Monoclonal antibodies as context-specific reagents: differential antigen reactivity of MAb 58K-9 during immunoprecipitation, immunoblotting, and immunofluorescence analyses.
Monoclonal antibodies (MAbs) are used as antigen-specific reagents in a variety of techniques. Usually it is assumed that antigen specificity shown by immunoprecipitation and immunoblotting will persist in immunofluorescence analyses. Here, we show that the behavior of MAbs is not always consistent with this assumption. Specifically, we demonstrate that a MAb (MAb 58K-9) preferentially interacts with the receptor-associated protein (RAP), a ubiquitous ER protein acting as a chaperone for the low density lipoprotein receptor-related protein (LRP) in immunoprecipitation and immunoblotting analyses. However, MAb 58K-9 does not recognize RAP in immunofluorescence studies. Interestingly, by this technique MAb 58K-9 exclusively detects an unrelated antigen peripherally associated with the cytosolic aspect of Golgi membranes. This reactivity was observed for primate but not rodent antigens. Our results indicate that MAbs that recognize a specific antigen by a single immuno-technique cannot be assumed to recognize the same antigen by other immuno-techniques, and that the identity of recognized antigens must be confirmed within each experimental context.


[The rank of data matrices in a spectrophotometric study of a ligand-biopolymer complex formation].
The dependences of concentrations of ligand-linear polymer complex species vs. the concentration of free ligand were calculated. The analysis of the data suggests that the variance of the property studied considerably depends on the parameters of ligand-polymer complex formation (cooperativity and the average number of monomers occupied by the ligand). The rank of the matrix of concentration fractions of species was estimated using the singular value decomposition by the method of principal component analysis. As the cooperativity increases, the rank matrix decreases from 4 to 2. An increase in the average number of monomers occupied by the ligand at high cooperativity leads to an increase in the matrix rank from 2 to 3.


Wireless Biological Electronic Sensors.
The development of wireless biological electronic sensors could open up significant advances for both fundamental studies and practical applications in a variety of areas, including medical diagnosis, environmental monitoring, and defense applications. One of the major challenges in the development of wireless bioelectronic sensors is the successful integration of biosensing units and wireless signal transducers. In recent years, there are a few types of wireless communication systems that have been integrated with biosensing systems to construct wireless bioelectronic sensors. To successfully construct wireless biological electronic sensors, there are several interesting questions: What types of biosensing transducers can be used in wireless bioelectronic sensors? What types of wireless systems can be integrated with biosensing transducers to construct wireless bioelectronic sensors? How are the electrical sensing signals generated and transmitted? This review will highlight the early attempts to address these questions in the development of wireless biological electronic sensors.


Molecular innovations in plant TIR-based immunity signaling.
A protein domain (Toll and Interleukin-1 receptor [TIR]-like) with homology to animal TIRs mediates immune signaling in prokaryotes and eukaryotes. Here, we present an overview of TIR evolution and the molecular versatility of TIR domains in different protein architectures for host protection against microbial attack. Plant TIR-based signaling emerges as being central to the potentiation and effectiveness of host defenses triggered by intracellular and cell-surface immune receptors. Equally relevant for plant fitness are mechanisms that limit potent TIR signaling in healthy tissues but maintain preparedness for infection. We propose that seed plants evolved a specialized protein module to selectively translate TIR enzymatic activities to defense outputs, overlaying a more general function of TIRs.


Molecular imaging of matrix metalloproteinases in atherosclerotic plaques.
Ischaemic stroke and myocardial infarction often result from the sudden rupture of an atherosclerotic plaque. The subsequent arterial thrombosis occluding the vessel lumen has been widely indicated as the crucial acute event causing peripheral tissue ischaemia. A complex cross-talk between systemic and intraplaque inflammatory mediators has been shown to regulate maturation, remodeling and final rupture of an atherosclerotic plaque. Matrix metalloproteinases (MMPs) are proteolytic enzymes (released by several cell subsets within atherosclerotic plaques), which favour atherogenesis and increase plaque vulnerability. Thus, the assessment of intraplaque levels and activity of MMP might be of pivotal relevance in the evaluation of the risk of rupture. New imaging approaches, focused on the visualisation of inflammation in the vessel wall and plaque, may emerge as tools for individualised risk assessment and prevention of events. In this review, we summarize experimental findings of the currently available invasive and noninvasive imaging techniques, used to detect the presence and activity of MMPs in atherosclerotic plaques.


Transcription regulation of human manganese superoxide dismutase gene.
The human MnSOD gene has a typical housekeeping gene promoter, but is highly inducible by various physical, chemical, and biological agents. Transcription factors SP-1 and AP-2 seem to have opposite roles in the transcriptional activity of the basal promoter. Whereas SP-1 plays a positive role, which is absolutely essential for transcription from the human MnSOD promoter, AP-2 appears to play a negative role in this process. An enhancer element is found in the promoter region of the human MnSOD gene. Several important enhancer elements are located in the second intron. The NF-kappa B site in the second intron is essential but not sufficient for high-level induction of MnSOD by cytokines. Although mutations in the regulatory elements may be partially responsible for the lack of induction of MnSOD in some cell types, differences in the degree of induction exist that cannot be accounted for by the defect in the DNA sequence. It is highly likely that this difference is due to the presence or absence of coactivator or suppressor proteins in the cells and may have a physiological role in the defense against oxidative stress.


Energy dependence of a scintillating fiber detector for preclinical dosimetry with an image guided micro-irradiator.
The dosimetry of preclinical micro-irradiators is challenging due to their millimetric beams and medium x-ray energy range. Plastic scintillator dosimeters (PSD) are good candidates for such a purpose as they provide a high spatial resolution although they show an energy dependence below 100 keV. The purpose of this study was to assess the energy dependence of a dedicated PSD (called DosiRat) for micro-irradiators dosimetry. The response of the PSD relative to air kerma was measured for different beam qualities (40-225 kV) with the X-RAD 225Cx irradiator. The corresponding energy spectra, determined by Monte Carlo simulations, allowed for correcting the differences in absorbed dose between the DosiRat material (polystyrene) and the air and therefore allowed to compare DosiRat intrinsic energy response to the Birks scintillation quenching model. The energy response of DosiRat was then assessed under preclinical conditions through percentage depth dose curves (PDD) and relative output factor (ROF) measurements in water for beam diameters ranging from 1 to 25 mm. DosiRat energy response showed a coefficient of variation of 23% from 40 to 225 kV, mainly explained by the mass energy-absorption coefficient variation between polystyrene and air. A remaining variation was shown to be caused by the quenching of the scintillation and was correctly reproduced by the Birks model (with kB = 10.27 mg MeV-1 cm-2). PDD and ROF measurements highlighted an energy response variation with depth and collimation up to 10%. A dose accuracy better than 1% was finally achieved with appropriate calibration and correction factors (CF), for beam collimations larger than the detector ([Formula: see text]2 mm diameter). DosiRat energy dependence was fully characterized in preclinical energy range and shown to be negligible with convenient calibration and corrections factors. It provided accurate dosimetry for medium energy (225 kV) and millimetric beams (down to 2.5 mm).


The French model of psychoanalytic training: Ethical conflicts.
Research on psychoanalytical education within the IPA may be clarified by reflecting on the ethic behind each of the three main models (Eitingonian, French and Uruguayan). In fact, the ethic underpinning psychoanalytical education, whatever the model, is confronted by irreducible conflicts between transmitting psychoanalysis by means of analytical experience or by means of academic teaching. Transmission by experience is essentially based on the ethic of psychoanalytic practice, which is difficult to regulate through institutional standards, whereas the academic aspect can be evaluated by objective and public criteria. The importance of both aspects and their relative weight in the training process depend on the conception of psychoanalysis underlying each model. This paper will look primarily at the French training model, the essentially analytical aspects of which favour the transmission of the very ethical foundations of psychoanalytic practice itself: the application of the method both as a working tool and as a tool of evaluation. It presupposes expanding the observation and analysis of transference beyond the framework of treatment to that of supervision. From this analysis, the paper will attempt to demonstrate how the French model proposes dealing with the inevitable conflicts between transmission by means of analysis and training by means of apprenticeship.


Endothelium and hyperviscosity.
The role of viscosity, and of interindividual variations in this parameter, in the pathophysiology of cardiovascular disease remain incompletely understood. Any speculation regarding the possible impact of "hemorheological" therapies is therefore even more complex. In the last years, the debate regarding the relationship between increased viscosity and atherogenesis has been opened again. While the traditional view postulates that an increased blood viscosity has invariably a negative impact on tissue perfusion and therefore should be considered as a risk factor (when not as a true disease), a more recent hypothesis has been formulated based on the observation that small increases in viscosity actually have vasodilatory effects, potentially improving tissue perfusion.


Patient dissatisfaction and institutional betrayal in the Canadian medical system: A qualitative study.
Individuals who struggle with chronic medical conditions frequently use medical services and may depend on the medical system to ensure their overall well-being. As a result, they may be at a greater risk of feeling betrayed by the medical system when their needs are not being met. The current study aimed to qualitatively assess patients' negative experiences with the medical system that may lead to feelings of institutional betrayal. A total of 14 Canadian adults struggling with various chronic conditions completed an online open-ended questionnaire. Results indicated that institutional betrayal is composed of doctor-level betrayal (inadequate medical care and lack of psychological support) as well as system-level betrayal. The findings are discussed in the context of betrayal trauma theory; specifically, patients' appraisals of their negative health care experiences may play a vital role when one is considering the impact of institutional betrayal on an individual's overall well-being.


Inhibition of lactate dehydrogenase A by microRNA-34a resensitizes colon cancer cells to 5-fluorouracil.
5-Fluorouracil (5-FU) chemotherapy is widely used in the treatment of advanced colon cancer. However, the development of resistance to 5-FU is a significant obstacle to successful treatment. MicroRNA-34a (miR-34a) has been reported to be downregulated in a number of tumor types and has also been shown to act as a tumor suppressor. However, the mechanisms underlying the biological effects of miR-34a in chemoresistance remain unclear. The present study showed that the expression of miR-34a is downregulated in 5-FU-resistant colon cancer cells. In addition, 5-FU-resistant colon cancer cells exhibited upregulation of lactate dehydrogenase A (LDHA) expression and activity compared with parental cells. Furthermore, LDHA was shown to be a direct target of miR-34a. Overexpression of miR-34a reduced the expression of LDHA, probably through binding to the 3' untranslated region, leading to the re-sensitization of 5-FU-resistant cancer cells to 5-FU. Additionally, overexpression of LDHA rendered colon cancer cells resistant to 5-FU, suggesting that the miR-34a-induced sensitization to 5-FU is mediated through the inhibition of LDHA. In conclusion, the current study showed that miR-34a is involved in sensitivity to 5-FU in part through its effects on LDHA expression. This indicates that miR-34a‑mediated inhibition of glucose metabolism may be a therapeutic target in patients with chemoresistant colon cancer.


