What have we learned from large drug treatment trials in COPD?
Although the development of effective treatments for patients with chronic obstructive pulmonary disease (COPD) has not been seen as a high priority, the past decade has seen a substantial increase in the number of clinical studies examining different treatments for this disease. Large studies are needed to adequately assess the effectiveness of treatment because of the chronic nature of the disease and the intermittent occurrence of some key outcomes such as exacerbations. Data from randomised controlled trials show that treatment improves exercise performance by increasing lung volume rather than changing expiratory flow. Although assessment of lung function remains the cornerstone of drug assessment, improvements in health status, the number of exacerbations and admissions to hospital are now recognised as important treatment outcomes. Randomised controlled trial data provide the best evidence for treatment efficacy, but results of these studies can be affected by differences in inclusion criteria and patient dropout during the study. Bronchodilator reversibility testing does not reliably define subgroups that will respond to a particular treatment. Carefully done and adequately powered clinical trials continue to inform, not only our views about treatment, but also our understanding of COPD and how it is best assessed and managed. Ensuring that these expensive studies are done objectively to the highest standard is an important goal for the next decade.


Lipoprotein(a) as a Risk Factor for Venous Thromboembolism: A Systematic Review and Meta-analysis of the Literature.
Elevated plasma levels of lipoprotein(a) (Lp(a)) are associated with increased cardiovascular risk in several clinical studies. However, there is a lack of data supporting a positive association between elevated Lp(a) levels and venous thromboembolism (VTE). Thus, we conducted a systematic review of the literature to better clarify its role as a risk factor for VTE. Medline and the Embase (up to May 2015) electronic databases were used to identify potentially eligible studies. Studies measuring Lp(a) values in adult patients with deep vein thrombosis and/or pulmonary embolism and in a population of patients without a VTE were selected. Studies on patients with major venous thromboembolic events occurring at other unusual site, case reports, and case series were excluded. The odds ratios (ORs) of the association between high values of Lp(a) and VTE and the weighted mean difference (WMD) in Lp(a) levels in cases and in controls were calculated using a random-effect model. Results were presented with 95% confidence interval (CI). Fourteen studies for a total of more than 14,000 patients were finally included in our analysis. Lp(a) was slightly but significantly associated with an increased risk of VTE (OR: 1.56, 95% CI: 1.36, 1.79; 10 studies, 13,541 patients). VTE patients had significantly higher Lp(a) values compared with controls (WMD: 14.46 mg/L, 95% CI: 12.14, 16.78; 4 studies, 470 patients). Lp(a) appeared to be significantly associated with increased risk of VTE. However, Lp(a) levels were only slightly increased in VTE patients compared with controls.


Measuring gambling participation.
There are many parallels between research on gambling and research on alcohol use, but a striking difference is the emphasis in the former area on problem gambling rather than the use of participation measures. We outline five topics that are underdeveloped as a consequence: (i) gambling participation and future problems; (ii) moderate gambling; (iii) separate measurement of exposures and harms; (iv) predictors of participation; and (v) natural history of participation. Challenges to the future development of gambling participation measures are discussed by reference to comparable difficulties in the field of alcohol use and some examples are given as to how progress could be made. Further development of measures will necessarily occur in the context of broader scientific aims. Some recent studies are highlighted that provide hope of gains in this area. We urge further progress to yield conceptually and operationally distinct indices of exposures and harms.


Animal-Assisted Therapy Improves Communication and Mobility among Institutionalized People with Cognitive Impairment.
Disorders of communication, social relationships, and psychomotricity are often characterized by cognitive impairment, which hinders daily activities and increases the risk of falls. This study aimed to evaluate the efficacy of an animal-assisted therapy (AAT) program in an institutionalized geriatric population with cognitive impairment. The variables evaluated included level of communication and changes in gait and/or balance. We performed a two-arm, parallel controlled, open-label, nonrandomized cluster clinical trial in two nursing home centers from an urban area. Patients in the two centers received 12 weekly sessions of physiotherapy, but the experimental group included AAT with a therapy dog. The study included a total of 46 patients (23 Control Group [CG], 23 Experimental Group [EG]) with a median age of 85.0 years. Of these, 32.6% had mild-moderate cognitive decline (Global Deterioration Scale of Reisberg [GDS] 2-4) and 67.4% severe cognitive decline (GDS 5-6). After the intervention, patients in the CG and EG showed a statistically significant improvement in all the response variables. When comparing both groups, no statistically significant differences were found in any of the Tinetti scale results (measuring gait and balance). However, the communication of patients in the EG, measured on the Holden scale, showed a statistically significant greater improvement postintervention than that of patients in the CG. AAT can be useful as a complementary, effective treatment for patients with different degrees of cognitive decline.


A paramyxovirus-vectored intranasal vaccine against Ebola virus is immunogenic in vector-immune animals.
Ebola virus (EBOV) causes outbreaks of a highly lethal hemorrhagic fever in humans. The virus can be transmitted by direct contact as well as by aerosol and is considered a potential bioweapon. Because direct immunization of the respiratory tract should be particularly effective against infection of mucosal surfaces, we previously developed an intranasal vaccine based on replication-competent human parainfluenza virus type 3 (HPIV3) expressing EBOV glycoprotein GP (HPIV3/EboGP) and showed that it is immunogenic and protective against a high dose parenteral EBOV challenge. However, because the adult human population has considerable immunity to HPIV3, which is a common human pathogen, replication and immunogenicity of the vaccine in this population might be greatly restricted. Indeed, in the present study, replication of the vaccine in the respiratory tract of HPIV3-immune guinea pigs was found to be restricted to undetectable levels. This restriction appeared to be based on both neutralizing antibodies and cellular or other components of the immunity to HPIV3. Surprisingly, even though replication of HPIV3/EboGP was highly restricted in HPIV3-immune animals, it induced a high level of EBOV-specific antibodies that nearly equaled that obtained in HPIV3-naive animals. We also show that the previously demonstrated presence of functional GP in the vector particle was not associated with increased replication in the respiratory tract nor with spread beyond the respiratory tract of HPIV3-naive guinea pigs, indicating that expression and functional incorporation of the attachment/penetration glycoprotein of this systemic virus did not mediate a change in tissue tropism.


The osseointegration technique for the rehabilitation of the exenterated orbit.
Osseointegrated implants can be used successfully to support maxillofacial prostheses. Six patients (aged 37 to 80 years; average age, 68 years) underwent orbital exenteration for eyelid neoplasms infiltrating into the orbit. Each patient underwent a two-stage osseointegration procedure in preparation for fitting an orbital prosthesis. In the first stage, three or four bone-anchored fixtures were placed into the bony orbital rim. Skin penetration and abutment placement were performed in the second-stage operation, performed 3 to 6 months later. Successful osseointegration was achieved in 17 (94.4%) of 18 fixtures in five nonirradiated patients. All five patients were successfully fitted with an implant-retained prosthesis. One patient who underwent irradiation lost all four fixtures that had been placed. This patient chose to wear a black patch rather than a conventional prosthesis held in with adhesive. Average follow-up was 16 months for the five patients who were successfully fitted with orbital prostheses (range, 6 to 30 months).


Inhibition of insect glutathione S-transferase (GST) by conifer extracts.
Insecticide synergists biochemically inhibit insect metabolic enzyme activity and are used both to increase the effectiveness of insecticides and as a diagnostic tool for resistance mechanisms. Considerable attention has been focused on identifying new synergists from phytochemicals with recognized biological activities, specifically enzyme inhibition. Jack pine (Pinus banksiana Lamb.), black spruce (Picea mariana (Mill.) BSP.), balsam fir (Abies balsamea (L.) Mill.), and tamarack larch (Larix laricina (Du Roi) Koch) have been used by native Canadians as traditional medicine, specifically for the anti-inflammatory and antioxidant properties based on enzyme inhibitory activity. To identify the potential allelochemicals with synergistic activity, ethanol crude extracts and methanol/water fractions were separated by Sephadex LH-20 chromatographic column and tested for in vitro glutathione S-transferase (GST) inhibition activity using insecticide-resistant Colorado potato beetle, Leptinotarsa decemlineata (Say) midgut and fat-body homogenate. The fractions showing similar activity were combined and analyzed by ultra pressure liquid chromatography-mass spectrometry. A lignan, (+)-lariciresinol 9'-p-coumarate, was identified from P. mariana cone extracts, and L. laricina and A. balsamea bark extracts. A flavonoid, taxifolin, was identified from P. mariana and P. banksiana cone extracts and L. laricina bark extracts. Both compounds inhibit GST activity with taxifolin showing greater activity compared to (+)-lariciresinol 9'-p-coumarate and the standard GST inhibitor, diethyl maleate. The results suggested that these compounds can be considered as potential new insecticide synergists.


Daylight photodynamic therapy for cutaneous leishmaniasis in a pediatric setting: A case report and literature review.
Cutaneous leishmaniasis (CL) is a vector-borne infection caused by the obligate intracellular parasites of the Leishmania genus. Children are more frequently affected due to increased exposure to sandflies and underdeveloped immune system. Currently, there is a lack of consensus on the most effective treatment approach for CL since most drugs are accompanied by numerous limitations, including adverse effects, toxicity, and onset of antimicrobial resistance phenomena. These limitations appear more relevant in the pediatric population, both for the treatment-related risks and for the reticence of the parents. Photodynamic therapy (PDT) has been increasingly employed in numerous inflammatory and infectious diseases, owing to its tissue selectivity and excellent cosmetic outcomes. On this topic, we report our experience with daylight-PDT (DL-PDT) therapy in a difficult-to-treat area like the facial region in a child with a six-month history of CL. Our case is paradigmatic of the potentiality of PDT to treat difficult lesions in a pediatric setting. However, its use has not yet been standardized either for the treatment of leishmania, with high variability in the number of sessions and time intervals. Specific protocols for pediatric patients should be better standardized in randomized clinical trials in order to provide clear indications for clinicians.


