-ligating or conventional preadjusted bracket.


Similarity and confidence in artificial grammar learning.
Three experiments examined the relationship between similarity ratings and confidence ratings in artificial grammar learning. In Experiment 1 participants rated the similarity of test items to study exemplars. Regression analyses revealed these to be related to some of the objective measures of similarity that have previously been implicated in categorization decisions. In Experiment 2 participants made grammaticality decisions and rated either their confidence in the accuracy of their decisions or the similarity of the test items to the study items. Regression analyses showed that the grammaticality decisions were predicted by the similarity ratings obtained in Experiment 1. Points on the receiver operating characteristics (ROC) curves for the similarity and confidence ratings were closely matched. These data suggest that meta-cognitive judgments of confidence are predicated on structural knowledge of similarity. Experiment 3 confirmed this by showing that confidence ratings to median similarity probe items changed according to the similarity of preceding items.


Interspecific and intraspecific variation in proximate, mineral, and fatty acid composition of milk in Old World fruit bats (Chiroptera: Pteropodidae).
We examine the effect of body mass on milk composition among Old World fruit bats, including Pteropus pumilus (0.175 kg), Pteropus rodricensus (0.265 kg), Pteropus hypomelanus (0.571 kg), and Pteropus vampyrus (1.133 kg). We describe intra- and interspecific differences in the proximate composition of milk among these four species and the minerals and fatty acids in the milk of the latter two species. There were no differences between species in the concentrations of dry matter, fat, or lactose in milk. However, there were significant, although small, differences in the protein content of milk among species, with protein being significantly greater in P. rodricensus than in P. pumilus and P. hypomelanus and protein being significantly less in P. hypomelanus than in P. rodricensus and P. vampyrus. There were no differences in mineral content between P. hypomelanus and P. vampyrus in milk minerals, but minor differences were evident in fatty acids 12:0, 14:0, 18:0, 18:1n11, and 18:2n6. Our findings suggest that milk composition is relatively constant across lactation for most proximate, mineral, and fatty acid components. We found a significant increase in dry matter and energy across lactation in the concentration of dry matter and energy in P. pumilus and fat in P. hypomelanus. In P. hypomelanus, we found a significant increase in the concentration of fatty acids 10:0 and 20:1n9 and a significant decrease in Iso15 and 20:1n7. No other differences associated with day of lactation were found. These findings suggest that milk composition is generally similar within the genus Pteropus, despite a 6.5-fold difference in body mass between species that we evaluated.


Qualitative radiology assessment of tumor response: does it measure up?
Our purpose was to assess whether a simpler qualitative evaluation of tumor response by computed tomography is as reproducible and predictive of clinical outcome as the Response Evaluation Criteria in Solid Tumors (RECIST) and World Health Organization (WHO) methods. This study was a two-reader retrospective evaluation in which qualitative assessment resulted in agreement in 21 of 23 patients with metastatic colorectal carcinoma (91.3%, kappa=0.78; 95% CI, 0.51-1.00). Hepatic metastases were classified as increased, decreased, or unchanged, compared with agreement in 20 of 23 patients (87.0%) for RECIST (kappa=0.62; 95% CI, 0.23-1.00) and WHO (kappa=0.67; 95% CI, 0.34-1.00) methods. Patients were placed into partial response, stable disease, and disease progression categories. Time to progression of disease was better predicted qualitatively than by RECIST or WHO. Our pilot data suggest that our qualitative scoring system is more reproducible and predictive of patient clinical outcome than the RECIST and WHO methods.


Tutor-demonstrated feedback in the mini-clinical evaluation exercise.
This article was migrated. The article was marked as recommended. Introduction: The mini-clinical evaluation exercise (mini-CEX) allows for assessment and subsequent feedback across a range of individual clinical cases, although much of what is assessed is left implicit. After the performance by a trainee, further observation of appropriate clinical performance by the tutor provides a strong standard of reference for effective learning. Methods: In each mini-CEX encounter one final year resident clinically assessed an unfamiliar patient in the presence of peers and a tutor. The tutor then demonstrated the appropriate clinical behaviour. Twelve peers rated the performance of the resident before and after the tutor-demonstrated feedback (TDF). The encounters with the first cases by each of the participants were completed one by one and the cycles of encounters with the next cases by each resident were similarly continued. Results: All 13 participants completed six mini-CEX encounters over one year with an overall response rate of 95% for peer-assessment ratings (PARs). There was a total of 1772 PAR forms each with seven parameters and 1772 peer-satisfaction rating (PSR). Reliability coefficients of PARs ranged from .77 to .93. The PARs decreased significantly in each of the six cases after TDF (p<0.001). Peers rated the performance more negatively after the TDF than before the TDF. The mean PSRs, however, increased significantly in each of the six cases after the TDF (p<0.05). The mean observation time of participants' performance decreased from 26 minutes in the initial two cases to 14.5 in the last two (p<0.001). Conclusion: The feasibility and the positive educational impact of the TDF were shown in our exploratory study. The novel concept of TDF in the mini-CEX as supervised learning events and formative assessment deserves further investigation.


