Reiki Reduces Burnout Among Community Mental Health Clinicians.
Clinicians working in community mental health clinics are at high risk for burnout. Burnout is a problem involving emotional exhaustion, depersonalization, and reduced personal accomplishment. Reiki is a holistic biofield energy therapy beneficial for reducing stress. The purpose of this study was to determine if 30 minutes of healing touch could reduce burnout in community mental health clinicians. We utilized a crossover design to explore the efficacy of Reiki versus sham Reiki, a pseudo treatment designed to mimic true Reiki, as a means to reduce symptoms of burnout. Subjects were randomized to whether they started with Reiki or sham. The Maslach Burnout Inventory-Human Services Survey (MBI-HSS) and the Measure Your Medical Outcome Profile Version 2 (MYMOP-2) were used as outcome measures. Multilevel modeling was used to represent the relations among variables. Reiki was statistically significantly better than sham Reiki in reducing burnout among community mental health clinicians (p=0.011). Reiki was significant in reducing depersonalization (p<0.001), but only among single people. Reiki reduced the primary symptom on the MYMOP also only among single people (p=0.03). The effects of Reiki were differentiated from sham Reiki. Reiki could be helpful in community mental health settings for the mental health of the practitioners.


Limb deformities and three-dimensional ultrasound.
To assess the ability of three-dimensional (3D) ultrasonography for improvement of antenatal detection of limb deformities. 347 patients were selected from a routine outpatient clinic or sent for supervision from other units because of suspected anomalies of fetal extremities. 3D ultrasound devices used in the study were Combison 530D and Voluson 530D MT (Kretztechnik, Zipf, Austria) with a 3-5 MHz annular array transducer for three-dimensional volume scanning. In 41 of 347 patients the initial diagnosis was suspected by two-dimensional sonography (gestational age 18-32 weeks). In 28 of 41 suspected cases the diagnosis of abnormalities was determined after examination by 3D sonography: 17/28 clubfoot, 3/28 hand-polydactily, 2/28 upper limb contractures, 1/28 lower limb contractures, 4/28 micromelia within the syndrome of skeletal dysplasia. In 13 of 41 suspected cases, normal anatomy was confirmed using 3D sonography. Three-dimensional sonography is the "method of choice" for the detection of an isolated defect of a single limb, developmental or positional deformations and minor defects of hands and feet. Surface-mode reconstruction of the complete limb and transparent-view reconstruction of the entire skeletal structure are effective technical advantages enabling a completely new visual perception of the unborn baby.


An update on the extraneuronal monoamine transporter (EMT): characteristics, distribution and regulation.
Biological membranes prevent transmembrane diffusion in the majority of organic molecules that bear net charges at physiological pH. Consequently, these compounds must use more or less specific membrane-bound transport systems to be imported into or exported from cells or organisms. The extraneuronal monoamine transporter (EMT) is a transmembranar transport system involved in the transfer of monoamine compounds across cell membranes. It was identified more than 30 years ago [1], its functional characteristics being thereafter described [review by 2]. The recent cloning of this transporter in man and rat reopened investigation and interest in this entity. EMT is a Na(+) and Cl(-)-independent, potential-dependent carrier, known to have a broad tissue distribution (eg. myocardium, vascular and non-vascular smooth muscle cells, glandular cells, placenta and CNS glial cells). According to its transport function and primary structure, EMT is included in the amphiphilic solute facilitator (ASF) family of transporters. Physiological substrates for EMT include the monoamines serotonin, dopamine, noradrenaline, adrenaline and histamine. Moreover, several xenobiotics including the neurotoxin 1-methyl-4-phenylpyridinium, clonidine, cimetidine and the K(+)-channel blocker tetraethylammonium interact with this transporter. The aim of this work is to review knowledge concerning EMT, making an update on its functional characteristics, physiological importance and regulation. A special emphasis will be given to very recent investigations concerning regulation of EMT by intracellular second messenger systems and the interaction of modulators of P-glycoprotein, the product of the multidrug resistance gene MDR1, with EMT.


One-health approach on the future application of snails: a focus on snail-transmitted parasitic diseases.
Snails are fascinating molluscs with unique morphological and physiological adaptive features to cope with various environments. They have traditionally been utilized as food and feed sources in many regions of the world. The future exploitation of alternative nutrient sources, like snails, is likely to increase further. Snails, however, also serve as an intermediate host for several zoonotic parasites. A category of parasitic infections, known as snail-transmitted parasitic diseases (STPDs), is harmful to humans and animals and is mainly driven by various trematodes, cestodes, and nematodes. The environment plays a crucial role in transmitting these parasites, as suitable habitats and conditions can facilitate their growth and proliferation in snails. In light of diverse environmental settings and biologically categorized snail species, this review evaluates the dynamics of significant STPDs of zoological importance. Additionally, possible diagnostic approaches for the prevention of STPDs are highlighted. One-health measures must be considered when employing snails as an alternative food or feed source to ensure the safety of snail-based products and prevent any adverse effects on humans, animals, and the environment.


Evaluation and classification of pulmonary arterial hypertension.
In early 2019, the 6th World Symposium on Pulmonary Hypertension (WSPH) released an updated document highlighting the advances in the last five years. During the quinquennial event many experts worked together to suggest new changes in the disease diagnosis and management. Since inception of the WSPH in 1973, this is the first time when the hemodynamic definition of pulmonary hypertension (PH) has been updated. These proceedings have re-defined the different hemodynamic types of PH that occur with the left heart disease along with introduction to the genetic testing as part of pulmonary arterial hypertension (PAH) evaluation. Objective of this review is to highlight the evaluation and diagnosis of PAH based on the proceedings of the 6th WSPH. Accurate early diagnosis and subsequent management of PH is necessary, as despite of treatment advances, survival remains suboptimal.


Equine bone marrow volume reduction, red blood cell depletion, and mononuclear cell recovery using the PrepaCyte-CB processing system.
Volume reduction and RBC depletion of equine bone marrow specimens are necessary processing steps for the immediate therapeutic use of bone marrow (BM)-derived mesenchymal stem cells (MSC), and for MSC expansion in culture. The purpose of the study was to evaluate the ability of the PrepaCyte-CB processing system to reduce volume, deplete RBC, and recover mononuclear cells (MNC) from equine BM specimens. One hundred and twenty mL of heparinized BM were obtained from each of 90 horses. A CBC was performed on the BM pre- and post-PrepaCyte-CB processing. Volume and RBC reduction, and total nucleated cell (TNC) and MNC recoveries were determined. Bone marrow volume was reduced from 120 mL to 21 mL with a median RBC depletion of 90.1% (range, 62.0-96.7%). The median preprocessing total TNC count was 2.2 × 10(9) (range, 0.46-7.9 × 10(9)) and the median postprocessing TNC count was 1.7 × 10(9) (range, 0.3-4.4 × 10(9); P < .0001), with a median recovery of 73.5% (range, 22.4-216.7%). The median preprocessing total MNC count was 0.9 × 10(9) (range, 0.1-4.7 × 10(9)) and median postprocessing total MNC count was 0.8 × 10(9) (range, 0.1-2.7 × 10(9); P = .06), with a median recovery of 83.7% (range, 15.4-413.9%). The PrepaCyte-CB processing system can be used to deplete both volume and RBC, and recover MNC from equine BM specimens. Further studies assessing the viability of MSC and the efficacy of MSC expansion after using the PrepaCyte-CB processing system are warranted.


Liquor licensing and community action in regional and remote Australia: a review of recent initiatives.
To review the effectiveness of community-based initiatives involving restrictions on alcohol availability in remote and regional locations in Australia, and to assess their implications for other communities or towns contemplating similar measures. Findings from evaluations of community initiatives in Tennant Creek (NT), Derby (WA), Halls Creek (WA), Elliott (NT) and Curtin Springs (NT) are compared with respect to impact on alcohol consumption, and on indicators of alcohol-related harm relating to public order, health and well-being, and economic activities. The extent of community support for these initiatives is also examined. Restrictions were found to have a modest but real impact on alcohol consumption, and a significant impact on indicators of alcohol-related harm, especially violence. Restrictions were also found to have widespread community support, often qualified by a belief that other measures were also required. Restrictions on availability are an effective means of reducing alcohol-related harm at a local level and, depending upon the processes involved in their introduction, are likely to have strong community support, provided that other measures are also pursued. Five issues are identified that are likely to arise in other settings where restrictions are contemplated. These are: the issue of representativeness ('who speaks for the community?'); selection of particular kinds of restrictions on availability; selection of most appropriate additional measures; universal vs. selective restrictions; and the role of liquor licensing authorities in imposing or facilitating restrictions.


Determinants of mortality in a combined cohort of 501 patients with HIV-associated Cryptococcal meningitis: implications for improving outcomes.
Cryptococcal meningitis (CM) is a leading cause of death in individuals infected with human immunodeficiency virus (HIV). Identifying factors associated with mortality informs strategies to improve outcomes. Five hundred one patients with HIV-associated CM were followed prospectively for 10 weeks during trials in Thailand, Uganda, Malawi, and South Africa. South African patients (n = 266) were followed for 1 year. Similar inclusion/exclusion criteria were applied at all sites. Logistic regression identified baseline variables independently associated with mortality. Mortality was 17% at 2 weeks and 34% at 10 weeks. Altered mental status (odds ratio [OR], 3.1; 95% confidence interval [CI], 1.7-5.9), high cerebrospinal fluid (CSF) fungal burden (OR, 1.4 per log10 colony-forming units/mL increase; 95% CI, 1.0-1.8), older age (>50 years; OR, 3.9; 95% CI, 1.4-11.1), high peripheral white blood cell count (>10 × 10(9) cells/L; OR, 8.7; 95% CI, 2.5-30.2), fluconazole-based induction treatment, and slow clearance of CSF infection were independently associated with 2-week mortality. Low body weight, anemia (hemoglobin <7.5 g/dL), and low CSF opening pressure were independently associated with mortality at 10 weeks in addition to altered mental status, high fungal burden, high peripheral white cell count, and older age. In those followed for 1 year, overall mortality was 41%. Immune reconstitution inflammatory syndrome occurred in 13% of patients and was associated with 2-week CSF fungal burden (P = .007), but not with time to initiation of antiretroviral therapy (ART). CSF fungal burden, altered mental status, and rate of clearance of infection predict acute mortality in HIV-associated CM. The results suggest that earlier diagnosis, more rapidly fungicidal amphotericin-based regimens, and prompt immune reconstitution with ART are priorities for improving outcomes.