Treatment of otitis media by transtympanic delivery of antibiotics.
Otitis media is the most common reason U.S. children receive antibiotics. The requisite 7- to 10-day course of oral antibiotics can be challenging to deliver in children, entails potential systemic toxicity, and encourages selection of antimicrobial-resistant bacteria. We developed a drug delivery system that, when applied once to the tympanic membrane through the external auditory canal, delivers an entire course of antimicrobial therapy to the middle ear. A pentablock copolymer poloxamer 407-polybutylphosphoester (P407-PBP) was designed to flow easily during application and then to form a mechanically strong hydrogel on the tympanic membrane. U.S. Food and Drug Administration-approved chemical permeation enhancers within the hydrogel assisted flux of the antibiotic ciprofloxacin across the membrane. This drug delivery system completely eradicated otitis media from nontypable Haemophilus influenzae (NTHi) in 10 of 10 chinchillas, whereas only 62.5% of animals receiving 1% ciprofloxacin alone had cleared the infection by day 7. The hydrogel system was biocompatible in the ear, and ciprofloxacin was undetectable systemically (in blood), confirming local drug delivery and activity. This fast-gelling hydrogel could improve compliance, minimize side effects, and prevent systemic distribution of antibiotics in one of the most common pediatric illnesses, possibly minimizing the development of antibiotic resistance.


Melanin-based coloration and host-parasite interactions under global change.
The role of parasites in shaping melanin-based colour polymorphism, and the consequences of colour polymorphism for disease resistance, remain debated. Here we review recent evidence of the links between melanin-based coloration and the behavioural and immunological defences of vertebrates against their parasites. First we propose that (1) differences between colour morphs can result in variable exposure to parasites, either directly (certain colours might be more or less attractive to parasites) or indirectly (variations in behaviour and encounter probability). Once infected, we propose that (2) immune variation between differently coloured individuals might result in different abilities to cope with parasite infection. We then discuss (3) how these different abilities could translate into variable sexual and natural selection in environments varying in parasite pressure. Finally, we address (4) the potential role of parasites in the maintenance of melanin-based colour polymorphism, especially in the context of global change and multiple stressors in human-altered environments. Because global change will probably affect both coloration and the spread of parasitic diseases in the decades to come, future studies should take into account melanin-based coloration to better predict the evolutionary responses of animals to changing disease risk in human-altered environments.


Brief presence of varicella-zoster vral DNA in mononuclear cells during relapses of multiple sclerosis.
A possible viral cause for multiple sclerosis (MS) has long been suspected. A progressive increase in MS has been reported in Mexico during the past 20 years; a conspicuous antecedent of varicella infection during childhood has been the most relevant finding in the medical history of patients with MS. To investigate the possible participation of varicella-zoster virus (VZV) in the etiopathogenesis of MS. We searched, by polymerase chain reaction (PCR), for VZV DNA in peripheral mononuclear cells of 82 patients with relapsing-remitting MS. Additionally, genes gD from herpes simplex viruses 1 and 2 were sought by PCR, as well as IgG and IgM serum antibodies to VZV. Viral DNA from the genes open reading frame (ORF)31, ORF62, ORF63, and ORF67 of VZV was found in mononuclear cells from 13 (87%) of 15 patients with MS who were tested during acute relapse. All patients who were tested during remission (n = 67) were negative for the DNA, including patients who were initially positive and were tested again after 2 months of remission. All control patients with a comprehensive variety of neurologic diseases (n = 100) and healthy controls (n = 20) also tested negative. All subjects were negative for herpes simplex viruses 1 and 2 DNA, and no differences were found in serum antibodies to VZV. The finding of genes of VZV in peripheral mononuclear cells, restricted to a brief period during clinical relapse of MS, suggests either its participation in the etiopathogenesis of MS or an epiphenomenon of viral activation simultaneous with the relapse of MS.


Effects of physical activity on melatonin levels in previously sedentary men and women.
The inverse association between physical activity and cancer risk may be mediated by higher melatonin levels. However, few studies have examined the effect of increased physical activity on melatonin levels. The parent study was a randomized controlled trial that randomized 51 men and 49 women to a 12-month moderate-to-vigorous aerobic exercise intervention ("exercisers") and 51 men and 51 women to a stretching control ("controls"). Participants were of ages 40 to 75 years, and previously sedentary. Levels of the principal urinary metabolite of melatonin, 6-sulfatoxymelatonin (aMT6s), corrected for creatinine levels, were measured in spot morning urine samples by immunoassay at baseline and 12 months. Changes in levels between exercisers and controls were compared using generalized estimating equations for linear regression. We observed no statistically significant difference in the change in aMT6s levels from baseline to 12 months in exercisers compared with controls (change in aMT6s levels: exercisers, +6.5%; controls, +13%; P = 0.66). There was no evidence of effect modification by age, sex, or body mass index. A 12-month moderate-intensity exercise intervention did not affect levels of aMT6s. Further research needs to focus on other potential mechanisms through which physical activity may reduce the risk of cancer. Cancer Epidemiol Biomarkers Prev; 23(8); 1696-9. ©2014 AACR.


Acquired nonaccommodative esotropia in childhood.
Acquired nonaccommodative esotropia (ANAET) in childhood is reported to occur infrequently and is often associated with an underlying neurologic or neoplastic disorder. The primary objective of this study was to ascertain the prevalence and clinical characteristics of this form of childhood esotropia. A cohort of all children younger than 11 years with esotropia from a predominantly rural Appalachian region was prospectively identified from August 1, 1995, through July 31, 1998. The age at onset, family history of strabismus, perinatal and medical history, ophthalmologic findings, and surgical results were reviewed for all patients with ANAET. Twenty-three (10.4%) of 221 consecutive children with esotropia were diagnosed with ANAET compared with 12 (5.4%) diagnosed with congenital esotropia. The median age at esotropia onset for the 23 children with ANAET was 31.4 months (range, 8-63 months) with a mean initial angle of esotropia of 24 PD. Although at least 2 children presented with diplopia, none of the 23 patients were known to have harbored intracranial tumors or other lesions of the central nervous system during the follow-up period. Fourteen of the 19 patients who underwent surgery attended follow-up visits for at least 6 months after their last surgical procedure: 13 were within 8 PD or less of orthotropia, whereas the final patient had persistent esotropia. Twelve of the 13 patients within 8 PD of orthotropia demonstrated some level of stereopsis, including 2 children with bifoveal fixation. Two (10.5 %) of the 19 operated patients later required a low hyperopic spectacle correction to control their deviation. ANAET was more prevalent than congenital esotropia in this cohort of children with esotropia. This clinically distinct form of strabismus typically begins between 1 and 5 years of age and appears to be infrequently associated with underlying disease. The angle of deviation is relatively small and early surgical correction is more likely to achieve bifoveal fixation for these patients than for those with congenital esotropia.


Magnetic resonance imaging in two patients with radiation myelopathy.
Findings of magnetic resonance imaging (MRI) in 2 patients with radiation myelopathy are reported. One patient developed Brown-Séquard syndrome 7 months after radiotherapy for carcinoma of the nasopharynx. MRI done 8 months after clinical manifestations revealed long segmental enlargement of the cervical cord, low signal intensity on T1-weighted images and high signal intensity on T2-weighted images of the cervical cord. The other patient presented with bulbar palsy and Brown-Séquard syndrome 4 years after radiotherapy for carcinoma of the nasopharynx. MRI done 1 month after clinical manifestations showed a high signal intensity area on T2-weighted images of the medulla oblongata and the higher cervical cord. These findings corresponded well with the neurological level. In a proper clinical setting, findings of MRI may be helpful in the diagnosis of radiation myelopathy.


Coronary Calcium Score for the Prediction of Asymptomatic Coronary Artery Disease in Patients With Ischemic Stroke.
Purpose: Many patients with ischemic stroke have concomitant coronary artery disease (CAD). However, it remains unclear which stroke patients should undergo evaluation for asymptomatic CAD, and which screening tools are appropriate. We investigated the role of coronary artery calcium (CAC) score as a screening tool for asymptomatic but severe CAD in acute stroke patients. We determined the selection criteria for CAC screening based on risk factors and cerebral atherosclerosis. Materials and Methods: The present study included consecutive patients with acute stroke who had undergone cerebral angiography and multi-detector computed tomography coronary angiography. Severe CAD was defined as left main artery disease or three-vessel disease. Enrolled patients were randomly assigned to two sets; a set for developing selection criteria and a set for validation. To develop selection criteria, we identified associated factors with severe CAD regarding clinical factors and cerebral atherosclerosis. CAD predictability of selection criteria with the CAC score was calculated. Results: Overall, 2,658 patients were included. Severe CAD was present in 360 patients (13.5%). CAC score was associated with CAD severity (P < 0.001). In the development set (N = 1,860), severe CAD was associated with age >65 years [odds ratio (95% confidence interval), 2.62 (1.93-3.55)], male sex (1.81 [1.33-2.46]), dyslipidemia (1.77 [1.25-2.61]), peripheral artery disease (2.64 [1.37-5.06]) and stenosis in the cervicocephalic branches, including the internal carotid (2.79 [2.06-3.78]) and vertebrobasilar arteries (2.08 [1.57-2.76]). We determined the combination of clinical and arterial factors as the selection criteria for CAC evaluation. The cut-off criterion was two or more elements of the selection criteria. The area under the curve (AUC) of the selection criteria was 0.701. The AUC significantly improved to 0.836 when the CAC score was added (P < 0.001). In the validation set (N = 798), the AUC of the selection criteria only was 0.661, and that of the CAC score was 0.833. The AUC of the selection criteria + CAC score significantly improved to 0.861(P < 0.001). Conclusion: The necessity for CAC evaluation could be determined based on the presence of risk factors and significant stenosis of the cervicocephalic arteries. CAC evaluation may be useful for screening for severe CAD in stroke patients.


Avoiding Breast Animation Deformity with Pectoralis-Sparing Subcutaneous Direct-to-Implant Breast Reconstruction.
There is renewed interest in sparing the pectoralis major muscle in implant breast reconstruction, placing the implant in the subcutaneous position. This advance is made reliable by improvements in mastectomy skin flap quality, evolution of technique, and increased awareness of breast animation deformity. This retrospective review presents 13 patients (23 breasts) reconstructed immediately with placing the definitive implant in the subcutaneous space without disruption of the underlying chest muscles. None of the 13 patients had breast animation deformity postoperatively. One patient experienced an early hematoma, and 3 patients had small seromas that resolved uneventfully. One morbidly obese patient undergoing chemotherapy using a nearby chest port experienced infection requiring implant removal. This study describes an early experience, demonstrates feasibility, and discusses patient selection considerations that are important because we continue to evolve breast reconstruction approaches.


