Genetic variants of the endothelial NO synthase gene (eNOS) may confer increased risk of sporadic congenital heart disease.
The endothelial NO synthase (eNOS) enzyme is expressed during the early stages of cardiogenesis and plays an important role in normal heart development. Genetic variations of eNOS G894T have been shown to influence individual susceptibility to some phenotypes of congenital heart disease (CHD) in different populations. We conducted a case-control study comprised of 945 CHD patients and 972 non-CHD individuals in a Chinese population. Two functional single nucleotide polymorphisms (SNPs) (T-786C: rs2070744 and G894T: rs1799983) and one tagging SNP (rs7830) were evaluated in our study, and we assessed their association with the risk of CHD. Compared with the rs7830 CC/AC genotypes, the eNOS rs7830 AA genotype showed a significantly increased risk of CHD (adjusted odds radio (OR) = 1.45, 95% confidence interval (CI = 1.13-1.85). A stratified analysis was performed and showed that the association between the rs7830 AA genotype and CHD risk was stronger in patients with perimembranous ventricular septal defects (adjusted OR = 1.62, 95%CI = 1.20-2.20). Our results suggest that the eNOS rs7830 polymorphism may contribute to the susceptibility of sporadic CHD in a Chinese population.


Screening of Cu4 O3 NPs efficacy and its anticancer potential against cervical cancer.
Cu4 O3 is the least explored copper oxide, and its nanoformulation is anticipated to have important therapeutic potential especially against cancer. The current study aimed to biosynthesize Cu4 O3 nanoparticles (NPs) using an aqueous extract of pumpkin seeds and evaluate its antiproliferative efficacy against cervical cells after screening on different cancer cell lines. The obtained NPs were characterized by different spectroscopic analyses, such as UV-vis, thermogravimetric, energy dispersive X-ray, and Fourier-transform infrared spectroscopy (FTIR). In addition, high-resolution transmission electron microscopes (HR-TEM) were used to observe the morphology of the biosynthesized NPs. The UV-vis spectra showed a peak at around 332 nm, confirming the formation of Cu4 O3 NPs. Moreover, FTIR and TAG analyses identified the presence of various bioactive phytoconstituents that might have worked as capping and stabilization agents and comparative stable NPs at very high temperatures, respectively. The HR-TEM data showed the spherical shape of Cu4 O3 NPs in the range of 100 nm. The Cu4 O3 NPs was screened on three different cancer cell lines viz., Hela, MDA-MB-231, and HCT-116 using cytotoxicity (MTT) reduction assay. In addition, Vero was taken as a normal epithelial (control) cell. The high responsive cell line in terms of least IC50 was further assessed for its anticancer potential using a battery of biological tests, including morphological alterations, induction of apoptosis/ROS generation, regulation of mitochondrial membrane potential (MMP), and suppression of cell adhesion/migration. Vero cells (control) showed a slight decline in % cell viability even at the highest tested Cu4 O3 NPs concentration. However, all the studied cancer cells viz., MDA-MB-231, HCT 116, and HeLa cells showed a dose-dependent decline in cell viability after the treatment with Cu4 O3 NPs with a calculated IC50 value of 10, 11, and 7.2 µg/mL, respectively. Based on the above data, Hela cells were chosen for further studies, that showed induction of apoptosis from 3.5 to 9-folds by three different staining techniques acridine orange/ethidium bromide (AO/EB), 4',6-diamidino-2-phenylindole (DAPI), and propidium iodide (PI). The enhanced production of reactive oxygen species (>3.5-fold), modulation in MMP, and suppression of cell adhesion/migration were observed in the cells treated with Cu4 O3 NPs. The current study obtained the significant antiproliferative potential of Cu4 O3 NPs against the cervical cancer cell line, which needs to be confirmed further in a suitable in vivo model. Based on our results, we also recommend the green-based, eco-friendly, and cost-effective alternative method for synthesizing novel nanoformulation.


A Comparative Study of Fibronectin Cleavage by MMP-1, -3, -13, and -14.
Fibronectin fragments are important for synovial inflammation and the progression of arthritis, and thus, identifying potential enzymatic pathways that generate these fragments is of vital importance. The objective of this study was to determine the cleavage efficiency of fibronectin by matrix metalloproteinases (MMP-1, MMP-3, MMP-13, and MMP-14). Intact human plasma fibronectin was co-incubated with activated MMPs in neutral or acidic pH for up to 24 hours at 37 °C. The size and distribution of fibronectin fragments were determined by Western blot analysis using antibodies that recognized the N-terminals of fibronectin. All MMPs were able to cleave fibronectin at neutral pH. MMP-13 and -14 had the highest efficiency followed by MMP-3 and -1. MMP-3, -13, and -14 generated 70-kDa fragments, a known pro-inflammatory peptide. Further degradation of fibronectin fragments was only found for MMP-13 and -14, generating 52-kDa, 40-kDa, 32-kDa, and 29-kDa fragments. Fibronectin fragments of similar size were also found in the articular cartilage of femoral condyles of normal bovine knee joints. At acidic pH (5.5), the activities of MMP-1 and -14 were nearly abolished, while MMP-3 had a greater efficiency than MMP-13 even though the activities of both MMPs were significantly reduced. These findings suggest that MMP-13 and -14 may play a significant role in the cleavage of fibronectin and the production of fibronectin fragments in normal and arthritic joints.


Children with Moderate Acute Malnutrition with No Access to Supplementary Feeding Programmes Experience High Rates of Deterioration and No Improvement: Results from a Prospective Cohort Study in Rural Ethiopia.
Children with moderate acute malnutrition (MAM) have an increased risk of mortality, infections and impaired physical and cognitive development compared to well-nourished children. In parts of Ethiopia not considered chronically food insecure there are no supplementary feeding programmes (SFPs) for treating MAM. The short-term outcomes of children who have MAM in such areas are not currently described, and there remains an urgent need for evidence-based policy recommendations. We defined MAM as mid-upper arm circumference (MUAC) of ≥11.0 cm and <12.5 cm with no bilateral pitting oedema to include Ethiopian government and World Health Organisation cut-offs. We prospectively surveyed 884 children aged 6-59 months living with MAM in a rural area of Ethiopia not eligible for a supplementary feeding programme. Weekly home visits were made for seven months (28 weeks), covering the end of peak malnutrition through to the post-harvest period (the most food secure window), collecting anthropometric, socio-demographic and food security data. By the end of the study follow up, 32.5% (287/884) remained with MAM, 9.3% (82/884) experienced at least one episode of SAM (MUAC <11 cm and/or bilateral pitting oedema), and 0.9% (8/884) died. Only 54.2% of the children recovered with no episode of SAM by the end of the study. Of those who developed SAM half still had MAM at the end of the follow up period. The median (interquartile range) time to recovery was 9 (4-15) weeks. Children with the lowest MUAC at enrolment had a significantly higher risk of remaining with MAM and a lower chance of recovering. Children with MAM during the post-harvest season in an area not eligible for SFP experience an extremely high incidence of SAM and a low recovery rate. Not having a targeted nutrition-specific intervention to address MAM in this context places children with MAM at excessive risk of adverse outcomes. Further preventive and curative approaches should urgently be considered.


Effect of various degrees of dystocia in dairy cattle on calf viability, milk production, fertility and culling.
The objective of this study was to analyse the effect of dystocia on calf viability, milk production, fertility and culling under field (farm) conditions in a retrospective case-control study. A total of 253 calvings that were attended by a veterinarian were classified into three categories: mild dystocia (MD), severe dystocia (SD) and caesarean section (CS). For every case of dystocia, a control was chosen from the same farm and matched for calving month, breed and parity. Cases and controls were compared concerning calf viability, milk production, fertility and culling. The proportion of stillbirths was significantly higher in cows with dystocia compared with their controls. However, the difference was lower in CS cows than in the other two groups. MD had no significant effects on milk production, fertility and culling. There were no obvious effects of SD on monthly test day milk production, while CS cows produced significantly less milk than their controls. Compared with controls, less SD (50.0% versus 61.6%) and CS cows (48.6% versus 70.3%) conceived until 200 days in milk and more CS cows (31.1% versus 16.2%) were culled in the same time interval. These data demonstrate the scope of economic and animal welfare opportunities associated with the reduction of dystocia by management and breeding.


Interleukin-33 expression is specifically enhanced in inflamed mucosa of ulcerative colitis.
Interleukin (IL)-33 is a cytokine belonging to the IL-1 family. IL-33 has been shown to elicit a Th2-like cytokine response in immune cells. In this study, we investigated IL-33 expression in the inflamed mucosa of patients with inflammatory bowel disease (IBD), and characterized the molecular mechanisms responsible for IL-33 expression in human colonic subepithelial myofibroblasts (SEMFs). IL-33 mRNA expression was determined by real-time polymerase chain reaction (PCR). IL-33 expression in the IBD mucosa was evaluated by immunohistochemical methods. IL-33 mRNA expression was significantly elevated in active lesions from patients with ulcerative colitis (UC), but was not detected in inactive lesions from UC patients or in lesions from patients with either active or inactive Crohn's disease. Colonic SEMFs were identified as a major source of IL-33 in the mucosa. IL-1β and tumor necrosis factor-α (TNF-α) significantly enhanced IL-33 mRNA and protein expression in isolated colonic SEMFs. IL-1β and TNF-α did not affect IL-33 expression in intestinal epithelial cell lines (HT-29 and Caco-2 cells). This IL-1β- and TNF-α-induced IL-33 mRNA expression was mediated by p42/44 mitogen activated protein kinase (MAPK) pathway-dependent activation of nuclear factor (NF)-κB and activator protein (AP)-1. IL-33, derived from colonic SEMFs, may play an important role in the pathophysiology of UC.


Acute effect of endothelins on intercellular communication of human embryonic stem cells.
Endothelin (ET) family comprises three isoforms, ET-1, ET-2 and ET-3 that bind to two receptors ET-A and ET-B. Upon hESC differentiation, ET-1 and ET-B are respectively up- and down-regulated, suggesting a potential role of ETs in hESC biology. Here we show expression of ET receptors in hESC and demonstrate that ET-1 and ET-2 inhibit gap junctional intercellular communication (GJIC), while ET-3 does not. Pre-incubation of the cell cultures with the two specific antagonists of ET-A and ET-B, BQ123 and BQ788 respectively, demonstrate that inhibition of GJIC by ETs is mediated by ET-A. Long-term treatment of hESC with ET-1 indicates no visible effect on hESC maintenance of pluripotency markers, as assessed by expression of the hESC markers Oct-4, GCTM-2 and TG-30. Altogether these data show that hESC are target cells of ETs.


