Arabidopsis group Ie formins localize to specific cell membrane domains, interact with actin-binding proteins and cause defects in cell expansion upon aberrant expression.
The closely related proteins AtFH4 and AtFH8 represent the group Ie clade of Arabidopsis formin homologues. The subcellular localization of these proteins and their ability to affect the actin cytoskeleton were examined. AtFH4 protein activity was identified using fluorimetric techniques. Interactions between Arabidopsis profilin isoforms and AtFH4 were assayed in vitro and in vivo using pull-down assays and yeast-2-hybrid. The subcellular localization of group Ie formins was observed with indirect immunofluorescence (AtFH4) and an ethanol-inducible green fluorescent protein (GFP) fusion construct (AtFH8). AtFH4 protein affected actin dynamics in vitro, and yeast-2-hybrid assays suggested isoform-specific interactions with the actin-binding protein profilin in vivo. Indirect immunofluorescence showed that AtFH4 localized specifically to the cell membrane at borders between adjoining cells. Expression of an AtFH8 fusion protein resulted in GFP localization to cell membrane zones, similar to AtFH4. Furthermore, aberrant expression of AtFH8 resulted in the inhibition of root hair elongation. Taken together, these data suggest that the group Ie formins act with profilin to regulate actin polymerization at specific sites associated with the cell membrane.


Molecular modeling of green fluorescent protein: structural effects of chromophore deprotonation.
Molecular dynamics (MD) simulations were carried out to study the conformational rearrangement induced by deprotonation of the fluorescent chromophore in GFP, as well as the associated changes in the hydrogen-bonding network. For both the structures with either a neutral or an anionic chromophore, it was found that the beta-barrel was stable and rigid, and the conformation of the chromophore was consistent with the available x-ray structure. The conformational change in Thr203 due to deprotonation was also found to be consistent with the three-state isomerization model. Although GFP is highly fluorescent, denatured-GFP is nonfluorescent, indicating that the environment of the protein plays an important role in its fluorescence behavior. Our MD simulations, which explore the effect of the protein shell on the conformation of the chromophore, find the flexibility of the central chromophore to be significantly restricted due to the rigid nature of the protein shell. The hydrogen-bonding between the chromophore and neighboring residues was also shown to contribute to the chromophore rigidity. In addition to the MD studies, quantum mechanics/molecular mechanics (QM/MM) ONIOM calculations were carried out to investigate the effect of the beta-barrel on the internal rotation in the chromophore. Along with providing quantitative values for torsional rotation barriers about the bridging bond in the chromophore, the ONIOM calculations also validate our MD force field parameters.


Immunogenicity and safety of a novel quadrivalent meningococcal conjugate vaccine (MenACWY-CRM) in healthy Korean adolescents and adults.
This phase III placebo-controlled study evaluated the immunogenicity and safety of MenACWY-CRM vaccination in healthy Korean adolescents and adults. Serum bactericidal activity with human complement (hSBA) was measured before and 1 month after vaccination against all four meningococcal serogroups. The IgG concentration specific for serogroup W capsular polysaccharide was measured in a subset of subjects in a post-hoc analysis. Adverse reactions were monitored throughout the study. Four hundred and fifty subjects were randomized 2:1 to receive MenACWY-CRM (N=297) or a saline placebo (N=153). MenACWY-CRM induced a good immune response against all four serogroups, with seroprotection rates (hSBA titers ≥8) of 79%, 99%, 98%, and 94% for serogroups A, C, W, and Y, respectively. Seroresponse rates were high for serogroups A, C, and Y, i.e. 76%, 86%, and 69%, respectively; the rate for serogroup W was 28%. MenACWY-CRM vaccine induced serum bactericidal antibodies against all four serogroups in a majority of subjects regardless of their baseline hSBA titers. MenACWY-CRM was generally well tolerated with most reactions being transient and mild to moderate in severity. Findings of this first study of a quadrivalent meningococcal polysaccharide conjugate vaccine in Korean adults and adolescents demonstrated that a single dose of MenACWY-CRM was well tolerated and immunogenic, as indicated by the percentages of subjects with hSBA titers ≥8 (79%, 99%, 98%, and 94% of subjects) and geometric mean titers (48, 231, 147, and 107) against serogroups A, C, W, and Y, respectively, at 1 month post-vaccination.


Extended Amygdala Neuropeptide Circuitry of Emotional Arousal: Waking Up on the Wrong Side of the Bed Nuclei of Stria Terminalis.
Sleep is fundamental to life, and poor sleep quality is linked to the suboptimal function of the neural circuits that process and respond to emotional stimuli. Wakefulness ("arousal") is chiefly regulated by circadian and homeostatic forces, but affective mood states also strongly impact the balance between sleep and wake. Considering the bidirectional relationships between sleep/wake changes and emotional dynamics, we use the term "emotional arousal" as a representative characteristic of the profound overlap between brain pathways that: (1) modulate wakefulness; (2) interpret emotional information; and (3) calibrate motivated behaviors. Interestingly, many emotional arousal circuits communicate using specialized signaling molecules called neuropeptides to broadly modify neural network activities. One major neuropeptide-enriched brain region that is critical for emotional processing and has been recently implicated in sleep regulation is the bed nuclei of stria terminalis (BNST), a core component of the extended amygdala (an anatomical term that also includes the central and medial amygdalae, nucleus accumbens shell, and transition zones betwixt). The BNST encompasses an astonishing diversity of cell types that differ across many features including spatial organization, molecular signature, biological sex and hormonal milieu, synaptic input, axonal output, neurophysiological communication mode, and functional role. Given this tremendous complexity, comprehensive elucidation of the BNST neuropeptide circuit mechanisms underlying emotional arousal presents an ambitious set of challenges. In this review, we describe how rigorous investigation of these unresolved questions may reveal key insights to enhancing psychiatric treatments and global psychological wellbeing.


Terminal Ileum Thickness During Maintenance Therapy Is a Predictive Marker of the Outcome of Infliximab Therapy in Crohn Disease.
Mucosal healing has been associated with long-term response to therapy for Crohn disease (CD). However, little is known about the significance of terminal ileum (TI) transmural thickness in predicting clinical outcomes. In this retrospective observational cohort study, we examined the association of an index ultrasonographic assessment of TI thickness during the maintenance phase and the subsequent clinical outcome of CD in a cohort of patients treated with infliximab (IFX). Treatment failure was defined as treatment discontinuation because of lack of efficacy, a need for dose escalation, or surgery. Clinical response was defined as treatment continuation in the absence of any of the aforementioned failure criteria. Sixty patients with CD receiving IFX therapy were included in the study. The patients were followed for a median of 16 months (5-24 months) after an index intestinal ultrasound. Thirty-eight patients (63.3%) maintained response to the therapy and 22 patients (36.6%) failed the treatment, with a mean follow up of 10.5 months (6.5-17 months) vs 9.25 months (1-10.25 months), respectively. On univariate analysis, the only variables differing between treatment response and failure were a TI thickness of 2.8 vs 5 mm (P < 0.0001) and an IFX trough level of 6.6 vs 3.9 µg/mL (P = 0.008).On multivariable analysis, only a small bowel thickness of ≥4 mm was associated with the risk of treatment failure (odds ratio, 2.9; 95% CI, 1.49-5.55; P = 0.002). Our findings suggest that transmural thickness of ≥4 mm can predict subsequent treatment failure in patients with CD treated using IFX, indicating transmural thickness <4 mm as a potential novel valuable therapeutic target.


Sarcoidosis patient: an unexpected reaction to carbonic anhydrase enzyme inhibitor.
Ocular diseases are very common in many of the systemic diseases such as sarcoidosis, and may sometimes be the presenting symptom of the disease. In this case report, we present an unusual reaction of the sarcoid granuloma to carbonic anhydrase enzyme inhibitors (CAIs), which was encountered in a patient with ocular sarcoidosis. This observation was taken after a 2-week interval between a CT scan orbits and an MRI orbits which showed a decrease in size from 4×3×4 cm to 2.5×2.5×2 cm, respectively. We suspected the dorzolamide CAI to have had a significant role in the reduction in size. It is suggested that acidotic changes that occur due to the effect of the carbonic anhydrase inhibitor causes electrolyte imbalance, intracellular as well as extracellular, which lead to the reduction in the size of the granuloma.


Coupling chemical oxidation and biostimulation: Effects on the natural attenuation capacity and resilience of the native microbial community in alkylbenzene-polluted soil.
Coupling chemical oxidation with bioremediation could be a cost-effective system to cope with soil and groundwater pollution. However, the effects of chemical oxidation on autochthonous microbial communities are scarcely known. A detailed analysis that considers both the efficiency of the two technologies and the response of the microbial communities was performed on a linear alkylbenzene-polluted soil and groundwater samples. The impacts of a modified Fenton's reaction (MFR) at various dosages and of permanganate on the microbiota over 4 weeks were assessed. The permanganate and MFR negatively affected microbial abundance and activity. However, the resilience of certain microbial populations was observed, with a final increase in potential hydrocarbon-degrading populations as determined by both the alkB gene abundance and the predominance of well-known hydrocarbon-degrading phylotypes such as Rhodococcus, Ochrobactrum, Acinetobacter and Cupriavidus genera as determined by 16S rRNA-based DGGE fingerprinting. The assessment of the chemical oxidant impact on autochthonous microbiota should be considered for the optimization of coupled field remediation technologies.


Organocatalytic Phosphonylation of in Situ Formed o-Quinone Methides.
A new class of Brønsted acid catalysts based on N-heterocyclic phosphorodiamidic acids (NHPAs) has been developed. The NHPA catalyst promotes phospha-Michael addition reaction of trialkylphosphites to in situ generated ortho-quinone methides (o-QMs) for the construction of diaryl phosphonates in moderate to excellent yields with 1.5 mol % catalyst. Diastereoselective synthesis of P-chiral phosphinate esters is achieved with the use of dialkyl phenylphosphonites.