Preferential expression of novel MUC1 tumor antigen isoforms in human epithelial tumors and their tumor-potentiating function.
The human MUC1 gene expresses at least 2 type 1 membrane proteins: MUC1/REP, a polymorphic high m.w. MUC1 glycoprotein often highly expressed in breast cancer tissues and containing a variable number of tandem 20 amino acid repeat units, and the MUC1/Y protein, which lacks this repeat array and, therefore, is not polymorphic. Despite their documented importance in signal transduction processes, the relative expression of the 2 isoforms in epithelial tumors is unknown. Using antibody reagents which recognize different MUC1 domains, the expression of these isoforms in malignant epithelial cells has been evaluated. A comparison of the amounts of the 2 isoforms revealed preferential expression of the novel MUC1/Y protein in breast cancer tissue samples. Furthermore, although the MUC1/REP protein is almost undetectable in HeLa cervical adenocarcinoma epithelial cells, the MUC1/Y isoform is extensively expressed in these cells. The presence of the MUC1/Y sequence as well as that of an additional tandem-repeat-array-lacking isoform, designated MUC1/X, were demonstrated by reverse transcriptase PCR amplification of RNA extracted from HeLa and ovarian carcinoma cells. It has been shown previously that the MUC1 cytoplasmic domain interacts with the SH2 domain containing GRB2 protein, which transduces signals to ras, a protein which in its activated form can lead to cell transformation. We present here data demonstrating that MUC1/Y isoform expression increases the tumorigenic potential of DA3 mouse mammary epithelial cells; in contrast, potentiation of tumorigenicity is not observed with MUC1/REP expression. Our studies thus demonstrate that expression of the MUC1 gene in epithelial tumors can give rise to substantial levels of MUC1 proteins devoid of the tandem repeat array, which are generated by alternative splicing mechanisms.


Predicting Acute Myocardial Infarction with a Single Blood Draw.
We desired to determine cardiac troponin (cTn) concentrations necessary to achieve a positive predictive value (PPV) of ≥75% for acute myocardial infarction (AMI) to justify immediate admission of patients to a monitored unit and, in general, early coronary angiography. In a prospective multicenter diagnostic study enrolling patients presenting to the emergency department with symptoms suggestive of AMI, final diagnoses were adjudicated by 2 independent cardiologists based on clinical information including cardiac imaging. cTn concentrations were measured using 5 different sensitive and high-sensitivity cTn (hs-cTn) assays in a blinded fashion at presentation and serially thereafter. The diagnostic end point was PPV for rule-in of AMI of initial cTn concentrations alone and in combination with early changes. Among 3828 patients, 616 (16%) had an AMI. At presentation, 7% to 14% of patients had cTnT/I concentrations associated with a PPV of ≥75%. Adding absolute or relative changes did not significantly further increase the PPV. PPVs increased from 46.5% (95% CI, 43.6-49.4) for hs-cTnT at presentation >14 ng/L to 78.9% (95% CI, 74.7-82.5) for >52 ng/L (P < 0.001), whereas PPVs in higher hs-cTnT strata remained largely unchanged [e.g., 82.4% (95% CI, 77.5-86.7) for >80 ng/L vs 83.9% (95% CI, 76.0-90.1) for >200 ng/L (P = 0.72)]. The addition of early changes in hs-cTnT further increased the PPV up to 60 ng/L, but not for higher concentrations. Serial sampling does not seem necessary for predicting AMI and concurrent decision-making in about 10% of patients, as it only marginally increases the PPV for AMI and not in a statistically or clinically significant way. NCT00470587.