An oligo-screening strategy to fill gaps found during shotgun sequencing projects.
During the course of projects to sequence human and nematode cosmids we encountered great difficulties generating contiguous sequence in regions with repetitive DNA (Alu repeats in humans and tandem or inverted repeats in the nematode). We have developed a simple and efficient strategy to fill gaps. By screening M13, plasmid or phagemid libraries with oligonucleotides flanking the gap, clones are identified that contiguate the cosmid sequence. Our method has been integrated into the GAP4 sequence assembly program. The strategy reduces both time and costs in large scale sequencing projects.


Single amino acid alteration between valine and isoleucine determines the distinct pyrabactin selectivity by PYL1 and PYL2.
Abscisic acid (ABA) is one of the most important phytohormones in plant. PYL proteins were identified to be ABA receptors in Arabidopsis thaliana. Despite the remarkably high degree of sequence similarity, PYL1 and PYL2 exhibit distinct responses toward pyrabactin, an ABA agonist. PYL1 inhibits protein phosphatase type 2C upon binding of pyrabactin. In contrast, PYL2 appears relatively insensitive to this compound. The crystal structure of pyrabactin-bound PYL1 revealed that most of the PYL1 residues involved in pyrabactin binding are conserved, hence failing to explain the selectivity of pyrabactin for PYL1 over PYL2. To understand the molecular basis of pyrabactin selectivity, we determined the crystal structure of PYL2 in complex with pyrabactin at 1.64 A resolution. Structural comparison and biochemical analyses demonstrated that one single amino acid alteration between a corresponding valine and isoleucine determines the distinct pyrabactin selectivity by PYL1 and PYL2. These characterizations provide an important clue to dissecting the redundancy of PYL proteins.


A Maternally Deposited Endosperm Cuticle Contributes to the Physiological Defects of transparent testa Seeds.
Mature dry seeds are highly resilient plant structures where the encapsulated embryo is kept protected and dormant to facilitate its ultimate dispersion. Seed viability is heavily dependent on the seed coat's capacity to shield living tissues from mechanical and oxidative stress. In Arabidopsis (Arabidopsis thaliana), the seed coat, also called the testa, arises after the differentiation of maternal ovular integuments during seed development. We recently described a thick cuticle tightly embedded in the mature seed's endosperm cell wall. We show here that it is produced by the maternal inner integument 1 layer and, remarkably, transferred to the developing endosperm. Arabidopsis transparent testa (tt) mutations cause maternally derived seed coat pigmentation defects. TT gene products encode proteins involved in flavonoid metabolism and regulators of seed coat development. tt mutants have abnormally high seed coat permeability, resulting in lower seed viability and dormancy. However, the biochemical basis of this high permeability is not fully understood. We show that the cuticles of developing tt mutant integuments have profound structural defects, which are associated with enhanced cuticle permeability. Genetic analysis indicates that a functional proanthocyanidin synthesis pathway is required to limit cuticle permeability, and our results suggest that proanthocyanidins could be intrinsic components of the cuticle. Together, these results show that the formation of a maternal cuticle is an intrinsic part of the normal integumental differentiation program leading to testa formation and is essential for the seed's physiological properties.


Multiple opiate binding sites in the central nervous system of the rabbit. Large predominance of a mu subtype in the cerebellum and characterization of a kappa subtype in the thalamus.
We have compared the binding characteristics of [3H]etorphine, a nonselective mu-, delta-, and kappa-opiate agonist, with those of [3H]Tyr-D-Ala-Gly-MePhe-NH(CH2)2OH ([3H]DAGO), a selective mu-agonist, in rabbit cerebellar and thalamic membranes. We have also examined the ability of various unlabeled opioid ligands to compete with the binding of [3H]etorphine in the two preparations. In cerebellar membranes, [3H]DAGO(Kd = 0.7 nM) labels slightly fewer sites than does [3H]etorphine (Kd = 0.06 nM): 0.18 versus 0.24 pmole/mg of protein. In addition, competition studies indicate that up to 75% of the [3H]etorphine binding sites in this preparation display (a) high apparent affinity for unlabeled DAGO and (b) higher apparent affinity for morphine, the prototypical mu-agonist, than for Tyr-D-Ala-Gly-Phe-D-Leu (DADL), a delta-agonist. Together, these results suggest that the rabbit cerebellum contains a very high proportion (0.7-0.8) of mu-opiate binding sites. In thalamic membranes, [3H]DAGO (Kd = 1.1 nM) labels considerably fewer sites than does [3H]etorphine (Kd = 0.08 nM): 0.09 versus 0.27 pmole/mg of protein. In this preparation, the competition curves of DAGO and of DADL resolve binding of [3H]etorphine into two components. The first component accounts for 40-50% of total binding and reflects the interaction of [3H]etorphine with mu-opiate binding sites. The second component (up to 50% of total binding) is unaffected in the presence of DADL at concentrations (1-10 microM) that rule out binding of [3H]etorphine to mu- and delta-opiate binding sites. It disappears readily in the presence of very low concentrations (Ki less than 1 nM) of benzomorphan opiates (bremazocine, cyclazocine, and ethylketocyclazocine) yet it is relatively insensitive to inhibition by mu- and delta-agonists. This second component may therefore reflect [3H]etorphine's interaction with a kappa-opiate binding site. The kappa-opiate binding site is assayed for as that site which binds [3H]etorphine (0.5 nM) in the presence of either DADL (2 microM) or 10 microM of another enkephalin: Tyr-D-Ser-Gly-Phe-Leu-Thr. We find that, in the rabbit central nervous system, the thalamus, followed by frontal cortex and caudate nucleus, shows the highest content of kappa-opiate binding sites.


Is the transtheoretical model and motivational interviewing approach applicable to the treatment of eating disorders? A review.
Research in eating disorders is reviewed examining the (1) utility of the Transtheoretical Model in predicting outcome, and (2) efficacy of Motivational Interviewing (MI). There were promising results showing significant relationships between initial stage of change and treatment outcome related to eating pathology (not including purging), body mass index, and some aspects of psychopathology. Of those treatment studies utilising a control group, there was little indication that using MI conferred significant treatment benefit, with the exception of improving motivation and binge eating for people with binge eating disorder and bulimia nervosa. Overall the content of the studies varied greatly with relation to: stage of change and outcome measures, format of MI, diagnostic groupings, age of participants, utilisation of other adjunctive treatments, sample size, presence of follow-up assessments, and study design. Few of the 9 studies examining the efficacy of MI could be considered to have robust methodology. It is recommended that future research using the Transtheoretical Model to predict outcome adopt more uniform methodology so that we can more specifically determine its applicability, and that well-designed treatment studies in eating disorder populations be conducted so that we develop a stronger evidence base from which to decide whether MI confers benefit.


The one-year attributable cost of post-stroke dysphagia.
With the recent emphasis on evidence-based practice and healthcare reform, understanding the cost of dysphagia management has never been more important. It is helpful for clinicians to understand and objectively report the costs associated with dysphagia when they advocate for their services in this economy. Having carefully estimated cost of illness, inputs are needed for cost-effectiveness analyses that help support the value of treatments. This study sought to address this issue by examining the 1-year cost associated with a diagnosis of dysphagia post-stroke in South Carolina. Furthermore, this study investigated whether ethnicity and residence differences exist in the cost of dysphagia post-stroke. Data on 3,200 patients in the South Carolina Medicare database from 2004 who had ICD-9 codes for ischemic stroke, 434 and 436, were retrospectively included in this study. Differences between persons with and without dysphagia post-stroke were compared with respect to age, gender, ethnicity, mortality, length of stay, comorbidity, rurality, discharge disposition, and cost to Medicare. Univariate analyses and a gamma-distributed generalized linear multivariable model with a log link function were completed. We found that the 1-year cost to Medicare for persons with dysphagia post ischemic stroke was $4,510 higher than that for persons without dysphagia post ischemic stroke when controlling for age, comorbidities, ethnicity, and proportion of time alive. Univariate analysis revealed that rurality, ethnicity, and gender were not statistically significantly different in comparisons of individuals with or without dysphagia post-stroke. Post-stroke dysphagia significantly increases post-stroke medical expenses. Understanding the expenditures associated with post-stroke dysphagia is helpful for optimal allocation and use of resources. Such information is needed to conduct cost-effectiveness studies.


Pregnancy associated plasma protein-A2: a novel biomarker for Down syndrome.
In an effort to improve prenatal screening for Trisomy 21, we evaluated pregnancy associated plasma protein-A2 (PAPP-A2) as a potential novel second trimester biomarker for Trisomy 21. Trisomy 21 and normal control mid-trimester placental samples were subjected to quantitative rt PCR analysis of seven genes we had previously found to be differentially expressed in Trisomy 21 placentae. The localization and differential expression of PAPP-A2 in second trimester placentae from normal and Trisomy 21 pregnancies was determined by immunohistochemistry. PAPP-A2 maternal serum protein levels in ten Trisomy 21 and ten diploid pregnancies were compared by Western blotting. Maternal serum PAPP-A2 levels were measured in 30 Down syndrome cases and 142 normal controls, using ELISA. Regression analysis was used to determine the correlation of PAPP-A2 with other existing markers of Trisomy 21. PAPP-A2 (aka PLAC 3) mRNA and protein expression were both increased in Down syndrome placentae as compared to diploid placentae. PAPP-A2 was also increased in maternal serum from Down syndrome pregnancies as compared to diploid pregnancies. PAPP-A2 expression correlated weakly with established markers. This work takes advantage of our previously performed systematic approach to the discovery of novel maternal serum biomarkers for Trisomy 21, using cDNA microarray analysis. Beginning with the validation of the microarray results, we have tracked PAPP-A2 overexpression in Down syndrome from placental mRNA to maternal serum protein. PAPP-A2 could serve as an additional maternal serum marker in prenatal screening for Trisomy 21.