No oculomotor plant, no final common path.
The assumption that there is an oculomotor plant, a fixed relationship between motoneuron firing rate and eye position, is disproved by brainstem recording studies showing that this relationship depends on which supernuclear subsystem determines firing rate. But it remains possible that there is a final common path (FCP), a fixed relationship between firing rate and muscle force. But then, brainstem recording studies predict that lateral rectus (LR) forces (and probably medial rectus (MR) forces, as well) will be higher in converged than in unconverged gaze for a given eye position. We recently measured these forces and found that they are slightly lower in convergence, disproving the FCP hypothesis. Thus, even the relationship between motoneuron firing rate and muscle force is under supernuclear control. What peripheral oculomotor articulations could vary the relationship of firing rate to muscle force?: (1) Actively movable EOM pulleys could alter oculorotary muscle force for a given oculorotory innervation by altering muscle lengths. (2) 'Outer' motoneurons may function as gamma efferents in conjunction with palisade endings and non-twitch global EOM fibers. (3) Complex nonlinear interactions likely arise among both parallel and serially connected muscle fibers.


Estimation of the coupling matrix elements for one-electron transfer systems.
A semiempirical method, based on a correlation of ninety-nine calculated or measured values, is suggested for estimating the coupling matrix elements H(12) needed in a Landau-Zener calculation. The procedure is applicable to one-electron transfer problems, such as charge transfer and ion-ion recombination calculations, and requires only a knowledge of the crossing distance R(x), and readily available atomic properties such as ionization potentials, electron affinities, and atomic energy levels. A discussion follows explaining the possible applications and limitations of the method.


The Current Landscape of Immune Checkpoint Blockade in Hepatocellular Carcinoma: A Review.
For more than a decade, sorafenib has been the only systemic treatment option for patients with advanced hepatocellular carcinoma (HCC). However, rapid progress over the past few years led to approval of other angiogenesis inhibitors and several immune checkpoint blockers (ICBs) that have been added to the treatment armamentarium for advanced HCC. Moreover, the recent success of a combination of bevacizumab with atezolizumab signals an important change in the front-line treatment of HCC. This review summarizes rapidly emerging clinical data on the promise and challenges of implementing ICBs in HCC and discusses the unmet need of biomarkers to predict response or resistance to therapy. Two strategies to target immunosuppression in tumors are also discussed: one proven (vascular endothelial growth factor pathway inhibition) and one currently under investigation (transforming growth factor-β pathway inhibition). The rationale and preliminary evidence on how their inhibition may reprogram the immunosuppressive milieu and enhance the efficacy of ICBs in HCC are reviewed. The recent successes and failures of angiogenesis inhibitors and ICBs, alone and in combination, have provided important insights into how to implement this novel systemic therapy in HCC and led to new avenues to enhance immunotherapy efficacy in this disease.


Radiotherapy outpatient review: a nurse-led clinic.
The study reported here was a descriptive account of the activities which take place at a traditional doctor-held radiotherapy review clinic. Comparison is made with a nurse-led clinic. Outcome measures, including number of interventions, consultation time, waiting time and degree of involvement with other support services were evaluated. Nurse consultations lasted significantly longer than those of the doctor, although waiting time was reduced. A greater number of referrals and liaison with other support services occurred and resulted in increased continuity of care. The results suggest that specialist nursing staff, given appropriate medical support, provide more effective care by using a holistic model for reviewing patients undergoing outpatient radiotherapy.


Influence of developmental hip dislocation on femoral head sphericity: experimental study in dogs.
The aim of this study was to assess the growth and the changes in the shape of a femoral head in dog puppies with dislocated and unstable hip joints. Ten 5-week-old, mountain shepherd puppies were divided into two subgroups. In five animals, the femoral head was dislocated, and in the other five dogs, femoral head was reduced after the dislocation procedure, making the joint congruent but unstable. After 4 weeks the animals were euthanised. The optical method of contour measurement of the femoral head sphericity was used. The shape of the dislocated femoral head differed from the normal one. The medial part was extended into beard-like form. The overgrowth of the femoral heads was significant and from 38% to 61% larger compared with the normal contralateral head. The unstable head of the second group did not differ in the shape nor in its size from the contralateral normal one. The large adaptive and deforming changes of shape in the dislocated femoral heads in our study show the pronounced malformation of the joint. The shape of beard-like deformity may be explained by the enlargement of contact surface between the medial part of the head and illiac bone. The restricting modelling role of the acetabulum, the minor vascular impairment during the surgery and the short time of follow-up were possible factors contributing to the minimal changes of the affected femoral head in the unstable but congruent hip joint.


Nitric oxide is used as an orthograde cotransmitter at identified histaminergic synapses.
1. Nitric oxide (NO) is produced by the enzyme nitric oxide synthase (NOS) and serves as an interneuronal messenger. Here, an identified histaminergic neuron (C2) in the mollusk Aplysia californica is shown to contain NOS with the use of NADPH diaphorase staining, suggesting that NO and histamine are used as cotransmitters by neuron C2. 2. The NOS containing neuron, C2, evokes a very slow excitatory postsynaptic potential in two of its identified postsynaptic follower neurons that are insensitive to H1 and H2 histamine receptor antagonists. 3. The very slow excitatory postsynaptic potential is blocked by inhibitors of NOS, such as nitroarginine methyl ester, and suppressed by the NO scavenger reduced hemoglobin. 4. Treatments with compounds that release NO, such as nitrosocysteine, mimic the membrane depolarization and the decrease in membrane conductance in the follower that are characteristic of the very slow excitatory postsynaptic potential induced normally by the presynaptic C2 neuron. 5. These results indicate that NO is used as an orthograde synaptic cotransmitter at synapses between histaminergic neuron C2 and its followers that receive the very slow excitatory postsynaptic potential.


Use of glycan microarrays to explore specificity of glycan-binding proteins.
Microarrays of defined glycans represent a high throughput approach to determining the specificity of lectins, or more generally glycan-binding proteins (GBPs). The utility of a glycan microarray is directly related to the number and variety of the glycans available on the printed surface for interrogation by GBPs. The Consortium for Functional Glycomics (CFG), funded by the National Institute of General Medical Sciences (NIGMS), has generated a glycan microarray available to the public as an investigator-driven resource, where hundreds of GBPs have been analyzed. Here we describe the methods generally used by the CFG to prepare glycan arrays and interrogate them with GBPs. We also describe our new approach to normalizing glycan microarray data derived from concentration-dependent analyses of GBP binding, and the application of this approach with the plant lectin Sambucus nigra agglutinin (SNA-I) and human galectin-8. The use of glycan microarrays with this approach readily generates a prediction of the glycan determinants required for high affinity binding by a GBP.


Effect of rat (beta)-interferon on intracellular levels of hydrolases in rat embryonal fibroblasts (Wistar strain).
The effect of rat beta-interferon (IFN) on the intracellular level of thiole and acid proteases and alkaline phosphatases was investigated. When nontransformed rat embryonal fibroblasts (REF) (Wistar strain) were treated with homologous IFN, a time independent decrease of hydrolase enzyme levels was observed. IFN treatment of transformed cells lead to a time dependent decrease of thiole proteases within 90 min. The values for acid proteases remained unchanged after the short term IFN treatment.


Synthesis of solid 2-pyridylzinc reagents and their application in Negishi reactions.
In search of alternatives to unstable or unreliable 2-pyridylboron reagents, we have explored two new varieties of solid, moderately air-stable 2-pyridylzinc reagents. Both reagents can be manipulated in air and are competent nucleophiles in Negishi cross-coupling reactions.


Membrane potential changes, cAMP stimulation and contraction in osteoblast-like UMR 106 cells in response to calcitonin and parathyroid hormone.
A multiphasic alteration in membrane potential occurs in response to calcitonin (CT), parathyroid hormone (PTH) and dibutyryl cAMP in the osteoblast-like UMR 106.01 and UMR 106.06 cell lines. This response consists of a small transient hyperpolarization, followed by a transient depolarization, followed by a long-term hyperpolarization. Experiments with channel blockers indicate that the depolarizing phase results from deactivation of K+ channels that are blocked by quinine but not by tetraethylammonium (TEA), and that the long-term hyperpolarization results from activation of K+ channels that are not blocked by quinine or by TEA. Correlating with the stimulation of intracellular cAMP by CT, a small percentage of the UMR 106.06 cells, but not UMR 106.01 cells, contract in response to CT. Both cell lines show a larger percentage of cells contracting in response to PTH than to CT.


[Central of mechanical fan: organization, safety and quality].
The headquarters of mechanical fans is the unit of the hospital with purpose of organizing ventilation resources promoting control and preventive maintenance and organizational of these equipments. The objective of this study was to elaborate a proposal of implantation of a headquarters of mechanical fans in an academical hospital, subsidized by the identification of the male nurse's scientific technical knowledge on the theme ventilation mechanics and for the detection of problems originating from of the decentralized administration of the fans. It is treated of exploratory descriptive study with quantitative approach, accomplished with 13 male nurses of ICU. The information was collected through structured interviews and submitted the descriptive analysis of the content. The results reveal that the male nurses possess several doubts, fact evidenced by 100% of the interviewees that mentioned the need of training courses gone back to the nursing attendance to the patient in ventilation mechanics. The situations described by the male nurses in the daily they demonstrate that the decentralization of the administration of the mechanical fans is shown ineffective as the organization, safety and quality. The proposal of implantation of a headquarters of fans appears for improvements in the attendance, in the formation of human resources and in the production of the knowledge. CONCLUSÕES: It is ended that the current profile can be changed through the breaking of institutional paradigms and of the institution of innovative practices that you/they will reinforce the purpose of a hospital of great load gone back to teaching, he/she researches and extension.


Weathering the Storm: How Superstorm Sandy Shaped the Response to COVID-19.
Catastrophic weather, like a pandemic, can wreak havoc on hospital operations. NYU Langone Health in New York City has experienced the extremes of both phenomena. In 2012, Superstorm Sandy severely damaged the system's core facilities, disabled operations, and forced the evacuation of more than 220 patients to other health systems. In 2020, the arrival of the novel coronavirus severely disrupted operations again-but this time, we were better prepared. Our experience in dealing with Superstorm Sandy taught us critical lessons that supported our readiness for COVID-19. Those lessons can be summed up as preparation, organization, and innovation.