Childhood Guillain-Barré syndrome. MR imaging in diagnosis and follow-up.
To evaluate the value of serially obtained spinal MR images in the diagnosis and clinical follow-up of childhood Guillain-Barré syndrome (GBS), and the time period for resolution of contrast enhancement during the course of illness. Contrast-enhanced spinal MR images of 11 pediatric patients with GBS were reviewed prospectively. In the first group of 6 patients, follow-up MR was performed three times after the 1st, 3rd and 6th months after onset of the disease. In the second group (the other 5 patients) who had not accepted serial MR examinations, control MR was repeated once after 1 year. Patients were graded clinically into five categories according to their initial clinical signs and symptoms and three grades (mild, moderate and severe) according to level of contrast enhancement on T1-weighted images to correlate the severity of enhancement with clinical findings and recovery. Ten of the 11 patients revealed contrast enhancement in varying degrees in the spinal nerve roots surrounding the conus medullaris and extending the length of the cauda equina. One patient showed no contrast enhancement. Anterior nerve roots enhanced more intensely than the posterior roots in 3 patients. Follow-up MR images in all patients revealed diminishing or resolving contrast enhancement in the nerve roots as the clinical symptoms improved. We suggest contrast-enhanced spinal MR imaging as a supplementary diagnostic modality in diagnosing GBS, especially when the clinical and electrophysiological findings are equivocal. Follow-up images may play a role in predicting the clinical course of the disease.


Investigation about forefoot pathology in Japan.
The author has been living in Japan for three years. From the beginning she got the impression that the feet of Japanese were different from those of Westerners. While attending at Japanese hospitals she never encountered patients with forefoot pathology. Therefore the digital and metatarsal formulas, plus the angle of Schuster, were investigated in Japanese and different values resulted. It is the author opinion that the foot of Japanese is a more developed one, from the philogenetic point of view. Moreover it is important that Japanese traditionally spend most of their time without shoes taking them off indoors.


Washout studies of 11C in rabbit thigh muscle implanted by secondary beams of HIMAC.
Heavy ion therapy has two definite advantages: good dose localization and higher biological effect. Range calculation of the heavy ions is an important factor in treatment planning. X-ray CT numbers are used to estimate the heavy ion range by looking up values in a conversion table which relates empirically photon attenuation in tissues to particle stopping power; this is one source of uncertainty in the treatment planning. Use of positron emitting radioactive beams along with a positron emission tomograph or a positron camera gives range information and may be used as a means of checking in heavy ion treatment planning. However, the metabolism of the implanted positron emitters in a living object is unpredictable because the chemical forms of these emitters are unknown and the metabolism is dependent on the organ species and may be influenced by many factors such as blood flow rate and fluid components present. In this paper, the washout rate of 11C activity implanted by injecting energetic 11C beams into thigh muscle of a rear leg of a rabbit is presented. The washout was found to consist of two components, the shorter one was about 4.2 +/- 1.1 min and the longer one ranged from 91 to 124 min. About one third of the implanted beta+ activity can be used for imaging and the rest was washed out of the target area.


Performance of three delignifying pretreatments on hardwoods: hydrolysis yields, comprehensive mass balances, and lignin properties.
In this work, three pretreatments under investigation at the DOE Bioenergy Research Centers (BRCs) were subjected to a side-by-side comparison to assess their performance on model bioenergy hardwoods (a eucalyptus and a hybrid poplar). These include co-solvent-enhanced lignocellulosic fractionation (CELF), pretreatment with an ionic liquid using potentially biomass-derived components (cholinium lysinate or [Ch][Lys]), and two-stage Cu-catalyzed alkaline hydrogen peroxide pretreatment (Cu-AHP). For each of the feedstocks, the pretreatments were assessed for their impact on lignin and xylan solubilization and enzymatic hydrolysis yields as a function of enzyme loading. Lignins recovered from the pretreatments were characterized for polysaccharide content, molar mass distributions, β-aryl ether content, and response to depolymerization by thioacidolysis. All three pretreatments resulted in significant solubilization of lignin and xylan, with the CELF pretreatment solubilizing the majority of both biopolymer categories. Enzymatic hydrolysis yields were shown to exhibit a strong, positive correlation with the lignin solubilized for the low enzyme loadings. The pretreatment-derived solubles in the [Ch][Lys]-pretreated biomass were presumed to contribute to inhibition of enzymatic hydrolysis in the eucalyptus as a substantial fraction of the pretreatment liquor was carried forward into hydrolysis for this pretreatment. The pretreatment-solubilized lignins exhibited significant differences in polysaccharide content, molar mass distributions, aromatic monomer yield by thioacidolysis, and β-aryl ether content. Key trends include a substantially higher polysaccharide content in the lignins recovered from the [Ch][Lys] pretreatment and high β-aryl ether contents and aromatic monomer yields from the Cu-AHP pretreatment. For all lignins, the 13C NMR-determined β-aryl ether content was shown to be correlated with the monomer yield with a second-order functionality. Overall, it was demonstrated that the three pretreatments highlighted in this study demonstrated uniquely different functionalities in reducing biomass recalcitrance and achieving higher enzymatic hydrolysis yields for the hybrid poplar while yielding a lignin-rich stream that may be suitable for valorization. Furthermore, modification of lignin during pretreatment, particularly cleavage of β-aryl ether bonds, is shown to be detrimental to subsequent depolymerization.


Discovering key residues of dengue virus NS2b-NS3-protease: New binding sites for antiviral inhibitors design.
The NS2B-NS3 protease is essential for the Dengue Virus (DENV) replication process. This complex constitutes a target for efficient antiviral discovery because a drug could inhibit the viral polyprotein processing. Furthermore, since the protease is highly conserved between the four Dengue virus serotypes, it is probable that a drug would be equally effective against all of them. In this article, a strategy is reported that allowed us to identify influential residues on the function of the Dengue NS2b-NS3 Protease. Moreover, this is a strategy that could be applied to virtually any protein for the search of alternative influential residues, and for non-competitive inhibitor development. First, we incorporated several features derived from computational alanine scanning mutagenesis, sequence, structure conservation, and other structure-based characteristics. Second, these features were used as variables to obtain a multilayer perceptron model to identify defined groups (clusters) of key residues as possible candidate pockets for binding sites of new leads on the DENV protease. The identified residues included: i) amino acids close to the beta sheet-loop-beta sheet known to be important in its closed conformation for NS2b ii) residues close to the active site, iii) several residues evenly spread on the NS2b-NS3 contact surface, and iv) some inner residues most likely related to the overall stability of the protease. In addition, we found concordance on our list of residues with previously identified amino acids part of a highly conserved peptide studied for vaccine development.


Chromosome speciation: humans, Drosophila, and mosquitoes.
Chromosome rearrangements (such as inversions, fusions, and fissions) may play significant roles in the speciation between parapatric (contiguous) or partly sympatric (geographically overlapping) populations. According to the "hybrid-dysfunction" model, speciation occurs because hybrids with heterozygous chromosome rearrangements produce dysfunctional gametes and thus have low reproductive fitness. Natural selection will, therefore, promote mutations that reduce the probability of intercrossing between populations carrying different rearrangements and thus promote their reproductive isolation. This model encounters a disabling difficulty: namely, how to account for the spread in a population of a chromosome rearrangement after it first arises as a mutation in a single individual. The "suppressed-recombination" model of speciation points out that chromosome rearrangements act as a genetic filter between populations. Mutations associated with the rearranged chromosomes cannot flow from one to another population, whereas genetic exchange will freely occur between colinear chromosomes. Mutations adaptive to local conditions will, therefore, accumulate differentially in the protected chromosome regions so that parapatric or partially sympatric populations will genetically differentiate, eventually evolving into different species. The speciation model of suppressed recombination has recently been tested by gene and DNA sequence comparisons between humans and chimpanzees, between Drosophila species, and between species related to Anopheles gambiae, the vector of malignant malaria in Africa.


Remote Consultations Versus Standard Face-to-Face Appointments for Liver Transplant Patients in Routine Hospital Care: Feasibility Randomized Controlled Trial of myVideoClinic.
Using technology to reduce the pressure on the National Health Service (NHS) in England and Wales is a key government target, and the NHS Long-Term Plan outlines a strategy for digitally enabled outpatient care to become mainstream by 2024. In 2020, the COVID-19 response saw the widespread introduction of remote consultations for patient follow-up, regardless of individual preferences. Despite this rapid change, there may be enduring barriers to the effective implementation of remote appointments into routine practice once the unique drivers for change during the COVID-19 pandemic no longer apply, to which pre-COVID implementation studies can offer important insights. This study aims to evaluate the feasibility of using real-time remote consultations between patients and secondary care physicians for routine patient follow-up at a large hospital in the United Kingdom and to assess whether patient satisfaction differs between intervention and usual care patients. Clinically stable liver transplant patients were randomized to real-time remote consultations in which their hospital physician used secure videoconferencing software (intervention) or standard face-to-face appointments (usual care). Participants were asked to complete postappointment questionnaires over 12 months. Data were analyzed on an intention-to-treat basis. The primary outcome was the difference in scores between baseline and study end by patient group for the three domains of patient satisfaction (assessed using the Visit-Specific Satisfaction Instrument). An embedded qualitative process evaluation used interviews to assess patient and staff experiences. Of the 54 patients who were randomized, 29 (54%) received remote consultations, and 25 (46%) received usual care (recruitment rate: 54/203, 26.6%). The crossover between study arms was high (13/29, 45%). A total of 129 appointments were completed, with 63.6% (82/129) of the questionnaires being returned. Patient satisfaction at 12 months increased in both the intervention (25 points) and usual care (14 points) groups. The within-group analysis showed that the increases were significant for both intervention (P<.001) and usual care (P=.02) patients; however, the between-group difference was not significant after controlling for baseline scores (P=.10). The qualitative process evaluation showed that-according to patients-remote consultations saved time and money, were less burdensome, and caused fewer negative impacts on health. Technical problems with the software were common, and only 17% (5/29) of patients received all appointments over video. Both consultants and patients saw remote consultations as positive and beneficial. Using technology to conduct routine follow-up appointments remotely may ease some of the resource and infrastructure challenges faced by the UK NHS and free up clinic space for patients who must be seen face-to-face. Our findings regarding the advantages and challenges of using remote consultations for routine follow-ups of liver transplant patients have important implications for service organization and delivery in the postpandemic NHS. ISRCTN Registry 14093266; https://www.isrctn.com/ISRCTN14093266. RR2-10.1186/s13063-018-2953-4.