Penn Healthy Diet survey: pilot validation and scoring.
Though diet quality is widely recognised as linked to risk of chronic disease, health systems have been challenged to find a user-friendly, efficient way to obtain information about diet. The Penn Healthy Diet (PHD) survey was designed to fill this void. The purposes of this pilot project were to assess the patient experience with the PHD, to validate the accuracy of the PHD against related items in a diet recall and to explore scoring algorithms with relationship to the Healthy Eating Index (HEI)-2015 computed from the recall data. A convenience sample of participants in the Penn Health BioBank was surveyed with the PHD, the Automated Self-Administered 24-hour recall (ASA24) and experience questions. Kappa scores and Spearman correlations were used to compare related questions in the PHD to the ASA24. Numerical scoring, regression tree and weighted regressions were computed for scoring. Participants assessed the PHD as easy to use and were willing to repeat the survey at least annually. The three scoring algorithms were strongly associated with HEI-2015 scores using National Health and Nutrition Examination Survey 2017-2018 data from which the PHD was developed and moderately associated with the pilot replication data. The PHD is acceptable to participants and at least moderately correlated with the HEI-2015. Further validation in a larger sample will enable the selection of the strongest scoring approach.


Effect of polar intermolecular interactions on the elastic constants of bent-core nematics and the origin of the twist-bend phase.
A molecular theory of both elastic constants and the flexoelectric coefficients of bent-core nematic liquid crystals has been developed taking into account dipole-dipole interactions as well as polar interactions determined by the bent molecular shape. It has been shown that if polar interactions are neglected, the elastic constants are increasing monotonically with the decreasing temperature. On the other hand, dipolar interactions between bent-core molecules may result in a dramatic increase of the bend flexocoefficient. As a result, the flexoelectric contribution to the bend elastic constant increases significantly, and the bend elastic constant appears to be very small throughout the nematic range and may vanish at a certain temperature. This temperature may then be identified as a temperature of the elastic instability of the bent-core nematic phase which induces a transition into the modulated phases with bend deformations like recently reported twist-bend phase. The temperature variation of the elastic constants is qualitatively similar to the typical experimental data for bent-core nematics.


In situ analysis of pancreatic islets in rats developing diabetes. Appearance of nonendocrine cells with surface MHC class II antigens and cytoplasmic insulin immunoreactivity.
Aberrant expression of MHC class II molecules on endocrine cells has been proposed to induce autoimmune reactions in thyroid and endocrine pancreas. The present study examines whether MHC class II positive insulin-containing islet cells occur at the onset of diabetes in rats, in analogy to the findings in man. At the onset of diabetes, both streptozotocin-treated and diabetes-prone BB rats exhibited numerous class II positive islet cells that presented ultrastructural features of monocytes and were surrounded by class II negative islet B cells. These class II positive cells were characterized by vacuoles that contained insulin immunoreactive granules and disrupted membranes. Similar cells also appeared positive for the monocyte marker OX-42. The presence of class II positive monocytes with insulin-containing vacuoles may indicate a removal of damage B cells by infiltrating leukocytes. A similar electron microscopical study in man will be necessary to distinguish the putative endocrine pancreatic B cells with aberrant class II expression from infiltrating nonendocrine class II positive cells with insulin-containing phagosomes.


Salinity source-induced changes in yield, mineral composition, phenolic acids and flavonoids in leaves of artichoke and cardoon grown in floating system.
Leaves of artichoke (Cynara cardunculus L. subsp. scolymus (L.) Hegi) and cardoon (Cynara cardunculus L. var. altilis DC) are traditionally used as herbal medicine. Moderate salt stress could enhance antioxidant activity and phytochemicals in leaves. The aim of this study was to evaluate the effect of chloride salts (NaCl, KCl and CaCl2) on biomass production, mineral composition, phenolic and flavonoid contents and antioxidant activity in leaves of artichoke and cardoon grown in a floating system. In both crops, NaCl and KCl treatments reduced biomass production, while similar values were recorded in CaCl2 and control treatments. In both crops, KCl treatment enhanced total phenolic and flavonoid contents, antioxidant activity and target polyphenols in leaves harvested at 48, 82 and 105 days after sowing (DAS), while leaf quality was improved by NaCl and CaCl2 treatments only at 82 and 105 DAS. Irrespective of salinity, leaves of cardoon had higher total phenolic and flavonoid contents, antioxidant activity and target polyphenols than those of artichoke. The results showed that application of KCl can be considered an effective way to produce high-quality leaves of artichoke and cardoon during the whole cropping cycle, although resulting in a 30% reduction in plant biomass.


Fatal familial insomnia: clinical and pathologic study of five new cases.
In 1986, we reported two anatomoclinical observations of a familial condition that we called "fatal familial insomnia" (FFI). We now present the pedigree as well as the clinical and neuropathologic findings in five new subjects. The pedigree includes 288 members from six generations. Men and women are affected in a pattern consistent with an autosomal dominant inheritance. The age of onset of the disease varies between 37 and 61 years; the course averages 13 months with a range of 7 to 25 months. Progressive insomnia (polygraphically proven in two cases); autonomic disturbances including hyperhidrosis, hyperthermia, tachycardia, and hypertension; and motor abnormalities including ataxia, myoclonus, and pyramidal dysfunction, were present in every case, but with variable severity and time of presentation. Sleep and autonomic disorders were the earliest signs in two subjects, motor abnormalities were dominant in one, and others had intermediate clinical patterns. Pathologically, all the cases had severe atrophy of the anterior ventral and mediodorsal thalamic nuclei. Other thalamic nuclei were less severely and inconsistently affected. In addition, most of the cases had gliosis of the cerebral cortex, a moderate degree of cerebellar atrophy with "torpedoes," and severe atrophy of the inferior olivary nuclei. One case also showed spongy degeneration of the cerebral cortex. We conclude that all the lesions were primary, and that FFI is a multisystem disease in which the different structures are primarily affected with different severity. The insomnia appears to correlate best with the major thalamic pathology. The possibility that FFI belongs to the group identified as prion diseases or diseases transmitted by unconventional agents is examined.


Quality control of coated antibodies: new, rapid determination of binding affinity.
A procedure is described for the determination of the affinity constant between a fluid-phase biotinylated antigen and a solid-phase monoclonal antibody. This procedure allows evaluation of the efficiency of an antibody as a coated tool for an immunoassay. For this purpose, the biotinylation of the antigen and its further quantitative measurement by streptavidin-peroxidase led to a single reversible interaction, the binding affinity of which greatly determines the quality of the assay. The free and bound fractions of the biotinylated antigen were obtained in wells coated with a low level of immobilized antibodies. At the equilibrium state, the free antigen present in the supernatant of these wells was further transferred to high level antibody coated wells which captured all the free antigen molecules. These molecules were quantified using a standard curve established with known concentrations of biotinylated antigen, also incubated in wells coated with the high level of antibody.


A New Benchmark for Consumer Visual Tracking and Apparent Demographic Estimation from RGB and Thermal Images.
Visual tracking and attribute estimation related to age or gender information of multiple person entities in a scene are mature research topics with the advent of deep learning techniques. However, when it comes to indoor images such as video sequences of retail consumers, data are not always adequate or accurate enough to essentially train effective models for consumer detection and tracking under various adverse factors. This in turn affects the quality of recognizing age or gender for those detected instances. In this work, we introduce two novel datasets: Consumers comprises 145 video sequences compliant to personal information regulations as far as facial images are concerned and BID is a set of cropped body images from each sequence that can be used for numerous computer vision tasks. We also propose an end-to-end framework which comprises CNNs as object detectors, LSTMs for motion forecasting of the tracklet association component in a sequence, along with a multi-attribute classification model for apparent demographic estimation of the detected outputs, aiming to capture useful metadata of consumer product preferences. Obtained results on tracking and age/gender prediction are promising with respect to reference systems while they indicate the proposed model's potential for practical consumer metadata extraction.


Hospital administrators' perceptions of pharmacy directors.
The results of a national survey of hospital administrators' perceptions of pharmacy directors are reported. A questionnaire was mailed to a random sample of 1000 hospital administrators nationwide. The questionnaire asked each administrator to rate the managerial skills of pharmacy directors in general and the pharmacy director at his or her institution on a 5-point scale. The value the administrator placed on clinical pharmacy services was investigated, as was the administrator's knowledge of the education of the pharmacy director and the reporting structure between administrator and pharmacy director. The net response rate was 47.8% (478 usable replies). The hospital administrators believed that pharmacy directors in general need to improve their relationship with the nursing staff, their basic managerial skills, and their communication skills. They believed that the pharmacy director at their own institution excelled at keeping up with progressive pharmacy practice, inventory management, and interacting with the pharmacy and therapeutics committee. The administrators indicated that it is very important for the pharmacy department to be involved in therapeutic drug monitoring and medication counseling and to be progressive in their offerings of services. Hospital administrators had a positive perception of the abilities of pharmacy directors but believed that there is still room for improvement.


Secondary prevention and health promotion after stroke: can it be enhanced?
The aim was to study if health outcome and secondary prevention were satisfactory 1 year after stroke and if nurse-led interventions 3 months after stroke could have impact. Design was a randomized controlled open trial in a 1-year population. Primary outcome was health status 1 year after stroke. One month after stroke, survivors were randomized into intervention group (IG) with follow-up by a specialist nurse (SN) after 3 months (n = 232), and control group (CG) with standard care (n = 227), all to be followed up 1 year after stroke. At the first follow-up, patients graded their health, replied to the EuroQol-5 Dimensions (EQ-5D) health outcome questions, health problems were assessed, and supportive counseling was provided in the IG. Health problems requiring medical interventions were primarily referred to a general practitioner (GP). One year after stroke, 391 survivors were followed up. Systolic blood pressure (BP) had decreased in IG (n = 194) from median 140 to 135 (P = .05), but about half were above the limit 139 in both groups. A larger proportion (22%) had systolic BP >155 in the CG (n = 197) than in the IG (14%; P = .05). In the IG, 62% needed referrals compared with the 75% in the CG (P = .009). Forty percent in the IG and 52.5% in the CG (P = .04) reported anxiety/depression. In the IG, 75% and 67% in the CG rated their general health as fairly good or very good (P = .05). Although nurse-led interventions could have some effect, the results were not optimal. A more powerful strategy could be closer collaboration between the SN and a stroke clinician, before referring to primary care.