Colour vision in ants (Formicidae, Hymenoptera).
Ants are ecologically one of the most important groups of insects and exhibit impressive capabilities for visual learning and orientation. Studies on numerous ant species demonstrate that ants can learn to discriminate between different colours irrespective of light intensity and modify their behaviour accordingly. However, the findings across species are variable and inconsistent, suggesting that our understanding of colour vision in ants and what roles ecological and phylogenetic factors play is at an early stage. This review provides a brief synopsis of the critical findings of the past century of research by compiling studies that address molecular, physiological and behavioural aspects of ant colour vision. With this, we aim to improve our understanding of colour vision and to gain deeper insights into the mysterious and colourful world of ants. This article is part of the theme issue 'Understanding colour vision: molecular, physiological, neuronal and behavioural studies in arthropods'.


[Analysis of the local synthesis of Toxoplasma gondii-specific immunoglobulins in the anterior eye chamber fluid. Report of two cases].
We reported two cases strongly suspected of ocular toxoplasmosis, in which a puncture of anterior eye chamber was performed for a final verification of clinical diagnosis. In both patients, fresh focal lesions of retinochoroiditis associated with old pigmented retinal scars were diagnosed. The 62-year-old women had a serological evidence of recently acquired T. gondii infection with an increasing IgG titre of a low avidity and a presence of specific IgM and IgA antibodies. In the 13-year-old girl, an immunological profile of the chronic infection was shown, confirmed by a high IgG avidity value. In the patient with a clinical picture of recurrent congenital toxoplasmosis, intraocular production of T. gondii-specific IgG was found in anterior eye chamber. The analysis of the specific antibodies synthesis in ocular fluid by the Western blot seems to be a valuable immunodiagnostic method for a final diagnosis of ocular toxoplasmosis.


Skin prick test responses and allergen-specific IgE levels as predictors of peanut, egg, and sesame allergy in infants.
Ninety-five percent positive predictive values (PPVs) provide an invaluable tool for clinicians to avoid unnecessary oral food challenges. However, 95% PPVs specific to infants, the age group most likely to present for diagnosis of food allergy, are limited. We sought to develop skin prick test (SPT) and allergen-specific IgE (sIgE) thresholds with 95% PPVs for challenge-confirmed food allergy in a large population-based cohort of 1-year-old infants with challenges undertaken irrespective of SPT wheal size or previous history of ingestion. HealthNuts is a population-based, longitudinal food allergy study with baseline recruitment of 1-year-old infants. Infants were recruited from council-run immunization sessions during which they underwent SPTs to 4 allergens: egg, peanut, sesame, and cow's milk/shrimp. Any infant with a detectable SPT response was invited to undergo oral food challenge and sIgE testing. Five thousand two hundred seventy-six infants participated in the study. Peanut SPT responses of 8 mm or greater (95% CI, 7-9 mm), egg SPT responses of 4 mm or greater (95% CI, 3-5 mm), and sesame SPT responses of 8 mm or greater (95% CI, 5-9 mm) had 95% PPVs for challenge-proved food allergy. Peanut sIgE levels of 34 kUA/L or greater (95% CI, 14-48 kUA/L) and egg sIgE levels of 1.7 kUA/L or greater (95% CI, 1-3 kUA/L) had 95% PPVs for challenge-proved food allergy. Results were robust when stratified on established risk factors for food allergy. Egg SPT responses and sIgE levels were poor predictors of allergy to egg in baked goods. These 95% PPVs, which were generated from a unique dataset, are valuable for the diagnosis of food allergy in young infants and were robust when stratified across a number of different risk factors.


Gompertz model with delays and treatment: mathematical analysis.
In this paper we study the delayed Gompertz model, as a typical model of tumor growth, with a term describing external interference that can reflect a treatment, e.g. chemotherapy. We mainly consider two types of delayed models, the one with the delay introduced in the per capita growth rate (we call it the single delayed model) and the other with the delay introduced in the net growth rate (the double delayed model). We focus on stability and possible stability switches with increasing delay for the positive steady state. Moreover, we study a Hopf bifurcation, including stability of arising periodic solutions for a constant treatment. The analytical results are extended by numerical simulations for a pharmacokinetic treatment function.


A closer look into the GL7 antigen: its spatio-temporally selective differential expression and localization in lymphoid cells and organs in human.
The GL7 epitope was originally described as part of a late lymphocyte activation antigen expressed in mouse and widely used since then as a marker of germinal center. Here we report on its differential expression by rat and human immune cells and lymphoid organs. Expression pattern of the GL7 epitope in rats is similar to that described earlier in mice, namely that GL7 antigen appears only on lymphocytes after 48h activation. In humans lymphocytes, but not the differentiated cells of myeloid origin, express this epitope. The GL7 epitope is up-regulated upon in vitro activation of primary T cells, while a slightly decreased expression is found on B lymphocytes. Fluorescent immunohistochemistry shows discrete location of GL7(hi) cells in human tonsil. GL7 antibody intensely stains CD19(+), IgD(+), IgM(low) B lymphocytes found at the margin of B cell follicles. The GL7 epitope is constitutively and highly raft-associated in human lymphoid cells. Strong neuraminidase- and partial papain-sensitivity of the GL7 epitope on human lymphocytes indicates a sialic acid-containing epitope linked either to one (or more) membrane protein(s) or to lipids. The lymphocyte-restricted GL7 epitope expression and the activation-dependent bi-directional change in the amount of the epitope suggest a functional role for GL7 epitope linked to carbohydrate-based immunoregulation.


The role of vancomycin in the treatment paradigm.
Vancomycin was introduced in the United States in 1956 as a possible treatment for infections due to penicillin-resistant Staphylococcus aureus, but it was not used widely because of toxicity and the nearly simultaneous development of semisynthetic antibiotics and cephalosporins. Thus, its main indication was the treatment of serious gram-positive infections in penicillin-allergic patients. For susceptible strains of S. aureus, vancomycin was more rapidly bactericidal than penicillin, nafcillin, or cefazolin, and, in a rabbit model of S. aureus endocarditis, sterilization of vegetations was more rapid with vancomycin. In clinical practice, however, nafcillin remained the treatment of choice for staphylococcal bacteremia, largely because it had failure rates of only 4%. With the appearance of methicillin-resistant S. aureus and coagulase-negative staphylococci, vancomycin became the drug of choice for these infections. Recently, the efficacy of vancomycin has been questioned because of vancomycin's increasing minimum inhibitory concentrations among staphylococci, poor tissue penetration, and apparently slower bacterial killing than previously was recognized.


Congenital unilateral pulmonary venous atresia: definitive diagnosis and treatment.
Three cases of unilateral right-sided pulmonary venous atresia were evaluated over an 18-year period. These bring the total number of cases to 25 in the literature. The clinical presentation of all these patients was similar and consisted of recurrent pulmonary infections, asthma-like symptoms, and exercise intolerance. The patients presented in 1982 (patient 1, a 12-year-old boy), 1994 (patient 2, a 9-year-old girl), and 1999 (patient 3, a 13-year-old boy). All patients were evaluated with a chest roentgenogram, and patients 1 and 2 had a ventilation and perfusion scan. Patients 1 and 3 also had cardiac catheterization and pulmonary angiography. Patient 2 had a magnetic resonance imaging study of the chest. Only patient 3 had wedge pulmonary angiography. Although a rare congenital defect, this diagnosis should be strongly suspected based on the typical clinical presentation and the preliminary studies, such as the chest roentgenogram and ventilation and perfusion scan. However, for definitive diagnosis, cardiac catheterization with wedge pulmonary angiography is necessary. Anastomosis of the atretic pulmonary veins to the left atrium is a theoretical consideration. However, this may not be feasible due to pulmonary venous anatomy or significant pulmonary dysfunction with pulmonary vascular changes. In these circumstances, we recommend performing pneumonectomy to remove the nidus for repeated bouts of pulmonary infections, to eliminate the left-to-right shunt, and to eliminate the dead space contributing to exercise intolerance.


Characterization of bla(CMY)-encoding plasmids among Salmonella isolated in the United States in 2007.
Salmonella enterica is one of the most common bacterial causes of foodborne illness, and nontyphoidal Salmonella is estimated to cause ∼1.2 million illnesses in the United States each year. Plasmids are mobile genetic elements that play a critical role in the dissemination of antimicrobial resistance determinants. AmpC-type CMY β-lactamases (bla(CMY)) confer resistance to extended-spectrum cephalosporins and β-lactam/β-lactamase inhibitor combinations and are commonly plasmid-encoded. A variety of plasmids have been shown to encode CMY β-lactamases and certain plasmids may be associated with particular Salmonella serotypes or environmental sources. In this study, we characterized bla(CMY) β-lactamase-encoding plasmids among Salmonella isolates. Isolates of Salmonella from specimens collected from humans in 2007 were submitted to the Centers for Disease Control and Prevention National Antimicrobial Resistance Monitoring System laboratory for susceptibility testing. Three percent (65/2161) of Salmonella isolates displayed resistance to ceftriaxone (minimum inhibitory concentration [MIC] ≥4 mg/L) and amoxicillin/clavulanic acid (MIC ≥32 mg/L), a combination associated with the presence of a bla(CMY) mechanism of resistance. Sixty-four (98.5%) isolates were polymerase chain reaction-positive for bla(CMY) genes. Transformation and conjugation studies showed that 95% (61/64) of the bla(CMY) genes were plasmid-encoded. Most of the bla(CMY)-positive isolates were serotype Typhimurium, Newport, Heidelberg, and Agona. Forty-three plasmids were replicon type IncA/C, 15 IncI1, 2 contained multiple replicon loci, and 1 was untypeable. IncI1 plasmids conferred only the bla(CMY)-associated resistance phenotype, whereas IncA/C plasmids conferred additional multi-drug resistance (MDR) phenotypes to drugs such as chloramphenicol, sulfisoxazole, and tetracycline. Most of the IncI1 plasmids (12/15) were sequence type 12 by plasmid multi-locus sequence typing. CMY β-lactamase-encoding plasmids among human isolates of Salmonella in the United States tended to be large MDR IncA/C plasmids or single resistance determinant IncI1 plasmids. In general, IncI1 plasmids were identified among serotypes commonly associated with poultry, whereas IncA/C plasmids were more likely to be identified among cattle/beef-associated serotypes.