[The clinical and social vulnerability of children with special healthcare needs].
Children with special healthcare needs (CSHCN) require increasing care by relatives and health services. Due to the 'invisibility' of this group, a qualitative studied was carried out to analyze and to discuss the dimension of physical care provided to CSHCN. The studied included the analyses of the patients' records, interviews of family caretakers in a university hospital in southern Brazil, and the application of a sensitive creative method. Findings showed that these children are clinically and socially vulnerable. Consequently, during home care, their caretakersf'ce challenges related to care complexity and to the fact of being women. The presence of the State is scarce, and follow-up nursing after discharge is absent. It is recommended that these caretakers are taught some nursing knowledge, and that actions are taken to improve prenatal care services as a preventive measure.


Ca2+ -dependent release of ATP from astrocytes affects herpes simplex virus type 1 infection of neurons.
Astrocytes provide metabolic support for neurons and modulate their functions by releasing a plethora of neuroactive molecules diffusing to neighboring cells. Here we report that astrocytes also play a role in cortical neurons' vulnerability to Herpes simplex virus type-1 (HSV-1) infection through the release of extracellular ATP. We found that the interaction of HSV-1 with heparan sulfate proteoglycans expressed on the plasma membrane of astrocytes triggered phospholipase C-mediated IP3 -dependent intracellular Ca2+ transients causing extracellular release of ATP. ATP binds membrane purinergic P2 receptors (P2Rs) of both neurons and astrocytes causing an increase in intracellular Ca2+ concentration that activates the Glycogen Synthase Kinase (GSK)-3β, whose action is necessary for HSV-1 entry/replication in these cells. Indeed, in co-cultures of neurons and astrocytes HSV-1-infected neurons were only found in proximity of infected astrocytes releasing ATP, whereas in the presence of fluorocitrate, an inhibitor of astrocyte metabolism, switching-off the HSV-1-induced ATP release, very few neurons were infected. The addition of exogenous ATP, mimicking that released by astrocytes after HSV-1 challenge, restored the ability of HSV-1 to infect neurons co-cultured with metabolically-inhibited astrocytes. The ATP-activated, P2R-mediated, and GSK-3-dependent molecular pathway underlying HSV-1 infection is likely shared by neurons and astrocytes, given that the blockade of either P2Rs or GSK-3 activation inhibited infection of both cell types. These results add a new layer of information to our understanding of the critical role played by astrocytes in regulating neuronal functions and their response to noxious stimuli including microbial agents via Ca2+ -dependent release of neuroactive molecules.


The role of emotion in musical improvisation: an analysis of structural features.
One of the primary functions of music is to convey emotion, yet how music accomplishes this task remains unclear. For example, simple correlations between mode (major vs. minor) and emotion (happy vs. sad) do not adequately explain the enormous range, subtlety or complexity of musically induced emotions. In this study, we examined the structural features of unconstrained musical improvisations generated by jazz pianists in response to emotional cues. We hypothesized that musicians would not utilize any universal rules to convey emotions, but would instead combine heterogeneous musical elements together in order to depict positive and negative emotions. Our findings demonstrate a lack of simple correspondence between emotions and musical features of spontaneous musical improvisation. While improvisations in response to positive emotional cues were more likely to be in major keys, have faster tempos, faster key press velocities and more staccato notes when compared to negative improvisations, there was a wide distribution for each emotion with components that directly violated these primary associations. The finding that musicians often combine disparate features together in order to convey emotion during improvisation suggests that structural diversity may be an essential feature of the ability of music to express a wide range of emotion.


Epidemiological and nutrition transition in developing countries: impact on human health and development.
Whereas common infectious and parasitic diseases such as malaria and the HIV/AIDS pandemic remain major unresolved health problems in many developing countries, emerging non-communicable diseases relating to diet and lifestyle have been increasing over the last two decades, thus creating a double burden of disease and impacting negatively on already over-stretched health services in these countries. Prevalence rates for type 2 diabetes mellitus and CVD in sub-Saharan Africa have seen a 10-fold increase in the last 20 years. In the Arab Gulf current prevalence rates are between 25 and 35% for the adult population, whilst evidence of the metabolic syndrome is emerging in children and adolescents. The present review focuses on the concept of the epidemiological and nutritional transition. It looks at historical trends in socio-economic status and lifestyle and trends in nutrition-related non-communicable diseases over the last two decades, particularly in developing countries with rising income levels, as well as the other extreme of poverty, chronic hunger and coping strategies and metabolic adaptations in fetal life that predispose to non-communicable disease risk in later life. The role of preventable environmental risk factors for obesity and the metabolic syndrome in developing countries is emphasized and also these challenges are related to meeting the millennium development goals. The possible implications of these changing trends for human and economic development in poorly-resourced healthcare settings and the implications for nutrition training are also discussed.


Regulation of Escherichia coli adenylate cyclase activity during hexose phosphate transport.
In Escherichia coli, cAMP levels vary with the carbon source used in the culture medium. These levels are dependent on the cellular concentration of phosphorylated EnzymeIIAglc, a component of the glucose-phosphotransferase system, which activates adenylate cyclase (AC). When cells are grown on glucose 6-phosphate (Glc6P), the cAMP level is particularly low. In this study, we investigated the mechanism leading to the low cAMP level when Glc6P is used as the carbon source, i.e. the mechanism preventing the activation of AC by phosphorylated EnzymeIIAglc. Glc6P is transported via the Uhp system which is inducible by extracellular Glc6P. The Uhp system comprises a permease UhpT and three proteins UhpA, UhpB and UhpC which are necessary for uhpT gene transcription. Controlled expression of UhpT in the absence of the regulatory proteins (UhpA, UhpB and UhpC) allowed us to demonstrate that (i) the Uhp regulatory proteins do not prevent the activation of AC by direct interaction with EnzymeIIAglc and (ii) an increase in the amount of UhpT synthesized (corresponding to an increase in the amount of Glc6P transported) correlates with a decrease in the cAMP level. We present data indicating that Glc6P per se or its degradation is unlikely to be responsible for the low cAMP level. It is concluded that the level of cAMP in the cell is determined by the flux of Glc6P through UhpT.


Rapid, highly discriminatory binary genotyping to demonstrate methicillin-resistant Staphylococcus aureus transmission in a tertiary care intensive care unit.
No previous studies of methicillin-resistant Staphylococcus aureus (MRSA) epidemiology in adult intensive care units (ICUs) have assessed the utility of rapid, highly discriminatory strain typing in the investigation of transmission events. Observational. A 22-bed medical-surgical adult ICU. Patients Those admissions MRSA-positive on initial screening and all admissions <48 hours in duration were excluded, leaving a cohort of 653 patients (median age, 61 years; APACHE-II, 19). We conducted this study of MRSA transmission over 1 year (August 1, 2011 to July 31, 2012) using a multiplex PCR-based reverse line blot (mPCR/RLB) assay to genotype isolates from surveillance swabs obtained at admission and twice weekly during ICU stays. MRSA prevalence and incidence rates were calculated and transmission events were identified using strain matching. Colonization pressure was calculated daily by summation of all MRSA cases. Of 1,030 admissions to ICU during the study period, 349 patients were excluded. MRSA acquisition occurred during 31 of 681 (4.6%) remaining admissions; 19 of 31(61%) acquisitions were genotype-confirmed, including 7 (37%) due to the most commonly transmitted strain. Moving averages of MRSA patient numbers on the days prior to a documented event were used in a Poisson regression model. A significant association was found between transmission and colonization pressure when the average absolute colonization pressure on the previous day was ≥3 (χ2=7.41, P=0.01). mPCR/RLB characterizes MRSA isolates within a clinically useful time frame for identification of single-source clusters within the ICU. High MRSA colonization pressure (≥3 MRSA-positive patients) on a given day is associated with an increased likelihood of a transmission event.


Glutamate and obesity - what is the link?
Many studies using metabolomics have tried to unravel the metabolic signature of obesity and understand the pathophysiology of this complex and heterogeneous disease. Circulating levels of the amino acid glutamate have been consistently associated with obesity and more specifically with measurements of abdominal fat accumulation. The purpose of this narrative review is to highlight recent studies documenting this association. Circulating glutamate concentrations have been positively correlated with measurements of central fat accumulation such as waist circumference and visceral adipose tissue area. Moreover, elevated glutamate levels have been linked to a higher prevalence of type 2 diabetes, cardiovascular diseases and nonalcoholic fatty liver disease. The association with adiposity is detected in early life, and genetic predisposition does not appear as a major driver. Glutamate levels reflect in vivo synthesis rather than dietary intake. However, interventions generating metabolic improvements such as incretin receptor agonist treatment or dietary improvements may reduce plasma levels of this amino acid. Recent findings confirm the consistent association between circulating glutamate and abdominal obesity and its cardiometabolic complications. The pathophysiological pathways underlying this phenomenon are still unclear. Furthermore, studies are needed to establish the usefulness of this analyte as a biomarker of abdominal obesity.


National medicines policies - a review of the evolution and development processes.
Continuous provision of appropriate medicines of assured quality, in adequate quantities, and at reasonable prices is a concern for all national governments. A national medicines policy (NMP) developed in a collaborative fashion identifies strategies needed to meet these objectives and provides a comprehensive framework to develop all components of a national pharmaceutical sector. To meet the health needs of the population, there is a general need for medicine policies based on universal principles, but nevertheless adapted to the national situation. This review aims to provide a quantitative and qualitative (describing the historical development) study of the development process and evolution of NMPs. The number of NMPs and their current status has been obtained from the results of the assessment of WHO Level I indicators. The policy formulation process is examined in more detail with case studies from four countries: Sri Lanka, Australia, former Yugoslav Republic of Macedonia and South Africa. The number of NMPs worldwide has increased in the last 25 years with the highest proportional increase in the last 5-10 years in high-income countries. Higher income countries seem to have more NMP implementation plans available and have updated their NMP more recently. The four case studies show that the development of a NMP is a complex process that is country specific. In addition, it demonstrates that an appropriate political window is needed for the policy to be passed (for South Africa and the FYR Macedonia, a major political event acted as a trigger for initiating the policy development). Policy-making does not stop with the official adoption of a policy but should create mechanisms for implementation and monitoring. The NMPs of the FYR Macedonia and Australia provide indicators for monitoring. To date, not all countries have a NMP since political pressure by national experts or non-governmental organizations is generally needed to establish a NMP. Case studies in four countries showed that the policy process is just as important as the policy document since the process must create a mechanism by which all stakeholders are brought together and a sense of collective ownership of the final policy may be achieved.