A flexible and conductive metallic paper-based current collector with energy storage capability in supercapacitor electrodes.
The development of flexible current collectors as an indispensable component in energy storage devices has been in strong demand for the ever-growing market of flexible and wearable electronics. Herein, flexible and conductive paper-based current collectors are fabricated by directly depositing a metallic Ni layer composed of spiny Ni nanospheres of 400 nm diameter on the surface of filter paper via electroless deposition. The metallic paper shows excellent electric and mechanical properties: the sheet resistance is 2.7 Ω cm-2 (R0 = 0.8 Ω cm-2) after 5000 bending cycles and the mass density is only 0.35 g cm-3. MnO2 is selected as an electrode active material to explore the role of flexible and conductive paper-based current collectors in supercapacitors. Electrochemical results reveal that the largest areal specific capacitance is 1095 mF cm-2 at 1 mA cm-2 and the excellent electrochemical performance can be attributed to the hierarchical porous fibre structure of paper and the lower contact resistance between the active material and the current collector. Note that the approach can be applied to an enlarged size of metallic conductive paper or textile, presenting a simple and feasible method to fabricate flexible current collectors in a large scale.


Distinct Drivers of Core and Accessory Components of Soil Microbial Community Functional Diversity under Environmental Changes.
It is a central ecological goal to explore the effects of global change factors on soil microbial communities. The vast functional gene repertoire of soil microbial communities is composed of both core and accessory genes, which may be governed by distinct drivers. This intuitive hypothesis, however, remains largely unexplored. We conducted a 5-year nitrogen and water addition experiment in the Eurasian steppe and quantified microbial gene diversity via shotgun metagenomics. Nitrogen addition led to an 11-fold increase in the abundance (based on quantitative PCR [qPCR]) of ammonia-oxidizing bacteria, which have mainly core community genes and few accessory community genes. Thus, nitrogen addition substantially increased the relative abundance of many core genes at the whole-community level. Water addition stimulated both plant diversity and microbial respiration; however, increased carbon/energy resources from plants did not counteract increased respiration, so soil carbon/energy resources became more limited. Thus, water addition selected for microorganisms with genes responsible for degrading recalcitrant soil organic matter. Accordingly, many other microorganisms without these genes (but likely with other accessory community genes due to relatively stable average microbial genome size) were selected against, leading to the decrease in the diversity of accessory community genes. In summary, nitrogen addition primarily affected core community genes through nitrogen-cycling processes, and water addition primarily regulated accessory community genes through carbon-cycling processes. Although both gene components may significantly respond as the intensity of nitrogen/water addition increases, our results demonstrated how these common global change factors distinctly impact each component.IMPORTANCE Our results demonstrated increased ecosystem nitrogen and water content as the primary drivers of the core and accessory components of soil microbial community functional diversity, respectively. Our findings suggested that more attention should be paid to certain components of community functional diversity under specific global change conditions. Our findings also indicated that microbial communities have adapted to nitrogen addition by strengthening the function of ammonia oxidization to deplete the excess nitrogen, thus maintaining ecosystem homeostasis. Because community gene richness is primarily determined by the presence/absence of accessory community genes, our findings further implied that strategies such as maintaining the amount of soil organic matter could be adopted to effectively improve the functional gene diversity of soil microbial communities subject to global change factors.


Global patterns of aegyptism without arbovirus.
The world's most important mosquito vector of viruses, Aedes aegypti, is found around the world in tropical, subtropical and even some temperate locations. While climate change may limit populations of Ae. aegypti in some regions, increasing temperatures will likely expand its territory thus increasing risk of human exposure to arboviruses in places like Europe, Northern Australia and North America, among many others. Most studies of Ae. aegypti biology and virus transmission focus on locations with high endemicity or severe outbreaks of human amplified urban arboviruses, such as dengue, Zika, and chikungunya viruses, but rarely on areas at the margins of endemicity. The objective in this study is to explore previously published global patterns in the environmental suitability for Ae. aegypti and dengue virus to reveal deviations in the probability of the vector and human disease occurring. We developed a map showing one end of the gradient being higher suitability of Ae. aegypti with low suitability of dengue and the other end of the spectrum being equal and higher environmental suitability for both Ae. aegypti and dengue. The regions of the world with Ae. aegypti environmental suitability and no endemic dengue transmission exhibits a phenomenon we term 'aegyptism without arbovirus'. We then tested what environmental and socioeconomic variables influence this deviation map revealing a significant association with human population density, suggesting that locations with lower human population density were more likely to have a higher probability of aegyptism without arbovirus. Characterizing regions of the world with established populations of Ae. aegypti but little to no autochthonous transmission of human-amplified arboviruses is an important step in understanding and achieving aegyptism without arbovirus.


Acceptance of mHealth among health professionals: a case study on anesthesia practitioners.
mHealth, the practice of medicine aided by mobile devices is a growing market. Although the offer on Anesthesia applications (Apps) is quite prolific, representative formal assessments on the views of anesthesia practitioners on its use and potential place in daily practice is lacking. This survey aimed thus to cross-assess the Belgian anesthesia population on the use of smartphone Apps and peripherals. The survey was exclusively distributed as an online anonymous questionnaire. Sharing took place via hyperlink forwarding by the Belgian Society for Anesthesia and Reanimation (BSAR) and by the Belgian Association for Regional Anesthesia (BARA) to all registered members. The first answer took place on 5 September 2018, the last on 22 January 2019. Three hundred forty-nine answers were obtained (26.9% corresponding to trainees, 73.1% to specialists). Anesthesiologists were positively confident that Apps and peripherals could help improve anesthesia care (57.0 and 47.9%, respectively, scored 4 or 5, in a scale from 0 to 5). Trainees were significantly more confident than specialists on both mobile Apps (71.2% and 51.8%, respectively; p = 0.001) and peripherals (77.7% and 45.1%, respectively; p = 0.09). The usefulness of Apps and Peripherals was rated 1 or below (on a 0 to 5 scale), respectively, by 9.5 and 14.6% of the total surveyed population, being specialists proportionally less confident in Smartphone peripherals than trainees (p = 0.008). Mobile apps are actively used by a significantly higher proportional number of trainees (67.0% vs. 37.3%, respectively; p = 0.000001). The preferred category of mobile Apps was dose-calculating applications (39.15%), followed by digital books (21. 1%) and Apps for active perioperative monitoring (20.0%). Belgian Anesthesia practitioners show a global positive attitude towards smartphone Apps and Peripherals, with trainees trending to be more confident than specialists. ClinicalTrials.gov database Identifier: NCT03750084. Retrospectively registered on 21 November 2018.


Reconstruction of the complete human cytomegalovirus genome in a BAC reveals RL13 to be a potent inhibitor of replication.
Human cytomegalovirus (HCMV) in clinical material cannot replicate efficiently in vitro until it has adapted by mutation. Consequently, wild-type HCMV differ fundamentally from the passaged strains used for research. To generate a genetically intact source of HCMV, we cloned strain Merlin into a self-excising BAC. The Merlin BAC clone had mutations in the RL13 gene and UL128 locus that were acquired during limited replication in vitro prior to cloning. The complete wild-type HCMV gene complement was reconstructed by reference to the original clinical sample. Characterization of viruses generated from repaired BACs revealed that RL13 efficiently repressed HCMV replication in multiple cell types; moreover, RL13 mutants rapidly and reproducibly emerged in transfectants. Virus also acquired mutations in genes UL128, UL130, or UL131A, which inhibited virus growth specifically in fibroblast cells in wild-type form. We further report that RL13 encodes a highly glycosylated virion envelope protein and thus has the potential to modulate tropism. To overcome rapid emergence of mutations in genetically intact HCMV, we developed a system in which RL13 and UL131A were conditionally repressed during virus propagation. This technological advance now permits studies to be undertaken with a clonal, characterized HCMV strain containing the complete wild-type gene complement and promises to enhance the clinical relevance of fundamental research on HCMV.


Functional Landscape of African Swine Fever Virus-Host and Virus-Virus Protein Interactions.
Viral replication fully relies on the host cell machinery, and physical interactions between viral and host proteins mediate key steps of the viral life cycle. Therefore, identifying virus-host protein-protein interactions (PPIs) provides insights into the molecular mechanisms governing virus infection and is crucial for designing novel antiviral strategies. In the case of the African swine fever virus (ASFV), a large DNA virus that causes a deadly panzootic disease in pigs, the limited understanding of host and viral targets hinders the development of effective vaccines and treatments. This review summarizes the current knowledge of virus-host and virus-virus PPIs by collecting and analyzing studies of individual viral proteins. We have compiled a dataset of experimentally determined host and virus protein targets, the molecular mechanisms involved, and the biological functions of the identified virus-host and virus-virus protein interactions during infection. Ultimately, this work provides a comprehensive and systematic overview of ASFV interactome, identifies knowledge gaps, and proposes future research directions.


OprF Impacts Pseudomonas aeruginosa Biofilm Matrix eDNA Levels in a Nutrient-Dependent Manner.
The biofilm matrix is composed of exopolysaccharides, eDNA, membrane vesicles, and proteins. While proteomic analyses have identified numerous matrix proteins, their functions in the biofilm remain understudied compared to the other biofilm components. In the Pseudomonas aeruginosa biofilm, several studies have identified OprF as an abundant matrix protein and, more specifically, as a component of biofilm membrane vesicles. OprF is a major outer membrane porin of P. aeruginosa cells. However, current data describing the effects of OprF in the P. aeruginosa biofilm are limited. Here, we identify a nutrient-dependent effect of OprF in static biofilms, whereby ΔoprF cells form significantly less biofilm than wild type when grown in media containing glucose or low sodium chloride concentrations. Interestingly, this biofilm defect occurs during late static biofilm formation and is not dependent on the production of PQS, which is responsible for outer membrane vesicle production. Furthermore, while biofilms lacking OprF contain approximately 60% less total biomass than those of wild type, the number of cells in these two biofilms is equivalent. We demonstrate that P. aeruginosa ΔoprF biofilms with reduced biofilm biomass contain less eDNA than wild-type biofilms. These results suggest that the nutrient-dependent effect of OprF is involved in the maintenance of P. aeruginosa biofilms by retaining eDNA in the matrix. IMPORTANCE Many pathogens form biofilms, which are bacterial communities encased in an extracellular matrix that protects them against antibacterial treatments. The roles of several matrix components of the opportunistic pathogen Pseudomonas aeruginosa have been characterized. However, the effects of P. aeruginosa matrix proteins remain understudied and are untapped potential targets for antibiofilm treatments. Here, we describe a conditional effect of the abundant matrix protein OprF on late-stage P. aeruginosa biofilms. A ΔoprF strain formed significantly less biofilm in low sodium chloride or with glucose. Interestingly, the defective ΔoprF biofilms did not exhibit fewer resident cells but contained significantly less extracellular DNA (eDNA) than wild type. These results suggest that OprF is involved in matrix eDNA retention in biofilms.