International association for the study of lung cancer (IASLC) lymph node map: radiologic review with CT illustration.
Accurate clinical or pretreatment stage classification of lung cancer leads to optimal treatment outcomes and improved prognostication. Such classification requires an accurate assessment of the clinical extent of regional lymph node metastasis. Consistent and reproducible regional lymph node designations facilitate reliable assessment of the clinical extent of regional lymph node metastasis. Regional lymph node maps, such as the Naruke lymph node map and the Mountain-Dresler modification of the American Thoracic Society lymph node map, were proposed for this purpose in the past. The most recent regional lymph node map to be published is the International Association for the Study of Lung Cancer (IASLC) lymph node map. The IASLC lymph node map supersedes all previous maps and should be used in tandem with the current seventh edition of the tumor, node, metastasis stage classification for lung cancer.


Laparoscopic-assisted resection of rectosigmoid carcinoma. Immediate and medium-term results.
The technique of laparoscopic-assisted resection of colorectal carcinoma has been established. However, whether such a procedure is beneficial to patients is uncertain. To review the immediate and medium-term results of laparoscopic-assisted resection in patients with rectosigmoid carcinoma. We attempted laparoscopic-assisted sigmoid colectomy or anterior resection in 50 patients with rectosigmoid carcinoma (ie, the study group). The results were compared with those of 50 matched patients who underwent conventional open resection in the immediate prelaparoscopic era (ie, the control group). The median follow-up times for the study and control groups were 32.8 and 39.1 months, respectively. The operating time was significantly longer (P < .001, Student t test), while the analgesic requirement was significantly less (P < .001, Mann-Whitney U test) and the duration of hospitalization was significantly shorter (P = .001, Mann-Whitney U test), in the study group than in the control group. The oncological clearance (ie, the number of lymph nodes removed and the distal resection margin), the complication rate, the disease-free rate, and the survival rate were comparable in the 2 groups. The immediate and medium-term results of laparoscopic-assisted resection of rectosigmoid carcinoma are promising.


The involvement of CYP1A2 and CYP3A4 in the metabolism of clozapine.
Clozapine (CLZ), an atypical neuroleptic with a high risk of causing agranulocytosis, is metabolized in the liver to desmethylclozapine (DCLZ) and clozapine N-oxide (CLZ-NO). This study investigated the involvement of different CYP isoforms in the formation of these two metabolites. Human liver microsomal incubations, chemical inhibitors, specific antibodies, and different cytochrome P450 expression systems were used. Km and Vmax values determined in human liver microsomes were lower for the demethylation (61 +/- 21 microM, 159 +/- 42 pmol min(-1) mg protein(-1) mean +/- s.d.; n = 4), than for the N-oxidation of CLZ (308 +/- 1.5 microM, 456 +/- 167 pmol min(-1) mg protein(-1); n = 3). Formation of DCLZ was inhibited by fluvoxamine (53 +/- 28% at 10 microM), triacetyloleandomycin (33 +/- 15% at 10 microM), and ketoconazole (51 +/- 28% at 2 microM) and by antibodies against CYP1A2 and CYP3A4. CLZ-NO formation was inhibited by triacetyloleandomycin (34 +/- 16% at 10 microM) and ketoconazole (51 +/- 13% at 2 microM), and by antibodies against CYP3A4. There was a significant correlation between CYP3A content and DCLZ formation in microsomes from 15 human livers (r=0.67; P=0.04). A high but not significant correlation coefficient was found for CYP3A content and CLZ-NO formation (r=0.59; P=0.09). Using expression systems it was shown that CYP1A2 and CYP3A4 formed DCLZ and CLZ-NO. Km and Vmax values were lower in the CYP1A2 expression system compared to CYP3A4 for both metabolic reactions. It is concluded that CYP1A2 and CYP3A4 are involved in the demethylation of CLZ and CYP3A4 in the N-oxidation of CLZ. Close monitoring of CLZ plasma levels is recommended in patients treated at the same time with other drugs affecting these two enzymes.


Percutaneous absorption of sunscreen agents from liquid paraffin: self-association of octyl salicylate and effects on skin flux.
This study provides an investigation of the availability of octyl salicylate (OS), a common sunscreen agent, from liquid paraffin and the effect of OS on skin permeability. A model membrane system to isolate the vehicle effect from membrane permeability has been developed. Partitioning of OS between liquid paraffin and aqueous receptor phases was conducted. Partition coefficients increased with increase in OS concentration. A range of OS concentrations in liquid paraffin was diffused across human epidermis and synthetic membranes into 4% bovine serum albumin in phosphate-buffered saline and 50% ethanol. Absorption profiles of OS obtained from silicone and low-density polyethylene (LDPE) membranes were similar to each other but higher than for the high-density polyethylene [HDPE (3 times)] membrane and human epidermis (15 times). The steady state fluxes and apparent permeability coefficients (Kp') obtained from the diffusion studies showed the same trends with all membranes, except for the HDPE membrane which showed greater increase in flux and Kp' at concentrations above 30%. IR spectra showed that several bands of OS were shifted with concentrations, and the molecular models further suggested that the main contribution to the self-association is from non-1,4 van der Waals interactions.


Effects of Enactment of Legislative (Public) Smoking Bans on Voluntary Home Smoking Restrictions: A Review.
The positive effects of worldwide increases in enactment of legislative bans on smoking in public areas have been well documented. Relatively little is known about the effects of such bans on voluntary home smoking behavior. Meanwhile, private spaces, such as homes, have replaced public spaces as the primary milieu of secondhand smoke exposure. A systematic search of peer-reviewed articles was conducted using multiple databases including Cochrane Library, Cinahl, Embase, Global Health, Health Star, Joanna Briggs, MEDLINE, PsycINFO, PAIS International, PubMed, and Web of Science. We examined peer-reviewed studies that considered the impact of legislation-based public smoking bans on enactment of private home smoking restrictions. Sixteen articles published between 2002 and 2014 were identified and included. Our results suggest overall positive effects post-legislative ban with the majority of studies demonstrating significant increases in home smoking restrictions. Studies focusing on smoking and nonsmoking samples as well as child populations are discussed in depth. Existing evidence indicates an overall significant positive effect post-legislative ban on voluntary home smoking restrictions. While disentangling these effects over space and time remains a challenge, scientific research has converged in dispelling any notion of significant displacement of smoking into the home. Policy makers, especially those in countries without existing public smoking legislation, can rest assured that these types of bans contribute to the minimization of tobacco-related harm. Findings converge in dispelling notions of displacement of smoking into the home as a consequence of legislative bans that prohibit smoking in public spaces. Evidence from the studies reviewed suggests that through their influence on social norms, legislative bans on smoking in public places may encourage citizens to establish voluntary home smoking restrictions, thus decreasing harm related to secondhand smoke.


Prevalence of extracranial carotid artery lesions at duplex in primary aldosteronism.
Renovascular hypertension and high renin hypertension were found to be associated with an excess prevalence of carotid artery atherosclerotic lesions and to a higher risk of stroke, respectively, as compared to low-to-normal renin hypertension. Primary aldosteronism, being characterized by hypertension and a chronically suppressed plasma renin activity, should be accompanied by a low prevalence of carotid artery lesions. To verify this hypothesis we investigated prospectively, by a high resolution duplex ultrasound technique, the prevalence of extracranial carotid artery lesions in a case-controlled study of 34 (22 women and 12 men, aged 22 to 76 years) patients with no history or symptoms of cerebrovascular disease. Primary aldosteronism was diagnosed in 17 patients; 12 had a surgically confirmed unilateral aldosterone-secreting adenoma; and 5 had idiopathic hyperaldosteronism. Each primary aldosteronism patient was individually matched with a control with primary hypertension for sex, race, age, body mass index, casual blood pressure levels, duration of hypertension, smoking, diabetes mellitus, total serum cholesterol, and triglycerides. After the matching, the two groups were similar in terms of demographic features and overall cardiovascular risk profile (all P = NS). However, plasma renin activity and aldosterone levels were significantly lower and higher, respectively, in primary aldosteronism than in primary hypertensive patients. In primary aldosteronism the overall prevalence of carotid artery lesions at duplex was 59%, not significantly different from that (53%) found in primary hypertensives. Thus, at variance with renovascular hypertension, primary aldosteronism is not associated with an excess prevalence of carotid artery lesions.


A Systematic Review on Serious Games in Attention Rehabilitation and Their Effects.
Attention is a basic and main mental task and can play an important role in the functioning of other brain abilities such as intelligence, memory, learning, and perception, and its deficit occurs in 80% of patients with traumatic brain injury. The use of game-based tools for rehabilitation is rapidly expanding. Cognitive rehabilitation via video games is an emerging hot topic in cognitive science. Serious games serve a specific purpose in addition to entertainment. They can be more engaging than exercises since they replace reward and motivation systems with real-world motivations as a complement for rehabilitation activities. This study was aimed at identifying and categorizing serious computer games used for attention rehabilitation and evaluating their effects. Six electronic databases (Scopus, PubMed, ISI, Embase, IEEE, and Cochrane) were searched in August 2021. The search strategy consisted of three main concepts of "serious game", "cognitive deficits", and "cognitive rehabilitation". The inclusion criteria were (1) journal articles, (2) English language, (3) being published in the last 10 years, (4) human participants, and (5) game-based intervention. In the 30 included studies, 22 unique games were utilized for attention rehabilitation. Lumosity (20%), Brain Age (Dr. Kawashima's Brain Training) (10%), and MoHRS (6.66%) were the most common games among the studies. There were (57%) casual, (23%) action, (10%) simulation, and (10%) multiple genres. Of the 47 tools used in the studies, 5 utilized cross-modal oddball attention tasks, 4 utilized game performance, 3 utilized the paced auditory serial additional test (PASAT), and the rest employed other tools. A total of 73 outcome measures were related to attention, 42 measures did not have significant results, 30 were significantly improved, 1 was significantly deteriorated, and 4 articles did not have any specific measures for attention evaluation. Thus, the results revealed the positive effect of serious games on attention. However, issues such as absence of scientific teams, the variety of the disorders that cause defects, the variety of criteria, differences in measurements, lack of long-term follow-up, insufficient RCT studies, and small sample sizes should be considered when designing, developing, and using game-based systems to prevent bias.


[The critical reading of scientific articles, an easy-to-use method].
Faced with the abundance of available literature, clinicians must follow a method in order to carry out a critical reading of scientific articles. This enables them to decide how relevant the selected article is to the issues specific to their area of work and to choose whether to undertake a basic critical reading or to embark on an active reading.