Can a Two Simple Stitches Method Provide Secure Fixation Strength in Biceps Tenodesis?: Biomechanical Evaluation of Various Suture Techniques.
The purpose of this study was to compare the initial fixation strength between four different suture methods for the long head of the biceps. Forty-eight fresh frozen porcine flexor hallucis longus tendons (mean width at suture site, 8.5 ± 0.9 mm) and phalanx bones were randomly assigned to one of the four arthroscopic biceps tenodesis techniques: simple stitch (SS), mattress suture (MS), lasso-loop (LL), and two simple stitches (2SS). A biceps tenodesis was performed according to the four techniques using all-suture type suture anchors (1.9-mm SUTUREFIX anchor with No. 1 ULTRABRAID sutures). Biomechanical evaluations were performed to test load to failure (N), stiffness (N/mm), stress (N/m2), and mode of failure. As for the SS, MS, LL, and 2SS, the mean load to failure was 50.9 ± 14.61 N, 82.3 ± 24.8 N, 116.2 ± 26.7 N, and 130.8 ± 22.5 N (p < 0.001), respectively; mean stiffness was 6.1 ± 1.3 N/mm, 6.7 ± 2.6 N/mm, 7.8 ± 1.4 N/mm, and 8.1 ± 4.2 N/mm, respectively (p = 0.258); and mean stress was 0.7 ± 0.3 N/m2, 1.4 ± 0.8 N/m2, 2.9 ± 0.7 N/m2, and 2.7 ± 0.8 N/m2, respectively (p < 0.001). All the failures happened by the suture cutting through the tendon along its longitudinal fibers. Neither the SS nor the MS method was enough to securely fix the biceps tendon with a significantly lower mechanical strength; however, the 2SS method showed similar initial fixation strength as the LL technique.


Oral manifestations and treatment of a child with Sjögren's syndrome.
Sjögren's syndrome (SS) is a chronic inflammatory systemic autoimmune disease affecting the exocrine, salivary and lacrimal glands. The condition occurs more often in adults and is rare in childhood. SS should be considered in the differential diagnosis of recurrent parotitis and keratoconjunctivitis sicca. Oral manifestations include early tooth decay and xerostomia. Treatment consists of operative dentistry and saliva management. This paper reports a case of a 10-year-old Brazilian boy with SS, stressing the oral manifestations, treatment conduct, clinical importance and need for an early diagnosis in order to improve the patient's quality of life.


[Autonomic Disorder in Multiple System Atrophy].
Multiple system atrophy (MSA) is a neurodegenerative disease characterized by both autonomic and motor dysfunction (cerebellar ataxia and/or parkinsonism). The former presents with symptoms associated with urological, cardiovascular, gastrointestinal, auditory, and respiratory disorders, among others. Therefore, multidisciplinary collaboration between neurologists and specialists from the aforementioned disciplines is extremely important for the management of these patients. Optimal management of autonomic disorders in patients with MSA can significantly maximize patients' quality of life.


Identification of valid reference genes for quantitative RT-PCR in Caragana microphylla under salt and drought stresses.
Caragana microphylla is a leguminosae plant and grows mainly in semi-arid areas of northwest China and Mongolia. However, the lack of studies on C. microphylla reference genes limits the accurate understanding of the molecular biology mechanisms in this crop under abiotic stresses. In this study, we selected nine candidate genes from salt-treated C. microphylla transcriptome data and evaluated their stability by using geNorm, NormFinder, BestKeeper, and RefFinder in salt and drought conditions. In addition, the relative expressions of Delta 1-pyrroline-5-carboxylate synthase 2 (P5CS2) and Catalase 2 (CAT2) were examined to confirm the stability of the candidate reference genes. As a results, glyceraldehyde-3-phosphate dehydrogenase C2 (GAPC2) and 26S proteasome regulatory subunit (RPN5) were the most stable in both salt and drought treatments. The relative expression of P5CS2 and CAT2 also showed more stable levels in normalization by GAPC2 and RPN5 than the most unstable gene, Ubiquitin 4 (UBQ4). Therefore, it is believed that these candidate reference genes selected and validated in our study could be used to study the molecular biological study of response to salt and drought stress in C. microphylla.


Family- and Person-Centered Interdisciplinary Telehealth: Policy and Practice Implications Following Onset of the COVID-19 Pandemic.
With the onset of the COVID-19 pandemic, telehealth was thrust to the forefront, becoming one of the most predominant forms of care almost overnight. Despite years of research, practice, and policymaking, tenets for providing telehealth in an interdisciplinary, family- and person-centered fashion, and across a wide breadth of settings remain underdeveloped. In addition, although telehealth has the potential to increase equity in care, it can also further exacerbate disparities. The current article discusses the opening created by the pandemic and provides recommendations for how to make permanent changes in telehealth policy and practice to allow for interdisciplinary, person- and family-centered care while also taking care to address issues of equity and ethics and privacy issues related to telehealth and remote monitoring. [Journal of Gerontological Nursing, 46(9), 9-13.].


The Interaction between Osmotic- and Pressure-induced Water Flow in Plant Roots.
This paper presents a general model for coupled solute and water flow through plant roots based on the thermodynamics of irreversible processes. The model explains in a straight-forward manner such experimentally observed phenomena as changes in root resistance, increased solute flux, and apparent negative resistance, which have been reported for root systems under the influence of a hydrostatic pressure gradient. These apparent anomalies are explained on the basis of the interaction between the osmotic and hydrostatic driving forces and the well known "sweeping away" or dilution effect. We show that with a constant hydraulic conductivity the only features necessary to explain these phenomena are some type of membrane or membranelike structure and a mechanism for actively accumulating solutes.


Neuromuscular fatigue thresholds of the vastus lateralis, vastus medialis and rectus femoris muscles.
The purpose of this study was to compare the power outputs associated with the onset of neuromuscular fatigue in the superficial muscles of the quadriceps femoris group by simultaneously monitoring the EMG activity from the vastus lateralis (VL), vastus medialis (VM) and rectus femoris (RF) muscles during a Physical Working Capacity at the Fatigue Threshold (PWCFT) test. Eleven adult males (mean +/- SD = 24 +/- 3y) served as subjects. There were no significant (p > 0.05) differences among the mean PWCFT values for the VL (226 +/- 58 W), VM (223 +/- 58 W) and RF (203 +/- 54 W) muscles. These findings indicates that the superficial muscles of the quadriceps femoris group generally responded as a unit with respect to the onset (threshold) of neuromuscular fatigue as measured by the PWCFT test.


Persistent tardive dyskinesia in bipolar patients.
The prevalence and outcome of persistent tardive dyskinesia (TD) was studied in 131 bipolar patients. There were 34 cases of persistent TD in the subgroup (n = 96) with a history of neuroleptic treatment (prevalence, 35.4%; 95% confidence interval, 25% to 45%); there were no cases of persistent TD in the subgroup (n = 35) without such treatment history. Except in one patient, signs of TD persisted in spite of lithium carbonate treatment in 23 patients (median duration, 16 months; range, five to 24 months), of whom 15 remained off of a neuroleptic regimen during the study period for a median duration of 14 months (range, four to 24 months). Using multiple regression analysis, two variables were found to predict the presence of persistent TD and account for 36% of the variance: longer cumulative duration of maintenance neuroleptic treatment and shorter duration of previous lithium carbonate treatment. There appears to be a significant risk of persistent TD among neuroleptic-treated bipolar patients. High-risk subgroups within this category need to be identified.


Inbreeding depression in a rare plant, Scabiosa canescens (Dipsacaceae).
Plants from a population of Scabiosa canescens, a locally rare species with a narrow ecological amplitude, were raised under uniform growth conditions to examine the phenotypic effects of one generation selfing and outcrossing. Particular attention was given to direct components of fitness (seedling biomass, rosette leaf number, head number, flower number per head), but two morphological characters (plant height, flower size) were also considered. Estimates of inbreeding depression (delta), adjusted for maternal effects and lack of balance, were compared and tested for significance using randomization and boostrap procedures. Inbreeding significantly depressed several characters during both early and late stages of the life cycle, with delta ranging from 0.14 (flower size) to 0.37 (seedling biomass). Based on these and other results, we propose that S. canescens is susceptible to inbreeding and that the genetic basis of inbreeding depression varies across life stages.


Design and synthesis of a short-chain bitistatin analogue for imaging thrombi and emboli.
Previously, we showed that labeled bitistatin analogues possessed excellent characteristics for imaging both deep-vein thrombosis and pulmonary embolism. We hypothesized that the N-terminal amino acid sequence of bitistatin, which is different from other disintegrins, likely interacts with the binding site of platelets to confer desirable properties to bitistatin for imaging. In this study, we present the design, synthesis, and initial biological testing of a short-chain analogue of the native 83-amino-acid bitistatin sequence. Our initial molecular modeling of the binding loop of bitistatin showed that the minimal sequence that represented the binding region was a cyclic 10 amino acid sequence cyclo[Cys-Arg-Ile-Ala-Arg-Gly-Asp-Trp-Asn-Cys(S)]. Systematic modeling of a truncated N-terminal sequence of bitistatin fused with the optimized binding region having a thioether sequence through a Gaba spacer ultimately yielded the 24-amino acid peptide, cyclo-[CH(2)CO-Arg-Ile-Ala-Arg-Gly-Asp-Trp-Asn-Cys(S-)]-Gaba-Gly-Asn-Glu-Ile-Leu-Glu-Gln-Gly-Glu-Asp-Ser-Asp-Ser-Lys-OH, 1. The peptide was then coupled to the hydrazino-nicotinic acid bifunctional chelating agent and the purified adduct labeled with (99m)Tc using tricine as a coligand. Binding of the unlabeled and labeled peptide to stimulated human platelets was assayed in vitro. The (99m)Tc labeling yield was > 90%. The in vitro binding assays showed that the IC(50) for inhibition of platelet aggregation was 3694 nM, while the Kd of the (99m)Tc labeled peptide was 185 nM, indicating moderate affinity for the receptor. The (99m)Tc-labeled peptide was able to identify sites of experimental thrombi and emboli in a canine model. The results suggest initial success in attempting to mimic the behavior of bitistatin for imaging thrombi and emboli.