Single-dose daclizumab induction therapy in patients with liver transplantation.
To investigate the efficacy and safety of a single-dose daclizumab induction therapy in orthotopic liver transplantation (OLTx). A retrospective study was made for 54 cases of OLTx in recent three years. The daclizumab group consisted of 23 cases of OLTx who received single-dose of 2 mg/kg intravenously after postoperative 24 hours. The control group consisted of the remaining 31 patients. Additional immunosuppressors included steroids, mycomphenolate mofetil, facrolimus or microemulsion cyclosporine used in all patients. Meta-statistical analysis was made for general data, incidence of acute rejection and infection, postoperative clinical course, complications and prognosis between two groups. Pretransplant demographies were not significantly different between two groups. In the induction group there were significantly less acute rejection episodes (5 of 23, 21.74 %) than those in the control group (12 of 31, 38.71 %), which were proved by pathologic diagnosis (P<0.05). The incidence of infection at the early stage was not significantly different between two groups. Induction therapy with single-dose of daclizumab is safe and effective and appears to be able to reduce the incidence of acute rejection.


Fetal maternal case management.
Multiple transfers, multiple specialists, and an unpredictable hospital course can result in ineffective communications among patients and the health care team. Complex care requirements easily become fragmented and lack coordination, thus overwhelming a family and even a well-intended multidisciplinary care delivery team. Because of this concern, the Greater Kansas City Fetal Board originated and implemented a new nursing case management model for high-risk obstetric and fetal patients. Fetal maternal case management crosses traditional hospital and provider boundaries with the pregnant patient. Case management allows parents to identify treatment options, and then create with specialists a plan of care. Case managers work with parents to identify support systems and explore parental preferences. The purpose of this article is to describe the fetal maternal case management model. Readers will better understand the need and value of such a program, as well as gain insight into how to facilitate fetal maternal case management.


Poly(adenylic acid) complementary DNA real-time polymerase chain reaction in pancreatic ductal juice in patients undergoing pancreaticoduodenectomy.
There is a need to develop methods of early diagnosis for pancreatic cancer. Pancreatic juice is easily collected by endoscopic retrograde cholangiopancreatography and may facilitate diagnosis using molecular markers. The aim of this work was to explore the feasibility of measurement of gene expression in RNA isolated from ductal juice. Intraoperative sampling of pancreatic juice was undertaken in 27 patients undergoing pancreaticoduodenectomy for suspected tumor. Total RNA was extracted and used as template for poly(adenylic acid) (poly[A]) polymerase chain reaction (PCR) to generate a globally amplified complementary DNA pool representative of all expressed messenger RNAs. Real-time PCR was performed for trefoil factor 2 (TFF2), carboxypeptidase B1 (CPB1), and kallikrein-related peptidase 3 (KLK3) in a subset of samples; all samples were normalized for 3 reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH], PSMB6, and beta-2-microglobulin [B2M]). The median volume of the pancreatic juice obtained was 1245 microL (range, 50-5000 microL). The RNA integrity number ranged from 1.9 to 10. Reverse transcriptase PCR was positive for pancreas-specific genes (TFF2 and CPB1) and negative for prostatic-specific antigen in all samples. These results demonstrate that RNA analysis of pancreatic juice is feasible using a combination of poly(A) PCR and real-time PCR. In addition, the poly(A) complementary DNA generated can be probed for multiple genes and is indefinitely renewable, thereby representing a molecular block of importance for future research.


[Effect of vasopressin and oxytocin on the dorsal root potentials of the isolated perfused spinal cord in rat pups].
The effect of vasopressin and oxytocin on the dorsal root potentials were studied in isolated spinal cord of 12-16 days old rats. It was shown that both neuropeptides evoked reversible depression of the dorsal root potential induced by stimulation of the neighbouring dorsal root. Application of vasopressin or oxytocin caused the reversible dose-dependent depolarization in the dorsal root. The depolarization was preserved in Ca2+-free manganese-containing solution. Functional significance of presynaptic effects of hypothalamic neuropeptides is discussed.


[Clear cell acanthoma. Ultrastructural and immunohistological study].
We report a new case of a clear-cell acanthoma with an unusual location (para-anal), which was studied by electronmicroscopic and immuno-histological methods, using monoclonal antibodies. The results of this study showed that: a) the inflammatory cellular infiltrate comprised a small number of T-suppressor/cytotoxic lymphocytes and a small number of Langerhans cells, b) Langerhans cells were present in the affected epidermis in reduced numbers (2.2 p. 100) when compared to normal epidermis (4-6 p. 100), c) there was a disorder of both keratinization and epidermal differentiation as was shown by the abnormal reactivity of the tumor to monoclonal antibodies KL1 and BL7, d) human papilloma viruses do not seem to be involved in the genesis of the tumour, e) tumoral cells did not show (secretory) eccrine differentiation, f) the pemphigus antigen was present in the intercellular space of the affected epidermis.


Schizosaccharomyces pombe MutSα and MutLα Maintain Stability of Tetra-Nucleotide Repeats and Msh3 of Hepta-Nucleotide Repeats.
Defective mismatch repair (MMR) in humans is associated with colon cancer and instability of microsatellites, that is, DNA sequences with one or several nucleotides repeated. Key factors of eukaryotic MMR are the heterodimers MutSα (Msh2-Msh6), which recognizes base-base mismatches and unpaired nucleotides in DNA, and MutLα (Mlh1-Pms1), which facilitates downstream steps. In addition, MutSβ (Msh2-Msh3) recognizes DNA loops of various sizes, although our previous data and the data presented here suggest that Msh3 of Schizosaccharomyces pombe does not play a role in MMR. To test microsatellite stability in S. pombe and hence DNA loop repair, we have inserted tetra-, penta-, and hepta-nucleotide repeats in the ade6 gene and determined their Ade+ reversion rates and spectra in wild type and various mutants. Our data indicate that loops with four unpaired nucleotides in the nascent and the template strand are the upper limit of MutSα- and MutLα-mediated MMR in S. pombe Stability of hepta-nucleotide repeats requires Msh3 and Exo1 in MMR-independent processes as well as the DNA repair proteins Rad50, Rad51, and Rad2FEN1 Most strikingly, mutation rates in the double mutants msh3 exo1 and msh3 rad51 were decreased when compared to respective single mutants, indicating that Msh3 prevents error prone processes carried out by Exo1 and Rad51. We conclude that Msh3 has no obvious function in MMR in S. pombe, but contributes to DNA repeat stability in MMR-independent processes.


The effects of mobile phones on apoptosis in cerebral tissue: an experimental study on rats.
The concern about mobile phone effects is increasing as the number of users increasing too. Different studies have different results, so this topic is still open to discussion. Aim of this report was to investigate the effects of the mobile phones on the Bcl-2 gene and p53 proteins in rat brains. In the study group of 10 rats; mobile phones that spread EMW at a frequency between 1900-2100 MHz and Specific Absorption Rate range between 0.005 W/kg and 0.288 W/kg (Dialing mode), 0.004 W/kg and 0.029 W/kg (Calling mode) were attached to rat ears for simulating usage in daily life for 7 times a day during 5 minutes (3 seconds dialing mode, 4 minutes and 47 seconds of calling mode) for a four week period. Sham group (n=10) rats were only immobilized without EMW exposure. Another group of rats (n=10) were counted as control without any application. immunohistopathological examination was performed for p53 and Bcl-2 expression. Immunohistopathological examinations revealed that the samples in the study group had more p53 and Bcl-2 positive stained cells and they were stained denser. In both evaluations, these differences between the study and control group were found statistically significant (p < 0.003); In Bcl-2 evaluation statistically significant difference was found between study and sham group to (p < 0.005); however, the p53 evaluation between the study and the sham group did not show any statistically significant difference (p > 0.005). Our results showed that the electro-magnetic waves emitted by the mobile phones may have effect on apoptosis. Besides, obtained data revealed that more realistic application of mobile phones during experiments is more important as expected.


Muscle activation and blood flow do not explain the muscle length-dependent variation in quadriceps isometric endurance.
We investigated the role of central activation in muscle length-dependent endurance. Central activation ratio (CAR) and rectified surface electromyogram (EMG) were studied during fatigue of isometric contractions of the knee extensors at 30 and 90 degrees knee angles (full extension = 0 degree). Subjects (n = 8) were tested on a custom-built ergometer. Maximal voluntary isometric knee extension with supramaximal superimposed burst stimulation (three 100-mus pulses; 300 Hz) was performed to assess CAR and maximal torque capacity (MTC). Surface EMG signals were obtained from vastus lateralis and rectus femoris muscles. At each angle, intermittent (15 s on 6 s off) isometric exercise at 50% MTC with superimposed stimulation was performed to exhaustion. During the fatigue task, a sphygmomanometer cuff around the upper thigh ensured full occlusion (400 mmHg) of the blood supply to the knee extensors. At least 2 days separated fatigue tests. MTC was not different between knee angles (30 degrees : 229.6 +/- 39.3 N.m vs. 90 degrees: 215.7 +/- 13.2 N.m). Endurance times, however, were significantly longer (P < 0.05) at 30 vs. 90 degrees (87.8 +/- 18.7 vs. 54.9 +/- 12.1 s, respectively) despite the CAR not differing between angles at torque failure (30 degrees: 0.95 +/- 0.05 vs. 90 degrees: 0.96 +/- 0.03) and full occlusion of blood supply to the knee extensors. Furthermore, rectified surface EMG values of the vastus lateralis (normalized to prefatigue maximum) were also similar at torque failure (30 degrees : 56.5 +/- 12.5% vs. 90 degrees : 58.3 +/- 15.2%), whereas rectus femoris EMG activity was lower at 30 degrees (44.3 +/- 12.4%) vs. 90 degrees (69.5 +/- 25.3%). We conclude that differences in endurance at different knee angles do not find their origin in differences in central activation and blood flow but may be a consequence of muscle length-related differences in metabolic cost.