The upregulation of matrix metalloproteinase-9 in inflamed human dental pulps.
The matrix metalloproteinases (MMPs) participate in a wide variety of extracellular matrix degradation. Detailed knowledge of MMPs may be important for understanding the pathogenesis of pulpal inflammation. The purpose of this study was to compare MMP-9 expression in clinically healthy human pulp and inflamed human pulp tissue specimens. We compared the levels of MMP-9 between clinically healthy pulp and inflamed pulp tissues by using the semi-quantitative reverse-transcriptase polymerase chain reaction analysis. In addition, immunohistochemistry was used to identify the in situ localization of MMP-9 expression in pulp specimens. MMP-9 mRNA gene was found to be increased in inflamed pulps as compared with clinically healthy pulp tissues (p < 0.05). The results from immunohistochemistry demonstrated that MMP-9 expression was significantly higher in the inflamed pulps than clinically healthy pulps (p < 0.05). MMP-9 stain was detected in the odontoblasts, fibroblasts, inflammatory infiltrates, and endothelial cells. Taken together, MMP-9 may play an important role in the pathogenesis of pulpal inflammation.


Formation of Organometallic Intermediate States in On-Surface Ullmann Couplings.
Possible origins of the formation of organometallic intermediates in on-surface Ullmann couplings have been investigated by surface tunneling microscopy (STM) and density functional theory (DFT) calculations. We consider the case of iodobenzenes on the coinage metals copper, silver, and gold. We found experimental evidence for the formation of surface vacancies and the presence of metal adatoms in these coupling reactions, which are taken as a hint for the reactive extraction of surface atoms by adsorbates. In a second step, we demonstrate by ab initio molecular dynamics calculations for aryl-iodides on copper that metal atoms can be pulled out of the surface to form metal-organic species. By contrast, a thermally activated provision of a metal atom from the surface to form an adatom is energetically unfavorable. Finally, we investigate the mechanism and energetics of the reactive extraction of surface metal atoms by means of (climbing-image) nudged elastic band density-functional theory calculations for iodobenzene on copper, silver, and gold, and analyze our results in the light of the experimental findings.


Association Between Bone Mineral Density and Depressive Symptoms in a Population-Based Sample.
This analysis was conducted to determine the relationship between bone mineral density (BMD) and depressive symptoms in a population-based cohort. Data were extracted from the second phase of the Dallas Heart Study (DHS-2), a large, multiethnic population sample in Dallas County, Texas, from September 1, 2007, to December 31, 2009. Depressive symptom severity was measured with the 16-item Quick Inventory of Depressive Symptomatology-Self Report (QIDS-SR₁₆), which is derived from DSM-IV major depressive disorder criteria. BMD was measured using dual-energy x-ray absorptiometry. Multiple linear regressions examined the relationship between QIDS-SR₁₆ score and BMD controlling for age, body mass index, sex, ethnicity, smoking status, alcohol use status, serum 25-hydroxyvitamin D concentration, antidepressant use, and physical activity as measured by total vigorous and moderate metabolic equivalents. Subgroup analyses explored differences related to age. QIDS-SR₁₆ score was not a significant predictor of either lumbar spine or total hip T-score (β = -0.01, P = .61 and β = -0.02, P = .39) in the overall population (n = 2,285). There was a significant negative interaction term between age and QIDS-SR₁₆ group (β = -0.01, P = .01). In participants aged 60 years or older (n = 465), QIDS-SR₁₆ score was a significant predictor of BMD at the lumbar spine and total hip (β = -0.14, P = .003 and β = -0.12, P = .006, respectively). QIDS-SR₁₆ score did not significantly predict BMD in the overall DHS-2 sample. There was, however, a significant association observed in participants aged ≥ 60 years. Results suggest that diagnosis and treatment of depressive symptoms may be of clinical importance in older individuals, a subgroup at high risk for osteoporosis and fractures.


Progress in meeting today's demands in genital herpes: an overview of current management.
Treatment of genital herpes requires accurate diagnosis, patient support, and effective treatment. Diagnosis is usually straightforward for classic presentations characterized by vesicular lesions but can be challenging for atypical presentations, which are more common. Diagnosis of asymptomatic infection requires access to molecular technology or type-specific serologic assays. Misconceptions about herpes simplex infection are common and patient education is essential. Patient concerns extend beyond disease frequency and severity-the psychological impact should not be underestimated. Antiviral therapy is relevant at all stages of infection. Acyclovir, valacyclovir, and famciclovir are effective and well tolerated for genital herpes treatment. Continuous suppressive therapy controls all symptoms of recurrent disease and helps to relieve disease complications. The prodrugs valacyclovir and famciclovir offer easier, less-frequent dosing than required for acyclovir. Valacyclovir achieves effective suppression when taken once a day. Interventions to prevent genital herpes transmission and to control the global problem are urgently required.


Physiological insights from genetic manipulation of the renin-angiotensin system.
The renin-angiotensin system is one of the most widely studied endocrine systems. It has an important role in the regulation of normal homeostasis, and disturbances in this system may be important in numerous pathological states. This review will focus on the major insights and important questions raised from gene targeting of this system.


On the decay of distractor-response episodes.
Distractor-to-distractor repetition effects can be explained by retrieval and/or inhibitory processes. Interestingly, the two accounts predict different effects from repeated distractors: Inhibition theories always predict benefits, whereas stimulus-response-retrieval theories predict an interaction of response repetition and distractor repetitions, resulting in benefits with response repetitions and costs with response changes. In the present experiment the time-course and the temporal separability of a stimulus-response episode on distractor-to-distractor repetitions were analyzed. The results showed that the interaction of response repetition and distractor repetitions was affected by a simple decay function. In addition, distractor repetition effects were affected by the temporal separability. In concert, the data yield evidence for retrieval-based explanations of distractor-to-distractor repetitions.


Implication of viral microRNAs in the genesis and diagnosis of Epstein-Barr virus-associated tumors.
The Epstein-Barr virus (EBV) is tightly associated with a variety of human tumors, including Burkitt lymphoma and acquired immune deficiency syndrome-related lymphoma of B-cell origin, as well as nasopharyngeal carcinoma and gastric cancer of epithelial origin. The virus latently infects the host cells and expresses proteins and non-coding RNAs to achieve malignancy. MicroRNAs (miRNAs or miRs) are small RNAs consisting of 19-25 nucleotides, which directly bind to the 3'-untranslated region of mRNAs to promote degradation and inhibit translation of mRNAs. EBV-encoded miRs are generated from two regions of the viral genome, within the apoptosis regulator BHRF1 gene locus and near the BamHI A region in a latency type-dependent manner. In addition, EBV-encoded miRs epigenetically regulate the expression of molecules that are effectors of the cell cycle progression, migration, apoptosis and innate immunity, serving a vital role in supporting viral replication and occurrence of EBV-associated tumors. The feasibility of using such miRs as biomarkers for the diagnosis and prognosis of EBV-associated tumors is currently under investigation.


Widespread ischemic brain lesions caused by vasculopathy associated with neurofibromatosis type 1.
We report the autopsy findings of a 63-year-old man with neurofibromatosis type 1 (NF1), in whom widespread ischemic brain lesions caused by vasculopathy associated with the disorder were observed. The patient, who had café au lait macules, axillary freckling, and neurofibromas, was inarticulate of speech, had difficulty in maintaining a sitting position, and was hyporeactive at the age of 57 years. He then developed autonomic dysfunction, followed by consciousness disturbance and status epilepticus. Repeated MRI studies disclosed multiple, ill-defined lesions in the brain and progressive cerebral atrophy. The histopathological features of the lesions were those of ischemia that had occurred with spatiotemporal variability in the brain. Characteristically, many arteries in the subarachnoid space manifested accumulation of cells in the intimal layer: this hyperplasia had resulted in narrowing and occlusion of the lumen. Immunoblotting demonstrated a marked decrease of neurofibromin, the NF1 product, which is known to act as a functional molecule in the normal process of vascular maintenance and repair. This case provides useful information about the pathomechanisms underlying central nervous system manifestations in patients with NF1.


[Importance of ultrasound in the evaluation of fetal status].
Ultrasound made it possible to discover endangered fetus on time, before its clinical manifestation. Evaluation of fetal vitality is possible during the whole pregnancy 1. early pregnancy (fetal heart action, fetal movement,...) 2. and 2. late pregnancy (placental maturity, amniotic fluid volume, cardiothocography,...). Contemporary prenatal medicine cannot be imagined without ultrasound and its quality is in a direct relationship with the level of ultrasound diagnostic utilisation.


Water droplets as template for next-generation self-assembled poly-(etheretherketone) with cardo membranes.
Next generation PEEK-WC membranes have been fabricated by using an innovative self-assembly technique. Patterned architectures have been achieved via a solvent-reduced and water-assisted process, resulting in honeycomb packed geometry. The membranes exhibit monodisperse pores with size and shape comparable to those left by templating water droplets. Influencing factors for the formation of self-assembled poly-(etheretherketone) with Cardo [PEEK-WC] membranes have been evaluated, identifying the critical parameters for nucleation, growth, and propagation of the droplet-mobile arrays through the overall films. Structure-transport relationships have been discussed according to the results achieved from the implementation of membrane distillation processes, yielding indication about the suitability of self-assembled PEEK-WC films to work as interfaces in contactor operations.


Hepatic Granulomas in Murine Visceral Leishmaniasis Caused by Leishmania chagasi
Protozoa belonging to the genus Leishmania cause the diverse forms of human leishmaniasis, which range from self-healing cutaneous ulcers to widely disseminated or disfiguring disease. Human and murine immune responses to species causing cutaneous and visceral leishmaniasis are considerably different. In murine models of cutaneous leishmaniasis the expansion of distinct CD4(+) T-cell subsets is associated with either disease prevention (TH1) or disease exacerbation (TH2). However, studies of murine visceral leishmaniasis have not shown expansion of disease-exacerbating cells during progressive disease. The latter studies have concentrated on immune responses in splenocytes from mice infected with visceralizing Leishmania species, even though the parasites are found primarily in the livers of infected mice. The histologic response to visceralizing Leishmania sp. in the liver is one of granuloma formation, which proceeds with similar kinetics in resistant C3H and susceptible BALB/c mice. We isolated hepatic granulomas from BALB/c mice infected for 6 weeks with the visceralizing parasite L. chagasi. Study of dispersed granuloma cells by FACS and in vitro cultivation with antigen showed differences between the responses of immune cells from the spleens and liver granulomas of these infected animals. A careful dissection of immune responses in hepatic granuloma cells from susceptible or resistant mice infected with L. chagasi may lead to a better understanding of murine immune responses to these important pathogens.