Early-life stress induces persistent alterations in 5-HT1A receptor and serotonin transporter mRNA expression in the adult rat brain.
Early-life experience plays a major role in the stress response throughout life. Neonatal maternal separation (MS) is an animal model of depression with an altered serotonergic response. We hypothesize that this alteration may be caused by differences in 5-HT1A receptor and serotonin transporter (SERT) mRNA expression in brain areas involved in the control of emotions, memory, and fear as well as in regions controlling the central serotonergic tone. To test this, Sprague-Dawley rats were subjected to MS for 3 h daily during postnatal days 2-12. As control, age matched rats were non-separated (NS) from their dams. When animals reached adulthood (11-13 weeks) brain was extracted and mRNA expression of 5-HT1A receptor in amygdala, hippocampus and dorsal raphé nucleus (DRN) and SERT in the DRN was analyzed through in situ hybridisation. Densitometric analysis revealed that MS increased 5-HT1A receptor mRNA expression in the amygdala, and reduced its expression in the DRN, but no changes were observed in the hippocampus in comparison to NS controls. Also, MS reduced SERT mRNA expression in the DRN when compared to NS rats. These results suggest that early-life stress induces persistent changes in 5-HT1A receptor and SERT mRNA expression in key brain regions involved in the development of stress-related psychiatric disorders. The reduction in SERT mRNA indicates an alteration that is in line with clinical findings such as polymorphic variants in individuals with higher risk of depression. These data may help to understand how early-life stress contributes to the development of mood disorders in adulthood.


Mild developmental delay due to ring chromosome 19 mosaicism.
Children with mild developmental delay without dysmorphic features do not often have identifiable underlying aetiological factors. We report on a 5-year-old girl with mild developmental delay and dysmorphic features which were previously unrecognized. She was found to have supernumerary ring chromosome 19 mosaicism which was the likely cause of her clinical problems. Her parents' chromosomes were normal. A careful examination for dysmorphic features should be done in all children with developmental delay. However, these may not be readily apparent in babies and very young children. Chromosomal analysis to identify a genetic cause and to offer genetic counselling should be considered in all such children unless the clinician is absolutely certain that there are no dysmorphic features.


Energetics of the Phase Transition in Free-Standing versus Supported Lipid Membranes.
We compared the thermodynamic behavior of supported and free-standing films of phospholipids with different chain length and showed that the change in free energy for the phase transition to a denser state was greater in free-standing than in supported membranes, with the differences being independent of the chain length. The presence of the support promoted a decrease in the energy gap between the coexisting phases of 1.7 kJ mol(-1), and this was a long-range effect affecting both leaflets, probably related with the impediment of out-of-plane motion of the film by the solid surface.


Anatomy of an endogenous antagonist: relationship between Agouti-related protein and proopiomelanocortin in brain.
Agouti-related protein (AGRP) is a recently discovered orexigenic neuropeptide that inhibits the binding and action of alpha-melanocyte-stimulating hormone derived from proopiomelanocortin (POMC) at the melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R) and has been proposed to function primarily as an endogenous melanocortin antagonist. To better understand the interplay between the AGRP and melanocortin signaling systems, we compared their nerve fiber distributions with each other by immunohistochemistry and their perikarya distribution with MC3R and MC4R by double in situ hybridization. Although deriving from distinct cell groups, AGRP and melanocortin terminals project to identical brain areas. Both AGRP and melanocortin neurons selectively express the MC3R, which provides a neuroanatomical basis for a dual-input circuit with biological amplification and feedback inhibition. These studies highlight a broader complexity in POMC-mediated behavior in the brain.


Applicability of extracellular electrical impedance tomography in monitoring respiratory tract inflammation.
The presence of persistent mild inflammation is widely considered to provide the immunopathogenic basis for bronchial, nasal, or sinusal inflammation between critical phases and in asymptomatic periods. Exhaled nitric oxide (NO) is currently the most reliable marker of rhinobronchial inflammation, but its routine assessment is difficult as the test is available only in highly specialized centers. The aim of this study was to evaluate the agreement between a new diagnostic method (extracellular electrical impedance tomography) and immunological and clinical symptom scores, anterior rhinomanometry, peak expiratory flow rate (PEFR), serum eosinophil cationic protein (ECP) level, and blood eosinophil count in the clinical monitoring of respiratory tract inflammation before and after treatment of asthma or rhinitis. Eighty-seven patients were studied; 73 had mild persistent asthma (PEFR > or = 20% below predicted) and 14 had rhinitis. At baseline (TO), the patients underwent a medical examination to record symptom scores, PEFR, anterior rhinomanometry, an extracellular electrolytic conductivity test (bioimpedance tomography), serum ECP level and blood eosinophil count. Appropriate treatment was prescribed, following the guidelines of the Global Initiative for Asthma and the Allergic Rhinitis and Its Impact on Asthma. After 21 days of therapy (T1), the patients were re-evaluated for the same parameters. This study demonstrates the good agreement (Cohen's kappa = 0.689) between the symptom scores of patients with rhinitis and the findings of extracellular tomography and very good agreement (kappa = 0.846) between symptom scores of asthma patients and extracellular tomography. These findings validate the use of this new technique for the real-time monitoring and adjustment of treatment in these clinical settings.


Luteolin Attenuates Allergic Nasal Inflammation via Inhibition of Interleukin-4 in an Allergic Rhinitis Mouse Model and Peripheral Blood From Human Subjects With Allergic Rhinitis.
Objectives: Luteolin is the active component of Perilla frutescens, an herb for the treatment of allergy in Asia. In this study, we aimed to investigate the effects and mechanisms of luteolin treatment. Methods: BALB/c mice sensitized with house dust mite (HDM) to induce allergic rhinitis (AR), and treated with dexamethasone or luteolin. In addition, mononuclear cells from peripheral blood (PBMC) of AR patients were co-cultured with dexamethasone or luteolin, and were re-stimulated with HDM. Results: Luteolin-treated mice had decreased allergic symptoms, and serum HDM-specific IgE when compared to the untreated group. Flow cytometric analyses of splenocytes and nasal lymphoid tissues from AR mice found that luteolin decreased CD4+ IL-4-secreting T cells when compared to those from vehicle treated AR mice. Histopathology sections showed reduced infiltration of eosinophils and decreased mucus secretion of mouse nasal epithelium. In the in vitro study, the results showed that luteolin reduced the percentage of CD4+ IL-4-secreting splenocytes expression was through reducing expression of pSTAT6 and GATA3. PBMCs from AR patients pretreated with luteolin could decrease percentage of CD4+ IL-4-secreting cells. Conclusion: Our study identified that luteolin attenuates allergic nasal inflammation via inhibition of IL-4 production, which supports the potential pharmaceutical application of luteolin treatment for AR.


Transesophageal echocardiographic diagnosis of ventricular noncompaction associated with an atrial septal aneurysm in a patient with dilated cardiomyopathy of unknown etiology.
The present report discusses a case of noncompaction of the ventricular myocardium in a 51-year-old woman with dilated cardiomyopathy of unknown etiology. Multiple transthoracic echocardiography examinations had failed to show myocardial noncompaction, but subsequently performed transesophageal echocardiography clearly demonstrated the characteristic findings of this unusual disease. Also, an atrial septal aneurysm was identified by transesophageal echocardiography. Patients with cardiomyopathy of unknown origin should be investigated to define the presence or absence of myocardial noncompaction, even if transthoracic echocardiography fails to show anatomical features of this disease. The present case is the first reported case of ventricular noncompaction associated with an atrial septal aneurysm as a congenital anomaly.


The role of decorated SDS micelles in sub-CMC protein denaturation and association.
We have combined spectroscopy, chromatography, calorimetry, and small-angle X-ray scattering (SAXS) to provide a comprehensive structural and stoichiometric description of the sodium dodecyl sulfate (SDS)-induced denaturation of the 86-residue alpha-helical bovine acyl-coenzyme-A-binding protein (ACBP). Denaturation is a multistep process. Initial weak binding of 1-3 SDS molecules per protein molecule below 1.3 mM does not perturb the tertiary structure. Subsequent binding of approximately 13 SDS molecules per ACBP molecule leads to the formation of SDS aggregates on the protein and changes in both tertiary and secondary structures. SAXS data show that, at this stage, a decorated micelle links two ACBP molecules together, leaving about half of the polypeptide chain as a disordered region protruding into the solvent. Further titration with SDS leads to the additional uptake of 26 SDS molecules, which, according to SAXS, forms a larger decorated micelle bound to a single ACBP molecule. At the critical micelle concentration, we conclude from reduced mobility and increased fluorescence anisotropy that each ACBP molecule becomes associated with more than one micelle. At this point, 56-60 SDS molecules are bound per ACBP molecule. Our data provide key structural insights into decorated micelle complexes with proteins, revealing a remarkable diversity in the different conformations they can stabilize. The data highlight that a minimum decorated micelle size, which may be a key driving force for intermolecular protein association, exists. This may also provide a structural basis for the known ability of submicellar surfactant concentrations to induce protein aggregation and fibrillation.


Accelerating wavefunction in density-functional-theory embedding by truncating the active basis set.
Methods where an accurate wavefunction is embedded in a density-functional description of the surrounding environment have recently been simplified through the use of a projection operator to ensure orthogonality of orbital subspaces. Projector embedding already offers significant performance gains over conventional post-Hartree-Fock methods by reducing the number of correlated occupied orbitals. However, in our first applications of the method, we used the atomic-orbital basis for the full system, even for the correlated wavefunction calculation in a small, active subsystem. Here, we further develop our method for truncating the atomic-orbital basis to include only functions within or close to the active subsystem. The number of atomic orbitals in a calculation on a fixed active subsystem becomes asymptotically independent of the size of the environment, producing the required O(N(0)) scaling of cost of the calculation in the active subsystem, and accuracy is controlled by a single parameter. The applicability of this approach is demonstrated for the embedded many-body expansion of binding energies of water hexamers and calculation of reaction barriers of SN2 substitution of fluorine by chlorine in α-fluoroalkanes.