Clinical pharmacological studies with the long-acting beta-adrenoceptor blocking drug bopindolol.
Bopindolol is a potent beta-adrenoceptor antagonist with mild intrinsic sympathomimetic activity that exhibits a long duration of action. The cardiac beta-adrenoceptor blockade produced by an oral dose of 1 mg bopindolol is of similar intensity as that seen after atenolol 100 mg p.o. or pindolol 10 mg p.o. Like other beta-adrenoceptor antagonists, bopindolol is effective in inhibiting resting and stimulated plasma renin activity. In contrast to compounds lacking ISA, bopindolol does not produce undesirable changes in plasma lipoprotein composition during chronic therapy. Its bioavailability after oral administration amounts to about 70%. In the studies reviewed, bopindolol was well tolerated even after an oral dose of 12 mg, which, as far as beta-adrenoceptor blockade is concerned, would be equivalent to a single oral dose of about 1.2 g of atenolol. The onset of action of bopindolol is relatively slow, a feature that to a certain extent might account for the good tolerance of the drug observed in experimental and therapeutic studies. The delayed onset of action of the drug has facilitated the elucidation of the relationship between plasma levels and pharmacological effects during the period immediately after oral administration. Analysis of the temporal relationship between drug plasma levels and beta-adrenoceptor blockade provides evidence suggesting that the cardiac beta adrenoceptors lie outside the plasma compartment.


Alterations in the characteristic size distributions of subcellular scatterers at the onset of apoptosis: effect of Bcl-xL and Bax/Bak.
Optical scatter imaging is used to estimate organelle size distributions in immortalized baby mouse kidney cells treated with 0.4 microM staurosporine to induce apoptosis. The study comprises apoptosis competent iBMK cells (W2) expressing the proapoptotic proteins Bax/Bak, apoptosis resistant Bax/Bak null cells (D3), and W2 and D3 cells expressing yellow fluorescent protein (YFP) or YFP fused to the antiapoptotic protein Bcl-x(L) (YFP-Bcl-x(L)). YFP expression is diffuse within the transfected cells, while YFP-Bcl-x(L) is localized to the mitochondria. Our results show a significant increase in the mean subcellular particle size from approximately 1.1 to 1.4 microm in both Bax/Bak expressing and Bax/Bak null cells after 60 min of STS treatment compared to DMSO-treated control cells. This dynamic is blocked by overexpression of YFP-Bcl-x(L) in Bax/Bak expressing cells, but is less significantly inhibited by YFP-Bcl-x(L) in Bax/Bak null cells. Our data suggest that the increase in subcellular particle size at the onset of apoptosis is modulated by Bcl-x(L) in the presence of Bax/Bak, but it occurs upstream of the final commitment to programmed cell death. Mitochondrial localization of YFP-Bcl-x(L) and the finding that micron-sized particles give rise to the scattering signal further suggest that alterations in mitochondrial morphology may underlie the observed changes in light scattering.


Structural comparison of bacterial and human iron-dependent phenylalanine hydroxylases: similar fold, different stability and reaction rates.
Structure determination of bacterial homologues of human disease-related proteins provides an efficient path to understanding the three-dimensional fold of proteins that are associated with human diseases. However, the precise locations of active-site residues are often quite different between bacterial and human versions of an enzyme, creating significant differences in the biological understanding of enzyme homologs. To study this hypothesis, phenylalanine hydroxylase from a bacterial source has been structurally characterized at high resolution and comparison is made to the human analog. The enzyme phenylalanine hydroxylase (PheOH) catalyzes the hydroxylation of l-phenylalanine into l-tyrosine utilizing the cofactors (6R)-l-erythro-5,6,7,8 tetrahydrobiopterin (BH(4)) and molecular oxygen. Previously determined X-ray structures of human and rat PheOH, with a sequence identity of more than 93%, show that these two structures are practically identical. It is thus of interest to compare the structure of the divergent Chromobacterium violaceum phenylalanine hydroxylase (CvPheOH) ( approximately 24% sequence identity overall) to the related human and rat PheOH structures. We have determined crystal structures of CvPheOH to high resolution in the apo-form (no Fe-added), Fe(III)-bound form, and 7,8-dihydro-l-biopterin (7,8-BH(2)) plus Fe(III)-bound form. The bacterial enzyme displays higher activity and thermal melting temperature, and structurally, differences are observed in the N and C termini, and in a loop close to the active-site iron atom.


Negative Ions Enhance Survival of Membrane Protein Complexes.
Membrane protein complexes are commonly introduced to the mass spectrometer solubilized in detergent micelles. The collisional activation used to remove the detergent, however, often causes protein unfolding and dissociation. As in the case for soluble proteins, electrospray in the positive ion mode is most commonly used for the study of membrane proteins. Here we show several distinct advantages of employing the negative ion mode. Negative polarity can yield lower average charge states for membrane proteins solubilized in saccharide detergents, with enhanced peak resolution and reduced adduct formation. Most importantly, we demonstrate that negative ion mode electrospray ionization (ESI) minimizes subunit dissociation in the gas phase, allowing access to biologically relevant oligomeric states. Together, these properties mean that intact membrane protein ions can be generated in a greater range of solubilizing detergents. The formation of negative ions, therefore, greatly expands the possibilities of using mass spectrometry on this intractable class of protein. Graphical Abstract .


Early Supported Discharge for Older Adults Admitted to Hospital with Medical Complaints: A Qualitative Study Exploring the Views of Stakeholders.
Early supported discharge (ESD) is well established as a model of health service delivery for people with stroke. Emerging evidence indicates that ESD also reduces the length of stay for older medical inpatients. There is a dearth of evidence exploring the views of stakeholders on ESD as a model of care for older medical inpatients. The overall aim of this study is to explore the views and perceptions of older adults, family carers and healthcare professionals on the potential role of ESD for older adults admitted to hospital with medical complaints. Purposeful sampling was used to recruit older adults and family carers for interview. For Healthcare Professionals (HCPs), snowball purposeful sampling was used. Phone interviews took place following a semi-structured interview guide. Focus groups were moderated by A-MM. Braun and Clarke's approach to thematic analysis was used. Ethical approval was granted by the HSE Mid-Western Area Regional Ethics Committee in November 2021 (REC Ref. 096/2021). Fifteen HCPs took part across three focus groups, with six older adults and two family members participating in one-to-one interviews. Three themes were identified: 1. Pre-ESD experiences of providing and receiving older adult inpatient care, 2. Navigating discharge procedures from acute hospital services, 3. A vision for more integrated model of care and a medical ESD team. This study provided insight into the current discharge experiences of older adult care in the acute setting, the potential role for ESD in this population and the key factors that would need to be considered for the running of an ESD service for older adults admitted to hospital with medical complaints. This research highlights the barriers and facilitators to ESD for older medical inpatients from the perspectives of key stakeholders. Given the adverse outcomes associated with prolonged hospital stay, these findings will help inform the development of a feasibility trial, examining patient and process outcomes for older adults admitted to hospital with medical complaints who receive an ESD intervention.


Carnitine palmitoyltransferase activities (EC 2.3.1.-) of rat liver mitochondria.
1. A continuously recording and sensitive fluorimetric assay is described for carnitine palmitoyltransferase. This assay has been applied to whole or disintegrated mitochondria and to soluble protein fractions. 2. When rat liver mitochondria had been disintegrated by ultrasound, the specific activity of carnitine palmitoyltransferase was 15-20m-units/mg of protein. Only one-fifth of this activity was assayable (with added substrates) before mitochondrial disintegration. 3. It is concluded that there are two carnitine palmitoyltransferase activities in rat liver mitochondria, of which one (type I) is relatively superficial in location and catalyses an acyl-group transfer between added CoA and carnitine, whereas the other (type II) is less superficial and catalyses an acyl-group transfer in unbroken mitochondria between added carnitine and intramitochondrial CoA. The existence of two distinct carnitine palmitoyltransferases was predicted by Fritz & Yue (1963). 4. In unbroken mitochondria, type I transferase is accessible to the inhibitor 2-bromostearoyl-CoA whereas the type II transferase is inaccessible. 5. A major part of the total carnitine palmitoyltransferase activity of rat liver mitochondria is membrane-bound and of type II. 6. These observations, when considered in conjunction with the penetration of mitochondria by CoASH or carnitine, indicate that the type II transferase is attached to the inner mitochondrial membrane.


Cardiac side effects of phenytoin and carbamazepine. A dose-related phenomenon?
Three patients with dystrophia myotonica and echocardiographic signs of subclinical cardiopathy had cardiac side effects during oral treatment with phenytoin sodium or carbamazepine. These side effects were dose related: ventricular tachycardia appeared at a toxic serum phenytoin level in one patient and disappeared as the concentration fell within the therapeutic range, and atrioventricular block grade 1 developed in two patients at low serum carbamazepine levels, its severity increasing with the drug level. Given the risk of dangerous side effects, cardiac status needs to be carefully assessed before administration of phenytoin or carbamazepine in the treatment of dystrophia myotonica.


Biophysical modeling of brain tumor progression: from unconditionally stable explicit time integration to an inverse problem with parabolic PDE constraints for model calibration.
A novel unconditionally stable, explicit numerical method is introduced to the field of modeling brain cancer progression on a tissue level together with an inverse problem (IP) based on optimal control theory that allows for automated model calibration with respect to observations in clinical imaging data. Biophysical models of cancer progression on a tissue level are in general based on the assumption that the spatiotemporal spread of cancerous cells is determined by cell division and net migration. These processes are typically described in terms of a parabolic partial differential equation (PDE). In the present work a parallelized implementation of an unconditionally stable, explicit Euler (EE(⋆)) time integration method for the solution of this PDE is detailed. The key idea of the discussed EE(⋆) method is to relax the strong stability requirement on the spectral radius of the coefficient matrix by introducing a subdivision regime for a given outer time step. The performance is related to common implicit numerical methods. To quantify the numerical error, a simplified model that has a closed form solution is considered. To allow for a systematic, phenomenological validation a novel approach for automated model calibration on the basis of observations in medical imaging data is developed. The resulting IP is based on optimal control theory and manifests as a large scale, PDE constrained optimization problem. The numerical error of the EE(⋆) method is at the order of standard implicit numerical methods. The computing times are well below those obtained for implicit methods and by that demonstrate efficiency. Qualitative and quantitative analysis in 12 patients demonstrates that the obtained results are in strong agreement with observations in medical imaging data. Rating simulation success in terms of the mean overlap between model predictions and manual expert segmentations yields a success rate of 75% (9 out of 12 patients). The discussed EE(⋆) method provides desirable features for image-based model calibration or hybrid image registration algorithms in which the model serves as a biophysical prior. This is due to (i) ease of implementation, (ii) low memory requirements, (iii) efficiency, (iv) a straightforward interface for parameter updates, and (v) the fact that the method is inherently matrix-free. The explicit time integration method is confirmed via experiments for automated model calibration. Qualitative and quantitative analysis demonstrates that the proposed framework allows for recovering observations in medical imaging data and by that phenomenological model validity.