Medical student selection and society: Lessons we learned from sociological theories.
The aim of this study was to show the interaction between the society, applicants and medical schools in terms of medical student selection. In this study, the trends to implement social factors in the selection process were highlighted. These social factors were explored through functionalism and conflict theories, each focusing on different categories of social factors. While functionalist theorists pay attention to diversity in the selection process, conflict theorists highlight the importance of socio-economic class. Although both theories believe in sorting, their different views are reflected in their sorting strategies. Both theories emphasize the importance of the person-society relationship in motivation to enter university. Furthermore, the impacts of social goals on the selection policies are derived from both theories. Theories in the sociology of education offer an approach to student selection that acknowledges and supports complexity, plurality of approaches and innovative means of selection. Medical student selection does not solely focus on the individual assessment and qualification, but it focuses on a social and collective process, which includes all the influences and interactions between the medical schools and the society. Sociological perspective of medical student selection proposes a model that envelops the individual and the society. In this model, the selection methods should meet the criteria of merit at the individual level, while the selection policies should aim at the society goals at the institutional level.


Bimanual adaptation: internal representations of bimanual rhythmic movements.
From tying your shoes and clipping your tie to the claps at the end of a fine seminar, bimanual coordination plays a major role in our daily activities. An important phenomenon in bimanual coordination is the predisposition toward mirror symmetry in the performance of bimanual rhythmic movements. Although learning and adaptation in bimanual coordination are phenomena that have been observed, they have not been studied in the context of adaptive control and internal representations-approaches that were successfully employed in the arena of reaching movements and adaptation to force perturbations. In this paper we examine the dynamics of the learning mechanisms involved when subjects are trained to perform a bimanual non-harmonic polyrhythm in a bimanual index finger tapping task. Subjects are trained in this task implicitly, using altered visual feedback, while their performance is continuously monitored throughout the experiment. Our experimental results indicate the existence of significant (p<<0.01) learning curves (i.e., error plots with significantly negative slopes) during training and aftereffects with a washout period after the visual feedback ceases to be altered. These results confirm the formation of internal representations in bimanual motor control. We present a simple, physiologically plausible, neural model that combines feedback and adaptation in the control process and which is able to reproduce key phenomena of bimanual coordination and adaptation.


Wheel running as a predictor of cocaine self-administration and reinstatement in female rats.
Avidity for behaviors mediated by nondrug rewards, such as novelty seeking or intake of sweets or fats, is predictive of enhanced vulnerability to the locomotor-activating and rewarding effects of drugs of abuse. The purpose of the present study was to determine whether avidity for wheel running was predictive of subsequent cocaine-induced locomotor activity, cocaine self-administration, and cocaine-seeking behavior in rats. Rats with high (HiR) and low (LoR) levels of wheel running were selected from an outbred sample of Wistar rats. These rats were first tested for their locomotor response to an acute injection of cocaine (10 mg/kg, i.p.). Subsequently, a multi-phase self-administration procedure was used to examine the effect of wheel running on the maintenance, extinction, and cocaine-induced reinstatement of cocaine-seeking behavior in HiR and LoR rats. The results indicate no significant differences between HiR and LoR rats in the cocaine-induced stimulation of locomotor activity. During maintenance, HiR rats self-administered more cocaine than LoR rats. While there were no group differences in saline self-administration behavior during extinction, HiR rats showed higher cocaine-induced reinstatement than LoR rats. Rats that were previously high responders to novelty (day 1 in locomotor track) also showed significantly higher reinstatement than low novelty responders. These results suggest that a propensity for wheel running is associated with increased vulnerability for cocaine self-administration and reinstatement and that HiR rats are more motivated than LoR rats to seek cocaine.


Oxalate metabolism by tobacco leaf discs.
The turnover rate of oxalate in leaf discs of Nicotiana tabacum, var Havana Seed, during photosynthesis was estimated to be 1 to 2 micromoles per gram fresh weight per hour. Radioactivity from the enzymic oxidation of [(14)C]oxalate rapidly appeared in neutral sugars (mainly sucrose), organic acids (mainly malate), and amino acids. Only 5% of the radioactivity was released to the atmosphere as (14)CO(2), and no formate or formaldehyde could be detected. The metabolism of oxalate was not increased by raising the O(2) concentration from 1% to 21% to 60%, nor was the formation of [(14)C]oxalate from [2-(14)C]glyoxylate changed under the same conditions as was previously observed in vitro (Havir 1983 Plant Physiol 71: 874-878). While oxalate is not an inert end product of the glycolate pathway, it contributes little to the formation of photorespiratory CO(2).


Neonatal 5-hydroxytryptamine depletion induces depressive-like behavior in adult rats.
The influence of neonatal serotonergic lesion on adult behavior in locomotor and depression models was studied in male Wistar rats. When 3-day-old rats were injected intracisternally with 5,7-dihydroxytryptamine (5,7-DHT), a marked depletion of brain 5-HT was observed when animals were killed 3 months after the treatment. Brain catecholamine content was generally not changed by neurotoxin treatment. The behavioral consequence of intracisternal 5,7-DHT administration to developing rats consisted in reduction of adult rats' activity in the forced swimming test. Both desipramine, and, to the lesser extent, fluoxetine, reversed 5,7-DHT-induced immobility of animals.


Vitamin D deficiency and primary hyperparathyroidism.
Vitamin D via its receptor has essential actions on parathyroid cells, inhibiting PTH secretion, and parathyroid cell proliferation. While the effects of vitamin D depletion in the pathogenesis of secondary hyperparathyroidism in elderly individuals or in the occurrence of parathyroid hyperplasia in patients with renal insufficiency are well established, the association between hypovitaminosis D and primary hyperparathyroidism (P-HPT) has only recently become appreciated. In different cohorts of patients with P-HPT, vitamin D deficiency has been recently associated with higher PTH levels, larger adenomas, and a more severe phenotype (including osteitis fibrosa cystica) as well as negative post-operative outcomes following parathyroidectomy. Despite current guidelines recommend measurement of serum 25OHD (25-hydroxy-cholecalciferol) in P-HPT and their repletion if the levels are <20 ng/ml, future well-designed trials of vitamin D supplementation in P-HPT patients with coexisting vitamin D deficiency are needed to evaluate the risk/benefit profile of this treatment.


T-wave ion mobility-mass spectrometry: basic experimental procedures for protein complex analysis.
Ion mobility (IM) is a method that measures the time taken for an ion to travel through a pressurized cell under the influence of a weak electric field. The speed by which the ions traverse the drift region depends on their size: large ions will experience a greater number of collisions with the background inert gas (usually N(2;)) and thus travel more slowly through the IM device than those ions that comprise a smaller cross-section. In general, the time it takes for the ions to migrate though the dense gas phase separates them, according to their collision cross-section (Omega). Recently, IM spectrometry was coupled with mass spectrometry and a traveling-wave (T-wave) Synapt ion mobility mass spectrometer (IM-MS) was released. Integrating mass spectrometry with ion mobility enables an extra dimension of sample separation and definition, yielding a three-dimensional spectrum (mass to charge, intensity, and drift time). This separation technique allows the spectral overlap to decrease, and enables resolution of heterogeneous complexes with very similar mass, or mass-to-charge ratios, but different drift times. Moreover, the drift time measurements provide an important layer of structural information, as Omega is related to the overall shape and topology of the ion. The correlation between the measured drift time values and Omega is calculated using a calibration curve generated from calibrant proteins with defined cross-sections(1). The power of the IM-MS approach lies in its ability to define the subunit packing and overall shape of protein assemblies at micromolar concentrations, and near-physiological conditions(1). Several recent IM studies of both individual proteins(2,3) and non-covalent protein complexes(4-9), successfully demonstrated that protein quaternary structure is maintained in the gas phase, and highlighted the potential of this approach in the study of protein assemblies of unknown geometry. Here, we provide a detailed description of IMS-MS analysis of protein complexes using the Synapt (Quadrupole-Ion Mobility-Time-of-Flight) HDMS instrument (Waters Ltd; the only commercial IM-MS instrument currently available)(10). We describe the basic optimization steps, the calibration of collision cross-sections, and methods for data processing and interpretation. The final step of the protocol discusses methods for calculating theoretical Omega values. Overall, the protocol does not attempt to cover every aspect of IM-MS characterization of protein assemblies; rather, its goal is to introduce the practical aspects of the method to new researchers in the field.


Primary structure of the P450 lanosterol demethylase gene from Saccharomyces cerevisiae.
We have sequenced the structural gene and flanking regions for lanosterol 14 alpha-demethylase (14DM) from Saccharomyces cerevisiae. An open reading frame of 530 codons encodes a 60.7-kDa protein. When this gene is disrupted by integrative transformation, the resulting strain requires ergosterol and, as expected, grows only in the absence of oxygen. The deduced amino acid sequence of 14DM includes a hydrophobic segment near the amino terminus which may be a transmembrane domain. The deduced sequence has been compared with those of eight other eukaryotic P450s, each from a different family within the P450 superfamily. These comparisons indicate that this yeast gene is the first member of a new P450 family, P450LI. The P450, designated P450LIA1, is more closely related to mammalian P450s than to the bacterial P450cam. In fact, both the yeast P450 and several mammalian P450s have equivalent alignment scores when each is compared with the bovine P450scc. Matrix comparisons of the amino acid sequence of this P450 with those of mammalian P450s reveal three conserved regions. The DNA region 5' to the structural 14DM gene includes poly(dA:dT) sequences and a repeating hexamer sequence.