Distinguishing sleep from wake with a radar sensor: a contact-free real-time sleep monitor.
This work aimed to evaluate whether a radar sensor can distinguish sleep from wakefulness in real time. The sensor detects body movements without direct physical contact with the subject and can be embedded in the roof of a hospital room for completely unobtrusive monitoring. We conducted simultaneous recordings with polysomnography, actigraphy, and radar on two groups: healthy young adults (n = 12, four nights per participant) and patients referred to a sleep examination (n = 28, one night per participant). We developed models for sleep/wake classification based on principles commonly used by actigraphy, including real-time models, and tested them on both datasets. We estimated a set of commonly reported sleep parameters from these data, including total-sleep-time, sleep-onset-latency, sleep-efficiency, and wake-after-sleep-onset, and evaluated the inter-method reliability of these estimates. Classification results were on-par with, or exceeding, those often seen for actigraphy. For real-time models in healthy young adults, accuracies were above 92%, sensitivities above 95%, specificities above 83%, and all Cohen's kappa values were above 0.81 compared to polysomnography. For patients referred to a sleep examination, accuracies were above 81%, sensitivities about 89%, specificities above 53%, and Cohen's kappa values above 0.44. Sleep variable estimates showed no significant intermethod bias, but the limits of agreement were quite wide for the group of patients referred to a sleep examination. Our results indicate that the radar has the potential to offer the benefits of contact-free real-time monitoring of sleep, both for in-patients and for ambulatory home monitoring.


Intestinal Cryptosporidiosis and the Profile of the CD4 Counts in a Cohort of HIV Infected Patients.
Cryptosporidium is an infectious enteric pathogen which is capable of causing life-threatening illnesses in immunocompromised patients. This prospective study was planned to know the frequency of intestinal cryptosporidiosis in HIV infected patients and its correlation with their immune status. Also, the conventional diagnostic methods were compared with the copro-antigen detection test by using Enzyme Linked Immuno Sorbent Assay (ELISA). This was a prospective cohort study. Three consecutive stool samples which were collected from 90 HIV seropositive patients and 50 seronegative controls were screened for cryptosporidiosis by wet mount, direct modified ZN (Ziehl Neelsen) staining, modified ZN staining with formol ether concentration and copro-antigen detection by ELISA. Their immune statuses were measured by CD4 + cell counting. Statistical Analytical Tests which were Used: Odds ratio, Chi square test, Fisher extract test. Cryptosporidiosis was detected in 15 HIV seropositive cases. 13 cases had CD4 cell counts of < 100 cells/ μL. The formol ether concentration technique resulted in an increased number of oocysts/oil immersion field in 8 cases. ELISA was positive in 2 cases which were shown to be negative by modified ZN staining. All the controls were negative for cryptosporidium. Cryptosporidiosis is an opportunistic infection in HIV infected people who present with diarrhoea. The wet mount technique, though it is simple and inexpensive, is insensitive for the detection of cryptosporidium. The conventional modified ZN staining and the modified ZN staining with concentration have a sensitivity and a specificity of 85.71% and 98.84% respectively. The copro antigen detection by ELISA which has a greater sensitivity and specificity, is a useful tool in epidemiological studies.


The aetiology of atypical bone health in individuals with Down syndrome.
Trisomy 21 (T21), more commonly known as Down syndrome (DS) is a genetic condition where every cell in the body has an additional copy of chromosome 21. Despite improvements in our management of DS-associated health risks, we still do not understand how T21 impacts human bone health. This is a critical area of research owing to increased life expectancy of people with DS, and the predisposition of individuals with DS to early-onset osteoporosis and osteopenia. We have conducted a scoping review using the methodological framework of Arksey and O'Malley (2005) which analysed the existing data on bone growth, development, maintenance and repair in T21 using the Medical Subject Headings (MeSH) terms: Trisomy 21, Down syndrome, Down's syndrome, bone development, bone growth, bone maintenance, fracture risk, osteoporosis, bone mineral density, bone strength, bone mineral content, bone formation, bone repair, osteoblast, osteoclast, osteocyte, osteomacs. A total of 31 papers were identified. After screening, 16 articles were included in full-text review. There was a total of eleven in vivo animal model studies identified and included in the scoping review. Of those eleven, ten revealed a difference in bone growth and development in animal models of DS, and two found that bone maintenance and repair in animal models of DS is reduced with both studies reporting an osteoporotic bone phenotype in male and female mice. All five studies that included human participants reported impacts on bone growth and development with reduced bone growth rates and delayed bone maturation in individuals with DS. At the time of review, there were no human studies directly investigating bone maintenance and repair in individuals with DS. We found documented evidence that T21 impacts bone growth and development, maintenance and repair in both animal models and human studies.


Unusual presentations, management and outcomes of gastric stromal tumors: a single-center case series.
Gastrointestinal stromal tumors (GISTs) are uncommon mesenchymal tumors of the gastrointestinal tract. This study explores the safety of laparoscopy and the long-term oncological outcome in gastroesophageal GIST treatment. A prospectively maintained single-institution database was examined. The variables collected were patient demographics and comorbidities, surgical access (laparoscopic/open), type of surgery, length of stay, and complications. A total of 69 patients underwent GIST resection between January 2011 and June 2018, of whom 56.5% were male; the median age was 68 years (interquartile range 60-76). The majority of patients (78.3%, n=54/69) had a laparoscopic resection. Median length of stay was 6 days in the laparoscopic group and 9 days in the open group (P=0.003). Most patients had wedge excision (n=57/69, 82.6%), while 12 patients (17.4%) required a gastrectomy (one a Merendino type). All patients had an R0 resection and 1 patient (1.4%) had a recurrence, despite having a low-risk grade original tumor with negative margins. Patients in the open group had a significantly higher proportion of patients with a high-risk tumor (50%) compared to the laparoscopic group (3.7%, P=0.001). The mean survival was 92.7 months (95% confidence interval 86.3-99.2). Survival was better in the laparoscopic group (100.4 months) compared with the open group (55.1 months, P<0.001). Laparoscopic gastric GIST resection is an oncologically safe alternative to open surgery and is associated with a shorter hospital stay with no difference in complication rates or recurrence rates.


Investigating Managers' Fine-Grained Evaluation Processes in Organizations: Exploring Two Dual-Process Perspectives.
The dual-process theory is a significant theory in both organizational theory and social psychology and two conjectures about this theory are considered in this manuscript; the default-interventionist vs. parallel-competitive account. Our research goal is to empirically investigate these two views. In concrete terms, by using event-related potentials (ERPs), we seek to study the fine-grained brain processes and self-reported feelings involved in managers' evaluations of target employees within an economic context (firing employees) vs. a social network context (excluding employees). Using the stereotype content model categories, each target employee has high (or low) warmth and high (or low) levels of competence. In the fine-grained ERP analysis of the brain process, we focus on three time windows of interest: novelty detection (N2) and goal violation detection (N400) at the unconscious level, and we then evaluate conscious emotional arousal (late positive potential, LPP). Finally, we focus on the self-reported feelings when having to fire or exclude target employees. As goal pursuit theory predicts, the brain dynamics and self-reported measures differ widely across the two organizational contexts; in concrete terms, processes at a later stage overrule early stages depending on the context. This implies that the data bespeaks more for the parallel-competitive account than the default-interventionist account. We discuss the implications of these findings for research in management and management practice.


Water column productivity and temperature predict coral reef regeneration across the Indo-Pacific.
Predicted increases in seawater temperatures accelerate coral reef decline due to mortality by heat-driven coral bleaching. Alteration of the natural nutrient environment of reef corals reduces tolerance of corals to heat and light stress and thus will exacerbate impacts of global warming on reefs. Still, many reefs demonstrate remarkable regeneration from past stress events. This paper investigates the effects of sea surface temperature (SST) and water column productivity on recovery of coral reefs. In 71 Indo-Pacific sites, coral cover changes over the past 1-3 decades correlated negative-exponentially with mean SST, chlorophyll a, and SST rise. At six monitoring sites (Persian/Arabian Gulf, Red Sea, northern and southern Galápagos, Easter Island, Panama), over half of all corals were <31 years, implying that measured environmental variables indeed shaped populations and community. An Indo-Pacific-wide model suggests reefs in the northwest and central Indian Ocean, as well as the central west Pacific, are at highest risk of degradation, and those at high latitudes the least. The model pinpoints regions where coral reefs presently have the best chances for survival. However, reefs best buffered against temperature and nutrient effects are those that current studies suggest to be most at peril from future ocean acidification.


Progressive human cone-rod dysfunction (dystrophy).
The author has classified progressive human cone-rod dysfunction into primary and secondary types. The primary type, identified by early ERG cone and usually also rod abnormalities, was further subdivided into types 1 and 2 based on, amoung other distinguishing characteristics, the extent of associated retinal pigment epithelial defects. Secondary cone-rod dysfunction apparently results from disease initially affecting the retinal pigment epithelium. Initially normal ERG findings and the presence of flecks characterize this (type 3) progressive cone-rod dysfunction.


HLA-DM is present in one-fifth the amount of HLA-DR in the class II peptide-loading compartment where it associates with leupeptin-induced peptide (LIP)-HLA-DR complexes.
HLA-DM has been shown in vitro to catalyze the release of invariant chain (Ii) derived peptides from the peptide-binding groove of class II molecules, thereby facilitating the binding of antigenic peptides. Previous studies showed that at steady state, the majority of DM resides in the class II peptide-loading compartment (IIPLC) where Ii dissociates from class II molecules and antigenic peptides are bound. Here we characterize the expression of DM in vivo in subcellular fractions containing the IIPLC. Using quantitative immunoblotting, we show that in the cell as a whole, class II molecules are expressed in 23-fold molar excess of DM. However, DM is concentrated in the IIPLC, where it is present in a considerably higher concentration relative to the class II molecules, in a molar ratio of 5DR:1 DM. This molar ratio of DM to DR in the IIPLC in vivo is consistent with the catalytic function proposed for DM from studies in vitro. We also provide both biochemical and genetic evidence that DM associates with complexes which contain Ii fragments and class II molecules in the IIPLC. Such complexes are only observed in leupeptin-treated cells in which Ii fails to be completely degraded and complexes containing the leupeptin-induced fragment of Ii (LIP) and class II molecules accumulate in the IIPLC. This observation is consistent with LIP-class II complexes being a substrate for DM in vivo and suggests that interactions of DM and LIP-class II are extremely transient under normal conditions.