Nitric oxide inhibits the cytochrome oxidase but not the alternative oxidase of plant mitochondria.
Oxygen consumption via the cytochrome pathway in isolated soybean (Glycine max [L.] Merr.) cotyledon mitochondria was inhibited by nitric oxide (NO) while respiration via the cyanide-insensitive alternative oxidase was not significantly affected. Inhibition of cytochrome pathway activity was rapidly reversible upon depletion of the added NO. NO production was also detected in solutions of NaNO2 plus ascorbate and the extent of cytochrome pathway inhibition was dependent on the NO2- concentration. Little inhibition of alternative pathway respiration was observed under similar conditions. The alternative oxidase may play a role in nitric oxide tolerance in higher plants and in organisms such as trypanosomes which contain a plant-like alternative oxidase.


[Dilemma of Environmental Health Research].
This article presents not only a brief overview of birth cohort studies focusing on environmental health in which the associations between health and environment were examined, but also a tentative plan to apply epidemiological data to benchmark dose calculation. According to the preceding studies, the checkpoints to be scrutinized when a result is not consistent with those of other researchers are as follows: (1) whether the study included all crucial confounders, (2) whether it included any exposure marker or confounder with a U-shaped dose-response curve, (3) whether the outcome measure was conducted by two or more examiners that might lead to measurement bias, (4) whether such examiners picked up information about exposure levels of the subjects before measuring the endpoints, and (5) whether subjects with different genetic factors were included in the analysis. In addition, (6) researchers conducting a children's study on developmental effects due to toxic substances must keep in mind that the impact of prenatal methylmercury exposure, independent of postnatal exposure, may continue for at least seven years. (7) When an environmental health research emphasizes to be population-based study, the levels of exposure to environmental chemical substances in developed countries with strict environmental regulations may be too low to examine a dose-response relationship for critical dose estimation. Such risk assessment should be carried out among the subjects with a wide range of exposure levels.


Role of 99mTc-IDA cholescintigraphy in evaluating biliary tract disorders.
Technetium-99m IDA cholescintigraphy has provided a new, noninvasive means of visualizing biliary tract function. It has become the procedure of choice in patients with suspected acute cholecystitis because of its ability to most accurately detect functional obstruction or patency of the cystic duct as opposed to ultrasound's ability to detect only anatomic changes such as the presence of calculi or a thickened gallbladder wall. These latter findings are more important in establishing the diagnosis of chronic cholecystitis where ultrasound shares a position of prime importance with the oral cholecystogram. Tc-99m IDA cholescintigraphy has also been particularly useful in evaluating bile leaks, biliary-enteric anastomosis patency and the post-cholecystectomy patient with recurrent pain. In the patient with cholestasis, ultrasound is usually the procedure of choice since it establishes whether or not ductal dilatation is present and frequently can determine the cause of obstruction. Cholescintigraphy has played an ancillary role in many cases by demonstrating the level of partial obstruction, but it does not have the anatomic resolution to visualize the cause of obstruction. Occasionally, in the evaluation of cholestasis, cholescintigraphy has proven to be the only modality which has identified the presence of acute common duct obstruction or localized intrahepatic ductal obstruction. All in all, Tc-99m IDA cholescintigraphy has had a dramatic impact upon hepatobiliary diagnosis.


Pediatric movement disorders.
Pediatric movement disorders are a heterogeneous group of symptoms that occur in the context of a large number of different neurological diseases. Accurate diagnosis and quantification of these disorders is essential for determining outcome, appropriate treatment, and criteria for inclusion in research trials. The purpose of this review is to summarize recent advances in diagnosis and treatment for childhood movement disorders. The ultimate goal is to discover new treatments that can lead to measurable improvement in functional outcome for affected children. In order to accomplish this goal, we must have consistent definitions and accurate measurements to determine the diagnosis and severity for each child in a clinic or research trial. Recent progress in defining childhood movement disorders has led to consensus definitions of different types of hypertonia. There has also been progress in the development of outcome measures that relate to meaningful functional performance in a variety of skill areas. Most exciting is the prospect of new treatments, and we survey the current non-medical, medical, and surgical therapies for childhood motor disorders. Although pediatric movement disorders are a complex and often poorly understood group of symptoms, recent work has shown that there is a possibility of defining, measuring, and ultimately treating these debilitating diseases.