Impaired detection of Mycobacterium tuberculosis immunity in patients using high levels of immunosuppressive drugs.
We have previously shown, in renal transplant recipients on maintenance immunosuppression, that a whole-blood assay was superior in detecting immunity towards purified protein derivative (PPD) compared with skin testing. As blood tests may have limitations during high-dose immunosuppression therapy, the present study was aimed at characterising the effect of high immunosuppressive drug levels on PPD-specific T-cell immunity. PPD-reactive CD4 T-cells from 13 renal transplant recipients were longitudinally quantified by the induction of cytokines using flow cytometry. To further address the effect of high and low maintenance immunosuppression, drug effects were studied in vitro and in 49 age-matched lung transplant recipients and 49 renal transplant recipients. Maintenance immunosuppression after renal transplantation did not affect PPD-specific T-cell detection (median T-cell frequencies 0.55% before and 0.46% >12 months after transplantation), whereas specific T-cell frequencies were significantly lower 3 months after transplantation (0.15%; p = 0.0002). Likewise, high-level maintenance immunosuppression after lung transplantation was associated with a significantly lower prevalence in PPD-specific T-cell reactivity compared with renal transplant recipients (16.7% versus 52.1%; p = 0.0005). In line with the observations made in vivo, calcineurin inhibitors analysed in vitro led to a dose-dependent decrease in antigen-specific T-cell reactivity. The flow cytometric assay is not adversely affected by low drug doses. In contrast, decreased levels of PPD-specific T-cells early after transplantation and low prevalence of PPD-reactivity in lung transplant recipients suggest a reduced sensitivity of in vitro testing during high-level immunosuppression.


The Role of Digitization in the Rapid Reproduction of an Obturator in a Frail Elderly Patient.
This case report describes the reproduction of the current obturator of a frail elderly patient using the digitization concept. A computed tomography scanner was used to scan the obturator. The virtual model was created using three-dimensional modeling software. The model was printed using a plastic with a low melting point with a fused deposition modeling technique in a three-dimensional printer and then conventionally processed. The definitive obturator was inserted and evaluated for fit, function, and esthetics and was successfully delivered at the second visit. The patient was satisfied with the oral function and esthetics of the reproduced obturator. This case report confirms the effectiveness of digitization in the reproduction of an obturator that enabled satisfactory oral function and esthetics.


Mutagenic activation of IQ, PhIP and MeIQx by hepatic microsomes from rat, monkey and man: low mutagenic activation of MeIQx in cynomolgus monkeys in vitro reflects low DNA adduct levels in vivo.
Cooked meat, poultry and fish contain a number of mutagenic and carcinogenic heterocyclic amines, including 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). In the present study we examined the capacity of hepatic microsomes from Fischer 344 rats, cynomolgus monkeys and humans to metabolically activate IQ, MeIQx and PhIP in vitro using the Ames Salmonella mutagenicity assay. The mutagenic activation of IQ was similar among the three species; however, there were significant differences among the species in the activation of PhIP and MeIQx. Liver microsomes from humans showed the greatest capacity to activate PhIP and MeIQx, followed by rats, and then monkeys. The largest differences between the species were observed when MeIQx was used as the mutagen. MeIQx-DNA adducts formed in vivo were then compared among rats and monkeys given MeIQx by gavage (20 mg/kg/day, 10 doses). 32P-Postlabeling analysis, carried out under intensification conditions, was used to examine MeIQx-DNA adducts in the liver, kidney, heart, colon and white blood cells. MeIQx-DNA adducts were highest in all tissues examined from male rats, followed by female rats, and much lower in monkeys. In the liver, the total MeIQx-DNA adduct levels of monkeys were approximately 19 and approximately 10 times lower than in male and female rats respectively. In extrahepatic tissues, the differences in MeIQx-DNA adduct levels between monkeys and rats were even greater. The results suggest that the low level of MeIQx-DNA adducts found in vivo in cynomolgus monkeys reflects a low capacity to activate MeIQx via the hepatic cytochrome P450 monooxygenase system.


[Spermatogenesis arrest].
Spermatogenesis arrest is a complex process of interruption in the differentiation of germinal cells of specific cellular type, which elicits an altered spermatozoa formation. In contrast, hypospermatogenesis is defined as a decrease in number of germ cells and its proportion. Factors identified intervening upon spermatogenesis arrest are: genetic, hormonal, growth factors, interaction between Sertoli and germ cells and ectoplasmic specialization integrity of spermatozoa. In addition, environmental toxic effects have shown to exert subletal and letal cellular damage with gene disruption. Hence in this work we review sperm physiology along with etiologic elements associated to spermatogenesis arrest delineating the most appropriate conduct for diagnosis and treatment.


Whole-Eye Perfusion Model for Screening of the Ocular Formulations via Confocal Laser Scanning Microscopy.
Various physiological, anatomical barriers make ocular drug delivery very challenging. Hence, better in vitro screening models are needed for rapid screening of the formulations. In this study, a simple whole-eye perfusion model was designed and its application was explored for screening targeted formulation across the full-thickness cornea using confocal laser scanning microscopy. PEG-cholecalciferol-based integrin targeted coumarin-6 micelles (TC6M) and non-targeted coumarin-6 micelles (NTC6M) were developed by solvent diffusion evaporation technique. The formulations NTC6M and TC6M had particles size 23.5 ± 5 nm and 28.5 ± 6 nm respectively and osmolality of 294-300 mOsml/Kg. The whole-eye perfusion model was developed using porcine eye. TC6M and NTC6M were instilled on the excised porcine eyes as well as in the eyes of NZW rabbits. Corneas were excised from the experimental eyes; coumarin-6 penetration across the corneas was analyzed using confocal microscope. Coumarin-6-loaded micelles had particle size below 50 nm. NTC6M formulations showed penetration to the deeper layers up to 500 μm porcine eyes and up to 50 μm in rabbit corneas. However, TC6M formulations exhibited superior retention, as higher fluorescent intensities were observed in upper layers up to 50 μm depth in the porcine eye and 20 μm depth in rabbit eye. Hence, applicability of whole-eye perfusion model in preliminary screening of the formulations was successfully demonstrated. Whole-eye perfusion model when combined with confocal microscopy has potential to be used as an efficient tool for rapid screening and optimization of various ophthalmic formulations.


DNA stretching in the nucleosome facilitates alkylation by an intercalating antitumour agent.
DNA stretching in the nucleosome core can cause dramatic structural distortions, which may influence compaction and factor recognition in chromatin. We find that the base pair unstacking arising from stretching-induced extreme minor groove kinking near the nucleosome centre creates a hot spot for intercalation and alkylation by a novel anticancer compound. This may have far reaching implications for how chromatin structure can influence binding of intercalator species and indicates potential for the development of site selective DNA-binding agents that target unique conformational features of the nucleosome.


Lack of increase in granulocyte colony-stimulating factor in psoriatic skin.
Previously, we showed an elevated level of pro-inflammatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) in psoriatic skin. Granulocyte (G)-CSF, which is also released from the infiltrating cells and epidermal keratinocytes, profoundly influences the biological activities of terminally differentiated neutrophils, in addition to its supporting effects on the proliferation and differentiation of progenitor cells of neutrophil lineage. We have carried out enzyme immunoassay for G-CSF in suction blister fluids and horny tissue extracts from psoriatic skin. Although some samples of the blister fluids and stratum corneum extracts showed G-CSF, there were no significant differences between the concentration in normal and psoriatic skin. These results suggest that, among CSFs, GM-CSF plays a more important role than G-CSF in the local immune responses in psoriasis.


Colour-Doppler ultrasound imaging as a laparoscopy substitute to count corpora lutea in superovulated sheep.
This study evaluated colour-Doppler ultrasound imaging (UI) as a substitute for laparoscopy to count the corpora lutea (CL) in superovulated sheep. Twenty-five Santa Ines ewes were superovulated three times at 21-day intervals. Corpora lutea were counted by colour-Doppler UI (CLDOPPLER ) 6 days after each superovulation and confirmed by laparoscopy (CLLAP ) 12 hr later. The mean number of CL was similar for both techniques (2.1 ± 2.5 vs. 2.1 ± 2.7 for CLDOPPLER and CLLAP , respectively) with a significant positive correlation (r = .94; r2 =.89). Colour-Doppler UI effectively evaluated the ovarian response in superovulated ewes and efficiently identified animals that did not respond to superovulation.


Evolutionary history of coastal tiger beetles in Japan based on a comparative phylogeography of four species.
To reveal the phylogeographical patterns of four species of coastal tiger beetles in Japan (Lophyridia angulata, Abroscelis anchoralis, Cicindela lewisii and Chaetodera laetescripta), we conducted phylogenetic and nested clade analysis (NCA) using the mitochondrial DNA sequences of two loci (COI and 16S rRNA), with specimens sampled from Japan and neighbouring countries. Abroscelis anchoralis and L. angulata have similar disjunct distributions in Japan. The NCA indicated past fragmentation involving three isolated areas of A. anchoralis. In contrast, local populations of L. angulata in Japan shared the same haplotype, indicating recent vicariance. Co-occurrence of haplotypes from several divergent clades in Japanese populations of Ch. laetescripta suggested ancient vicariance and subsequent intermixing of local populations. The tree topology of C. lewisii, with shallow branches and little geographical segregation of haplotypes between Japan and Korea or within Japan, suggested that the Japanese population was segregated from the Korean population only recently. Restricted gene flow, with isolation by distance, was inferred for various geographical associations of haplotypes for coastal tiger beetles in the NCA. Based on these phylogeographical patterns, coupled with a molecular clock approach, the evolutionary history of four species of coastal tiger beetles was deduced, with the additional consideration of the competitive relationships among those species. We also discuss the conservation of highly localized A. anchoralis populations in Japan, using the concept of evolutionarily significant units.


Rational In Silico Design of an Organic Semiconductor with Improved Electron Mobility.
Organic semiconductors find a wide range of applications, such as in organic light emitting diodes, organic solar cells, and organic field effect transistors. One of their most striking disadvantages in comparison to crystalline inorganic semiconductors is their low charge-carrier mobility, which manifests itself in major device constraints such as limited photoactive layer thicknesses. Trial-and-error attempts to increase charge-carrier mobility are impeded by the complex interplay of the molecular and electronic structure of the material with its morphology. Here, the viability of a multiscale simulation approach to rationally design materials with improved electron mobility is demonstrated. Starting from one of the most widely used electron conducting materials (Alq3 ), novel organic semiconductors with tailored electronic properties are designed for which an improvement of the electron mobility by three orders of magnitude is predicted and experimentally confirmed.