Mutations in COCH that result in non-syndromic autosomal dominant deafness (DFNA9) affect matrix deposition of cochlin.
The COCH gene mutated in autosomal dominant sensorineural deafness (DFNA9) encodes cochlin, a major constituent of the inner ear extracellular matrix. Sequence analysis of cochlin from DFNA9 patients identified five distinct single-amino-acid mutations within a conserved region (the LCCL domain) of cochlin. To define the molecular basis of DFNA9, we have generated myc-tagged wild-type and mutant cochlins and explored their behavior in transient transfection systems. Western blotting of cell lysates and culture media indicates that wild-type and mutant cochlins are synthesized and secreted in similar amounts. Immunofluorescent staining confirms that all are detected within the endoplasmic reticulum and the Golgi complex of transfected cells. Our findings suggest that COCH mutations are unlikely to cause abnormalities in secretion and suggest that extracellular events might cause DFNA9 pathology. In agreement, we show that wild-type cochlin accumulates in extracellular deposits that closely parallel the matrix component fibronectin, whereas mutant cochlins vary in the amount and pattern of extracellular material. Whereas some mutants exhibit an almost normal deposition pattern, some show complete lack of deposition. Our results suggest that DFNA9 results from gene products that fail to integrate correctly into the extracellular matrix. The partial or complete penetrance of integration defects suggests that DFNA9 pathology may be caused by multiple molecular mechanisms, including compromised ability of cochlin to self-assemble or to form appropriate complexes with other matrix components.


Population-based estimate of the contribution of TP53 mutations to subgroups of early-onset breast cancer: Australian Breast Cancer Family Study.
Although germline TP53 mutations have been identified in women with breast cancer from families meeting Li-Fraumeni criteria, their contribution to breast cancer per se is not well known, but is thought to be minimal. We aimed to determine the prevalence of germline TP53 mutations in subgroups of early-onset breast cancer. Germline TP53 mutation status was assessed by DNA sequencing, screening for heterozygous single-nucleotide polymorphisms, and Multiplex Ligation-Dependent Probe Amplification analyses. From an Australian population-based series of invasive breast cancers, we studied (a) 52 women diagnosed before age 30 years unselected for family history [very early-onset (VEO)] and (b) 42 women diagnosed in their 30s with two or more first- or second-degree relatives with breast or ovarian cancer [early-onset family history (EO-FH)]. Of the VEO group, two (4%) had a mutation: G13203A (exon 6 missense) in a 24-year-old and a large 5,338-bp genomic deletion in a 26-year-old. Neither had a family cancer history that met Li-Fraumeni criteria. Of the EO-FH group, three (7%) had a mutation: T13240G (a known intron 5 splicing mutation) in a 36-year-old from a classic Li-Fraumeni family; G12299A (exon 4 missense) in a 33-year-old from a Li-Fraumeni-like family; and 14058delG (exon 7 frame-shift) in a 39-year-old with a family cancer history that did not meet Li-Fraumeni criteria. Germline TP53 mutations play a larger role in early-onset breast cancer than previously thought, and in this context, can be evident outside clinically defined Li-Fraumeni families.


Chondromalacia of the knee: evaluation with a fat-suppression three-dimensional SPGR imaging after intravenous contrast injection.
Twenty-one MRI studies with a fat-suppression three-dimensional spoiled gradient-recalled echo in a steady state (3D SPGR) pulse sequence after intravenous contrast injection were evaluated to assess the accuracy in depicting chondromalacia of the knee. On the basis of MR images, chondromalacia and its grade were determined in each of five articular cartilage regions (total, 105 regions) and then the results were compared to arthroscopic findings. The sensitivity, specificity, and accuracy of MRI were 70%, 99%, and 93%, respectively. MR images depicted 7 of 11 lesions of arthroscopic grade 1 or 2 chondromalacia, and seven of nine lesions of arthroscopic grade 3 or 4 chondromalacia. The cartilage abnormalities in all cases appeared as focal lesions with high signal intensity. Intravenous contrast-injection, fat-suppression 3D SPGR imaging showed high specificity in excluding cartilage abnormalities and may be considered as an alternative to intra-articular MR arthrography when chondromalacia is suspected.


The flow cytometer model markedly affects measurement of ex vivo whole blood platelet-bound P-selectin expression in patients with chest pain: are we comparing apples with oranges.
Surface expression of P-selectin is known to be a marker of platelet activation in patients with acute coronary syndromes. However, direct comparisons of flow cytometer data may be obscured by differences in methodology, artifactual platelet activation during washing procedures, choice of antibodies, absence of control measurements, and possible observer bias due to unblinded data collection. We sought to test the hypothesis that the model of flow cytometer represents another variable affecting P-selectin measurements. Platelet P-selectin in whole blood was measured by FACScan (Becton Dickinson, Inc., San Diego, CA, USA) or EPICS XL (Coulter Corporation, Hialeah, FL, USA) flow cytometry in 338 patients presenting with chest pain to the emergency departments of three community hospitals as part of a multicenter diagnostic trial. Platelet expression of P-selectin (% of cell positivity) was consistently higher for each discharge diagnosis when measured with FACScan flow cytometer (13.2+/-4.1 for myocardial infarction, 10.0+/-3.6 for unstable angina, 9.9+/-3.5 for heart failure, 4.7+/-0.1 for gastrointestinal illness, and 6.3+/-0.7 for patients with noncardiac chest pain) when compared with results obtained from the EPICS XL instrument (2.4+/-0.2, 2.5+/-0.2, 2.5+/-0.1, 1.8+/-0.1, and 2.3+/-0.1 respectively, p=0.0001 for all groups). This study reveals marked discrepancies in the level of platelet P-selectin measurement based exclusively on the model of flow cytometer used. If P-selectin is to become a diagnostic tool for differentiating an etiology of chest pain, standardized measurements must be defined for each model of flow cytometer.


Expression of a finger millet transcription factor, EcNAC1, in tobacco confers abiotic stress-tolerance.
NAC (NAM, ATAF1-2, and CUC2) proteins constitute one of the largest families of plant-specific transcription factors and have been shown to be involved in diverse plant processes including plant growth, development, and stress-tolerance. In this study, a stress-responsive NAC gene, EcNAC1, was isolated from the subtracted stress cDNA library generated from a drought adapted crop, finger millet, and characterized for its role in stress-tolerance. The expression analysis showed that EcNAC1 was highly induced during water-deficit and salt stress. EcNAC1 shares high amino acid similarity with rice genes that have been phylogenetically classified into stress-related NAC genes. Our results demonstrated that tobacco transgenic plants expressing EcNAC1 exhibit tolerance to various abiotic stresses like simulated osmotic stress, by polyethylene glycol (PEG) and mannitol, and salinity stress. The transgenic plants also showed enhanced tolerance to methyl-viologen (MV) induced oxidative stress. Reduced levels of reactive oxygen species (ROS) and ROS-induced damage were noticed in pot grown transgenic lines under water-deficit and natural high light conditions. Root growth under stress and recovery growth after stress alleviation was more in transgenic plants. Many stress-responsive genes were found to be up-regulated in transgenic lines expressing EcNAC1. Our results suggest that EcNAC1 overexpression confers tolerance against abiotic stress in susceptible species, tobacco.


Modeling Tomato Ripening Based on Carotenoid Raman Spectroscopy: Experimental Versus Kinetic Model.
This work reports on a combined experimental and theoretical investigation on the carotenoid Raman signal in several tomato fruits during their postharvest time evolution and ripening. Both resonant (180°) backscattering and polarized (90°) Raman scattering were used to monitor the most prominent bands of carotenoid (lycopene and β-carotene) evolution in different tomato varieties. Relevant findings of the present investigations were that while the depolarization ratio of the ν1 band hardly changed with time, the Raman ν1 band intensity did change showing a similar pattern for all tomatoes investigated. Indeed, all cases investigated revealed a rise of the carotenoid signal coincident with the onset of the turning stage of the fruit ripening, a pronounced maximum of the Raman signal followed by a post-maximum decline at the red ripening stage. A kinetic model has been developed to describe the time evolution of the observed Raman signatures based on the rate coefficient of the carotenoid synthesis and the time evolution of the scattering coefficient of the fruit. The model describes satisfactorily the tomato evolution through the distinct ripening stages providing new insight on the assessment of the postharvest fruit control and quality.


Bilateral Continuous Erector Spinae Plane Block Contributes to Effective Postoperative Analgesia After Major Open Abdominal Surgery: A Case Report.
The erector spinae plane (ESP) block is a regional anesthetic technique involving local anesthetic injection in a paraspinal plane deep to the erector spinae muscle. Originally described for thoracic analgesia when performed at the T5 transverse process, the ESP block can provide abdominal analgesia if performed at lower thoracic levels because the erector spinae muscles extend to the lumbar spine. A catheter inserted into this plane can extend analgesic duration and can be an alternative to epidural analgesia. In this report, we describe using bilateral ESP catheters inserted at the T8 level to provide effective perioperative analgesia for major open lower abdominal surgery.


Computational intelligence aspects for defect classification in aeronautic composites by using ultrasonic pulses.
Production of carbon fiber reinforced polymers (i.e., one of the basic material of the modern airplanes) is an elaborate process unfree from faults and problems. Errors during the manufacturing or the plies' overlapping, in fact, can cause particular flaws in the resulting material, so compromising its same integrity. Within this framework, ultrasonic tests could be useful to characterize the presence of defect, depending on its dimensions. On the contrary, the requirement of a perfect state for used polymers is unavoidable in order to assure both transport reliability and passenger safety. Therefore, a real-time approach able to recognize and classify the defect starting from the measured ultrasonic echoes could be very useful in industrial applications. The ill-posedness of the so defined process induce a regularization method. In this paper, an heuristic approach is proposed for this aim. Particularly, the proposed method is based on the use of support vector machines. Obtained results assure good performances of the implemented classifier, with very interesting applications.