Longitudinal assessment of mental development in infants with nonsyndromic craniosynostosis with and without cranial release and reconstruction.
The effect of cranial release and reconstruction on the mental development of infants with nonsyndromic craniosynostosis was evaluated. Longitudinal assessment of mental development for infants before and after cranial release and reconstruction and for infants not undergoing surgical treatment was obtained by using the mental scale of the Bayley Scales of Infant Development. Severity of anatomic craniofacial deformity, perinatal medical risk factors, and age at time of surgery also were investigated. None of the infants displayed mental retardation [Mental Development Index (MDI) score < 70] before or after cranial release and reconstruction. Scores ranged from borderline retardation to very superior following a normal distribution. Severity of anatomic craniofacial deformity and perinatal risk factors were unrelated to mental development. Cranial release and reconstruction did not affect mental development positively or negatively but did result in improvement of the original craniofacial deformity.


Simultaneous stable carbon isotopic analysis of wine glycerol and ethanol by liquid chromatography coupled to isotope ratio mass spectrometry.
A novel procedure was established for the simultaneous characterization of wine glycerol and ethanol (13)C/(12)C isotope ratio, using liquid chromatography/isotope ratio mass spectrometry (LC-IRMS). Several parameters influencing separation of glycerol and ethanol from wine matrix were optimized. Results obtained for 35 Spanish samples exposed no significant differences and very strong correlations (r = 0.99) between the glycerol (13)C/(12)C ratios obtained by an alternative method (gas chromatography/isotope ratio mass spectrometry) and the proposed new methodology, and between the ethanol (13)C/(12)C ratios obtained by the official method (elemental analyzer/isotope ratio mass spectrometry) and the proposed new methodology. The accuracy of the proposed method varied from 0.01 to 0.19 per thousand, and the analytical precision was better than 0.25 per thousand. The new developed LC-IRMS method it is the first isotopic method that allows (13)C/(12)C determination of both analytes in the same run directly from a liquid sample with no previous glycerol or ethanol isolation, overcoming technical difficulties associated with complex sample treatment and improving in terms of simplicity and speed.


External or internal fixation in the treatment of non-reducible distal radial fractures?
We have previously shown in a randomized study that in the first year after treatment, open reduction and internal fixation resulted in better grip strength and forearm rotation than closed reduction and bridging external fixation. In the present study, we investigated whether this difference persists over time. The 50 patients included in the original study (mean age 53 years, 36 women) were sent a QuickDASH questionnaire and an invitation to a radiographic and clinical examination after a mean of 5 (3-7) years. All 50 patients returned the QuickDASH questionnaire and 45 participated in the clinical and radiographic examination. In the internal fixation group, the grip strength was 95% (SD 12) of the uninjured side and in the external fixation group it was 90% (SD 21) of the uninjured side (p = 0.3). QuickDASH score, range of motion, and radiographic parameters were similar between the groups. The difference originally found between internal and external fixation in distal radial fractures at 1 year regarding grip strength and range of motion was found to diminish with time. At 5 years, both groups had approached normal values.


Coronal root canal morphology of permanent two-rooted mandibular first molars with novel 3D measurements.
To measure the coronal root canal morphology of permanent mandibular first molars using 3D coordinates for more precise conservative endodontic cavity preparation. In total, 57 cone beam computed tomography (CBCT) images of sound mandibular molars with fully formed apices without previous endodontic treatment were taken from 33 patients aged 16 to 75 years. The CBCT machine (MCT-1[EX-2F], J. Morita Manufacturing Corp, Kyoto, Japan) provided 14-bit greyscale images with the voxel size of 0.125 mm. All images were taken at 80 kV and 5.0 mA, with a 17-s exposure time, which were then reconstructed in 3D models and viewed by an endodontist. The gender of the patients and the tooth position, and number of root canals, were recorded. The landmarks of coronal root canals were determined, then the distribution of landmarks, maximum curvature in the axial direction and curvature directions in the horizontal direction of coronal root canals were measured. Distributions of landmarks were analysed using a spatial statistics method. Data about curvature were compared using a t test. Overall, the distribution of root canal orifices and the centre of the canal primary curve were more centralized than other landmarks. The landmarks were located more mesiobuccally to the centre of the occlusal plane of mandibular first molars. Specifically, the measurements of the maximum curvature of coronal root canals in the axial direction were as follows: in 3-canal teeth, the average angles of curvatures were 22°,23°,15° for mesiobuccal (MB), mesiolingual (ML) and distobuccal (DB) canals, respectively; in 4-canal teeth, the average angles of curvatures were 24°,26°,15°,22°for MB, ML, DB and distolingual (DL) canals, respectively. The degrees of coronal root canal curvatures in the horizontal direction were as follows: in 3-canal teeth, the average angles of curvatures were -8°,47°,-2° for MB, ML and DB canals, respectively; in 4-canals teeth, the average angles were -11°,50°,-28°,45° for MB, ML, DB and DL canals, respectively. Statistically, there was no reliable side or sex difference in any of the measurements (P > 0.05). In permanent two-rooted mandibular first molars, the coronal canals clustered in the direction of the mesiobuccal side of the occlusal surface of the teeth.


GuidAge study: a 5-year double blind, randomised trial of EGb 761 for the prevention of Alzheimer's disease in elderly subjects with memory complaints. i. rationale, design and baseline data.
Primary and secondary prevention strategies for Alzheimer's disease (AD) are urgently needed. We have initiated a five-year prospective prevention study involving patients spontaneously reporting memory complaints. The primary objective is to determine the effect of treatment with EGb 76 on the rate of conversion from memory complaints to AD using survival analysis. Ambulatory patients aged at least 70 years who spontaneously reported a memory complaint during a GP or memory centre consultation were eligible for inclusion. Patients with major objective memory impairment or clinically relevant symptoms of anxiety and depression were excluded. Subjects were randomised to receive either EGb 761 120mg bid or matching placebo. Participants undergo an annual visit at a memory centre, where a series of neuropsychological tests are administered to assess cognitive function (Grober and Buschke, Trail-Making and controlled oral word association tests) and cognitive status (MMS and CDR). Functional status is evaluated with the Instrumental Activities of Daily Living questionnaire. The primary outcome is the transition to a diagnosis of AD (DSM-IV and NINCDS-ADRDA criteria), determined at the annual memory centre visit. A total of 4066 patients were screened for participation, of whom 2854 fulfilled the eligibility criteria and were entered into the study. Their mean age was 76.8+/-4.4 years and 66.7% were female. The mean MMSE score was 27.8+/-1.7 and 55.5% presented a CDR score of 0.5. This study will enable us to evaluate the efficacy of EGb761 in the prevention of AD, and to assess the usefulness of various baseline characteristics as predictors of conversion to AD in this population.


Fast and Accurate Detection of COVID-19 Along With 14 Other Chest Pathologies Using a Multi-Level Classification: Algorithm Development and Validation Study.
COVID-19 has spread very rapidly, and it is important to build a system that can detect it in order to help an overwhelmed health care system. Many research studies on chest diseases rely on the strengths of deep learning techniques. Although some of these studies used state-of-the-art techniques and were able to deliver promising results, these techniques are not very useful if they can detect only one type of disease without detecting the others. The main objective of this study was to achieve a fast and more accurate diagnosis of COVID-19. This study proposes a diagnostic technique that classifies COVID-19 x-ray images from normal x-ray images and those specific to 14 other chest diseases. In this paper, we propose a novel, multilevel pipeline, based on deep learning models, to detect COVID-19 along with other chest diseases based on x-ray images. This pipeline reduces the burden of a single network to classify a large number of classes. The deep learning models used in this study were pretrained on the ImageNet dataset, and transfer learning was used for fast training. The lungs and heart were segmented from the whole x-ray images and passed onto the first classifier that checks whether the x-ray is normal, COVID-19 affected, or characteristic of another chest disease. If it is neither a COVID-19 x-ray image nor a normal one, then the second classifier comes into action and classifies the image as one of the other 14 diseases. We show how our model uses state-of-the-art deep neural networks to achieve classification accuracy for COVID-19 along with 14 other chest diseases and normal cases based on x-ray images, which is competitive with currently used state-of-the-art models. Due to the lack of data in some classes such as COVID-19, we applied 10-fold cross-validation through the ResNet50 model. Our classification technique thus achieved an average training accuracy of 96.04% and test accuracy of 92.52% for the first level of classification (ie, 3 classes). For the second level of classification (ie, 14 classes), our technique achieved a maximum training accuracy of 88.52% and test accuracy of 66.634% by using ResNet50. We also found that when all the 16 classes were classified at once, the overall accuracy for COVID-19 detection decreased, which in the case of ResNet50 was 88.92% for training data and 71.905% for test data. Our proposed pipeline can detect COVID-19 with a higher accuracy along with detecting 14 other chest diseases based on x-ray images. This is achieved by dividing the classification task into multiple steps rather than classifying them collectively.


Inducibility of the 70 kD heat shock protein in peripheral blood monocytes is decreased in human acute respiratory distress syndrome and recovers over time.
The heat shock/stress proteins (HSP), and, in particular, the inducible, cytosolic Hsp70, represent an extremely conserved response to many different cellular injuries, including reactive oxygen species (ROS). Hsp70 has been shown to confer to cells and tissues protection against the deleterious effects of ROS or cytokines, both in vitro and in animal models of acute respiratory distress syndrome (ARDS). We hypothesized that Hsp70 expression levels in peripheral blood monocytes (PBM) of patients with ARDS, would correlate with disease severity. We prospectively included 13 patients with previous ARDS (50 +/- 17 yr; range, 20 to 76 yr), nine ventilated patients with non-ARDS/ALI disease (45 +/- 20 yr; range, 19 to 76 yr), and 14 healthy volunteers (45 +/- 20 yr; range, 22 to 77 yr). PBM activation state was evaluated according to their membrane expression of CD16, and oxidative status according to plasma lipid peroxidation products. Both baseline expression and Hsp70 inducibility (after in vitro heat shock) were examined in PBM, using flow cytometric analysis. We found that basal expression of Hsp70 in PBM was similar for patients and control subjects, whereas Hsp70 inducibility- a reflection of the ability to mount a stress response-was significantly reduced in the patients with ARDS (p = 0. 02). Among all correlation analyses we considered between Hsp70 inducibility on the one hand, clinical and laboratory biomarkers for disease severity and outcome in the patients with ARDS on the other, only the duration of ventilatory support was significant (p < 0.003). As an approach to distinguish between disease and ventilation, we also analyzed a group of, ventilated patients without ARDS. Our results indicate that in patients with ARDS, Hsp70 inducibility in PBM is decreased, but it recovers over time with duration of ventilatory support.