Aptamers as both drugs and drug-carriers.
Aptamers are short nucleic acid oligos. They may serve as both drugs and drug-carriers. Their use as diagnostic tools is also evident. They can be generated using various experimental, theoretical, and computational techniques. The systematic evolution of ligands by exponential enrichment which uses iterative screening of nucleic acid libraries is a popular experimental technique. Theory inspired methodology entropy-based seed-and-grow strategy that designs aptamer templates to bind specifically to targets is another one. Aptamers are predicted to be highly useful in producing general drugs and theranostic drugs occasionally for certain diseases like cancer, Alzheimer's disease, and so on. They bind to various targets like lipids, nucleic acids, proteins, small organic compounds, and even entire organisms. Aptamers may also serve as drug-carriers or nanoparticles helping drugs to get released in specific target regions. Due to better target specific physical binding properties aptamers cause less off-target toxicity effects. Therefore, search for aptamer based drugs, drug-carriers, and even diagnostic tools is expanding fast. The biophysical properties in relation to the target specific binding phenomena of aptamers, energetics behind the aptamer transport of drugs, and the consequent biological implications will be discussed. This review will open up avenues leading to novel drug discovery and drug delivery.


The B-subunit of DNA polymerase alpha-primase associates with the origin recognition complex for initiation of DNA replication.
The B-subunit (p70/Pol12p) of the DNA polymerase alpha-primase (Polalpha-primase) complex is thought to have a regulatory role in an early stage of S phase. We generated a panel of fission yeast thermosensitive mutants of the B-subunit (termed Spb70) to investigate its role in initiation of DNA replication by genetic and biochemical approaches. Here, we show that the fission yeast Spb70 genetically interacts and coprecipitates with origin recognition complex proteins Orp1/Orc1 and Orp2/Orc2 and primase coupling subunit Spp2/p58. A fraction of Spb70 associates with Orp2 on chromatin throughout the cell cycle independent of the other subunits of Polalpha-primase. Furthermore, primase Spp2/p58 subunit preferentially associates with the unphosphorylated Orp2, and the association requires Spb70. Mutations in orp2+ that abolish or mimic the Cdc2 phosphorylation of Orp2 suppress or exacerbate the thermosensitivity of the spb70 mutants, respectively, indicating that an unphosphorylated Orp2 promotes an Spb70-dependent replication event. Together, these results indicate that the chromatin-bound B-subunit in association with origin recognition complex mediates recruiting Polalpha-primase complex onto replication origins in G1 pre-Start through an interaction with primase Spp2/p58 subunit. Our results thus suggest a role for the recruited Polalpha-primase in the initiation of both leading and lagging strands at the replication origins.


Neuromuscular monitoring, muscle relaxant use, and reversal at a tertiary teaching hospital 2.5 years after introduction of sugammadex: changes in opinions and clinical practice.
Sugammadex was introduced to Royal Perth Hospital in early 2011 without access restriction. Two departmental audits (26-page online survey and 1-week in-theatre snapshot audit) were undertaken to investigate the change of beliefs and clinical practice related to the use of neuromuscular blocking agents at the Royal Perth Hospital since this introduction. Results were compared with data from 2011. We found that, in the 2.5 years since introduction of Sugammadex, more anesthetists (69.5 versus 38%) utilized neuromuscular monitoring, and aminosteroidal neuromuscular blocking agents were used in 94.3% of cases (versus 77% in 2011). Furthermore, 53% of anesthetists identified with a practice of "deeper and longer" intraoperative paralysis of patients. All 71 patients observed during the 5-day in-theatre audit were reversed with Sugammadex. Since the introduction of Sugammadex, 69% (n = 20) of respondents felt it provided "faster turnover," less postoperative residual neuromuscular blockade (n = 23; 79%), and higher anesthetist satisfaction (n = 17; 59%). 45% (n = 13) of colleagues reported that they would feel professionally impaired without the unrestricted availability of Sugammadex, and 1 colleague would refuse to work in a hospital without this drug being freely available. In clinical practice Sugammadex was frequently (57%) mildly overdosed, with 200 mg being the most commonly administered dose.


High-throughput multiparameter analysis of individual mitochondria.
Mitochondria are one of the most important organelles responsible for cellular energy metabolism and apoptosis regulation. However, single-mitochondrion analysis is challenging, because of their small sizes and the low content of organelle constituents. Here, we report the development of a sensitive and versatile platform for high-throughput multiparameter analysis of individual mitochondria. Employing specific fluorescent staining with a laboratory-built high-sensitivity flow cytometer (HSFCM), we demonstrate the simultaneous detection of side scatter, cardiolipin, and mitochondria DNA (mtDNA) of a single mitochondrion. Simultaneous measurements of side scatter, porin, and cytochrome c of individual mitochondria are reported for the first time. Correlation analysis among multiple attributes on an organelle-by-organelle basis could provide a more definitive assessment of the purity, structure integrity, and apoptosis-related proteins of isolated mitochondria than bulk measurement. This work represents a significant advancement in single-mitochondrion analysis. We believe that the HSFCM holds great potential for studying apoptotic signal transduction pathways at the single-mitochondrion level.


Novel examination for evaluating medical student clinical reasoning: reliability and association with patients seen.
Medical students learn clinical reasoning, in part, through patient care. Although the numbers of patients seen is associated with knowledge examination scores, studies have not demonstrated an association between patient problems and an assessment of clinical reasoning. To examine the reliability of a clinical reasoning examination and investigate whether there was association between internal medicine core clerkship students' performance on this examination and the number of patients they saw with matching problems during their internal medicine clerkship. Students on the core internal medicine clerkship at the Uniformed Services University students log 11 core patient problems based on the Clerkship Directors in Internal Medicine curriculum. On a final clerkship examination (Multistep), students watch a scripted video encounter between physician and patient actors that assesses three sequential steps in clinical reasoning: Step One focuses on history and physical examination; Step Two, students write a problem list after viewing additional clinical findings; Step Three, students complete a prioritized differential diagnosis and treatment plan. Each Multistep examination has three different cases. For graduating classes 2010-2012 (n = 497), we matched the number of patients seen with the problem most represented by the Multistep cases (epigastric pain, generalized edema, monoarticular arthritis, angina, syncope, pleuritic chest pain). We report two-way Pearson correlations between the number of patients students reported with similar problems and the student's percent score on: Step One, Step Two, Step Three, and Overall Test. Multistep reliability: Step 1, 0.6 to 0.8; Step 2, 0.41 to 0.65; Step 3, 0.53 to 0.78; Overall examination (3 cases): 0.74 to 0.83. For three problems, the number of patients seen had small to modest correlations with the Multistep Examination of Analytic Ability total score (r = 0.27 for pleuritic pain, p < 0.05, n = 81 patients; r = 0.14 for epigastric pain, p < 0.05, n = 324 patients; r = 0.19 for generalized edema, p < 0.05, n = 118 patients). DISCUSSION or Although a reliable assessment, student performance on a clinical reasoning examination was weakly associated with the numbers of patients seen with similar problems. This may be as a result of transfer of knowledge between clinical and examination settings, the complexity of clinical reasoning, or the limits of reliability with patient logs and the Multistep.


Trends in the incidence of ischaemic stroke in young adults between 1985 and 2011: the Dijon Stroke Registry.
Recent data have suggested that stroke incidence in young people may be rising. In this population-based study, we aimed to determine whether the incidence of stroke in people aged <55 years old had changed over the last three decades. All cases of first-ever stroke (ischaemic stroke, spontaneous intracerebral haemorrhage, and undetermined stroke) occurring in Dijon, France, from 1985 to 2011 were prospectively collected from a population-based registry. Incidence rates were calculated and temporal trends were analysed by age groups and stroke subtypes using a Poisson regression to estimate incidence rate ratios (IRR). Risk factors and premorbid treatments were analysed. Over the 27-year study period, 4506 patients were recorded (53% women, mean age 74.6±14.4, 10.1% aged <55 years). An increase in overall stroke incidence was noted, as was a rise in ischaemic stroke in individuals aged <55 years (IRR 1.308; 95% CI 0.982 to 1.741, p=0.066 for period 1994-2002 vs period 1985-1993, and IRR 1.697; 95% CI 1.340 to 2.150, p<0.001 for period 2003-2011 vs period 1994-2002), which was consistent for men and women. In these young patients, smoking was the most frequent risk factor (43%). Multiple factors may account for the increased incidence of ischaemic stroke in people aged <55 years including changes in vascular risk factors, better awareness of the disease and treatment options in the population and among practitioners leading to more frequent referrals for specialised care, and improvements in stroke diagnosis. Stroke prevention must be encouraged even in young adults.


Liver mitochondria from manganese-deficient and pallid mice: function and ultrastructure.
Oxidative phosphorylation was studied in isolated liver mitochondria from manganese-deficient mice and in those from a mutant strain, pallid. In mitochondria from manganese-deficient mice, ratios of adenosine triphosphate formed to oxygen consumed were normal, but oxygen uptake was reduced. Electron microscopy of these mitochondria revealed ultrastructural abnormalities including elongation and reorientation of cristae. No biochemical or structural abnormalities were found in mitochondria from pallid mice.


Epoxy Resin-Based Root Canal Sealer Penetration into Dentin Tubules Does not Improve Root Filling Dislodgement Resistance.
This study sought to evaluate the effect of the penetration of an epoxy resin-based root canal sealer into dentin tubules on the force required to dislodge the root canal filling. Sixty extracted human central incisors with single canals were decoronated, instrumented, and filled with gutta-percha and AH Plus sealer labeled with 0.1% rhodamine B dye. The roots were further sectioned horizontally at 3, 6, and 8 mm from the apex. The coronal surfaces of the resulting 180 slices were evaluated using confocal laser scanning microscopy to measure the amount of sealer that penetrated into the dentin tubules. To quantify the force required to dislocate the root filling material, the root fillings of the slices were subjected to a dislodgement resistance test (push-out). Spearman's rho correlation test was further used to test the correlation between the push-out bond strength and sealer penetration into the dentin tubules (P<0.05). No significant correlation was observed between sealer penetration into the dentin tubules and the force required to dislodge the root canal filling (P=0.626). Following the results of this study, the penetrating ability of the AH Plus sealer into dentin tubules has no correlation with the force required to dislodge the root canal filling.


Will current clinical trials answer the most important questions about prostate adenocarcinoma?
Despite a heightened focus of the medical and research community on prostate cancer, many important questions about this disease remain unanswered. These include questions about the possible prevention of prostate cancer, as well as the optimal treatment approaches for localized, locally advanced, metastatic, and hormone-refractory disease. A whole host of prospective, well-designed clinical trials are currently in progress that should answer many of these questions. This review briefly explores some of these unresolved issues and describes ongoing trials designed to address them.