Influences of the substituents on the M-M bonding in Cp4Al4 and Cp2M2X2 (M = B, Al, Ga; Cp = C5H5, X = halogen).
Although the geometries of CpAl4 (Cp* = C5Me5) and Cp4Al4 (Cp = C5H5) are similar, CpAl4 is more stable than Cp4Al4. CpAl2I2 is the first complex involving an Al(ii)-Al(ii) bond to be supported by Cp-type ligands. In this work, the stability of CpAl4 and Cp4Al4 (Cp = C5H5), the nature of M-M bonding in Cp2M2X2 (M = B, Al, and Ga), and the influences of the X atom on the M-M bonds have been analyzed and compared within the framework of the atoms in molecules (AIM) theory, electron localization function (ELF), energy decomposition analysis (EDA), and natural bond orbital (NBO) analysis. The calculated results show that CpAl4 is more stable than Cp4Al4 because of HH interactions between the methyl groups on the same and different Cp rings and not because of the Al-Al bonds. In Cp2M2X2, the B-B bond is stronger than the Al-Al and Ga-Ga bonds. The B-B bond is most consistent with covalent bonding, whereas the Al-Al and Ga-Ga bonds are more consistent with metallic bonding. The strengths of the B-B bond increase in the sequence X = F, Cl, Br, and I, whereas the Al-Al and Ga-Ga bonds decrease in the sequence X = F, Cl, Br, and I. The different change tendencies arise from the different M-M bonds and the orbital interactions between atoms X and M.


Regulation of translation factors during hindlimb unloading and denervation of skeletal muscle in rats.
In the rat, denervation and hindlimb unloading are two commonly employed models used to study skeletal muscle atrophy. In these models, muscle atrophy is generally produced by a decrease in protein synthesis and an increase in protein degradation. The decrease in protein synthesis has been suggested to occur by an inhibition at the level of protein translation. To better characterize the regulation of protein translation, we investigated the changes that occur in various translation initiation and elongation factors. We demonstrated that both hindlimb unloading and denervation produce alterations in the phosphorylation and/or total amount of the 70-kDa ribosomal S6 kinase, eukaryotic initiation factor 2 alpha-subunit, and eukaryotic elongation factor 2. Our findings indicate that the regulation of these protein translation factors differs between the models of atrophy studied and between the muscles evaluated (e.g., soleus vs. extensor digitorum longus).


A Cross-Sectional Study on Central Sensitization and Autonomic Changes in Fibromyalgia.
Fibromyalgia is a multi-symptomatic disorder characterized by generalized pain. The pathophysiology of fibromyalgia is supposedly an interplay between central nervous system hyper-responsiveness, autonomic dysfunction, and peripheral pain. In this cross-sectional study, the objective was to assess central sensitization and autonomic activity in patients with fibromyalgia compared with control. Fifty adults diagnosed with fibromyalgia by the modified American College of Rheumatology 2010 criteria and an equal number of age- and sex-matched controls participated in the study in an urban tertiary care hospital. Central sensitization was assessed by history and by evidence of increased prefrontal cortical activity as measured by cortical oxygenation using functional near-infrared spectroscopy. Autonomic activity was assessed by heart rate variability, electrodermal activity, and deep breathing test in three physiological states: rest, sympathetic stress (cold pressor test), and deep breathing. Mann-Whitney U-test, paired t-test, Wilcoxon test, and Friedman test with Bonferroni a priori were used to analyze the data. Cortical activity was significantly higher in the fibromyalgia group than control. There was no significant difference in autonomic activity between the fibromyalgia and control groups. In the fibromyalgia group, variable degrees of sympathetic hyperactivity and normal parasympathetic activity were observed. Central sensitization may be playing a primary role in the pathophysiology of generalized pain in fibromyalgia.


The Configuration of Copolymer Ligands on Nanoparticles Affects Adhesion and Uptake.
Nanoparticles (NPs) are promising carriers for targeted drug delivery, photodynamic therapy, and imaging probes. A fundamental understanding of the dynamics of polymeric NP targeting to bilayer membranes is important to enhance the design of NPs for higher adhesion, binding percentage, and efficiency. In this study, dissipative particle dynamics simulations are applied to investigate the adhesion and uptake processes of the rod, spherical, and striped NPs to cell membranes. It is observed that the striped ligands can prevent NPs from rotating even in active rotation. We further optimize striped NP to a more stabilized structure. Uptake processes of NPs with different configurations are thoroughly investigated in our simulations and among which Janus NP are indicated to improve the penetration rate to 100%. These findings provide better insight into patterned NP design and may help fabricate new NPs for biomedical applications.


Predictors of opinions on prison smoking bans: Analyses of survey data from Scottish staff and prisoners.
Policy-makers and practitioners need to understand characteristics associated with support for smoking restrictions to identify both potential allies and groups requiring particular support/targeted communication in the face of restrictions. Using data from prison staff and prisoners, we explored the structure and correlates of opinions relating to prison smoking bans. Questionnaires were completed by staff (online, N=1271; 27% return) and prisoners (paper-based, N=2512; 34%) in all 15 Scottish prisons in 2016-17. At that time, prisoners could smoke in their own cells and during outdoor recreation; staff smoking was prohibited anywhere on prison grounds. Staff and prisoner questionnaires included identical/very similar questions about opinions on smoking in prisons and prison smoking bans, own smoking behaviour, health and sociodemographic details. We also measured in every prison fine particulate matter (PM2.5) as a proxy for secondhand smoke (SHS) levels. Principal components analysis identified two factors: 'Positive about bans' (higher scores among staff) and 'Bans will be difficult' (higher scores among prisoners). In multivariable analyses, 'Positive about bans' was associated with: not smoking (both staff and prisoners), better general health, more respiratory symptoms and working in an operational role among staff; and no asthma, more sensory symptoms, higher educational level and status/release date among prisoners. 'Bans will be difficult' was associated with: fewer sensory symptoms and lower prison SHS levels among staff and being a smoker among prisoners. In smoker-only analyses, heavier smokers were less positive about bans and more likely to believe bans will be difficult. Results suggest it is possible to be positive about prison smoking bans whilst also recognising and/or concerned about potential operational difficulties, and that these opinions are associated with several characteristics additional to smoker status. Support for future prison bans may be stronger if staff have access to objective SHS exposure measures.


[Studies on Functional Molecules Based on Peptide Chemistry].
Studies on functional molecules starting from syntheses of cysteine-containing peptides and protein are described. Starting from evaluation of a cysteine specific side-reaction, a specific reaction for disulfide-bond formation was developed. The reaction made it possible to independently construct a disulfide bridge without effecting the existing disulfide bonds, which resulted in a unique approach for the synthesis of human insulin by site-specific disulfide bond formation. In a series of studies on sulfur-containing amino acids, another cysteine related un-natural amino acid, α-methyl cysteine, was used for the total syntheses of natural products containing a unique thiazorine/thiazole ring system. Chloroimidazolidium coupling reagent developed by us was effective for the successive couplings of the α-methyl cysteine residues. Based on these synthetic studies, design and evaluation of protease inhibitors were then studied, since a stereo-specific synthesis of the key structure is crucial to make the inhibitor an effective functional molecule in the interactions with its target protease. As the target proteases, β-site amyloid precursor protein cleaving enzyme 1 (BACE1) and chymotrypsin-like protease of severe acute respiratory syndrome (SARS 3CL protease) were selected: the former is a crucial enzyme for amyloid β production and the latter is an essential enzyme for the re-construction of SARS corona virus in host cells. Structure optimization procedure of the respective inhibitors are described based on X-ray crystal structure analyses of the inhibitor-protease complex.


Authenticity and Clinical Decision-Making.
Two articles in the January-February 2022 issue of the Hastings Center Report concern authenticity as an aid to clinical decision-making. The lead article, by Matilda Carter, intervenes in a debate concerning decision-making on behalf of people with advance directives instructing that, should they develop dementia, they not be given life-sustaining treatment and who now have the illness and are unable to make informed medical decisions. Carter develops her position on the authority an advance directive should hold in such cases by arguing that the only principle needed for substitute decision-making is that of determining authenticity. In the second article, Ryan Nelson et al. identify pediatric authenticity as an important decision-making standard, especially in caring for children too young to assent to treatment.


Genetic analysis of Père David's x red deer interspecies hybrids.
Interspecies hybrids provide unique opportunities for fundamental genetic analyses and for genetic improvement of farmed deer. We have bred F1 hybrids by artificial insemination of red deer hinds (Cervus elaphus) with semen from Père David's deer (Elaphurus davidianus). The male and female F1 Père David deer x red deer hybrids were fertile and in matings with red deer have produced over 300 viable backcross hybrids. DNA was collected from the backcross progeny as an international reference panel for gene linkage mapping and to investigate associations between segregating species-specific genetic markers and phenotypic traits. We have measured a range of phenotypic traits in the backcross hybrids and red deer. Several traits appear suitable for genetic analysis using mapped genetic markers, including gestation length, growth rate, live weight, head morphometrics, and tail length. Typically these traits show a large difference between Père David's deer and red deer and a high variance in the backcross so that many individuals have a phenotype outside the range observed in red deer.


Hemostatic activation in spontaneous intracerebral hemorrhage.
There is no in-depth information available on the changes in hemostatic systems in patients in the acute phase of spontaneous intracerebral hemorrhage (ICH). This study was conducted to assess the relationships between the changes in hemostatic systems and clinical parameters in patients in acute-phase ICH. The records of 358 patients admitted within 6 hours of onset of ICH were reviewed to examine the relationships between changes in hemostatic systems and computed tomographic findings and clinical parameters. -The white blood cell counts and the levels of thrombin-antithrombin complex, plasmin-antiplasmin complex, and D-dimer in patients with intraventricular extension (IVE) or subarachnoid hemorrhage (SAH) were significantly (P<0.05) higher than those in patients without IVE or SAH. Most of the hemostatic system parameters in patients without IVE or SAH showed no significant differences compared with normal subjects. Multiple linear regression analysis revealed that the levels of thrombin-antithrombin complex significantly increased with an increase in the amount of SAH (P<0.001) and IVE (P<0.001). The levels of thrombin-antithrombin complex were not significantly associated with the volume of intraparenchymal hematoma. The level of the complex, however, was significantly (P<0.001) and independently associated with the presence of IVE or SAH (multiple logistic regression analysis). The systemic activation of hemostatic systems in ICH patients seems to take place only when blood reaches the subarachnoid space. The intraparenchymal hematoma itself seems unlikely to activate hemostatic systems in peripheral blood, although the hematoma is expected to cause local activation of hemostatic systems.