Tricuspid regurgitation due to blunt chest trauma. Report of a case and review of the literature.
A 25-year-old female developed high-grade atrioventricular block and markedly elevated central venous pressure after sustaining a crushing injury to the chest while driving a car. An echocardiographic examination with color Doppler revealed severe tricuspid regurgitation due to a torn papillary muscle. An extensive review of the literature showed the following: 1) correct diagnosis is often delayed because of coexisting multisystem involvement and the subtleness of abnormal physical signs, 2) identification of abnormally elevated right atrial pressure with a prominent "v" wave, and characteristic electrocardiogram appeared to be the key to early diagnosis, and 3) the final diagnosis may be confirmed by echocardiography with Doppler and/or cardiac catheterization. The role of echocardiographic examination with color Doppler technique deserves special emphasis because the final diagnosis can be easily reached during the acute phase at the bedside noninvasively.


Transcatheter Valve-in-Valve Implantation for Degenerated Mitral or Tricuspid Bioprosthetic Valves: A Heath Technology Assessment.
Bioprosthetic valves used to treat mitral or tricuspid valve disease can be expected to deteriorate and eventually fail after 10 to 15 years. For patients who are considered inoperable or high-risk for surgery, medical management (i.e., drug therapy, the current standard of care in Ontario) does not significantly alter the course of valvular heart disease or improve degenerated bioprosthetic valves. An alternative for these patients is transcatheter mitral or tricuspid valve-in-valve implantation (TMViV or TTViV). We conducted a health technology assessment of transcatheter valve-in-valve implantation for adults with degenerated mitral or tricuspid bioprosthetic valves who are considered inoperable or high-risk for surgery, which included an evaluation of effectiveness, safety, the budget impact of publicly funding TMViV or TTViV, and patient preferences and values. We leveraged a previously published systematic review, supplementing the work with two new registry studies identified during the development of this report. We assessed the risk of bias of each included study using the Downs and Black checklist and the quality of the body of evidence according to the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) Working Group criteria. As the earlier systematic review did not identify any cost-effectiveness studies, we conducted a hand search of the grey literature using targeted websites to identify relevant cost-effectiveness studies. We analyzed the budget impact of publicly funding transcatheter valve-in-valve implantation for adults with degenerated mitral or tricuspid bioprostheses who are considered inoperable or high-risk for surgery in Ontario. To contextualize the potential value of TMViV and TTViV, we spoke with people who had experience with heart valve replacement or who were awaiting heart valve replacement. We included 19 studies in the clinical evidence review. No studies compared TMViV or TTViV to medical management (standard care). TMViV was associated with in-hospital, 30-day and 1-year mortality rates of 0% to 5%, 0% to 15%, and 14% to 27%, respectively (GRADE: Very low). TTViV was associated with 30-day and 1-year mortality rates of 0% to 3% and 0% to 14%, respectively (GRADE: Very low). Patients experienced functional improvement related to their heart failure symptoms after TMViV or TTViV. Compared to before the intervention, both TMViV and TTViV were associated with a decrease in the number of patients with New York Heart Association class III or IV symptoms in hospital and at 30-day follow-up (GRADE: Low). We identified no relevant cost-effectiveness studies from our targeted search. The annual budget impact of publicly funding TMViV and TTViV in Ontario over the next 5 years ranges from an additional $0.35 million in year 1 to a cost saving of $0.19 million in year 5, for a total cost saving of $0.33 million. The people we spoke to who had bioprosthetic heart valve failure reported the negative effects of valvular heart disease and described their positive perceptions of transcatheter valve-in-valve implantation. They valued the minimally invasive nature of transcatheter procedures and the quick recovery time. TMViV or TTViV may reduce mortality, but the evidence is very uncertain. TMViV or TTViV may improve heart failure symptoms. We estimated that publicly funding TMViV and TTViV in Ontario would result in a cost saving of $0.33 million over the next 5 years. People with valvular heart disease reported their preference for a minimally invasive transcatheter procedure with a quick recovery time.


[Does laparoscopic colonic carcinoma surgery satisfy the radicality criteria of open surgery?].
Minimal invasive surgery is applicable to almost all colorectal operations, with major benefit for the patient. Technically even cancer operations can be performed. However, in laparoscopic assisted colorectal surgery the question of radicality outweighs that of performability and patient comfort. From a prospective series of 88 laparoscopic colorectal operations, 36 were for carcinoma. 34 patients who underwent conventional surgery were matched with regard to age, sex, type of operation and tumor stage (TNM, grading) to compare the two techniques. The two interventions followed exactly the same guidelines. We compared the length of the fixed specimens, the number of resected lymph nodes and the need for blood transfusions. Postoperative complications were noted and follow-up was 3-12 months. The data obtained showed no difference between the two treatment groups, with a slight trend towards laparoscopic surgery with regard to number of resected lymph nodes. A portside metastasis was observed 9 months after an initial tumor stage T4N1M1. The close relation between the portside and infiltration of the adenocarcinoma into the abdominal wall together with a drain placed through this whole was suspected to be the cause of this complication. We conclude that laparoscopic assisted colorectal surgery for cancer is equal to open operation regarding radicality of resection. Long term results are mandatory to determine the value of minimal invasive surgery in the field of oncology.


Sleep in older adults with Alzheimer's disease.
Sleep in older adults with Alzheimer's disease can often be extremely fragmented, thus disturbing normal sleep-wake rhythms. This poor-quality sleep is challenging for caregivers and frequently results in admitting older adults to long-term care institutions. Many variables, such as genes, medications, depression, and environmental factors, influence sleep in persons with Alzheimer's disease. Interventions, such as exposure to light, melatonin therapy, sleep hygiene, and physical activity, are commonly used to treat or to prevent sleep problems. Neuroscience nurses can play an important role in assessing sleep characteristics and related factors, educating caregivers and staff, providing family members with emotional support, and conducting research.


Use of amplitude and frequency transformations to generate adiabatic pulses of wide bandwidth and low RF power deposition.
An amplitude transformation of adiabatic pulses stated previously [Baum et al., Phys. Rev. A 32, 3435-3447 (1985)] is generalized using the Bloch equations. Both amplitude and frequency transformations are used to create new adiabatic pulses of wide bandwidth and low RF power deposition. Several adiabatic pulses (including the tanh/tan pulse used for construction of BIR-4, SSAP, and BISEP pulses) are transformed into new pulses. These new pulses are demonstrated numerically and experimentally to operate at a significantly lower RF power level while maintaining the same performance or over a wider bandwidth while using the same RF power. They are expected to be useful for in vivo NMR experiments, especially for applications involving wide frequency dispersion and pulse sequences composed of many adiabatic half-passage pulses.


Intravesical capsaicin in neurologic impaired patients with detrusor hyperreflexia.
Capsaicin is known to be neurotoxic for C-fiber afferents. We investigated the intravesical application of capsaicin in the treatment of detrusor hyperreflexia (DH) in seven patients (ages 23-52) with neurologic impairment. The patients were evaluated with both ice-water cystometry and formal video-urodynamic studies. Four biweekly courses of intravesical capsaicin treatment were administered using increasing concentrations (100 microM, 500 microM, 1 mM, and 2 mM). Treatment effect was monitored using a bladder diary and urodynamic evaluation one month after capsaicin treatment. Prior to treatment, six of the seven patients demonstrated a positive ice-water test and DH. Two patients were not able to complete the study due to discomfort attributed to capsaicin. Five of the seven patients completed the four courses of increasingly concentrated capsaicin. Three patients noted symptomatic improvement while two did not. The mean urodynamic bladder capacity significantly increased from 124 +/- 39 ml pre-capsaicin to 231 +/- 62 ml one month post-capsaicin in the three patients with symptomatic improvement (p < 0.05). Urodynamic testing revealed that one of the six patients with a positive ice-water test lost that response after intravesical capsaicin. Intravesical capsaicin is a novel and promising treatment for detrusor hyperreflexia in neurologically impaired patients.


Targeted neonatal echocardiography training: a survey of trainees in a region of England.
Targeted neonatal echocardiography (TNE) is an important skill to advise diagnosis and management. Training in TNE is currently optional for neonatal subspecialty (grid) trainees and accessing training is often challenging. We disseminated a survey, asking neonatal grid trainees for their views on TNE training. 48 out of 91 trainees (53%) completed the survey. 96% of trainees (n=48) wanted to learn TNE with similar numbers eager to access a formal training package, using a variety of teaching media. Identified barriers to TNE training included time, access to supervision and the perceived complexity of the skill. These findings will influence the design and delivery of a regional TNE training programme.


Development of negative ion extractor in the high-power and long-pulse negative ion source for fusion application.
High power and long-pulse negative ion extractor, which is composed of the plasma grid (PG) and the extraction grid (EXG), is newly developed toward the neutral beam injector for heating and current drive of future fusion machines such as ITER, JT-60 Super Advanced and DEMO reactor. The PG is designed to enhance surface production of negative ions efficiently by applying the chamfered aperture. The efficiency of the negative ion production for the discharge power increased by a factor of 1.3 against that of the conventional PG. The EXG is also designed with the thermal analysis to upgrade the cooling capability for the long pulse operation of >1000 s required in ITER. Though the magnetic field for electron suppression is reduced to 0.75 of that in the conventional EXG due to this upgrade, it was experimentally confirmed that the extracted electron current can be suppressed to the allowable level for the long pulse operation. These results show that newly developed extractor has the high potential for the long pulse extraction of the negative ions.


Spray Drying for the Encapsulation of Oils-A Review.
The application of the spray drying technique in the food industry for the production of a broad range of ingredients has become highly desirable compared to other drying techniques. Recently, the spray drying technique has been applied extensively for the production of functional foods, pharmaceuticals and nutraceuticals. Encapsulation using spray drying is highly preferred due to economic advantages compared to other encapsulation methods. Encapsulation of oils using the spray drying technique is carried out in order to enhance the handling properties of the products and to improve oxidation stability by protecting the bioactive compounds. Encapsulation of oils involves several parameters-including inlet and outlet temperatures, total solids, and the type of wall materials-that significantly affect the quality of final product. Therefore, this review highlights the application and optimization of the spray drying process for the encapsulation of oils used as food ingredients.