The effect of test difficulty on the sensitivity of speech discrimination tests.
The sensitivity of speech discrimination tests is defined as being large if a small change in experimental conditions causes a large and repeatable change in test score. The aim of this paper is to investigate ways by which test sensitivity can be maximized. The effect of test difficulty on sensitivity is examined by the use of a model based on signal detection theory and subjective similarity rating concepts. For several sets of assumptions about the parameters underlying the perceptual processes, the model indicates that test difficulty should be in the range 73% to 87% to achieve the maximum sensitivity with speech tests using between four and eight response foils. The model results also indicate that the sensitivity increases with the number of response foils. When the variation of test reliability with test difficulty is taken into account, the test difficulty which maximizes the sensitivity/reliability trade-off lies in the range 85% to 90%. It is argued that irrespective of the optimum test difficulty, a maximally sensitive test will contain items equal to one another in difficulty.


Detection of high-frequency energy changes in sustained vowels produced by singers.
The human voice spectrum above 5 kHz receives little attention. However, there are reasons to believe that this high-frequency energy (HFE) may play a role in perceived quality of voice in singing and speech. To fulfill this role, differences in HFE must first be detectable. To determine human ability to detect differences in HFE, the levels of the 8- and 16-kHz center-frequency octave bands were individually attenuated in sustained vowel sounds produced by singers and presented to listeners. Relatively small changes in HFE were in fact detectable, suggesting that this frequency range potentially contributes to the perception of especially the singing voice. Detection ability was greater in the 8-kHz octave than in the 16-kHz octave and varied with band energy level.


Anti-biofilm properties and immunological response of an immune molecule lectin isolated from shrimp Metapenaeus monoceros.
The study is carried out to understand the antimicrobial and immunological response of a potential immune molecule lectin, MmLec isolated from haemolymph of Speckled shrimp, Metapenaeus monoceros. MmLec was purified using mannose coupled Sepharose CL-4B affinity chromatography, which was further subjected on SDS-PAGE to ascertain the distribution of their molecular weight. Sugar binding specificity assay was conducted at various pH and temperatures to investigate the binding affinity of MmLec towards the specific carbohydrate molecule. Functional analysis of immune molecule MmLec included haemagglutination assays performed using human erythrocytes and yeast agglutination activity against Saccharomyces cerevisiae which, were analyzed using light microscopy. In order to study the antimicrobial activity, two Gram-negative (Vibrio parahaemolyticus and Aeromonas hydrophila) and two Gram-positive (Staphylococcus aureus and Enterococcus faecalis) bacteria were treated with purified MmLec. Moreover, these bacterial species were also treated at different concentration of the MmLec to speculate the antibiofilm properties of MmLec which was analyzed under Light Microscopy and Confocal Laser Scanning Microscopy. In addition, other functional characterization of MmLec showed the uniqueness of MmLec in agglutination of human erythrocyte as well as the cells of yeast Saccharomyces cerevisiae. Also, the phenoloxidase activity and encapsulation assay was evaluated. MTT assay displayed that MmLec are potent in anticancer activity. The study will help to understand the immunological interference and antimicrobial nature of MmLec which would be supportive in establishing a potential therapeutic tool and to develop better and novel disease control strategies in shrimp and farmed aquaculture industries as well as in health management.


Longitudinal scRNA-seq analysis in mouse and human informs optimization of rapid mouse astrocyte differentiation protocols.
Macroglia (astrocytes and oligodendrocytes) are required for normal development and function of the central nervous system, yet many questions remain about their emergence during the development of the brain and spinal cord. Here we used single-cell/single-nucleus RNA sequencing (scRNA-seq/snRNA-seq) to analyze over 298,000 cells and nuclei during macroglia differentiation from mouse embryonic and human-induced pluripotent stem cells. We computationally identify candidate genes involved in the fate specification of glia in both species and report heterogeneous expression of astrocyte surface markers across differentiating cells. We then used our transcriptomic data to optimize a previous mouse astrocyte differentiation protocol, decreasing the overall protocol length and complexity. Finally, we used multi-omic, dual single-nuclei (sn)RNA-seq/snATAC-seq analysis to uncover potential genomic regulatory sites mediating glial differentiation. These datasets will enable future optimization of glial differentiation protocols and provide insight into human glial differentiation.


Effect of pulsed radiofrequency on rat sciatic nerve chronic constriction injury: a preliminary study.
Pulsed radiofrequency (PRF) application to the dorsal root ganglia can reduce neuropathic pain (NP) in animal models, but the effect of PRF on damaged peripheral nerves has not been examined. We investigated the effect of PRF to the rat sciatic nerve (SN) on pain-related behavior and SN ultrastructure following chronic constriction injury (CCI). The analgesic effect was measured by hindpaw mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL). Twenty rats with NP induced by ligating the common SN were then randomly divided into a PRF treatment group and a sham group. The contralateral SN served as a control. The MWT and TWL were determined again 2, 4, 6, 8, 10, 12, and 14 days after the PRF or sham treatment. On day 14, ipsilateral and contralateral common SNs were excised and examined by electron microscopy. Ipsilateral MWT was significantly reduced and TWL significantly shorter compared to the contralateral side 14 days after CCI (both P = 0.000). In the PRF group, MWT was significantly higher and TWL significantly longer 14 days after the PRF treatment compared to before PRF treatment (both P = 0.000), while no such difference was observed in the sham group (P > 0.05). Electron microscopy revealed extensive demyelination and collagen fiber formation in the ipsilateral SN of sham-treated rats but sparse demyelination and some nerve fiber regrowth in the PRF treatment group. Hyperalgesia is relieved, and ultrastructural damage ameliorated after direct PRF treatment to the SN in the CCI rat model of NP.


Disparities in urban neighborhood conditions: evidence from GIS measures and field observation in New York City.
Although many low-income urban areas are highly walkable by conventional measures such as population density or land use mix, chronic diseases related to lack of physical activity are more common among residents of these areas. Disparities in neighborhood conditions may make poor areas less attractive environments for walking, offsetting the advantages of density and land use mix. This study compared poor and nonpoor neighborhoods in New York City, using geographic information systems measures constructed from public data for US census tracts within New York City (N=2,172) as well as field observation of a matched-pair sample of 76 block faces on commercial streets in poor and nonpoor neighborhoods. Poor census tracts had significantly fewer street trees, landmarked buildings, clean streets, and sidewalk cafes, and higher rates of felony complaints, narcotics arrests, and vehicular crashes. The field observation showed similar results. Improving aesthetic and safety conditions in poor neighborhoods may help reduce disparities in physical activity among urban residents.


The effect of continuous intrusive force on human pulpal blood flow.
The purpose of this study was to examine the effect of continuous intrusive force application on human pulpal blood flow (PBF). Recordings were made of 13 vital upper left central incisors in 13 healthy participants (experimental group, n = 8; control group, n = 5) who had clinically healthy tooth crowns and periodontal tissues. PBF was recorded by means of a laser Doppler flow meter (LDF) with an opaque rubber dam applied to the teeth. The basal blood flow in the pulp (BBFP) was compared during three observation periods: (1) before orthodontic archwire engagement; (2) during wire engagement (control group: no application of force; experimental group: continuous intrusive force of 0.5 N); and (3) after the removal of the wire. In the experimental group, brief intrusive forces (magnitude 0.5, 1.0, and 2.0 N) were applied to the incisal edge of the examined teeth to determine their effect on acute changes in PBF. The results obtained were as follows: (1) the BBFP in the experimental group was significantly reduced during the period of continuous intrusive force application (P < 0.05), which was followed by recovery after the removal of the wire. (2) Brief intrusive force produced a significant reduction of PBF (P < 0.05), but the reduction rate (percentage) did not differ significantly during the observation periods. The results indicate that the measurement technique shown in this study could detect PBF change produced by continuous orthodontic force application.


Antiproliferative activities of lesser galangal (Alpinia officinarum Hance Jam1), turmeric (Curcuma longa L.), and ginger (Zingiber officinale Rosc.) against acute monocytic leukemia.
Acute monocytic leukemia (AML M5 or AMoL) is one of the several types of leukemia that are still awaiting cures. The use of chemotherapy for cancer management can be harmful to normal cells in the vicinity of the target leukemia cells. This study assessed the potency of the extracts from lesser galangal, turmeric, and ginger against AML M5 to use the suitable fractions in neutraceuticals. Aqueous and organic solvent extracts from the leaves and rhizomes of lesser galangal and turmeric, and from the rhizomes only of ginger were examined for their antiproliferative activities against THP-1 AMoL cells in vitro. Lesser galangal leaf extracts in organic solvents of methanol, chloroform, and dichloromethane maintained distinctive antiproliferative activities over a 48-h period. The turmeric leaf and rhizome extracts and ginger rhizome extracts in methanol also showed distinctive anticancer activities. The lesser galangal leaf methanol extract was subsequently separated into 13, and then 18 fractions using reversed-phase high-performance liquid chromatography. Fractions 9 and 16, respectively, showed the greatest antiproliferative activities. These results indicate that the use of plant extracts might be a safer approach to finding a lasting cure for AMoL. Further investigations will be required to establish the discriminatory tolerance of normal cells to these extracts, and to identify the compounds in these extracts that possess the antiproliferative activities.


A deep-learning model for transforming the style of tissue images from cryosectioned to formalin-fixed and paraffin-embedded.
Histological artefacts in cryosectioned tissue can hinder rapid diagnostic assessments during surgery. Formalin-fixed and paraffin-embedded (FFPE) tissue provides higher quality slides, but the process for obtaining them is laborious (typically lasting 12-48 h) and hence unsuitable for intra-operative use. Here we report the development and performance of a deep-learning model that improves the quality of cryosectioned whole-slide images by transforming them into the style of whole-slide FFPE tissue within minutes. The model consists of a generative adversarial network incorporating an attention mechanism that rectifies cryosection artefacts and a self-regularization constraint between the cryosectioned and FFPE images for the preservation of clinically relevant features. Transformed FFPE-style images of gliomas and of non-small-cell lung cancers from a dataset independent from that used to train the model improved the rates of accurate tumour subtyping by pathologists.