Low CAIX expression and absence of VHL gene mutation are associated with tumor aggressiveness and poor survival of clear cell renal cell carcinoma.
We attempted to describe, in a series of clear cell renal cell carcinoma (RCC), the relationship between CAIX expression, VHL gene mutations, tumor characteristics and outcome. Radical nephrectomy was performed in 100 patients. Genomic DNA was extracted from frozen tumor samples. Four amplimers covering the whole coding sequence of the VHL gene were synthesized by PCR and sequenced. The monoclonal antibody M75 was used to evaluate CAIX protein expression immunohistochemically. VHL mutations were identified in 58 patients (58%) and high CAIX expression (>85%) was observed in 78 (78%). Tumors with VHL mutation showed higher CAIX expression than those without (p = 0.02). Low CAIX expression and absence of VHL mutation were associated with a more advanced tumors e.g., higher T stages and presence of metastases. VHL mutation and high CAIX expression predicted longer progression-free survival (p = 0.037) and disease-specific survival (p = 0.001), respectively. In combination, they defined three prognostic groups (p = 0.002): (i) good prognosis, defined as VHL mutation and high CAIX (2-year survival: 86%), (ii) intermediate prognosis with either VHL mutation or high CAIX (69%), and (iii) poor prognosis with no VHL mutation and low CAIX (45%, median survival 18 months). CAIX expression, but not VHL mutational status, was an independent prognostic factor in multivariate analysis. Taken together, CAIX expression and VHL mutational status are able to stratify patients with clear cell RCC into distinct groups with regards to clinicopathological variables and prognosis, with low CAIX expression and absence of VHL mutation being associated with a poor clinicopathological phenotype and diminished survival.


Intensity of factor VIII treatment and the development of inhibitors in non-severe hemophilia A patients: results of the INSIGHT case-control study.
Essentials Research suggests that intensive treatment episodes may increase the risk to develop inhibitors. We performed an international nested case-control study with 298 non-severe hemophilia A patients. Surgery and a high dose of factor VIII concentrate were associated with increased inhibitor risk. Physicians need to review arguments for factor VIII dose and elective surgery extra critically. Background Inhibitor development is a major complication of treatment with factor VIII concentrates in hemophilia. Findings from studies among severe hemophilia A patients suggest that intensive treatment episodes increase the risk of developing inhibitors. Objectives We set out to assess whether intensive treatment is also associated with an increased risk of inhibitor development among non-severe hemophilia A patients. Patients/Methods We performed a nested case-control study. A total of 75 inhibitor patients (cases) and 223 control patients were selected from 2709 non-severe hemophilia A patients (FVIII:C, 2-40%) of the INSIGHT cohort study. Cases and controls were matched for date of birth and cumulative number of exposure days (EDs) to FVIII concentrates. Conditional logistic regression was used to calculate both unadjusted and adjusted odds ratios (aOR); the latter were adjusted for a priori specified confounders. Results Peak treatment of 5 or 10 consecutive EDs did not increase inhibitor risk (aOR, 1.0; 95% confidence interval (CI), 0.4-2.5; and aOR, 1.8; CI, 0.6-5.5, respectively). Both surgical intervention (aOR, 4.2; CI, 1.7-10.3) and a high mean dose (> 45 IU kg-1 /ED) of FVIII concentrate (aOR, 7.5; CI, 1.6-35.6) were associated with an increased inhibitor risk. Conclusions Our findings suggest that high-dose FVIII treatment and surgery increase the risk of inhibitor development in non-severe hemophilia A. Together with the notion that non-severe hemophilia A patients are at a lifelong risk of inhibitor development, we suggest that in the future physicians will review the arguments for the FVIII dose and elective surgery extra critically.


In situ preparation and properties of polyvinyl alcohol/synthetic ribbon-like nanocellulose composites.
This study presents a novel approach in which a dual network (DN) composite, comprising polyvinyl alcohol (PVA) and ribbon-like nanocellulose (RC), was synthesized in one step using the volume exclusion effect involved in enzyme-catalyzed cellulose synthesis. Additionally, the impact of PVA as a crowding reagent during enzymatic catalysis on the in situ formation of nanocellulose and its resulting aspect ratio was explored. In contrast, the other two composites were created by incorporating enzyme-catalyzed synthetic block cellulose (BC) and its acid-hydrolyzed regenerated disc-shaped cellulose (DC) into the PVA. Subsequently, the mechanism by which three distinct types of nanocellulose, varying in morphology and size, was explored to elucidate their contributions to enhancing the properties of PVA. The results demonstrated that PVA/RC outperformed PVA/BC and PVA/DC. The elevated aspect ratio and intricate network structure of RCs not only significantly bolster the mechanical robustness of PVA/RC, leading in an 86.40 % surge in tensile strength and a remarkable 277.03 % rise in tensile modulus in comparison to pure PVA, but also induce a slight enhancement in elongation at break. Moreover, the thermal stability and biodegradability of PVA/RC was enhanced. Collectively, this study introduces an innovative strategy for the efficient fabrication of biodegradable composites with enhanced properties.


The contribution of collision cascades to sputtering and radiation damage.
Measurement of the velocity distribution of sputtered atoms has tested theories of sputtering and radiation damage. Experiments measuring the time of flight of sputtered atoms in precisely defined directions from the emitting surface, from which the related velocity and energy distributions may be deduced, are described. The energy distribution from polycrystalline targets often fits the form E/(E + Eb)3 derived from a theoretical model in which the bombarding ions initiate collision cascades that eject atoms through the surface. It is assumed that atoms are bound to the surface by a binding force normal to the surface, represented as a binding energy, in the formula. It is shown how this theory relates to that of radiation damage. Departures from the formula seem to correlate with high-energy density in cascades and/or low values of Eb/kT0, with T0 the target temperature. A second component then appears in the energy spectrum approximated by [formula: see text], where deltaT is an effective local temperature rise induced by cascades. The velocity distributions from single crystals are strongly affected by both the direction of ion incidence, indicative of ion channelling, and by emission in directions close to simple crystal axes, indicative of momentum focusing within cascades. Models of the cascade region, and the local heating it causes, have been deduced from sputtering experiments and have advanced our understanding of defect structures caused by radiation damage. Momentum focusing processes are active in creating interstitial-vacancy pairs in both radiation damage and sputtering and their properties have been deduced from these experiments. It is shown how the study of sputtering has enhanced the understanding of radiation damage.


A diet rich in conjugated linoleic acid and butter increases lipid peroxidation but does not affect atherosclerotic, inflammatory, or diabetic risk markers in healthy young men.
Intake of conjugated linoleic acid (CLA) has been demonstrated to beneficially affect risk markers of atherosclerosis and diabetes in rats. CLA is naturally found in milk fat, especially from cows fed a diet high in oleic acid, and increased CLA intake can occur concomitantly with increased milk fat intake. Our objective was to investigate the effect of CLA as part of a diet rich in butter as a source of milk fat on risk markers of atherosclerosis, inflammation, diabetes type II, and lipid peroxidation. A total of 38 healthy young men were given a diet with 115 g/d of CLA-rich fat (5.5 g/d CLA oil, a mixture of 39.4% cis9, trans11 and 38.5% trans10, cis12) or of control fat with a low content of CLA in a 5-wk double-blind, randomized, parallel intervention study. We collected blood and urine before and after the intervention. The fatty acid composition of plasma triacylglycerol, cholesterol esters, and phospholipids reflected that of the intervention diets. The CLA diet resulted in increased lipid peroxidation measured as an 83% higher 8-iso-prostaglandin F2alpha concentration compared with the control, P < 0.0001. We observed no other significant differences in the effect of the interventions diets. In conclusion, when given as part of a diet rich in butter, a mixture of CLA isomers increased lipid peroxidation but did not affect risk markers of cardiovascular disease, inflammation, or fasting insulin and glucose concentrations.


Immunosuppression-induced defective lymphocyte proliferation to murine cytomegalovirus is prolonged following the cessation of immunosuppression.
Defective murine cytomegalovirus (MCMV) antigen-specific proliferation, induced by treatment of MCMV-infected mice with either antilymphocyte globulin (ALG), prednisolone, or both agents, was eventually restored following the cessation of immunosuppression. At 100 and 278 days following the end of immunosuppressive therapy splenic lymphocytes from infected and subsequently immunosuppressed mice responded significantly in vitro to soluble MCMV antigen after having lost this response immediately upon initiation of immunosuppression. Circulating specific antibodies and mitogen-induced blast transformation were comparable between infected mice and infected mice that also were immunosuppressed. At 278 days following the cessation of immunosuppression splenocytes from infected mice that had been treated with ALG yielded greatly increased background proliferation. Nylon-wool adherence was used to obtain enriched populations of T cells, and B cells and monocytes from MCMV-infected mice. While T cells alone did not respond in vitro to MCMV antigen, recombining B cells and monocytes with the T cells reconstituted in vitro proliferation. Defective lymphocyte proliferation to MCMV for an extended period of time following the end of immunosuppressive therapy indicated a prolonged inability to respond to an active MCMV infection. Identification of the cellular basis for the proliferation defect might lead to the development of effective immunotherapy.


Dermal fibroblast-like cells reprogrammed directly from adipocytes in mouse.
In deep burns, early wound closure is important for healing, and skin grafting is mainly used for wound closure. However, it is difficult to achieve early wound closure in extensive total body surface area deep burns due to the lack of donor sites. Dermal fibroblasts, responsible for dermis formation, may be lost in deep burns. However, fat layers composed of adipocytes, lying underneath the dermis, are retained even in such cases. Direct reprogramming is a novel method for directly reprograming some cells into other types by introducing specific master regulators; it has exhibited appreciable success in various fields. In this study, we aimed to assess whether the transfection of master regulators (ELF4, FOXC2, FOXO1, IRF1, PRRX1, and ZEB1) could reprogram mouse adipocytes into dermal fibroblast-like cells. Our results indicated the shrinkage of fat droplets in reprogrammed mouse adipocytes and their transformation into spindle-shaped dermal fibroblasts. Reduced expression of PPAR-2, c/EBP, aP2, and leptin, the known markers of adipocytes, in RT-PCR, and enhanced expression of anti-ER-TR7, the known anti-fibroblast marker, in immunocytochemistry, were confirmed in the reprogrammed mouse adipocytes. The dermal fibroblast-like cells, reported here, may open up a new treatment mode for enabling early closure of deep burn wounds.