Photosensitivity of gallium-doped silica core fiber to 193 nm ArF excimer laser.
Grating inscription in a Ga-doped silica core fiber (~5 wt. % Ga) has been demonstrated using ArF (193 nm) and KrF (248 nm) excimer lasers. In a comparative study with germanosilicate fiber with similar Ge concentration, a Ga-doped silica core fiber shows greater photosensitivity to an ArF excimer laser due to the higher absorbance in the region of 190-195 nm. In addition, the photosensitivity of a Ga-doped silica core fiber has been greatly enhanced with hydrogenation. Ga-doped fibers are potential photosensitive fibers for fiber Bragg grating production with an ArF excimer laser.


Antioxidant effects of taurine, vitamin C, and vitamin E on oxidative damage in hippocampus caused by the administration of 3-nitropropionic acid in rats.
The administration of 3-nitropropionic acid increases reactive oxygen species (ROS). Antioxidant defense mechanisms buffer these ROS converting them into non-damaging compounds. Taurine and vitamins C and E are antioxidants that play a role in the defense against cellular damage. This study examines the antioxidant effect of taurine, vitamin C, and vitamin E on acute hippocampal damage caused by 3-NP. Animals treated with 3-NP increased lipid peroxidation levels and astrocytic damage in the hippocampus. Administration of taurine, vitamin C, and vitamin E partially protected from oxidative damage, indicate that while all substances had antioxidant effects, only taurine showed morphological protection in surviving cells.


Basin management inspiration from impacts of alternating dry and wet conditions on water production and carbon uptake in Murray-Darling Basin.
The impacts of alternating dry and wet conditions on water production and carbon uptake at different scales remain unclear, which limits the integrated management of water and carbon. We quantified the response of runoff efficiency (RE) and plant water-use efficiency (PWUE) to a typical shift from dry to wet episode of 2003-2014 in Australia's Murray-Darling basin using good and specific data products for local application, including Australian Water Availability Project, Penman-Monteith-Leuning Evapotranspiration V2 product, MODIS MCD12Q1 V6 Land Cover Type and MODIS MOD17A3 V055 GPP product. The results show that there are significant power function relationships between RE and precipitation for basin and all ecosystems, while the PWUE had a negative quadratic correlation with precipitation and satisfied the significance levels of 0.05 for basin and the ecosystems except the grassland and cropland. The shrubs can achieve the best water production and carbon uptake under dry conditions, while the evergreen broadleaf trees and evergreen needleleaf trees can obtain the best water production and carbon uptake in wet conditions, respectively. These findings help integrated basin management for balancing water resource production and climate change mitigation.


Alveolar-capillary membrane permeability for early prediction of response of inhaled steroid on patients with chronic obstructive pulmonary disease.
Glucocorticosteroid reversibility-testing is undertaken over a period of 3 months to identify whether patients with chronic obstructive pulmonary disease (COPD) would benefit from long-term inhaled corticosteroids. This study assessed whether alveolar-capillary membrane (a/c) permeability testing can be used as an early alternative test method for the same purpose. Fourteen patients with severe and symptomatic moderate COPD (group S) were prescribed inhaled steroid 800 microg/day for 3 months. Before inhalation and 4 weeks after inhalation therapy, forced expiratory volume in 1 s (FEV(1)) and a/c permeability using (99m)Tc-DTPA were performed. FEV(1) was recorded again at the end of the third month. Another 10 patients with COPD of comparable severity (group B) prescribed with inhaled bronchodilators were examined and studied as controls. In group S, the permeability decreased in eight patients (group D) and increased in six patients (group I). No significant change was noted in FEV(1) at the end of the first month. However, seven patients in group D showed significant improvement in FEV(1) at the end of the third month, whereas in patients in group I no significant changes were observed. In group B, no significant change in a/c permeability was observed, although the FEV(1) increased by 12-17%. With steroid inhalation, the a/c permeability at 4 weeks predicts future changes in lung functions. Long-term inhaled corticosteroids are likely to be useful if permeability decreases. This test, which needs further validation, appears to provide much earlier prediction of response than glucocorticoid reversibility testing.


Seroepidemiology of Toxoplasma gondii infection in Tepehuanos in Durango, Mexico.
The epidemiology of Toxoplasma gondii infection is known to depend on a number of behavioral factors, including contact with animals and eating habits. The epidemiology in specific populations (i.e., in indigenous ethnic groups) is not well studied. We therefore investigated the presence of anti-Toxoplasma IgG and IgM antibodies in 156 Tepehuanos, an indigenous ethnic group in Durango State in northern Mexico, using ELISAs. Of the 156 Tepehuanos (mean age 31.03±16.71 years), 35 (22.4%) had anti-Toxoplasma IgG antibodies. IgG levels of 14-99, 100-150, and >150 IU/mL were found in 15 (42.9%), 1 (2.8%), and 19 (54.3%) anti-Toxoplasma IgG-positive Tepehuanos, respectively. Also, 15 (9.6%) subjects had anti-Toxoplasma IgM antibodies. Male Tepehuanos aged 31-50 years showed a significantly higher frequency of exposure to T. gondii than female Tepehuanos of the same age. Seroprevalence of anti-T. gondii IgM antibodies was significantly higher in Tepehuanos without education than those with 7-12 years of education. Tepehuanos suffering from frequent headaches had significantly higher seroprevalences of anti-T. gondii IgG and IgM antibodies than those without this clinical feature. In addition, Tepehuanos suffering from hearing impairment had significantly higher seroprevalence of anti-T. gondii IgM antibodies than those without this clinical feature. Logistic regression analysis did not show an association of T. gondii exposure with any behavioral characteristics, including contact with cats or eating habits. The seroprevalence of T. gondii exposure in Tepehuanos is comparable to that reported in other populations in North America; however, seroprevalence in the largely rural Tepehuanos is higher than that reported in a nearby urban center. Toxoplasma may be impacting the health of Tepehuanos. This is the first report of T. gondii infection in Tepehuanos, and the results should prove useful for the design of preventive measures.


Survival benefit with capecitabine/docetaxel versus docetaxel alone: analysis of therapy in a randomized phase III trial.
In a large phase III trial of 511 patients with anthracycline-pretreated advanced/metastatic breast cancer, capecitabine/docetaxel combination therapy was shown to have significantly superior efficacy compared with single-agent docetaxel, including superior progression-free and overall survival and objective response rate. An updated survival analysis with >/= 27 months follow-up shows that patients receiving combination therapy maintained significantly superior survival (hazard ratio [HR], 0.777 [95% CI, 0.645-0.942]; P < 0.01; median survival, 14.5 months vs. 11.5 months) compared with those receiving single-agent docetaxel. Following the failure of docetaxel monotherapy, 35% of patients did not receive additional cytotoxic chemotherapy. Among patients randomized to single-agent docetaxel, only those given poststudy single-agent capecitabine had significantly prolonged survival compared with those given any other poststudy chemotherapy (HR, 0.500; P = 0.0046; median survival, 21.0 months vs. 12.3 months, respectively). By contrast, poststudy vinorelbine-containing chemotherapy did not affect survival following progression on single-agent docetaxel compared with other poststudy chemotherapy regimens (HR, 1.014; P = 0.94; median survival, 13.5 months vs. 12.6 months, respectively). Among patients randomized to combination therapy, discontinuing docetaxel of capecitabine has a similar effect on survival (HR, 0.720; P = 0.20; median survival, 15.8 months vs. 18.3 months, respectively). Median survival was 18.3 months in patients who discontinued docetaxel and continued to receive capecitabine versus 15.8 months in patients who discontinued capecitabine and continued to receive docetaxel, with a trend toward improved survival in patients continuing to receive capecitabine. Although this is a retrospective analysis, these data suggest that the sequential administration of docetaxel followed by capecitabine is associated with prolonged survival in patients who are candidates for sequential single-agent therapy.


The effects of litter carrying on rifle shooting.
This study investigated whether the use of a shoulder harness would affect shooting accuracy after patient litter carrying. Two- and four-person teams, 12 male and 9 female soldiers, fired at targets before and after (1) a 15-minute bout of rapid, short litter carries and lifts, and (2) a moderate speed 30-minute litter carry with and without a harness for both types of carries. Shooting accuracy was 10% poorer (p < 0.05) after the 15-minute bout (mean +/- SD = 8.9 +/- 1.9 mm) than after the 30-minute carry (8.1 +/- 1.7 mm). Four-person teams using litter-carriage harnesses had 17% tighter shot groups (45.5 +/- 30.4 mm2) (p < 0.05) than four-person teams that did not use harnesses (54.5 +/- 26.1 mm2) and two-person teams with (56.3 +/- 29.1 mm2) or without harnesses (54.9 +/- 30.7 mm2). The harness can potentially improve shooting accuracy after litter carrying.


A 29-gene signature associated with NOX2 discriminates acute myeloid leukemia prognosis and survival.
The molecular complexity displayed in acute myeloid leukemia (AML) hinders patient stratification and treatment decisions. Previous studies support the utility of using specific gene panels for this purpose. Focusing on two salient features of AML, the production of reactive oxygen species (ROS) by NADPH oxidases (NOX) and metabolism, we aimed to identify a gene panel that could improve patient stratification. A pairwise comparison of AML versus healthy gene expression revealed the downregulation of four members of the NOX2 complex including CYBB (coding for NOX2) in AML patients. We analyzed the expression of 941 genes related to metabolism and found 28 genes with expression correlated to CYBB. This panel of 29 genes (29G) effectively divides AML samples according to their prognostic group. The robustness of 29G was confirmed by 6 AML cohort datasets with a total of 1821 patients (overall accuracies of 85%, 78%, 80%, 75%, 59% and 83%). An expression index (EI) was developed according to the expression of the selected discriminatory genes. Overall Survival (OS) was higher for low 29G expression index patients than for the high 29G expression index group, which was confirmed in three different datasets with a total of 1069 patients. Moreover, 29G can dissect intermediate-prognosis patients in four clusters with different OS, which could improve the current AML stratification scheme. In summary, we have found a gene signature (29G) that can be used for AML classification and for OS prediction. Our results confirm NOX and metabolism as suitable therapeutic targets in AML.