Future research and therapeutic applications of human stem cells: general, regulatory, and bioethical aspects.
There is much to be investigated about the specific characteristics of stem cells and about the efficacy and safety of the new drugs based on this type of cells, both embryonic as adult stem cells, for several therapeutic indications (cardiovascular and ischemic diseases, diabetes, hematopoietic diseases, liver diseases). Along with recent progress in transference of nuclei from human somatic cells, as well as iPSC technology, has allowed availability of lineages of all three germ layers genetically identical to those of the donor patient, which permits safe transplantation of organ-tissue-specific adult stem cells with no immune rejection. The main objective is the need for expansion of stem cell characteristics to maximize stem cell efficacy (i.e. the proper selection of a stem cell) and the efficacy (maximum effect) and safety of stem cell derived drugs. Other considerations to take into account in cell therapy will be the suitability of infrastructure and technical staff, biomaterials, production costs, biobanks, biosecurity, and the biotechnological industry. The general objectives in the area of stem cell research in the next few years, are related to identification of therapeutic targets and potential therapeutic tests, studies of cell differentiation and physiological mechanisms, culture conditions of pluripotent stem cells and efficacy and safety tests for stem cell-based drugs or procedures to be performed in both animal and human models in the corresponding clinical trials. A regulatory framework will be required to ensure patient accessibility to products and governmental assistance for their regulation and control. Bioethical aspects will be required related to the scientific and therapeutic relevance and cost of cryopreservation over time, but specially with respect to embryos which may ultimately be used for scientific uses of research as source of embryonic stem cells, in which case the bioethical conflict may be further aggravated.


Autonomic neurosurgery: from microvascular decompression to image guided stimulation.
The paper reviews mechanisms underlying autonomic disorders, with a focus on cardiovascular dysfunction. Neurosurgical approaches are described for medically refractory hypertension and orthostatic hypotension. After review of microvascular decompression of the rostral ventrolateral medulla, stereotactic CT and MRI guided deep brain stimulation of the periaqueductal grey matter (PAG) is evaluated. Results are presented from patient studies showing reductions in blood pressure with ventral PAG stimulation and increases in blood pressure with dorsal PAG stimulation. A rationale for the treatment of autonomic disorders by neurosurgical intervention is discussed.


Recommendations for the design of therapeutic trials for neonatal seizures.
Although seizures have a higher incidence in neonates than any other age group and are associated with significant mortality and neurodevelopmental disability, treatment is largely guided by physician preference and tradition, due to a lack of data from well-designed clinical trials. There is increasing interest in conducting trials of novel drugs to treat neonatal seizures, but the unique characteristics of this disorder and patient population require special consideration with regard to trial design. The Critical Path Institute formed a global working group of experts and key stakeholders from academia, the pharmaceutical industry, regulatory agencies, neonatal nurse associations, and patient advocacy groups to develop consensus recommendations for design of clinical trials to treat neonatal seizures. The broad expertise and perspectives of this group were invaluable in developing recommendations addressing: (1) use of neonate-specific adaptive trial designs, (2) inclusion/exclusion criteria, (3) stratification and randomization, (4) statistical analysis, (5) safety monitoring, and (6) definitions of important outcomes. The guidelines are based on available literature and expert consensus, pharmacokinetic analyses, ethical considerations, and parental concerns. These recommendations will ultimately facilitate development of a Master Protocol and design of efficient and successful drug trials to improve the treatment and outcome for this highly vulnerable population.


[Physiopathology and nutritional care of patients with gestational diabetes].
Maternal risks include the development of diabetes after pregnancy, as well as having an infant with macrosomia, with elevated risk of developing obesity and diabetes in childhood. The main goal of treatment is to maintain an adequate glycemic control during pregnancy and guarantee the recommended weight gain. The first treatment strategy is diet therapy, however, some women need insulin therapy to achieve adequate glycemic control. The risk of diabetic fetopathy decreases when maintaining postprandial glycemic levels within normal ranges. These levels are directly associated with the amount and type of carbohydrates consumed during meals. So, nutrition therapy should be an integral part of gestational diabetes treatment. Nutrition therapy includes a complete nutrition assessment, an individual food plan that meets energy and protein requirements for pregnancy (in obese women never lesser than 1,700 kcal/day), in which lipids and carbohydrates may provide lesser than 40 and between 40 and 45% of total energy intake. Education about food groups that provide carbohydrates, portion sizes and how to achieve an equal carbohydrate distribution throughout the day should be provided. Orientation about eating healthy fats and increasing the consumption of high-fiber foods should also be included. This approach requires that treatment of women with gestational diabetes should be provided by a multidisciplinary team, including nutrition specialists.