Identification of thyroglobulin in orbital tissues of patients with thyroid-associated ophthalmopathy.
Thyroid-associated ophthalmopathy (TAO) is thought to have an autoimmune pathogenesis because of its association with autoimmune thyroid disease, in particular with Graves' disease. Nevertheless, the nature of the autoimmune reaction is unclear, and a target orbital autoantigen has not been conclusively identified. A widely discussed hypothesis is that antigens constitutively shared by the thyroid and orbital tissues are targets of an autoimmune reaction. It has been also postulated that a thyroid-soluble antigen, namely thyroglobulin (Tg), is transported to orbital tissues through the lymphatics, where it accumulates and elicits autoimmune damages in susceptible individuals. Here we have investigated whether Tg is present in orbital tissues from patients with TAO. Retrobulbar tissue specimens were obtained from three patients with Graves' disease and TAO who underwent decompressive orbitotomy, and at autopsy from two patients with no thyroid or eye disease. All patients with TAO had been previously treated with radioiodine to control Graves' hyperthyroidism. Western blot analysis with a monoclonal anti-Tg antibody showed the presence of intact Tg, both in soluble and membrane-associated fractions of orbital tissue extracts from the patients with TAO, in amounts estimated to range from approximately 320 to approximately 900 pg/microg of tissue protein. In contrast, Tg was not detected in orbital tissue extracts from patients with no thyroid or eye disease. Tg was also demonstrated in orbital tissue extracts from two of three patients with TAO by enzyme-linked immunosorbent assay (ELISA), in amounts estimated to range from approximately 450 to approximately 1000 pg/microg of protein. In addition, Tg in orbital tissue extracts from patients with TAO was immunoprecipitated by a rabbit anti-Tg antibody, suggesting that it retained its native conformation. An anti-thyroxine (T4) antibody captured in solid-phase Tg from orbital tissue extracts, showing that it contained thyroid hormone residues and had therefore originated in the thyroid. Tg-anti-Tg immune complexes were not found in orbital tissues, suggesting that if an autoimmune reaction to Tg occurs in TAO, it is likely to be cell mediated.


Attractive forces between uncharged hydrophobic surfaces: direct measurements in aqueous solution.
Long, double-chained alkylammonium acetate surfactants are soluble in water and, under suitable conditions, adsorb onto sheets of muscovite mica, forming an electrically neutral, hydrophobic surface. Attractive forces measured between such surfaces are 10 to 100 times stronger than expected from van der Waals theory over distances D up to about 10 nanometers. The forces decay exponentially [with a force proportional to exp(-D/1.4)] instead of following the power-law behavior of continuum theory. The results of these and earlier experiments indicate that the strength of these attractive forces depends critically on the degree of hydrophobicity of the surface and is due to the long-range influence of the surface on the structure of water. In addition, for very hydrophobic surfaces, the cavitation effects on pulling the surfaces apart are described.


Angiotensin II sustains brain inflammation in mice via TGF-beta.
The renin-angiotensin-aldosterone system (RAAS) is a key hormonal system regulating blood pressure. However, expression of RAAS components has recently been detected in immune cells, and the RAAS has been implicated in several mouse models of autoimmune disease. Here, we have identified Ang II as a paracrine mediator, sustaining inflammation in the CNS in the EAE mouse model of MS via TGF-beta. Ang II type 1 receptors (AT1Rs) were found to be primarily expressed in CNS-resident cells during EAE. In vitro, astrocytes and microglia responded to Ang II treatment by inducing TGF-beta expression via a pathway involving the TGF-beta-activating protease thrombospondin-1 (TSP-1). TGF-beta upregulation in astrocytes and microglia during EAE was blocked with candesartan (CA), an inhibitor of AT1R. Treatment of EAE with CA ameliorated paralysis and blunted lymphocyte infiltration into the CNS, outcomes that were also seen with genetic ablation of AT1Ra and treatment with an inhibitor of TSP-1. These data suggest that AT1R antagonists, frequently prescribed as antihypertensives, may be useful to interrupt this proinflammatory, CNS-specific pathway in individuals with MS.


Mode of inhibition of electron transport by orthophenanthroline in chromatophores and reaction centers of Rhodopseudomonas sphaeroides.
In Rhodopseudomonas sphaeroides, light causes the transfer of electrons from bacteriochlorophyll to ubiquinone in the photochemical reaction centers. Electrons from this "primary" ubiquinone move on to a secondary ubiquinone. We have studied the manner in which o-phenanthroline inhibits the transfer of electrons from primary to secondary ubiquinone in chromatophores (intracytoplasmic membrane fragments) and isolated reaction centers of Rp. sphaeroides. The formation of anionic semiquinones, both primary and secondary, is signaled by an absorption band at 450 nm and by band shifts of bacteriopheophytin and bacteriochlorophyll in the near infrared. The pattern of band shifts is different for primary and secondary semiquinone, allowing us to distinguish which quinone has become semireduced. This point was established for isolated reaction centers [Vermeglio, A. & Clayton, R. K. (1977) Biochim. Biophys. Acta 461, 159-165] and is confirmed here for chromatophores. By adding o-phenanthroline at various times during a sequence of actinic light flashes and monitoring the resulting optical absorbance changes, we have found that o-phenanthroline inhibits electron transfer from primary semiquinone to secondary quinone if the latter is in its oxidized form, but not if the latter is semireduced. Our findings can explain the decay kinetics of delayed fluorescence from bacteriochlorophyll in Rp. sphaeroides as measured by R. P. Carithers and W. W. Parson [(1976) Biochim. Biophys. Acta 440, 215-232].


Fluorescent Staining of Silicone Micro- and Nanopatterns for Their Optical Imaging.
Performance of engineered surfaces can be enhanced by making them hydrophobic or superhydrophobic via coating them with low-surface-energy micro- and nanopatterns. However, the wetting phenomena of particularly irregular shape and spacing (super)hydrophobic patterns such as polysiloxane coatings are not yet fully understood from a microscopic perspective. Here, we show a new method to collect 3D confocal images from irregular polysiloxane micro- and nanorods from a single rod resolution to discuss their wetting response over long liquid/solid interaction times and quantify the length and diameter of these rods. To collect such 3D confocal images, fluorescent dye containing water droplets were left on our superhydrophobic and hydrophobic polysiloxane coated surfaces. Then their liquid/solid interfaces were imaged at different staining scenarios: (i) using different fluorescent dyes, (ii) when the droplets were in contact with surfaces, or (iii) after the droplets were taken away from the surface at the end of staining. Using such staining strategies, we could resolve the micro- and nanorods from root to top and determine their length and diameter, which were then found to be in good agreement with those obtained from their electron microscopy images. 3D confocal images in this paper, for the first time, present the long-time existence of more than one wetting state under the same droplet in contact with surfaces, as well as external and internal three-phase contact lines shifting and pinning. In the end, these findings were used to explain the time-dependent wetting kinetics of our surfaces. We believe that the proposed imaging strategy here will, in the future, be used to study many other irregular patterned (super)antiwetting surfaces to describe their wetting theory, which is today impossible due to the complicated surface geometry of these irregular patterns.


Predicting the unpredictable: issues for output-based funding in psychiatric services.
In this study a number of important issues relating to diagnosis related groups and psychiatry are highlighted. Retrospective analysis of admissions to a rural psychiatric admission unit was undertaken utilising two differing approaches to statistical analysis. Results revealed that diagnosis alone is not highly predictive of cost. However, when linked to other variables, greater explanation in variation of length of stay was achieved. Data from social variables yielded sufficiently interesting results to warrant further investigation. The authors conclude by reporting the resultant groups from the classification analysis and offer some suggestions for further research.


Transfer of Problem Solving Skills from Touchscreen to 3D Model by 3- to 6-Year-Olds.
Although much published research purports that young children struggle to solve problems from screen-based media and to transfer learning from a virtual to a physical modality, Huber et al. (2016)'s recent study on children solving the Tower of Hanoi (ToH) problem on a touchscreen app offers a clear counter example. Huber et al. (2016) reported that children transferred learning from media to the physical world. As this finding arguably differs from that of prior research in this area, the current study tests whether the Huber et al. (2016) results could be replicated. Additionally, we extended the scope of the Huber et al. (2016) work by testing a broader age range, including children as young as 3 years, and using a culturally distinct participant pool. The results of the current study verified Huber et al.'s (2016) conclusion that 4- to 6-year-old children are capable of transferring the ToH learning from touchscreen devices to the physical version of the puzzle. Children under 4 years of age, in contrast, showed little ability to improve at the ToH problem regardless of the practice modality-suggesting that a different problem-solving task is required to probe very young children's ability to learn from touchscreen apps.


Regulation of protein synthesis and degradation in L8 myotubes. Effects of serum, insulin and insulin-like growth factors.
We have examined the regulation of protein turnover in rat skeletal myotubes from the L8 cell line. We measured protein synthesis by the rates of incorporation of radiolabelled tyrosine into protein in the presence of a flooding dose of non-radioactive tyrosine. We monitored degradation of proteins labelled with radioactive tyrosine by the release of acid-soluble radioactivity into medium containing excess nonradioactive tyrosine. Extracellular tyrosine pools and intracellular tyrosyl-tRNA equilibrate rapidly during measurements of protein synthesis, and very little reutilization of the radiolabelled tyrosine occurs during degradation measurements. Measured rates of protein synthesis and degradation are constant for several hours, and changes in myotube protein content can be accurately predicted by the measured rates of protein synthesis and degradation. Most of the myotube proteins labelled with radioactive tyrosine for 2 days are degraded, with half-lives (t1/2) of approx. 50 h. A small proportion (less than 2.5%) of the radiolabelled proteins are degraded more rapidly (t1/2 less than 10 h), and, at most, a small proportion (less than 15%) are degraded more slowly (t1/2 greater than 50 h). A variety of agents commonly added to primary muscle cell cultures or to myoblast cell lines (18% Medium 199, 1% chick-embryo extract, antibiotics and antifungal agents) had no effect on rates of protein synthesis or degradation. Horse serum, fetal bovine serum and insulin stimulate protein synthesis and inhibit the degradation of long-lived proteins without affecting the degradation of short-lived proteins. Insulin-like growth factors (IGF)-1 and -2 also stimulate protein synthesis and inhibit protein degradation. The stimulation of protein synthesis and the inhibition of protein degradation are of similar magnitude (a maximum of approx. 2-fold) and display similar sensitivities to a particular anabolic agent. Insulin stimulates protein synthesis and inhibits protein degradation only at supraphysiological doses, whereas IGF-1 and -2 are effective at physiological concentrations. These and other findings suggest that IGFs may be important regulators of skeletal muscle growth during the fetal and early neonatal periods.