Gene Expression Analysis of Neurons and Astrocytes Isolated by Laser Capture Microdissection from Frozen Human Brain Tissues.
Different cell types and multiple cellular connections characterize the human brain. Gene expression analysis using a specific population of cells is more accurate than conducting analysis of the whole tissue homogenate, particularly in the context of neurodegenerative diseases, where a specific subset of cells is affected by the different pathology. Due to the difficulty of obtaining homogenous cell populations, gene expression in specific cell-types (neurons, astrocytes, etc.) has been understudied. To leverage the use of archive resources of frozen human brains in studies of neurodegenerative diseases, we developed and calibrated a method to quantify cell-type specific-neuronal, astrocytes-expression profiles of genes implicated in neurodegenerative diseases, including Parkinson's and Alzheimer's diseases. Archive human frozen brain tissues were used to prepare slides for rapid immunostaining using cell-specific antibodies. The immunoreactive-cells were isolated by Laser Capture Microdissection (LCM). The enrichment for a particular cell-type of interest was validated in post-analysis stage by the expression of cell-specific markers. We optimized the technique to preserve the RNA integrity, so that the RNA was suitable for downstream expression analyses. Following RNA extraction, the expression levels were determined digitally using nCounter Single Cell Gene Expression assay (NanoString Technologies®). The results demonstrated that using our optimized technique we successfully isolated single neurons and astrocytes from human frozen brain tissues and obtained RNA of a good quality that was suitable for mRNA expression analysis. We present here new advancements compared to previous reported methods, which improve the method's feasibility and its applicability for a variety of downstream molecular analyses. Our new developed method can be implemented in genetic and functional genomic research of neurodegenerative diseases and has the potential to significantly advance the field.


Capsule endoscopy of the small bowel.
Capsule endoscopy (CE) is a first line small bowel investigative modality which provides more sensitive mucosal imaging than comparators. It is a non-invasive, non-irradiating tool well tolerated by patients. The risk of retention of the capsule can be minimised by ensuring luminal patency using the Agile patency device. Research continues into how to minimise missed pathology and variability in the identification of pathology or interpretation of images. The consensus is that bowel preparation using laxatives improves visibility and diagnostic yield. Research includes the development of image recognition software, both to eliminate sequentially identical images to improve viewing speed and to select or enhance images likely to represent pathology. However, careful reading by experienced capsule endoscopists remains the benchmark. This should be performed at a speed comfortable to the viewer, probably at a maximum of 15 frames per second. Some prior experience of endoscopy appears to be helpful for novice capsule endoscopists and formal training on a hands-on training course seems to improve pathology recognition, for novices and for those with CE experience.


Microangiopathic haemolytic anaemia and systemic vasculitis.
Two cases of systemic vasculitis complicated by microangiopathic haemolytic anaemia (MAHA) are described: this association has not previously been reported. Both patients had atypical presentations of their primary disease, one with parotitis and one with a Guillain-Barré syndrome. Other causes of MAHA were excluded and a possible link with macromolecular von Willebrands factor is speculated upon.


Complexation of phenol and thiophenol by amine N-oxides: isothermal titration calorimetry and ab initio calculations.
To develop a new solvent-impregnated resin (SIR) system for removal of phenols from water, the complex formation of dimethyldodecylamine N-oxide (DMDAO), trioctylamine N-oxide (TOAO), and tris(2-ethylhexyl)amine N-oxide (TEHAO) with phenol (PhOH) and thiophenol (PhSH) is studied. To this end we use isothermal titration calorimetry (ITC) and quantum chemical modeling (on B3LYP/6-311G(d,p)-optimized geometries: B3LYP/6-311+G(d,p), B3LYP/6-311++G(2d,2p), MP2/6-311+G(d,p), and spin component scaled (SCS) MP2/6-311+G(d,p); M06-2X/6-311+G(d,p)//M06-2X/6-311G(d,p), MP2 with an extrapolation to the complete basis set limit (MP2/CBS), as well as CBS-Q). The complexes are analyzed in terms of structural (e.g., bond lengths) and electronic elements (e.g., charges). Furthermore, complexation and solvent effects (in benzene, toluene, and mesitylene) are investigated by ITC measurements, yielding binding constants K, enthalpies ΔH(0), Gibbs fre energies ΔG(0), and entropies ΔS(0) of complex formation, and stoichiometry N. The ITC measurements revealed strong 1:1 complex formation between both DMDAO-PhOH and TOAO-PhOH. The binding constant (K=1.7-5.7×10(4) M(-1)) drops markedly when water-saturated toluene was used (K=5.8×10(3) M(-1)), and π-π interaction with the solvent is shown to be relevant. Quantum mechanical modeling confirms formation of stable 1:1 complexes with linear hydrogen bonds that weaken on attachment of electron-withdrawing groups to the amine N-oxide moiety. Modeling also showed that complexes with PhSH are much weaker than those with PhOH, and in fact too weak for ITC determination. CBS-Q incorrectly predicts equal or even higher binding enthalpies for PhSH than for PhOH, which invalidates it as a benchmark for other calculations. Data from the straightforward SCS-MP2 method without counterpoise correction show very good agreement with the MP2/CBS values.


TSC2 c.1864C>T variant associated with mild cases of tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is an autosomal dominantly inherited disorder with variable expressivity associated with hamartomatous tumors, abnormalities of the skin, and neurologic problems including seizures, intellectual disability, and autism. TSC is caused by pathogenic variants in either TSC1 or TSC2. In general, TSC2 pathogenic variants are associated with a more severe phenotype than TSC1 pathogenic variants. Here, we report a pathogenic TSC2 variant, c.1864C>T, p.(Arg622Trp), associated with a mild phenotype, with most carriers meeting fewer than two major clinical diagnostic criteria for TSC. This finding has significant implications for counseling patients regarding prognosis. More patient data are required before changing the surveillance recommendations for patients with the reported variant. However, consideration should be given to tailoring surveillance recommendations for all pathogenic TSC1 and TSC2 variants with documented milder clinical sequelae. © 2017 Wiley Periodicals, Inc.


Phosphatidylcholine synthesis: differing patterns in soybean and carrot.
The methylation steps in the biosynthesis of phosphatidylcholine by tissue culture preparations of carrot (Daucus carota L.) and soybean (Glycine max), and by soybean leaf discs, have been studied. Preparations were incubated with tracer concentrations of l-[(3)H(3)C]methionine and the kinetics of appearance of radioactivity in phosphomethylethanolamine, phosphodimethylethanolamine, phosphocholine, phosphatidylmethylethanolamine, phosphatidyldimethylethanolamine, phosphatidylcholine, methylethanolamine, dimethylethanolamine, and choline followed at short incubation times. With soybean (tissue culture or leaves), an initial methylation utilizes phosphoethanolamine as substrate, forming phosphomethylethanolamine. The latter is converted to phosphatidylmethylethanolamine, which is successively methylated to phosphatidyldimethyethanolamine and to phosphatidylcholine. With carrot, again, an initial methylation is of phosphoethanolamine. Subsequent methylations occur at both the phospho-base and phosphatidyl-base levels. Both of these patterns differ qualitatively from that previously demonstrated in Lemna (SH Mudd, AH Datko 1986 Plant Physiol 82: 126-135) in which all three methylations occur at the phospho-base level. For soybean and carrot, some added contribution from initial methylation of phosphatidylethanolamine has not been excluded. These results, together with those from similar experiments carried out with water-stressed barley leaves (WD Hitz, D Rhodes, AD Hanson 1981 Plant Physiol 68: 814-822) and salinized sugarbeet leaves (AD Hanson, D Rhodes 1983 Plant Physiol 71: 692-700) suggest that in higher plants some, perhaps all, phosphatidylcholine synthesis occurs via a common committing step (conversion of phosphoethanolamine to phosphomethylethanolamine) followed by a methylation pattern which differs from plant to plant.


Methodology for the assessment of lung protection. Human pulmonary artery endothelial cell preservation using haemaccel.
This investigation was designed to show an original methodology for the assessment of lung preservation and to analyze the efficacy of a low potassium polygelin solution (haemaccel [HM]) on isolated human pulmonary artery endothelial cells. The effects of HM were compared with those of low potassium dextran (LPD), Belzer (University of Wisconsin [UWS]), and Euro-Collins solutions. The viability of the endothelial cultures was assessed by means of both total protein content and recovery of metabolic cellular function expressed as the protein synthesis rate after 6 hr and 16 hr of incubation at 10 degrees C. Our results failed to show any significant difference in the total protein content for HM, LPD, and UWS, both after 6 and 16 hr of incubation; however, the Euro-Collins-preserved sample revealed a significant drop in this parameter as early as 6 hr after the start. This finding was regarded as a clear indication of cellular cytotoxicity. In contrast, the metabolism recovery capacity of the cells varied significantly between HM and UWS at 6 hr and among HM, LPD, and UWS at 16 hr; at 6 hr, however, no significant difference was observed between HM and LPD. In conclusion, HM appears to exert a more significant effect on human pulmonary artery endothelial cell metabolism recovery than do the other fluids, thus suggesting its suitability as a long-term pulmonary perfusate.


Household Transmission of SARS-CoV-2: A Prospective Longitudinal Study Showing Higher Viral Load and Increased Transmissibility of the Alpha Variant Compared to Previous Strains.
We studied the secondary attack rate (SAR), risk factors, and precautionary practices of household transmission in a prospective, longitudinal study. We further compared transmission between the Alpha (B.1.1.7) variant and non-Variant of Concern (non-VOC) viruses. From May 2020 throughout April 2021, we recruited 70 confirmed COVID-19 cases with 146 household contacts. Participants donated biological samples eight times over 6 weeks and answered questionnaires. SARS-CoV-2 infection was detected by real-time RT-PCR. Whole genome sequencing and droplet digital PCR were used to establish virus variant and viral load. SARS-CoV-2 transmission occurred in 60% of the households, and the overall SAR for household contacts was 50%. The SAR was significantly higher for the Alpha variant (78%) compared with non-VOC viruses (43%) and was associated with a higher viral load. SAR was higher in household contacts aged ≥40 years (69%) than in younger contacts (40-47%), and for contacts of primary cases with loss of taste/smell. Children had lower viral loads and were more often asymptomatic than adults. Sleeping separately from the primary case reduced the risk of transmission. In conclusion, we found substantial household transmission, particularly for the Alpha variant. Precautionary practices seem to reduce SAR, but preventing household transmission may become difficult with more contagious variants, depending on vaccine use and effectiveness.