Vibrational biospectroscopy coupled with multivariate analysis extracts potentially diagnostic features in blood plasma/serum of ovarian cancer patients.
Despite numerous advances in "omics" research, early detection of ovarian cancer still remains a challenge. The aim of this study was to determine whether attenuated total reflection Fourier-transform infrared (ATR-FTIR) or Raman spectroscopy could characterise alterations in the biomolecular signatures of human blood plasma/serum obtained from ovarian cancer patients compared to non-cancer controls. Blood samples isolated from ovarian cancer patients (n = 30) and healthy controls (n = 30) were analysed using ATR-FTIR spectroscopy. For comparison, a smaller cohort of samples (n = 8) were analysed using an InVia Renishaw Raman spectrometer. Resultant spectra were pre-processed prior to being inputted into principal component analysis (PCA) and linear discriminant analysis (LDA). Statistically significant differences (P < 0.001) were observed between spectra of ovarian cancer versus control subjects for both biospectroscopy methods. Using a support vector machine classifier for Raman spectra of blood plasma, a diagnostic accuracy of 74% was achieved, while the same classifier showed 93.3% accuracy for IR spectra of blood plasma. These observations suggest that a biospectroscopy approach could be applied to identify spectral alterations associated with the presence of insidious ovarian cancer.


Protozoa and the decline of Rhizobium populations added to soil.
A fall in Rhizobium abundance occurred in nonsterile soil inoculated with large numbers of the root-nodule bacteria, but many of the rhizobia still survived. No such decline was evident in sterile soil. Protozoa feeding on these bacteria were isolated from soil and other environments. As the abundance of Rhizobium meliloti and a cowpea Rhizobium strain in soil decreased, the protozoan density increased. The inability of the predators to eliminate their prey from soil was not the result of the presence of organisms feeding on the protozoa because many rhizobia survived in sterile soil inoculated with the prey and cultures of individual protozoa, nor was it the result of the rapid multiplication of the bacteria to replace those consumed because survivors were still numerous in essentially organic matter free soil in which the bacteria did not grow appreciably. The lack of elimination also was not associated with a protective effect of soil particles because survivors were still abundant in solutions inoculated with protozoa and bacteria. It is suggested that the size of the prey population diminishes until a density is attained at which the energy used by the predator in hunting for the survivors equals that obtained from the feeding.


General practice - a fertile lagoon in the ocean of medical knowledge.
General practitioners (GPs) often find that linear, deductive knowledge does not provide a sufficient map for clinical management. But experience, accompanied by enduring familiarity with individual patients, may offer unique complementary skills to interpret a patient's symptoms and navigate skilfully through diagnosis, treatment, follow-up and prevention.In this article, we draw attention to the nature of this tacit knowing that is executed by many GPs every day. We argue that the nonlinear, unpredictable complexity of this domain nurtures a particular logic of clinical knowing. This kind of knowledge is not intuition and can to some extent be intersubjectively accessible. We substantiate and discuss how and why general practice research can contribute to knowledge development by transforming reflection-in-action to reflection-on-action.We briefly present some concepts for reflection-on-action of clinical knowing in general practice. The VUCA model (volatility, uncertainty, complexity, ambiguity) embraces dynamic and confusing situations in which agile work (adaptive, flexible and responsive behaviour and cognitive creativity) is assumed to be an appropriate response. Using such perspectives, we may sharpen our gaze and apply reflexivity and analytic elaboration to interpret unique incidents and experiences and appreciate the complexity of general practice. In this way, exploratory research can fertilize general practice and offer innovation to the entire domain of clinical knowledge.


Activation of protein kinase C in human neutrophils attenuates agonist-stimulated rises in cytosolic free Ca2+ concentration by inhibiting bivalent-cation influx and intracellular Ca2+ release in addition to stimulating Ca2+ efflux.
Stimulation of fura-2-loaded human neutrophils with formylmethionyl-leucyl-phenylalanine (FMLP) or ionomycin elevated the cytosolic free Ca2+ concentration, [Ca2+], to a maintained elevated level. Activation of protein kinase C (C-kinase) with phorbol 12-myristate 13-acetate, 4 beta-phorbol 12,13-didecanoate or dioctanoylglycerol caused decreases in [Ca2+]i from this level. 4 alpha-Phorbol didecanoate, which does not activate C-kinase, had no effect. These results confirm previous reports that C-kinase activation decreases neutrophil [Ca2+]i by stimulating removal of Ca2+ from the cytosol. Further experiments showed that activation of C-kinase attenuated the component of the FMLP-stimulated [Ca2+]i rise that was dependent on external Ca2+. C-kinase activation also inhibited FMLP-stimulated entry of the quenching cation, Mn2+, used as an indicator of bivalent-cation entry. In contrast, C-kinase activation caused only a partial inhibition of FMLP-stimulated release of Ca2+ from intracellular stores. 4 alpha-Phorbol didecanoate was ineffective in inhibiting Ca2+ entry, Mn2+ entry and intracellular Ca2+ release. Addition of FMLP also stimulated a decrease in the ionomycin-elevated [Ca2+]i, and this effect was blocked by staurosporine, a protein kinase inhibitor. These results show that, in addition to stimulating Ca2+ efflux, C-kinase activation in neutrophils inhibits FMLP-stimulated entry of bivalent cations, and partially inhibits intracellular release of Ca2+. Further, FMLP itself can modulate [Ca2+]i by activation of C-kinase.


A new vulnerability to BET inhibition due to enhanced autophagy in BRCA2 deficient pancreatic cancer.
Pancreatic cancer is one of the deadliest diseases in human malignancies. Among total pancreatic cancer patients, ~10% of patients are categorized as familial pancreatic cancer (FPC) patients, carrying germline mutations of the genes involved in DNA repair pathways (e.g., BRCA2). Personalized medicine approaches tailored toward patients' mutations would improve patients' outcome. To identify novel vulnerabilities of BRCA2-deficient pancreatic cancer, we generated isogenic Brca2-deficient murine pancreatic cancer cell lines and performed high-throughput drug screens. High-throughput drug screening revealed that Brca2-deficient cells are sensitive to Bromodomain and Extraterminal Motif (BET) inhibitors, suggesting that BET inhibition might be a potential therapeutic approach. We found that BRCA2 deficiency increased autophagic flux, which was further enhanced by BET inhibition in Brca2-deficient pancreatic cancer cells, resulting in autophagy-dependent cell death. Our data suggests that BET inhibition can be a novel therapeutic strategy for BRCA2-deficient pancreatic cancer.


Development of theophylline floating microballoons using cellulose acetate butyrate and/or Eudragit RL 100 polymers with different permeability characteristics.
The objective of the present investigation was to design a sustained release floating microcapsules of theophylline using two polymers of different permeability characteristics; Eudragit RL 100 (Eu RL) and cellulose acetate butyrate (CAB) using the oil-in-oil emulsion solvent evaporation method. Polymers were used separately and in combination to prepare different microcapsules. The effect of drug-polymer interaction was studied for each of the polymers and for their combination. Encapsulation efficiency, the yield, particle size, floating capability, morphology of microspheres, powder X-ray diffraction analysis (XRD), and differential scanning calorimetry (DSC) were evaluated. The in vitro release studies were performed in PH 1.2 and 7.4. The optimized drug to polymer ratios was found to be 4:1 (F(2)) and 0.75:1 (F'(2)) with Eu RL and CAB, respectively. The best drug to polymer ratio in mix formulation was 4:1:1 (theophylline: Eu RL: CAB ratio). Production yield, loading efficiencies, and particle size of F(2) and F'(2) were found to be 59.14% and 45.39%, 73.93% and 95.87%, 372 and 273 micron, respectively. Microsphere prepared with CAB showed the best floating ability (80.3 ± 4.02% buoyancy) in 0.1 M HCl for over 12 h. The XRD and DSC showed that theophylline in the drug loaded microspheres was stable and in crystaline form. Microparticles prepared using blend of Eu RL and CAB polymers indicated more sustained pattern than the commercial tablet (P<0.05). Drug loaded floating microballoons prepared of combination of Eu RL and CAB with 1:1 ratio were found to be a suitable delivery system for sustained release delivery of theophylline which contained lower amount of polymer contents in the microspheres.


[The management of acute hepatitis C and needlestick accidents with hepatitis-C virus positive blood].
Acute hepatitis C is rarely diagnosed, in part because of its usually subclinical course. Infection with the hepatitis C virus (HCV) has a high chronicity rate, 70-90%. The risk of infection after a needlestick accident with HCV positive blood is 3-10%. There are no efficacious preventive measures regarding HCV infection but treatment with the antiviral drug interferon alpha during the acute phase of the disease has shown to significantly reduce the risk of subsequent chronic infection. It is advised to evaluate individuals who were exposed to infected blood by a needlestick accident regarding HCV transmission, and to offer interferon treatment to them in case they become HCV positive, as demonstrated with a positive serum HCV-RNA test.


Ocean acidification modifies biomolecule composition in organic matter through complex interactions.
The main source of marine organic carbon (OC) is autotrophic production, while heterotrophic degradation is its main sink. Increased anthropogenic CO2 release leads to ocean acidification and is expected to alter phytoplankton community composition, primary production rates and bacterial degradation processes in the coming decades with potential consequences for dissolved and particulate OC concentration and composition. Here we investigate effects of increased pCO2 on dissolved and particulate amino acids (AA) and carbohydrates (CHO), in arctic and sub-arctic planktonic communities in two large-scale mesocosm experiments. Dissolved AA concentrations responded to pCO2/pH changes during early bloom phases but did not show many changes after nutrient addition. A clear positive correlation in particulate AA was detected in post-bloom phases. Direct responses in CHO concentrations to changing pCO2/pH were lacking, suggesting that observed changes were rather indirect and dependent on the phytoplankton community composition. The relative composition of AA and CHO did not change as a direct consequence of pCO2 increase. Changes between bloom phases were associated with the prevailing nutrient status. Our results suggest that biomolecule composition will change under future ocean conditions but responses are highly complex, and seem to be dependent on many factors including bloom phase and sampling site.