Echocardiographic features of aortic valve endocarditis in a dog, a cow, and a horse.
A horse, a cow, and a dog with aortic valve vegetative endocarditis were studied by M-mode echocardiography. Echocardiographic abnormalities of the aortic valve, mitral valve, and left ventricle were observed. These features were identical to those reported in human beings with aortic valve endocarditis. Abnormalities associated with aortic valve endocarditis included irregular thickening of the valve, multiple linear echoes in the aortic root, diastolic prolapse of the aortic vegetation, and diastolic fluttering of a torn aortic valve. Some of these features were found in each animal. The consequences of aortic regurgitation observed by echocardiography were left ventricular dilation, diastolic fluttering of the mitral valve, premature closure of the mitral valve, and left ventricular hyperkinesia.


Near-IR emissive chlorin-bacteriochlorin energy-transfer dyads with a common donor and acceptors with tunable emission wavelength.
Design, synthesis, and optical properties of a series of novel chlorin-bacteriochlorin energy transfer dyads are described. Each dyad is composed of a common red-absorbing (645-646 nm) chlorin, as an energy donor, and a different near-IR emitting bacteriochlorin, as an energy acceptor. Each bacteriochlorin acceptor is equipped with a different set of auxochromes, so that each of them emits at a different wavelength. Dyads exhibit an efficient energy transfer (≥0.77) even for chlorin-bacteriochlorin pairs with large (up to 122 nm) separation between donor emission and acceptor absorption. Excitation of the chlorin donor results in relatively strong emission of the bacteriochlorin acceptor, with a quantum yield Φf range of 0.155-0.23 in toluene and 0.12-0.185 in DMF. The narrow, tunable emission band of bacteriochlorins enables the selection of a series of three dyads with well-resolved emissions at 732, 760, and 788 nm, and common excitation at 645 nm. Selected dyads have been also converted into bioconjugatable N-succinamide ester derivatives. The optical properties of the described dyads make them promising candidates for development of a family of near-IR fluorophores for simultaneous imaging of multiple targets, where the whole set of fluorophores can be excited with the common wavelength, and fluorescence from each can be independently detected.


Convergence dynamics of stochastic Cohen-Grossberg neural networks with unbounded distributed delays.
This paper addresses the issue of the convergence dynamics of stochastic Cohen-Grossberg neural networks (SCGNNs) with white noise, whose state variables are described by stochastic nonlinear integro-differential equations. With the help of Lyapunov functional, semi-martingale theory, and inequality techniques, some novel sufficient conditions on pth moment exponential stability and almost sure exponential stability for SCGNN are given. Furthermore, as byproducts of our main results, some sufficient conditions for checking stability of deterministic CGNNs with unbounded distributed delays have been established. Especially, even when the spectral radius of the coefficient matrix is greater than 1, in some cases our theory is also effective.


Robot-assisted repair of ureteroileal anastomosis strictures: initial cases and literature review.
Ureteroileal anastomosis strictures are well-known complications of ileal conduit urinary diversion that occur in 4% to 8% of patients. Open surgical repair is the standard definitive treatment with minimally invasive, endoscopic approaches developed to prevent the need for major surgery when possible. Robot-assisted surgery has been applied to most primary urologic procedures, but the role of this surgery in the management of complications is undefined. We report our experience with two cases of robotic repair of ureteroileal anastomotic strictures after robot-assisted cystectomy, the first such cases to our knowledge, and review the literature regarding management of these strictures. Two patients underwent robot-assisted ureteroileal anastomosis revision for left-sided strictures of 1 and 6 cm in length after failed endoscopic management. Three ports were used in the first and four in the second procedure. The diseased segment was identified, and the healthy end of the ureter anastomosed to a new site on the conduit with a temporary stent. In the second case, the conduit was mobilized and brought to the end of the ureter for a tension-free anastomosis because of the length of the stricture. Both patients were discharged on the first postoperative day without complications and are without recurrence after nearly 2 and 3 years since the robotic procedure. Minimally invasive definitive revision of ureteroileal anastomotic strictures is feasible with a robotic surgical approach. The advantages of robotic instrumentation allowed successful repair in two patients after previous robot-assisted cystectomy and avoided major open surgery.


Differential staining of actin in metaphase spindles with 7-nitrobenz-2-oxa-1,3-diazole-phallacidin and fluorescent DNase: is actin involved in chromosomal movement?
The distribution and polymerization state of actin in metaphase rat kangaroo cells was studied by fluorescence microscopy. Formaldehyde-fixed, acetone-extracted cells were labeled with either of two types of actin probes. The first, 7-nitrobenz-2-oxa-1,3-diazole-phallacidin, has high affinity for F actin and does not bind monomeric G actin. The second was a conjugate of DNase I labeled with either tetramethylrhodamine or fluorescein. DNase binds with high affinity to G actin and with lesser affinity to F actin. The polymerization state of actin was deduced by comparing the fluorescence distribution of the phallacidin derivative with that of the fluorescent DNase. The results indicate that the pole-to-chromosome region of the metaphase spindle contains G actin but little if any conventional F actin. F actin is found concentrated in a diffuse distribution outside the spindle region in metaphase cells and returns to the interzone area between the chromosomes by early telophase. These results exclude spindle models for chromosomal movement that require more than about five F actin filaments per chromosome, support the hypothesis that F actin is involved in force generation for cell cleavage, and are not inconsistent with the possibility that actin outside the spindle may be involved in chromosomal movement.


Fast-moving bat ears create informative Doppler shifts.
Many animals have evolved adept sensory systems that enable dexterous mobility in complex environments. Echolocating bats hunting in dense vegetation represent an extreme case of this, where all necessary information about the environment must pass through a parsimonious channel of pulsed, 1D echo signals. We have investigated whether certain bats (rhinolophids and hipposiderids) actively create Doppler shifts with their pinnae to encode additional sensory information. Our results show that the bats' active pinna motions are a source of Doppler shifts that have all attributes required for a functional relevance: (i) the Doppler shifts produced were several times larger than the reported perception threshold; (ii) the motions of the fastest moving pinna portions were oriented to maximize the Doppler shifts for echoes returning from the emission direction, indicating a possible evolutionary optimization; (iii) pinna motions coincided with echo reception; (iv) Doppler-shifted signals from the fast-moving pinna portion entered the ear canal of a biomimetic pinna model; and (v) the time-frequency Doppler shift signatures were found to encode target direction in an orderly fashion. These results indicate that instead of avoiding or suppressing all self-produced Doppler shifts, rhinolophid and hipposiderid bats actively create Doppler shifts with their own pinnae. These bats could hence make use of a previously unknown nonlinear mechanism for the encoding of sensory information, based on Doppler signatures. Such a mechanism could be a source for the discovery of sensing principles not only in sensory physiology but also in the engineering of sensory systems.


Mitochondrial respiratory dysfunction in familiar parkinsonism associated with PINK1 mutation.
In the present study mitochondrial respiratory function of fibroblasts from a patient affected by early-onset parkinsonism carrying the homozygous W437X nonsense mutation in the PINK1 gene has been thoroughly characterized. When compared with normal fibroblasts, the patient's fibroblast mitochondria exhibited a lower respiratory activity and a decreased respiratory control ratio with cellular ATP supply relying mainly on enhanced glycolytic production. The quantity, specific activity and subunit pattern of the oxidative phosphorylation complexes were normal. However, a significant decrease of the cellular cytochrome c content was observed and this correlated with a reduced cytochrome c oxidase in situ-activity. Measurement of ROS revealed in mitochondria of the patient's fibroblasts enhanced O(2)(*-) and H(2)O(2) production abrogated by inhibition of complex I. No change in the glutathione-based redox buffering was, however, observed.


Symptomatic bacteriuria due to Ureaplasma and Mycoplasma in adults.
314 women, and 52 men were screened during 18 months, for the presence of Ureaplasma urealyticum and Mycoplasma hominis in a midstream specimens of urine. All the patients were adults with symptoms of urinary tract infection who consulted a physician at a Primary Health Center "José Castro Villagrana" in Tlalpan, D.F. México. Ureaplasma urealyticum was isolated in 54 (17.1%) of the women and 8 (15.4%) of the men. Only Escherichia coli and Gardnerella vaginalis were isolated more frequently in both groups, 36.6% and 25.4%, respectively in women and 58.5% and 30.8% in men Mycoplasma hominis was isolated in 12 (3.8%) in women and 4 (7.7%) in men. The bacteriuria due to fastidious bacteria or Ureaplasmas that is not detected by routine culture methods are relatively common and would not necessarily respond to antibiotic treatment for conventional urinary tract pathogens.


Lipid droplets hypertrophy: a crucial determining factor in insulin regulation by adipocytes.
Lipid droplets (LDs) hypertrophy in adipocytes is the main cause of energy metabolic system dysfunction, obesity and its afflictions such as T2D. However, the role of adipocytes in linking energy metabolic disorders with insulin regulation is unknown in humans. Human adipocytes constitutively synthesize and secrete insulin, which is biologically functional. Insulin concentrations and release are fat mass- and LDs-dependent respectively. Fat reduction mediated by bariatric surgery repairs obesity-associated T2D. The expression of genes, like PCSK1 (proinsulin conversion enzyme), GCG (Glucagon), GPLD1, CD38 and NNAT, involved in insulin regulation/release were differentially expressed in pancreas and adipose tissue (AT). INS (insulin) and GCG expression reduced in human AT-T2D as compared to AT-control, but remained unchanged in pancreas in either state. Insulin levels (mRNA/protein) were higher in AT derived from prediabetes BB rats with destructed pancreatic β-cells and controls than pancreas derived from the same rats respectively. Insulin expression in 10 human primary cell types including adipocytes and macrophages is an evidence for extrapancreatic insulin-producing cells. The data suggest a crosstalk between AT and pancreas to fine-tune energy metabolic system or may minimize the metabolic damage during diabetes. This study opens new avenues towards T2D therapy with a great impact on public health.