DNP Preparation in Supporting Critical Skills for Magnet® Program Directors.
Magnet® organizations are required to identify a Magnet program director (MPD), who plays a critical role in attaining and maintaining Magnet designation. The evidence is sparse related to qualifications or recommended educational levels of those assuming an MPD role. In this article, the author discusses how one of the DNP Essentials, clinical scholarship and the use of analytical methods for evidence-based practice, is reported to prepare nurses for the MPD role.


[Isolation and characteristics of intracellular nuclease isoforms from Serratia marcescens].
Two enzyme forms were isolated from the commercial preparation of extracellular endonuclease of Serratia marcescens strain B10 M1. The chromatographic and electrophoretic properties, isoelectric points and N-terminal amino acid residues are different for both enzymes. At the final step of the purification procedure including ion-exchange chromatography on phospho- and DEAE-cellulose columns the yields of nucleases Sm1 and Sm2 were 13% and 25%, respectively. No significant differences were found in the specific activities of nucleases Sm1 and Sm2 (3.6 x 10(6) and 4.0 x 10(6) un. act./mg of protein). A comparative analysis of tryptic nuclease hydrolysate peptides was carried out. The amino acid sequences of some polypeptide segments of the proteins were determined. The structural similarity of the enzyme was established and the amino terminal regions of the proteins were identified. The localization of the disulfide bonds in the molecules of the both nucleases was determined. The similarity of nucleases Sm1 and Sm2 strain B10 M1 to S. marcescens endonucleases obtained from other strains was demonstrated.


Diffusion MRI with concurrent magnetic field monitoring.
Diffusion MRI is compromised by unknown field perturbation during image encoding. The purpose of this study was to address this problem using the recently described approach of concurrent magnetic field monitoring. Magnetic field dynamics were monitored during the echo planar imaging readout of a common diffusion-weighted MRI sequence using an integrated magnetic field camera setup. The image encoding including encoding changes over the duration of entire scans were quantified and analyzed. Field perturbations were corrected by accounting for them in generalized image reconstruction. The impact on image quality along with geometrical congruence among different diffusion-weighted images was assessed both qualitatively and quantitatively. The most significant field perturbations were found to be related to higher-order eddy currents from diffusion-weighting gradients and B0 field drift as well as gradual changes of short-term eddy current behavior and mechanical oscillations during the scan. All artifacts relating to dynamic field perturbations were eliminated by incorporating the measured encoding in image reconstruction. Concurrent field monitoring combined with generalized reconstruction enhances depiction fidelity in diffusion imaging. In addition to artifact reduction, it improves geometric congruence and thus facilitates image combination for quantitative diffusion analysis.


A Comparison of the Differences in Postoperative Chronic Pain Between Video-Assisted and Robotic-Assisted Approaches in Thoracic Surgery.
Background and objective In the last decade, there has been significant evolution in thoracic surgery with the advent of robotic surgery. In this study, we aimed to evaluate the incidence of postoperative chronic pain (for six months and beyond) in robotic and video-assisted approaches to analyze the long-term effects of the two different techniques. Methods This was a retrospective study involving 92 patients who underwent various thoracic operations between six months and two years preceding the study. Patients were classified into two groups based on the type of surgery: video-assisted (VATS) (n=51), and robotic-assisted (RATS) (n=41) thoracoscopic Surgery. We employed the EuroQol (EQ-5D-5L) questionnaire to assess the utility values in terms of five quality-of-life measures (self-care, pain/discomfort, mobility, anxiety/depression, and usual activities). Results In the VATS group, the median age was 68 years while it was 57 years in the RATS group (p=0.001). A higher proportion of patients in the VATS group had anatomical lung resection (lobectomy) compared to the RATS group: 61.2 vs. 41.6% respectively (p=0.005). However, the groups were well-matched on other patient characteristics such as relevant past medical history, underlying disease pathology, and final disease staging (if malignant), with no significant differences between groups observed regarding these traits. In the VATS group, 62.7% of patients were pain-free at the time of the questionnaire-based evaluation compared to 51.2% in the RATS group. Additionally, 25.5% vs. 39% of patients had mild pain in the VATS and RATS groups respectively. Neither of these differences was statistically significant. Conclusion Patients who undergo RATS are known to have better recovery and less pain compared to those who have VATS in the immediate postoperative period. However, our results did not find RATS to be superior to VATS in terms of long-term pain. Additionally, robotic surgery is associated with higher hospital costs. In light of these findings, further comparative studies between the two approaches are recommended, while strategies to reduce postoperative pain and financial cost should continue to be explored.