Sigma(2)-receptor regulation of dopamine transporter via activation of protein kinase C.
The elucidation of the mechanisms underlying sigma(2)-receptor activation and signal transduction is crucial to the understanding of sigma(2)-receptor function. Previous studies in our laboratory have demonstrated sigma(2)-receptor-mediated regulation of the dopamine transporter (DAT) as measured by amphetamine-stimulated release of [(3)H]dopamine (DA) from both rat striatal slices and PC12 cells. The regulation of the DAT in the PC12 cell model was dependent upon activation of Ca(2+)/calmodulin-dependent kinase II. We have now studied the second messenger systems involved in sigma(2)-receptor-mediated regulation of amphetamine-stimulated [(3)H]DA release in rat striatal slices, including Ca(2+)/calmodulin-dependent kinase II, protein kinase C, and sources of calcium required for the enhancement of release produced by sigma(2)-receptor activation. The Ca(2+)/calmodulin-dependent kinase II inhibitors 1-[N,O-bis-(5-isoquionolinesulfonyl)]-N-methyl-L-tyrosyl-4-phenylpiperazine and N-[2-[[[3-(4'-chlorophenyl)-2-propenyl]methylamino]methyl]phenyl]-N-(2-hydroxyethyl)-4'-methoxy-benzenesulfonamide phosphate did not significantly affect the (+)-pentazocine-mediated enhancement of amphetamine-stimulated [(3)H]DA release. However, we found that an inhibitor of protein kinase C, 3-[1-[3-(dimethylamino)propyl]-1H-indol-3-yl)-1H-pyrrole-2,5-dione, blocks the (+)-pentazocine-mediated enhancement in rat striatal slices. The protein kinase C activator phorbol 12-myristate 13-acetate, but not the inactive isophorbol 4 alpha,9 alpha,12 alpha,13 alpha,20-pentahydroxytiglia-1,6-dien-3-one, enhanced the amphetamine-stimulated [(3)H]DA release comparable to the enhancement seen by (+)-pentazocine alone. Additionally, the L-type voltage-dependent calcium channel inhibitor nitrendipine or prior treatment with thapsigargin, but not the N-type voltage-dependent calcium channel omega-conotoxin MVIIA, attenuated the (+)-pentazocine-mediated enhancement. Together, these data suggest that activation of sigma(2)-receptors results in the regulation of DAT activity via a calcium- and protein kinase C-dependent signaling mechanism.


Transcallosal Control of Bilateral Actions.
The corpus callosum is an important neural structure for controlling and coordinating bilateral movements of the upper limbs; however, there remains a substantial lack of knowledge regarding its association with lower limb control. We argue that transcallosal structure is an integral neural mechanism underlying control of the lower limbs and callosal degradation is a key contributor to mobility declines.


[Social failure: the concept and a method for its clinical assessment].
The social problematic one of the patient often conditions the medical therapeutic attitude, being able to force the hospital admission, to be able to guarantee its attention. In spite of its frequency, we do not have objective procedures that they allow us to quantify and to value its severity. 1 degree: To define the concept of social insufficiency (SI). 2 degrees: To enunciate a formula that allows us to quantify it objectively. 3 degrees: To make a practical validation of the procedure. We did not find in the bibliographical revision, a similar descriptive term neither procedures in order to quantify it. For that reason, we elaborated our own scale of social valuation, applying it within a protocol that also includes clinical aspects, valuations functional physical, mental and cognitive. The SI was considered like the quotient, between the functional capacity of the patient and the degree of social support. The SI could be defined as the incapacity of the socioeconomic and familiar surroundings of a patient, in order to guarantee the satisfy all the necessities derived from their pathological state, views from an integral perspective. The relationship between functional capacity (determined by means of the scale of the Red Cross of Valuation of Incapacity) and social support (evaluated by means of our scale) in quotient form, could be used to detect and quantify the SI objectively, being in our opinion a simple and practical procedure. We described to its frequency and severity, as well as the clinical and epidemiological characteristics to whom it is associated with greater statistical significance to that the existence in our means. The SI is a frequent problem, being necessary to establish their concept and forms of quantification. We propose a formal definition. The relationship between functional capacity and social support, expressed in quotient form, could be used can serve to value the SI objectively.


Inhibition of Host Arginase Activity Against Staphylococcal Bloodstream Infection by Different Metabolites.
Staphylococcus aureus is a notorious bacterial pathogen that often causes soft tissue and bloodstream infections and invariably garners resistance mechanisms against new antibiotics. Modulation of the host immune response by metabolites is a powerful tool against bacterial infections, but has not yet been used against S. aureus infections. In this study, we identified four metabolite biomarkers: L-proline, L-isoleucine, L-leucine, and L-valine (PILV), through a metabolomics study using animal models of S. aureus bloodstream infection. The exogenous administration of each metabolite or of PILV showed anti-infective effects, and a higher protection was achieved with PILV in comparison to individual metabolites. During the staphylococcal infection, the expression of most host arginase and nitric oxide synthase (NOS) isozymes was simultaneously induced in mouse liver, kidney, and blood samples. However, the induction of arginase isozymes was dramatically stronger than that of NOS isozymes. This elevated arginase activity was inhibited by the metabolite biomarkers thus killing S. aureus, and PILV exhibited the strongest inhibition of arginase activity and bacterial inhibition. The suppression of arginase activity also contributed to the metabolite-mediated phagocytic killing of S. aureus in mouse and human blood. Our findings demonstrate the metabolite-mediated arginase inhibition as a therapeutic intervention for S. aureus infection.


Correlating Neuroimaging and CNVs Data: 7 Years of Cytogenomic Microarray Analysis on Patients Affected by Neurodevelopmental Disorders.
The aim of this study was to evaluate the relationship between neurodevelopmental disorders, brain anomalies, and copy number variations (CNVs) and to estimate the diagnostic potential of cytogenomical microarray analysis (CMA) in individuals neuroradiologically characterized with intellectual developmental disorders (IDDs) isolated or associated with autism spectrum disorders (ASDs) and epilepsy (EPI), all of which were identified as a "synaptopathies." We selected patients who received CMA and brain magnetic resonance imaging (MRI) over a 7-year period. We divided them into four subgroups: IDD, IDD + ASD, IDD + EPI, and IDD + ASD + EPI. The diagnostic threshold of CMA was 16%. The lowest detection rate for both CMA and brain anomalies was found in IDD + ASD, while MRI was significantly higher in IDD and IDD + EPI subgroups. CMA detection rate was significantly higher in patients with brain anomalies, so CMA may be even more appropriate in patients with pathological MRI, increasing the diagnostic value of the test. Conversely, positive CMA in IDD patients should require an MRI assessment, which is more often associated with brain anomalies. Posterior fossa anomalies, both isolated and associated with other brain anomalies, showed a significantly higher rate of CMA positive results and of pathogenic CNVs. In the next-generation sequencing era, our study confirms once again the relevant diagnostic output of CMA in patients with IDD, either isolated or associated with other comorbidities. Since more than half of the patients presented brain anomalies in this study, we propose that neuroimaging should be performed in such cases, particularly in the presence of genomic imbalances.


[Endonasal microsurgery].
Endoscopic surgery of the sinuses is a valuable procedure for evaluating allergic or infectious chronic recurrent sinusitis. It allows assessment of the ostium and the state of the mucosa. Its complications are exceptional (rhinorrhoea, meningitis, orbital trauma), but require a detailed preoperative examination in order to reveal variations in sinus anatomy. The role of preoperative CT is therefore to analyse the type and the extent of the disease and individual anatomical variants. It is difficult to predict the depth and angle of introduction of the endoscope on the basis of the CT scan, which is performed in the coronal plane with variable angle. Ideally, only lateral scans would allow guidance of the introduction of the endoscope (hence the value of MRI or 3-D reconstructions).


Efficacy of vaginally applied estrogen, testosterone, or polyacrylic acid on vaginal atrophy: a randomized controlled trial.
Vaginal atrophy is a common chronic condition among postmenopausal women that can affect their quality of life. Recent studies have evaluated new treatment alternatives for vaginal atrophy; however, few therapeutic options have been thoroughly evaluated. This study aimed to compare the effectiveness and adverse effects of estrogen, testosterone, polyacrylic acid, and placebo lubricant for the treatment of postmenopausal women with vaginal atrophy. We conducted a randomized clinical trial with 80 postmenopausal women aged between 40 and 70 years who were being followed up at the Menopause Clinic of CAISM UNICAMP between November 2011 and January 2013. Women were randomly assigned to topical vaginal treatment with estrogen, testosterone, polyacrylic acid, and placebo lubricant, three times a week for 12 weeks. We used the vaginal maturation index, pH, vaginal health score, vaginal flora, laboratory tests, and ultrasound to evaluate changes of vaginal atrophy at baseline and after 6 and 12 weeks of treatment. After a 12-week treatment with topical estrogen and testosterone compared with the lubricant, an increased percentage of participants had vaginal pH less than 5, increased vaginal score, and an increase in the number of lactobacilli. Treatment with topical estrogen improved the vaginal maturation index and showed increased levels of estradiol in three women. No changes were observed in the endometrial evaluation of all treatment groups. After a 12-week treatment with testosterone and estrogen compared with placebo lubrication, there was a significant improvement in vaginal trophism in postmenopausal women with vaginal atrophy.


Hemoptysis in HIV-infected patients.
To define the causes and clinical features of hemoptysis in HIV-infected patients. Retrospective analysis of inpatient medical records for patients with HIV infection and hemoptysis. A large New York City teaching hospital and tertiary referral center. Fifty HIV-infected adult patients with 51 episodes of hemoptysis. The incidence of hemoptysis was 1.9%. A definite or presumptive etiology was identified for most (78%) episodes, with infection being the leading cause (80% of episodes with identified etiology). Hemoptysis was most often (40% of episodes with a definite or presumptive etiology) attributed to bacterial pneumonia. Mycobacterium tuberculosis was infrequently the cause of hemoptysis (6%). Mortality attributed to hemoptysis was 8%. In most patients, hemoptysis was mild (77%) and resolved (78%) without specific treatment. Our retrospective series of HIV-infected patients hospitalized for short-term care in New York City suggests that hemoptysis in this group has diverse causes but usually is infectious in etiology and most often due to bacterial pneumonia. Bleeding is typically mild and resolves without specific treatment, but can occasionally be fatal. Differential diagnosis should be approached with specific reference to this patient population and its unique features.


Thymic cyst in the differential diagnosis of paediatric cervical masses.
Thymic cyst in the differential diagnosis of paediatric cervical masses. Cervical ectopic thymic tissue is rarely reported. However, it should be included in the differential diagnosis of neck masses, especially in children. We present a case of a 7-year-old male with a soft, asymptomatic right-sided cervical mass. The patient underwent complete excision of a 3 x 4 cm cyst, which was derived from a mass of soft tissue and was in close relation with the carotid sheath. Histological examination of the specimen showed a thymic cyst. The child's post-operative course was unremarkable and follow-up appointment six months later showed complete recovery. The embryological development, clinical presentation and management of ectopic thymic lesions are discussed together with a brief review of the literature.