[Treatment of severe haemolytic-uraemic syndrome by dialysis (author's transl)].
Since 1973 haemodialysis was performed on 30 children with severe haemolytic-uraemic syndrome. Serial measurements were made of blood pressure, blood picture, renal function, complement values, excretion of fibrinogen split-products and beta2-microglobulin in urine. Of 22 children in the acute severe stage two died, in one case treatment having been started too late, in the other as a result of fulminating pneumococcal pneumonia. Twenty children survived without residual symptoms and with normal renal function. Of six children with severe progressive haemolytic-uraemic syndrome two developed terminal renal failure. In one child a renal transplantation has since been performed. One child requires chronic dialysis after nephrectomy. Three children died as a result of arterial hypertension, one of yeast septicaemia. Of two children with recurrent haemolytic-uraemic syndrome one developed terminal renal failure which was successfully treated by renal transplantation. One child died in a hypertensive crisis. The overall death rate of the group was 23%, in the group with the acute severe haemolytic-uraemic syndrome it was 9%. The results suggest that haemodialysis significantly improves the prognosis of severe haemolytic-uraemic syndrome.


Persistence of common alleles in two related populations or species.
Mathematical studies are conducted on three problems that arise in molecular population genetics. (1) The time required for a particular allele to become extinct in a population under the effects of mutation, selection, and random genetic drift is studied. In the absence of selection, the mean extinction time of an allele with an initial frequency close to 1 is of the order of the reciprocal of the mutation rate when 4Nv less than 1, where N is the effective population size and v is the mutation rate per generation. Advantageous mutations reduce the extinction time considerably, whereas deleterious mutations increase it tremendously even if the effect on fitness is very slight. (2) Mathematical formulae are derived for the distribution and the moments of extinction time of a particular allele from one or both of two related populations or species under the assumption of no selection. When 4Nv less than 1, the mean extinction time is about half that for a single population, if the two populations are descended from a common original stock. (3) The expected number as well as the proportion of common neutral alleles shared by two related species at the tth generation after their separation are studied. It is shown that if 4Nv is small, the two species are expected to share a high proportion of common alleles even 4N generations after separation. In addition to the above mathematical studies, the implications of our results for the common alleles at protein loci in related Drosophila species and for the degeneration of unused characters in cave animals are discussed.


Deep Sequencing of T-cell Receptor DNA as a Biomarker of Clonally Expanded TILs in Breast Cancer after Immunotherapy.
In early-stage breast cancer, the degree of tumor-infiltrating lymphocytes (TIL) predicts response to chemotherapy and overall survival. Combination immunotherapy with immune checkpoint antibody plus tumor cryoablation can induce lymphocytic infiltrates and improve survival in mice. We used T-cell receptor (TCR) DNA sequencing to evaluate both the effect of cryoimmunotherapy in humans and the feasibility of TCR sequencing in early-stage breast cancer. In a pilot clinical trial, 18 women with early-stage breast cancer were treated preoperatively with cryoablation, single-dose anti-CTLA-4 (ipilimumab), or cryoablation + ipilimumab. TCRs within serially collected peripheral blood and tumor tissue were sequenced. In baseline tumor tissues, T-cell density as measured by TCR sequencing correlated with TIL scores obtained by hematoxylin and eosin (H&E) staining. However, tumors with little or no lymphocytes by H&E contained up to 3.6 × 106 TCR DNA sequences, highlighting the sensitivity of the ImmunoSEQ platform. In this dataset, ipilimumab increased intratumoral T-cell density over time, whereas cryoablation ± ipilimumab diversified and remodeled the intratumoral T-cell clonal repertoire. Compared with monotherapy, cryoablation plus ipilimumab was associated with numerically greater numbers of peripheral blood and intratumoral T-cell clones expanding robustly following therapy. In conclusion, TCR sequencing correlates with H&E lymphocyte scoring and provides additional information on clonal diversity. These findings support further study of the use of TCR sequencing as a biomarker for T-cell responses to therapy and for the study of cryoimmunotherapy in early-stage breast cancer. Cancer Immunol Res; 4(10); 835-44. ©2016 AACR.


Hypercalcaemia as a prodromal feature of indolent Pneumocystis jivorecii after renal transplantation.
Following renal transplantation, hypercalcaemia is frequently caused by persisting hyperparathyroidism. Unregulated extrarenal 1,25-dihydroxyvitamin D (1,25(OH)(2)D) synthesis, which is well recognized as a cause of hypercalcaemia in granulomatous diseases, may also occur after kidney transplantation. This mechanism is also likely to be responsible for hypercalcaemia reported during treatment of cytomegalovirus and associated with acute symptomatic pneumocystis jivorecii pneumonia (PCP). Hypercalcaemia as a prodromal feature of indolent PCP has not been described. We report a renal transplant recipient who developed hypercalcaemia 30 months post-transplant due to extrarenal production of 1,25(OH)(2)D. Two months later, PCP was diagnosed and hypercalcaemia resolved after initiation of treatment.


Bioaerosols from the land application of biosolids in the desert southwest USA.
This study evaluated bioaerosol emissions during land application of Class B biosolids in and around Tucson, Arizona, to aid in developing models of the fate and transport of bioaerosols generated from the land application of biosolids. Samples were collected for 20 min at distances between 2 m and 20 m downwind of point sources, using an SKC BioSampler impinger. A total of six samples were collected per sampling event, which consisted of a biosolid spray applicator applying liquid biosolids to a cotton field. Each application represented one exposure. Samples were collected in deionised water amended with peptone and antifoam agent. Ambient weather conditions were also monitored every 10 min following initiation of sampling. Concurrently with downwind samples, background (ambient) air samples were collected to compensate for any ambient airborne microorganisms. In addition, biosolids samples were collected for analysis of target indicator and pathogenic organisms. Soil samples were also collected and analysed. Significant numbers of heterotrophic plate count (HPC) bacteria were found in air samples collected during the biosolid application process. These could have arisen from soil particles being aerosolised during the land application process. Aerosolised soil may contribute significantly to the amount of aerosolised microorganisms. Soil particles may be able to more readily aerosolise, due to their low density, small particle size and low mass. Aerosolised HPC bacteria found during biosolids land application were similar to those found during normal tractor operation on non-biosolids applied fields. Coliforms and coliphages were not routinely detected even though they were found to be present in the biosolids at relatively high concentrations, 10(6) and 10(4)/g (dry weight) of biosolids respectively. This could be due to the die-off rate of aerosolised Gram-negative bacteria or sorption to the solid portion of the biosolids. Low numbers of aerosolised coliphages may likewise be due to sorption phenomena. We theorise that only organisms in the aqueous phase of the biosolids were available to desorb and be aerosolised. Animal viruses, which were not detected in the biosolids, were likewise not detected in the aerosol samples. Clostridium perfringens was detected in only a small percent of aerosol samples although it was detected during all weather conditions; other microorganisms were detected during more favourable environmental conditions (relative humidity >10%). Despite the fact that many of these organisms were present in the biosolids at significant concentrations, their presence in bioaerosols generated during the land application of biosolids was limited to only a small percentage of samples. Bacteria as well as viruses may sorb to biosolids, which contain a high percentage of organic matter, and desorption during land application of biosolids may not readily take place; therefore, these microorganisms may not be readily aerosolised.


A lymphoblastoid response of human foetal lymphocytes to ultraviolet-irradiated herpes simplex virus.
Cultures of foetal lymphocytes were exposed to u.v.-irradiated herpes simplex virus (HSV). The cells responded with increased 6-3H-thymidine incorporation, the formation of clumps of enlarged lymphoblastoid cells and cell division. This response was first detected 3 to 4 days after exposure to virus material and was shown to be virus-dose dependent. The ability to stimulate foetal cells was considerably more u.v. resistant than infectivity. Two isolates of HSV type 2 (4663 and 37174), which had a high "transforming" ability, produced large numbers of non-infectious particles (particle: infectivity ratios in excess of 10(4)). The cells, which responded to u.v.-irradiated HSV with blastoid transformation, were associated with the non-E-rosetting (T-cell-depleted) subpopulation.


Mechanisms for the adverse effects of late gestational increases in maternal cortisol on the heart revealed by transcriptomic analyses of the fetal septum.
We have previously shown in sheep that 10 days of modest chronic increase in maternal cortisol resulting from maternal infusion of cortisol (1 mg/kg/day) caused fetal heart enlargement and Purkinje cell apoptosis. In subsequent studies we extended the cortisol infusion to term, finding a dramatic incidence of stillbirth in the pregnancies with chronically increased cortisol. To investigate effects of maternal cortisol on the heart, we performed transcriptomic analyses on the septa using ovine microarrays and Webgestalt and Cytoscape programs for pathway inference. Analyses of the transcriptomic effects of maternal cortisol infusion for 10 days (130 day cortisol vs 130 day control), or ∼25 days (140 day cortisol vs 140 day control) and of normal maturation (140 day control vs 130 day control) were performed. Gene ontology terms related to immune function and cytokine actions were significantly overrepresented as genes altered by both cortisol and maturation in the septa. After 10 days of cortisol, growth factor and muscle cell apoptosis pathways were significantly overrepresented, consistent with our previous histologic findings. In the term fetuses (∼25 days of cortisol) nutrient pathways were significantly overrepresented, consistent with altered metabolism and reduced mitochondria. Analysis of mitochondrial number by mitochondrial DNA expression confirmed a significant decrease in mitochondria. The metabolic pathways modeled as altered by cortisol treatment to term were different from those modeled during maturation of the heart to term, and thus changes in gene expression in these metabolic pathways may be indicative of the fetal heart pathophysiologies seen in pregnancies complicated by stillbirth, including gestational diabetes, Cushing's disease and chronic stress.