Problem of track offset in optical disk systems.
In an optical disk drive, it is well known that a tilt of the disk causes an offset in the tracking-error signal (TES). One effect of disk tilt is the introduction of a dc component to the TES, which can be largely corrected by operation of the tracking system at the midpoint between the maximum and the minimum values of the open-loop TES. However, this method of correcting for the dc shift in the TES does not correct for the effect of coma in the focused spot, which leads to track offset. The track offset of a system is defined as the distance between the peak irradiance in the focused spot and the center of the groove when the tracking system is operating at the midpoint between the maximum and the minimum values of the open-loop TES in the presence of disk tilt. Calculations are performed that show the dependence of track offset on various system parameters, including track pitch, wavelength, and numerical aperture and rim intensity of the objective lens, and on the regions of the beam used to generate the TES. The track offsets for several beam-segmentation schemes are calculated for a digital versatile disk that uses push-pull and differential phase tracking. It is shown that for differential phase tracking the value of track offset depends on the mark length.


Diabetic neuropathy.
Diabetic neuropathy is not a single entity but manifests as several different clinical syndromes. It is likely that different pathophysiological mechanisms contribute to the development of the neuropathy, including metabolic alterations, microvascular changes, and inflammatory changes. A summary of the underlying pathophysiological mechanisms and clinical aspects of different diabetic neuropathies are outlined in this article.


Methylphenidate-induced improvements of various measures of attention in adults with attention deficit hyperactivity disorder.
The present study examined the effect of the stimulant medication methylphenidate (MPH) on attentional functioning of adults with ADHD. Sixteen adults with a diagnosed ADHD without comorbidity were assessed twice, at baseline off MPH and following MPH treatment. The assessment battery consisted of reaction time tasks of low complexity, including measures of alertness--subdivided into tonic and phasic alertness, vigilance, divided attention, flexibility and such aspects of selective attention as including focused attention, inhibition and integration of sensory information. In addition, 16 healthy participants who were matched to adults with ADHD according to sex, age, education level and intellectual functions were also assessed twice using the same test battery. The results of the present study suggest that adults with ADHD off stimulant medication are seriously impaired in various components of attention including vigilance, divided attention, selective attention and flexibility. These impairments of attention were observed primarily in regard to reaction time and its variability. Treatment of adults with ADHD using individually tailored doses of MPH has a positive effect on measures of alertness, vigilance, selective attention, divided attention and flexibility. However, even on MPH adults with ADHD displayed considerable deficits in vigilance and integration of sensory information. The present findings indicate that adults with ADHD are not differentially impaired in attentional processes but may suffer from a more global deficit of attention. Although MPH treatment has been found to be effective in the treatment of the attention deficit of adults with ADHD, additional treatment appears to be necessary.


Tracheoesophageal Prosthesis Malfunction - A Case Report.
Tracheoesophageal prosthesis (TEP) is an artificial connection between the trachea and esophagus allowing air into the upper esophagus from the trachea thereby vibrating it. TEPs give patients who lose their vocal cords to laryngectomies a tracheoesophageal voice. A potential complication of this is silent aspiration of gastric content. We present a case of a 69-year-old female with a TEP placed after a laryngectomy for laryngeal cancer who presented to the hospital with shortness of breath and hypoxia. She was initially treated for a presumed diagnosis of chronic obstructive pulmonary disease (COPD) and congestive heart failure (CHF) exacerbations but continued to be hypoxic despite aggressive medical management. Further evaluation revealed silent aspirations as a consequence of TEP malfunction. Through our case report we urge clinicians to consider this differential diagnosis, as the clinical presentation of silent aspiration among patients with a TEP can be easily mistaken for a COPD exacerbation. A large number of patients with TEPs are smokers with underlying COPD. TEPs give patients who lose their vocal cords to laryngectomies a tracheoesophageal voice.A potential complication of TEPs is silent aspiration around or through the TEP, causing coughing and even recurrent aspiration pneumonitis or pneumonia.Patients with TEPs typically have extensive smoking histories and underlying COPD or CHF which can have exacerbations that are similar in presentation.


One-step RT-PCR for detection of Zika virus.
Zika virus (ZIKV) is an emerging mosquito-borne flavivirus circulating in Asia and Africa. Human infection induces an influenza-like syndrome that is associated with retro-orbital pain, oedema, lymphadenopathy, or diarrhea. Diagnosis of Zika fever requires virus isolation and serology, which are time consuming or cross-reactive. To develop a one-step RT-PCR assay to detect ZIKV in human serum. An assay targeting the envelope protein coding region was designed and evaluated for its specificity, detection limit, repeatability, and capacity to detect ZIKV isolates collected over a 40-year period from various African countries and hosts. The assay's detection limit and repeatability were respectively 7.7pfu/reaction and 100% in serum and L-15 medium; none of 19 other flaviviruses tested were detected. The assay is rapid, sensitive, and specific to detect ZIKV in cell culture or serum, but needs to be validated for diagnosis using clinical samples.


PbS Nanocrystal Emission Is Governed by Multiple Emissive States.
Lead chalcogenide colloidal nanocrystals (NCs) are promising materials for solution processable optoelectronics. However, there is little agreement on the identity and character of PbS NC emission for different degrees of quantum confinement-a critical parameter for realizing applications for these nanocrystals. In this work, we combine ensemble and single NC spectroscopies to interrogate preparations of lead sulfide NCs. We use solution photon correlation Fourier spectroscopy (S-PCFS) to measure the average single NC linewidth of near-infrared-emitting PbS quantum dots and find it to be dominated by homogeneous broadening. We further characterize PbS NCs using temperature-dependent linear and time-resolved emission spectroscopy which demonstrate that a kinetically accessed defect state dominates room temperature emission of highly confined emitting NCs. These experiments, taken together, demonstrate that the linewidth and Stokes shift of PbS NCs are the result of emission from two states: a thermally accessed defect-with an energetically pinned charge carrier-and an inhomogeneously broadened band-edge state.


Myofibril Assembly in Cultured Mouse Neonatal Cardiomyocytes.
The formation of myofibrils was analyzed in neonatal mouse cardiomyocytes grown in culture and stained with fluorescent antibodies directed against myofibrillar proteins. The cardiomyocyte cultures also were exposed to siRNA probes to test the role of nonmuscle myosin IIB expression in the formation of myofibrils. In culture, new myofibrils formed in the spreading cell margins surrounding contractile myofibrils previously assembled in utero. Observations indicated that assembly of mature myofibrils occurred in three-stages, as previously reported in cultured mouse skeletal muscle. Premyofibrils, characterized by minisarcomeres with nonmuscle myosin IIB and muscle-specific alpha-actinin bound to actin filaments, formed in the first stage; followed by nascent myofibrils, the second stage when muscle myosin II and titin were first detected. In the mature myofibril stage muscle myosin II filaments aligned in periodic A-Bands; late assembling proteins, including myomesin and telethonin, were integrated in the sarcomeres, and nonmuscle IIB was absent from the sarcomeres. Treatment of the cultured neonatal cardiomyocytes with gene-specific siRNAs for nonmuscle myosin IIB, led to a marked decrease in the formation of premyofibrils, and subsequently of mature myofibrils. Anat Rec, 301:2067-2079, 2018. © 2018 Wiley Periodicals, Inc.


Frequency and relevance of anterior cerebral artery embolism caused by mechanical thrombectomy of middle cerebral artery occlusion.
Spread of thrombus material in previously unaffected vessels is a potential hazard of mechanical thrombectomy, but it has not yet been investigated in detail, to our knowledge. Our purpose was to evaluate the frequency and relevance of these events in mTE of M1 occlusions. We retrospectively reviewed all patients treated for isolated M1 occlusion between January 2008 and July 2012. Angiographic images were analyzed to assess emboli in anterior cerebral artery branches induced by mTE and associated devices. Recanalization attempts in the ACA were reported as well as technical success and adverse events of rescue therapies. ACA infarcts on follow-up imaging served as a surrogate for clinical relevance. ACA infarcts were quantified volumetrically and assessed visually for involvement of motor or supplementary motor areas. New ACA emboli occurred in 12 of 105 (11.4%) M1 recanalization procedures and were caused by a stent-retriever in 11 intances. Attempts to recanalize the ACA were made in 6 patients and were deemed technically successful in 5 with no adverse events. We detected 6 (5.7%) new infarcts on follow-up imaging with an average volume of 26.9 cm(3). Involvement of motor or supplementary motor areas was seen in 4 (3.8%) cases. Three patients developed ACA infarcts despite successful endovascular ACA recanalization. The frequency of ACA emboli in mTE of M1 occlusions is relevant, causing ACA infarcts in 5.7% of patients; 3.8% of emboli were likely to hamper motor-function recovery. Endovascular recanalization of major ACA branches reduced the incidence of infarcts with no adverse events.


Mechanistic studies on millerite chlorination with ammonium chloride.
Millerite (NiS) is the main source for metallurgical production of nickel worldwide. To improve the extraction rate of nickel, chlorination is usually carried out, as the resulting nickel chloride (NiCl2) can easily dissolve in water and be separated. Although molecular chlorine (Cl2) can be used as the chlorination reagent, greener reagents such as ammonium chloride (NH4Cl) are preferable from a process design perspective. However, the efficiency of NH4Cl as a chlorination reagent must be further improved for this process to be viable for industrial applications, and mechanistic understanding is imperative to this end. Here, we performed extensive density functional theory (DFT) calculations to elucidate the chlorination mechanism of NiS by exploring three possible pathways. We first considered the direct chlorination of NiS by Cl2, which was suggested to form by the reaction between NH4Cl and SO3 catalyzed by a metal oxide. Alternatively, NH4Cl was found to react favorably with the partially or fully oxidized NiS surface in the presence of oxygen (O2). During the oxidation of NiS, sulfur dioxide (SO2) may form. Furthermore, sulfur or oxygen vacancy was predicted to form during the chlorination of NiS or NiO with NH4Cl. Based on the available experimental evidence and our computational results, three possible mechanisms for the chlorination of NiS using NH4Cl as the chlorination reagent in the presence of O2 were proposed.