In vitro study of SDF-1α-loaded injectable and thermally responsive hydrogels for adipose stem cell therapy by SDF-1/CXCR4 axis.
Stem cell-based approaches have become a promising therapeutic strategy for treating ischemic diseases. The aim of this study was to develop injectable hydrogel systems for the local release of stromal cell-derived factor-1α (SDF-1α) to recruit adipose stem cells (ASCs) that express CXCR4 to achieve stem cell therapy and therapeutic angiogenesis. Thermoresponsive and injectable chitosan (CS)/β-glycerophosphate disodium salt pentahydrate (βGP) hydrogels with different concentrations of hyaluronic acid (HA) were designed and fabricated to achieve local and sustained release of SDF-1α for ASC recruitment. Herein, the material structures, physical properties, gelation temperature, and gelation time of hydrogels with different compositions were determined. The incorporation of 0.9% HA in CS-based hydrogels not only enhanced the gelation time but also increased the strength of the hydrogels. In addition, the results revealed that the thermoresponsive and injectable CS/βGP/HA hydrogels showed good biocompatibility. In addition, the in vitro release profiles showed that the hydrogels achieved sustained release of SDF-1α over 7 days and enhanced ASC migration. The results revealed that the hydrogels with HA enhanced the sustained release effect compared with the hydrogel without HA, indicating that the HA content regulated the physical and release properties of the injectable hydrogels. Therefore, thermoresponsive and injectable CS/βGP/HA hydrogels may provide an alternative for treating ischemic diseases via SDF-1/CXCR4 axis for ASC recruitment and retention.


A structural basis for sequence comparisons. An evaluation of scoring methodologies.
A residue-exchange matrix has been derived that is suitable for comparison of amino acid sequences. This matrix is based on the tabulation of 207,795 amino acid replacements observed in 65 homologous sets of structurally aligned three-dimensional structures (235 proteins). The majority of the data is from structural comparisons where there is between 15 and 40% sequence identity. As a result, a scoring matrix such as the one devised here should provide a sensitive basis for the comparison of amino acid sequences and the search for homologous sequences in amino acid databases. In order to assess the value of this matrix we have made a comparative analysis with 12 other published scoring matrices that have been used for the alignment of protein amino acid sequences. We find that the matrix derived here is among the better performers in terms of alignment significance, detection of homologous sequences and the accuracy of alignments.


Treatment of cutaneous and/or soft tissue manifestations of corticosteroids refractory chronic graft versus host disease (cGVHD) by a total nodal irradiation (TNI).
The management of corticosteroids refractory chronic graft versus host disease (cGVHD) remains controversial. Retrospective analysis of patients treated at the Integrated Center of Oncology by total nodal irradiation (TNI) was performed to evaluate its therapy potency. TNI delivers a dose of 1 Gy in a single session. The delimitation of the fields is clinical (upper limit: external auditory meatus; lower limit: mid-femur). No pre-therapeutic dosimetry scanner was necessary. Evaluation of the efficacy was by clinical measures at 6 months after the treatment. Twelve patients were treated by TNI between January 2010 and December 2013. TNI was used in second-line treatment or beyond. The median time between allograft and TNI was 31.2 months, and the median time between the first manifestations of cGVHD and TNI was about 24.2 months. Of the 12 patients, nine had a clinical response at 6 months (75%), including five complete clinical responses (41.6%). Five patients could benefit from a reduction of corticosteroid doses. Three patients had hematologic toxicity. TNI could be considered as an option for the treatment of a cutaneous and/or soft tissues corticosteroids refractory cGVHD. However, prospective randomized and double-blind trials remain essential to answer the questions about TNI safety and effectiveness.


Trabecular Bone Microarchitecture Improvement Is Associated With Skeletal Nerve Increase Following Aerobic Exercise Training in Middle-Aged Mice.
Advancing age is associated with bone loss and an increased risk of osteoporosis. Exercise training improves bone metabolism and peripheral nerve regeneration, and may play a critical role in osteogenesis and increase in skeletal nerve fiber density. In this study, the potential positive role of aerobic exercise training in bone metabolism and skeletal nerve regeneration was comprehensively evaluated in 14-month-old male C57BL/6 mice. The mice were divided into two groups: no exercise (non-exercise group) and 8-weeks of aerobic exercise training (exercise group), with six mice in each group. Dual-energy X-ray absorptiometry and micro-computed tomography showed that femoral and tibial bone parameters improved after aerobic exercise training. Greater skeletal nerve fiber density was also observed in the distal femoral and proximal tibial periostea, measured and analyzed by immunofluorescence staining and confocal microscopy. Pearson correlation analysis revealed a significant association between skeletal nerve densities and trabecular bone volume/total volume ratios (distal femur; R 2 = 0.82, p < 0.05, proximal tibia; R 2 = 0.59, p = 0.07) in the exercise group; while in the non-exercise group no significant correlation was found (distal femur; R 2 = 0.10, p = 0.54, proximal tibia; R 2 = 0.12, p = 0.51). Analysis of archival microarray database confirmed that aerobic exercise training changed the microRNA profiles in the mice femora. The differentially expressed microRNAs reinforce the role of aerobic exercise training in the osteogenic and neurogenic potential of femora and tibiae. In conclusion, 8-weeks of aerobic exercise training positively regulate bone metabolism, an effect that paralleled a significant increase in skeletal nerve fiber density. These findings suggest that aerobic exercise training may have dual utility, both as a direct stimulator of bone remodeling and a positive regulator of skeletal nerve regeneration.


Prevalence of pathogenic Leptospira spp. in sheep in a sheep-only abattoir in New Zealand.
To determine the prevalence of the two most commonly diagnosed pathogenic Leptospira spp. serovars, Hardjobovis and Pomona, in sheep in a sheep-only abattoir in New Zealand, and to determine the prevalence of kidneys which were leptospire culture-positive collected from sheep seropositive or seronegative to the microscopic agglutination test (MAT). A repeated cross-sectional observational study was conducted of serological and kidney culture prevalences of Leptospira borgpetersenii serovar Hardjobovis and Leptospira interrogans serovar Pomona. Lines of sheep and individual sheep were systematically randomly selected at a sheep-only abattoir during 18 May 2004 to November 2004 and 06 December 2004 to 14 June 2005. Additionally, a cross-sectional study examined prevalences in a purposively selected line of sheep from a flock with clinical evidence of an outbreak of leptospirosis. In the study population of 15,855 sheep of which 2,758 were sampled, 5.7 (95% CI=4.9-6.7)% were seropositive to one or both serovars; 44.2 (95% CI=34.6-54.2)% of 95 lines of sheep and 44.9 (95% CI=35.0-55.3)% of 89 farms showed serological evidence of infection. The serological prevalence of serovar Hardjobovis was significantly higher than that of serovar Pomona both at line (33% and 4%, respectively) and individual (5% and 1%, respectively) levels. A low but persistent seroprevalence of Hardjobovis throughout both years suggested low-level endemicity to this serovar, whereas Pomona infections appeared to be sporadic. Leptospires were isolated from kidneys of 8/37 (22%) Hardjobovis- and 1/6 (17%) Pomona-seropositive, and 5/499 (1%) seronegative animals. Of the animals purposively sampled from a farm with a clinical outbreak of leptospirosis, all kidneys from the 13 seropositive animals were culture-positive, indicating a high risk of exposure of meat workers in outbreak situations. Kidneys of MAT-seropositive sheep were 21.7 (95% CI=7.6-61.9) times more likely to test culture-positive than kidneys from animals with negative MAT titres. In general, the results indicated that 13/1,000 sheep slaughtered were potentially shedding leptospires. The study demonstrated the presence of a definite risk of occupational exposure of meat workers in a sheep-only slaughterhouse to the two most commonly diagnosed pathogenic Leptospira spp. serovars in New Zealand.


Trypanosomatid isolates from Honduras: differentiation between Trypanosoma cruzi and Trypanosoma rangeli.
With the aim of identifying and differentiating Trypanosoma cruzi from Trypanosoma rangeli, culture epimastigotes from 30 Honduran trypanosomatid isolates were analyzed by susceptibility to complement lysis, reactivity to lectins, reactivity to monoclonal antibodies specific for T. cruzi, and isoenzymatic electrophoretic patterns. Using these four methodologies, 27 of the 30 trypanosomatid isolates, as well as 5 clones, were identified as T. cruzi, whereas the remaining three isolates were classified as T. rangeli. None of the isolates presented mixed trypanosome species. Results indicate that both trypanosomatid species circulate in Honduras and that any of the four methods employed may be used to reliably differentiate T. cruzi from T. rangeli.


Rapid extraction of watermelon seed proteins using microwave and its functional properties.
Solid food industry waste like watermelon seed is an excellent source of value-added components such as proteins, oil, and carbohydrate. In the present study, protein extraction was carried out using microwave energy from defatted watermelon seeds (DWS), containing 50% of proteins. Microwave-assisted extraction (MAE) was optimized with different parameters, namely, solid to solvent ratio (1:10-1:40), pH (7-10), microwave power (30 W, 50 W, 70 W), extraction time (30 s-8 min) and moisture content or pre-leaching effect. Maximum protein recovery was achieved with 50 W microwave power, solid to solvent ration of 1:30, and pH 10 in 2 minutes of microwave irradiation time. MAE gave higher yield in less time compared to conventional extraction. SDS-PAGE confirmed the molecular weight of watermelon seed proteins (WSP) in the range of 25-250 kDa. A comparative study showed 90% protein recovery with MAE in 2 min with 1:30 (w/v) solid to solvent ratio, whereas ultrasound gave 87% in 9 min with 1:50 (w/v) ratio and batch 72% in 25 min with 1:70 (w/v) ratio. Watermelon seed proteins obtained from MAE method possess excellent functional properties with reference to conventional extraction method indicating its application in food products.


The magnitude of innovation and its evolution in social animals.
Innovative behaviour in animals, ranging from invertebrates to humans, is increasingly recognized as an important topic for investigation by behavioural researchers. However, what constitutes an innovation remains controversial, and difficult to quantify. Drawing on a broad definition whereby any behaviour with a new component to it is an innovation, we propose a quantitative measure, which we call the magnitude of innovation, to describe the extent to which an innovative behaviour is novel. This allows us to distinguish between innovations that are a slight change to existing behaviours (low magnitude), and innovations that are substantially different (high magnitude). Using mathematical modelling and evolutionary computer simulations, we explored how aspects of social interaction, cognition and natural selection affect the frequency and magnitude of innovation. We show that high-magnitude innovations are likely to arise regularly even if the frequency of innovation is low, as long as this frequency is relatively constant, and that the selectivity of social learning and the existence of social rewards, such as prestige and royalties, are crucial for innovative behaviour to evolve. We suggest that consideration of the magnitude of innovation may prove a useful tool in the study of the evolution of cognition and of culture.