Evaluation of Abelmoschus moschatus extracts for antioxidant, free radical scavenging, antimicrobial and antiproliferative activities using in vitro assays.
Abelmoschus moschatus Medik. leaves and seeds are considered as valuable traditional medicine. The aromatic seeds of this plant are aphrodisiac, ophthalmic, cardio tonic, antispasmodic and used in the treatment of intestinal complaints and check queasiness. To give a scientific basis for traditional usage of this medicinal plant, the seed and leaf extracts were evaluated for their antioxidant, free radical scavenging, antimicrobial and antiproliferative activities. In this study, antioxidant, antimicrobial and antiproliferative activities of A. moschatus extracts were evaluated in a series of in vitro assay involving free radicals, reactive oxygen species and their IC50 values were also determined. The antioxidant activities of the seed and leaf extracts of A. moschatus were determined by total antioxidant, DPPH, and ferrous reducing antioxidant property (FRAP) methods. In addition, the antiproliferative activity was also evaluated using colorectal adenocarcinoma and retinoblastoma human cancer cell lines. Moreover, six bacterial reference strains, two gram-positive (Bacillus subtilis and Staphylococcus aureus), four gram-negative (Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Salmonella enterica paratyphi) and one fungal strain (Candida albicans) were used to evaluate its antimicrobial activity. The results from this study showed that the antioxidant activities of A. moschatus as determined by the total phenol, flavonoids, total antioxidant and FRAP methods were higher in leaf than that of the seed extracts. On the other hand, the aqueous overnight seed extract (AMS-I) has shown significant radical scavenging activity as in 1, 1- Diphenyl-2-picrylhydrazyl (DPPH), hydrogen peroxide, hydroxyl radical, superoxide and lipid peroxidation as compared to other seed and leaf extracts. The AMS-I and AML-IV have shown activity against six and seven microorganisms respectively. Simulteneously, AMS-IV and AML-IV have demonstrated potential antiproliferative activity against two human cell lines - Colorectal adenocarcinoma (COLO-205) and retinoblastoma (Y79). The seed and leaf extracts of A. moschatus possess significant antioxidant activity and could serve as free radical inhibitors or scavenger, or substitute, probably as primary antioxidants. The plant possesses moderate antibacterial activity against bacterial strains used in this study. Hydroalcoholic seed and leaf extracts also exhibited antiproliferative activity against two human cancer cell lines. A. moschatus may therefore, be a good candidate for functional foods as well as pharmaceutics.


Nestin expression in embryonic and adult human teeth under normal and pathological conditions.
Nestin is an intermediate filament most related to neurofilaments and expressed predominantly in the developing nervous system and muscles. In the present study we examined the in vivo distribution of nestin in human teeth during embryonic development and in permanent teeth under normal and pathological conditions. The results show that nestin is first expressed at the bell stage and that its distribution is restricted in pulpal cells located at the cusp area of the fetal teeth. In young permanent teeth, nestin is found only in functional odontoblasts, which produce the hard tissue matrix of dentin. Expression is progressively down-regulated and nestin is absent from older permanent teeth. In carious and injured teeth, nestin expression is up-regulated in a selective manner in odontoblasts surrounding the injury site, showing a link between tissue repair competence and nestin up-regulation under pathological conditions. In an in vitro assay system of human dental pulp explants, nestin is up-regulated after local application of bone morphogenic protein-4. A similar effect is seen in cultures of primary pulp cells during their differentiation into odontoblasts. Taken together, these results suggest that nestin plays a potential role in odontoblast differentiation during normal and pathological conditions and that bone morphogenic protein-4 is involved in nestin up-regulation.


Posttranslational modifications of FERREDOXIN-NADP+ OXIDOREDUCTASE in Arabidopsis chloroplasts.
Rapid responses of chloroplast metabolism and adjustments to photosynthetic machinery are of utmost importance for plants' survival in a fluctuating environment. These changes may be achieved through posttranslational modifications of proteins, which are known to affect the activity, interactions, and localization of proteins. Recent studies have accumulated evidence about the crucial role of a multitude of modifications, including acetylation, methylation, and glycosylation, in the regulation of chloroplast proteins. Both of the Arabidopsis (Arabidopsis thaliana) leaf-type FERREDOXIN-NADP(+) OXIDOREDUCTASE (FNR) isoforms, the key enzymes linking the light reactions of photosynthesis to carbon assimilation, exist as two distinct forms with different isoelectric points. We show that both AtFNR isoforms contain multiple alternative amino termini and undergo light-responsive addition of an acetyl group to the α-amino group of the amino-terminal amino acid of proteins, which causes the change in isoelectric point. Both isoforms were also found to contain acetylation of a conserved lysine residue near the active site, while no evidence for in vivo phosphorylation or glycosylation was detected. The dynamic, multilayer regulation of AtFNR exemplifies the complex regulatory network systems controlling chloroplast proteins by a range of posttranslational modifications, which continues to emerge as a novel area within photosynthesis research.


Endoluminal repair of aneurysms containing ostia of essential branch arteries: an experimental model.
To examine experimentally the feasibility of transfemoral endoluminal repair of aneurysms containing the ostia of essential branch arteries. In a canine model (n = 4), suprarenal aortic aneurysms were created by suturing an artificial patch onto an anterior arteriotomy. Following a 2-week recovery period, the dogs underwent endovascular exclusion of their aneurysms using an aortic stent-graft with separate renal artery branch grafts. Outcome was evaluated using angiography, intravascular ultrasound (IVUS), Doppler flow, invasive pressure monitoring, and autopsy, respectively. Successful creation and subsequent endovascular exclusion of the aneurysm using aortic stent-grafts and separate bilateral renal artery stent-grafts was achieved in all trials. Angiographically, all aneurysms were excluded from aortic flow and all renal arteries were patent at completion of the procedure. With IVUS, good graft apposition and absence of perigraft flow were demonstrated in all animals. Mean pressure in the aneurysmal sac at completion of the procedure was 40 +/- 7 mmHg, compared to a mean systemic blood pressure of 105 +/- 8 mmHg (p < 0.05). At autopsy, no gross intimal damage was seen in the aorta or the renal arteries, and intact aortic grafts and branch grafts without twisting, coiling, or kinking were found in all trials. In an acute animal model, suprarenal aortic aneurysms can be excluded from the circulation with preservation of renal flow using an endoluminally placed aortic stent-graft with separate branch grafts.


A Self-powered Neural Stimulator Based on Programmable Triboelectric Nanogenerators.
Modulation of peripheral nerve is an emerging field for neuroprosthesis and bioelectronic medicine. With the developing neural interfacing technology that directly communicates with peripheral nerves, several powering schemes have been investigated for long-term use of implantable devices such as wireless and conversion of human body energy. But due to the limitations such as energy conversion efficiency and complexity, none of these methods can fully replace the current battery-based neuroprosthetic systems. This study proposes a new scheme based on programmable triboelectric nanogenerators for self-powered neural stimulations. The device can generate current pulses of more than 100 V by slightly shaking the device. The capability of neural stimulation is validated by sciatic nerve stimulation. Furthermore, the shaking frequency can control the measured kicking force of the rat leg. This prototype can be further minimized and optimized for a fully implantable self-powered/wireless neuroprosthetic system.


Management patterns of gastric polyps in the United States.
Recent guidelines on endoscopic sampling recommend complete gastric polyp removal for solitary fundic polyps >10 mm, hyperplastic polyps >5 mm and all adenomatous polyps. We aim to describe endoscopic approach to polyps in the time period prior to the American Society of Gastrointestinal Endoscopy (ASGE) guidelines and to identify opportunities for clinical practice improvements. Retrospective review of the Clinical Outcome Research Initiative (CORI) database, including all oesophagogastroduodenoscopies (OGDs). Reviewers grouped interventions during procedures based on instruments used for polyp sampling by forceps or snare polypectomy. Logistic regression estimated the effect of variables of interest on method of polypectomy. Of 783 037 OGDs reported in the CORI database, 25 670 (3.3%) described gastric polyps and met the inclusion criteria. Mean gastric polyp size was 6.5±4.9 mm, and 46.2% and 14.5% were located in the corpus and antrum, respectively. Polyps in the forceps group were smaller than polyps in the snare group (5.7±4.0 mm vs 9.3±6.4 mm, respectively, p<0.001). We identified 1056 polyps (41.3%) >10 mm that only underwent forceps biopsy. Forceps were used more frequently in the gastric fundus. Snare polypectomy was underused in gastric polyps, per current ASGE guidelines. Anatomical location and endoscopic features of polyps were important predictors of the approach to gastric polypectomy.


Becoming an empowered parent. How do parents successfully take up their role as a collaborative partner in their child's specialist care?
The purpose of this article is to offer an improved understanding of how parents of children with long-term disabilities are empowered to successfully take up their role as decision-making partners in the design and delivery of the care of their child. The intention is to stimulate dialogue, encourage reflection and provide practical suggestions for health professionals working with children and their families. The reported findings are from a study which was guided by a constructivist grounded theory methodology. This involved an iterative process of repeated cycles of data collection and analysis, which comprised 12 semi-structured, in-depth interviews with 14 parents of children accessing paediatric services within a single National Health Service Trust. A novel model, explaining how the power im/balance and the perceived state of the therapeutic relationship influence how successfully a parent takes up their position in the collaborative partnership, is presented and discussed. It is suggested that by thoughtfully addressing the traditional hierarchy that exists within healthcare, health professionals might facilitate the development of a 'truly' therapeutic relationship, which can help promote parental empowerment.


Microtubule C-Terminal Tails Can Change Characteristics of Motor Force Production.
Control of intracellular transport is poorly understood, and functional ramifications of tubulin isoform differences between cell types are mostly unexplored. Motors' force production and detachment kinetics are critical for their group function, but how microtubule (MT) details affect these properties--if at all--is unknown. We investigated these questions using both a vesicular transport human kinesin, kinesin-1, and also a mitotic kinesin likely optimized for group function, kinesin-5, moving along either bovine brain or MCF7(breast cancer) MTs. We found that kinesin-1 functioned similarly on the two sets of MTs--in particular, its mean force production was approximately the same, though due to its previously reported decreased processivity, the mean duration of kinesin-1 force production was slightly decreased on MCF7 MTs. In contrast, kinesin-5's function changed dramatically on MCF7 MTs: its average detachment force was reduced and its force-velocity curve was different. In spite of the reduced detachment force, the force-velocity alteration surprisingly improved high-load group function for kinesin-5 on the cancer-cell MTs, potentially contributing to functions such as spindle-mediated chromosome separation. Significant differences were previously reported for C-terminal tubulin tails in MCF7 versus bovine brain tubulin. Consistent with this difference being functionally important, elimination of the tails made transport along the two sets of MTs similar.


[Sports diets and oral health].
During competition and exercise sessions, (elite) sportsmen may need additional carbohydrates, resulting in an increased number of food intakes during the day. An increased number of food intakes may induce an increased caries risk. Additionally, almost all carbohydrate-containing sportdrinks and sportpowders have a high acidity level. Where low body weights are essential for the competition, sportsmen are at risk for disordered eating or eating disorders. Dentists should be aware of oral health risks of sportsmen and should advise them properly.