Increased T-regulatory cells within lymphocyte follicles in moderate COPD.
Lymphoid follicles in the lung parenchyma are a characteristic feature of chronic obstructive pulmonary disease (COPD). There are reports of altered CD4 T-regulatory cell numbers in COPD lungs, but the location of these cells within COPD lung tissue specific follicles has not been investigated. The presence of CD4(+)FOXP3(+) T-regulatory cells was assessed in surgically resected lung tissue from 12 COPD patients, 11 smokers with normal lung function and seven nonsmokers by combined immunofluorescence and immunohistochemistry. Organised lymphoid follicles were observed in all three groups of patients, as well as lymphoid clusters lacking organisation. The percentage of CD4 cells that were T-regulatory cells were significantly increased (p = 0.02) within COPD (16%) follicles compared with smokers (10%) and nonsmokers (8%). In contrast, there was no change (p>0.05) in the percentage of T-regulatory cells in clusters or the subepithelium between groups. Lymphoid follicles in COPD patients have increased T-regulatory cells. Therefore, T-regulatory activity may be altered within COPD lymphoid follicles.


Saccade-related information in the superior temporal motion complex: quantitative functional mapping in the monkey.
Although the role of the motion complex [cortical areas middle temporal (V5/MT), medial superior temporal (MST), and fundus of the superior temporal (FST)] in visual motion and smooth-pursuit eye movements is well understood, little is known about its involvement in rapid eye movements (saccades). To address this issue, we used the quantitative 14C-deoxyglucose method to obtain functional maps of the cerebral cortex lying in the superior temporal sulcus of rhesus monkeys executing saccades to visual targets and saccades to memorized targets in complete darkness. Fixational effects were observed in MT-foveal, FST, the anterior part of V4-transitional (V4t), and temporal-occipital areas. Saccades to memorized targets activated areas V5/MT, MST, and V4t, which were also activated for saccades to visual targets. Regions activated in the light and in the dark overlapped extensively. In addition, saccades to visual targets activated areas FST and the intermediate part of the polysensory temporal-parietal-occipital area. Cortical activity related to visually guided saccades could be explained, at least in part, by visual motion. Because only oculomotor signals can account for the equally robust activations induced by memory saccades in complete darkness, we suggest that areas V5/MT, MST, and V4t receive and/or process saccade-related oculomotor information.


The impact of financial crisis on coronary artery disease burden in Greece.
Economic crisis poses an immense threat to public health worldwide and has been linked to cardiovascular morbidity and mortality. Greece is facing a distinctive recession over the recent years. However, the exact impact on coronary artery disease (CAD) burden has not been adequately addressed. Demographic, clinical, and angiographic data of 3895 hospitalized patients were retrospectively studied. Patients were classified into two groups: those before crisis (2006-2007, n = 1228) and those during crisis (2011-2015, n = 2667). All data before and during crisis were compared. During crisis, patients presented with less acute coronary syndrome (ACS - 45.5% vs. 39.9%, p < 0.001). Subsequently, there were more patients without CAD (23.7% vs. 35.1%, p < 0.001) or one-vessel disease (20.5% vs. 23%, p < 0.001). The prevalence of traditional risk factors decreased significantly or remained stable except obesity (26.3% vs. 31.4%, p = 0.002). A significant increase in the examined females (23.6% vs. 26.7%, p = 0.04) was also observed. The burden of CAD in Greece was partially affected during the financial crisis. Even though the incidence of ACS was decreased, more women and more patients with no- or single-vessel disease were referred for cardiac catheterization. In addition, the prevalence of traditional risk factors for CAD did not increase except obesity confirming the "obesity paradox." It seems that the impact of traditional risk factors for CAD is not an immediate process and is somewhat related to living conditions or other exogenous and social factors.