Asthma variability in patients previously treated with beta2-agonists alone.
According to national asthma guidelines, asthma severity can be classified as intermittent, mild, moderate, or severe on the basis of lung function, symptoms, nighttime awakenings, and exacerbations. Although it is widely believed that patients might not remain consistently in any given severity category over time, few studies have examined this directly. We sought to assess the variability in disease severity-control among patients with persistent asthma who have not yet received an asthma maintenance treatment. We performed an analysis of asthma severity-control over time in placebo-treated patients (n = 85) from 2 randomized, double-blind, 12-week clinical trials in patients with asthma previously receiving beta(2)-agonists alone. Asthma severity-control was assessed on the basis of morning percent predicted peak expiratory flow, albuterol use, and symptoms. At baseline, all patients met the criteria for moderate or severe persistent asthma (mean FEV(1) of 64% of predicted value or albuterol use and symptoms on 4.7 and 6.0 days per week, respectively). The mean percentage of treatment weeks that patients met all criteria for intermittent, mild, moderate, and severe asthma were 9%, 14%, 71%, and 6%, respectively. On the basis of morning peak expiratory flow, patients were classified as having intermittent-mild, moderate, or severe disease on 52%, 41%, and 7% of days, respectively. With regard to days per week with albuterol use or asthma symptoms, patients spent 59% and 45% of weeks, respectively, in the intermittent and mild categories. Asthma control cannot be adequately assessed in many patients by using discrete point-in-time assessments of lung function, short-acting beta-agonist use, or asthma symptoms. This might lead to underestimation of disease severity and contribute to inadequate therapy and, ultimately, asthma morbidity.


Cathepsin L in secretory vesicles functions as a prohormone-processing enzyme for production of the enkephalin peptide neurotransmitter.
Multistep proteolytic mechanisms are essential for converting proprotein precursors into active peptide neurotransmitters and hormones. Cysteine proteases have been implicated in the processing of proenkephalin and other neuropeptide precursors. Although the papain family of cysteine proteases has been considered the primary proteases of the lysosomal degradation pathway, more recent studies indicate that functions of these enzymes are linked to specific biological processes. However, few protein substrates have been described for members of this family. We show here that secretory vesicle cathepsin L is the responsible cysteine protease of chromaffin granules for converting proenkephalin to the active enkephalin peptide neurotransmitter. The cysteine protease activity was identified as cathepsin L by affinity labeling with an activity-based probe for cysteine proteases followed by mass spectrometry for peptide sequencing. Production of [Met]enkephalin by cathepsin L occurred by proteolytic processing at dibasic and monobasic prohormone-processing sites. Cellular studies showed the colocalization of cathepsin L with [Met]enkephalin in secretory vesicles of neuroendocrine chromaffin cells by immunofluorescent confocal and immunoelectron microscopy. Functional localization of cathepsin L to the regulated secretory pathway was demonstrated by its cosecretion with [Met]enkephalin. Finally, in cathepsin L gene knockout mice, [Met]enkephalin levels in brain were reduced significantly; this occurred with an increase in the relative amounts of enkephalin precursor. These findings indicate a previously uncharacterized biological role for secretory vesicle cathepsin L in the production of [Met]enkephalin, an endogenous peptide neurotransmitter.


Visuomotor integration of relevant and irrelevant angry and fearful facial expressions.
Our brain codes the features of perceptual events in a distributed fashion, raising the question of how information belonging to one event is processed without any interference of features from other events. Hommel (1998) suggested the "event file" concept to elucidate these mechanisms: an episodic memory trace "binding" together perceptual features and actions related to an object. Using a similar paradigm, we designed a pilot experiment and four additional experiments to investigate whether emotion, similarly than perceptual features, could bind with a motor response when the emotion was relevant and irrelevant for the task. Few studies have revealed this to be the case. We investigated how angry and fearful faces expressed by avatars and humans might be subject to a binding phenomenon. Our results show that at least three degrees of visuomotor binding seem to coexist: one implicating the relevant feature of the task with a strong effect on behavior, another implicating the location with a smaller behavioral effect (even if not task related), and a third implicating non-task-related features with behavioral effects only under specific conditions in which emotion could play a role. Our adaptation of Hommel's paradigm showed that emotional percepts can be subject to visuomotor binding effects even if the emotion is not task related confirming the important role of emotional information for the central nervous system. These findings offer new perspective in the investigation of the emotion-action binding at the neuronal level.


Parathyroid hormone-related protein as a common target molecule in specific immunotherapy for a wide variety of tumor types.
Parathyroid hormone-related protein (PTH-rP) has been considered to be responsible for malignancy-associated hypercalcemia and is thought to participate in pathological changes in bone metastases of cancer. In this study, we determined whether or not PTH-rP could be a common target molecule in specific immunotherapy for patients with a wide variety of tumor types. Various types of tumor cell lines were examined for PTH-rP expression at the mRNA and protein levels by RT-PCR, flow cytometry, and immunocytochemistry. We also determined whether or not cancer-reactive cytotoxic T lymphocytes (CTLs) could be induced from the peripheral blood mononuclear cells (PBMCs) of HLA-A24+ patients with gastric, colon, renal, or cervical cancer by in vitro stimulation with two PTH-rP peptides. As a result, PTH-rP mRNA was expressed in the majority of gastric, breast, lung, colon, cervical, and renal cancer cell lines. Expression of the protein was confirmed by both flow cytometry and immunocytochemistry. Furthermore, PTH-rP peptide-specific and cancer-reactive CTLs were successfully generated from the PBMCs of HLA-A24+ patients with different tumor types using in vitro stimulation with either the PTH-rP(102-111) or PTH-rP(110-119) peptide. These findings indicate that PTH-rP could be a common target molecule in specific immunotherapy for patients with a wide variety of tumor types, particularly bone metastases.


Role of the SphK-S1P-S1PRs pathway in invasion of the nervous system by SARS-CoV-2 infection.
Global spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still ongoing. Before an effective vaccine is available, the development of potential treatments for resultant coronavirus disease 2019 (COVID-19) is crucial. One of the disease hallmarks is hyper-inflammatory responses, which usually leads to a severe lung disease. Patients with COVID-19 also frequently suffer from neurological symptoms such as acute diffuse encephalomyelitis, brain injury and psychiatric complications. The metabolic pathway of sphingosine-1-phosphate (S1P) is a dynamic regulator of various cell types and disease processes, including the nervous system. It has been demonstrated that S1P and its metabolic enzymes, regulating neuroinflammation and neurogenesis, exhibit important functions during viral infection. S1P receptor 1 (S1PR1) analogues including AAL-R and RP-002 inhibit pathophysiological responses at the early stage of H1N1 virus infection and then play a protective role. Fingolimod (FTY720) is an S1P receptor modulator and is being tested for treating COVID-19. Our review provides an overview of SARS-CoV-2 infection and critical role of the SphK-S1P-SIPR pathway in invasion of SARS-CoV-2 infection, particularly in the central nervous system (CNS). This may help design therapeutic strategies based on the S1P-mediated signal transduction, and the adjuvant therapeutic effects of S1P analogues to limit or prevent the interaction between the host and SARS-CoV-2, block the spread of the SARS-CoV-2, and consequently treat related complications in the CNS.


The use of diatomite microcolumns for the chromatographic separation of steroids prior to radioimmunoassay.
Since most anti-steroid antibodies are not usually specific enough to permit direct radioimmunoassay of these compounds, chromatographic purification is required to achieve reliability. We have used Celite microcolumns for the separation of a wide range of steroids prior to radioimmunoassay. Depending on the polarity of the steroids to be separated, various mixtures of solvents are used as stationary and mobile phases. This system is rapid, easy, economical, and reliable.


Pressor effects elicited by stimulation within the medullary raphe nuclei of the guinea pig (Cavia porcellus).
The medullary raphe nuclei are involved in central autonomic regulation. In all species investigated, electrical stimulation of the raphe nuclei causes cardiovascular responses, although, these changes vary between species. The present study was designed to investigate the participation of these nuclei in cardiovascular regulation in the guinea pig. We studied the effects on arterial blood pressure (BP) and heart rate (HR) of electrical stimulation (isolated cathodal square wave pulses for 10 s at 100 Hz, 40-100 microA and 1-ms pulse duration) within the medullary raphe nuclei in urethane-anesthetized (1.2 g/kg, i.p.) guinea pigs (400-600 g, either sex). Electrical stimulation of the same sites was performed on a group of paralyzed (Flaxedil, 1 mg/kg, i.v.) and artificially ventilated animals. Stimulation sites were histologically defined and maps of the stimuli were obtained for the effect of electrical stimulation on arterial blood pressure. In another series of experiments L-glutamate (0.2 M) was microinjected (75 to 150 nl) into the nucleus raphe obscurus. Electrical stimulation of the raphe nuclei produced predominantly pressor responses (delta = +15 to +100 mmHg; 43% of the stimulated sites). Hypotension (delta = -10 to -25 mmHg, 24% of the stimulated sites), biphasic responses (2%) or no change in BP (31%) were evoked from fewer stimulation sites. Pressor responses were also predominant in paralyzed animals (delta = +15 to +95 mmHg; 47% of the stimulated sites), and after microinjection of L-glutamate into the raphe obscurus (A = +20 to +45 mmHg). The present results demonstrate that in the guinea pig the stimulation of these nuclei evokes mainly pressor responses. These responses are similar to those obtained in the rat and hamster but opposite to those observed in the cat and rabbit.


A simple method to evaluate groundwater vulnerability in urbanizing agricultural regions.
A methodology to evaluate groundwater vulnerability was developed and tested in a case study in the Central Valleys of the state of Oaxaca, Mexico, a region known for intensive agricultural activities and poor water management policies. An analysis was conducted to create and evaluate scenarios reflecting anthropogenic and natural stressors on groundwater using an analytical hierarchy process (AHP) and geographic information systems. Uncertainty in the vulnerability model was assessed using a Monte Carlo analysis. Five indices (abstraction (Abs), pollution (Po), runoff (Ru), groundwater recharge (Re), and marginalization (Ma)) were selected after an evaluation of the effects of population growth, climatology, hydrogeological features, and social marginalization on access to groundwater. Abstraction, pollution, and recharge rates are the main drivers of groundwater vulnerability, accounting for 87% of the vulnerability. The analysis revealed that the proposed model generates consistent results and contains low uncertainty. It also showed that more than 50% of the region's groundwater is moderately, and the vulnerability has become increasingly with abstraction, reduced recharge, and pollution (the most sensitive indices), indicating that groundwater in the Central Valleys is under great stress. Pollution and abstraction of groundwater resources are expected to rise in the more vulnerable areas, which will increase water crises and reduce access to water in rural communities. The approach and the indicators establish a baseline for the management and protection of water resources in developing countries where high-resolution data are lacking.