Electrospinning and Additive Manufacturing: Adding Three-Dimensionality to Electrospun Scaffolds for Tissue Engineering.
The final biochemical and mechanical performance of an implant or scaffold are defined by its structure, as well as the raw materials and processing conditions used during its fabrication. Electrospinning and Additive Manufacturing (AM) are two contrasting processing technologies that have gained popularity amongst the fields of medical research i.e., tissue engineering, implant design, drug delivery. Electrospinning technology is favored for its ability to produce micro- to nanometer fibers from polymer solutions and melts, of which, the dimensions, alignment, porosity, and chemical composition are easily manipulatable to the desired application. AM, on the other hand, offers unrivalled levels of geometrical freedom, allowing highly complex components (i.e., patient-specific) to be built inexpensively within 24 hours. Hence, adopting both technologies together appears to be a progressive step in pursuit of scaffolds that better match the natural architecture of human tissues. Here, we present recent insights into the advances on hybrid scaffolds produced by combining electrospinning (melt electrospinning excluded) and AM, specifically multi-layered architectures consisting of alternating fibers and AM elements, and bioinks reinforced with fibers prior to AM. We discuss how cellular behavior (attachment, migration, and differentiation) is influenced by the co-existence of these micro- and nano-features.


Triptans for the management of migraine.
Migraine is a chronic, recurrent, disabling condition that affects millions of people in the US and worldwide. Proper acute care treatment for migraineurs is essential for a full return of function and productivity. Triptans are serotonin (5-HT)(1B/1D) receptor agonists that are generally effective, well tolerated and safe. Seven triptans are available worldwide, although not all are available in every country, with multiple routes of administration, giving doctors and patients a wide choice. Despite the similarities of the available triptans, pharmacological heterogeneity offers slightly different efficacy profiles. All triptans are superior to placebo in clinical trials, and some, such as rizatriptan 10 mg, eletriptan 40 mg, almotriptan 12.5 mg, and zolmitriptan 2.5 and 5 mg are very similar to each other and to the prototype triptan, sumatriptan 100 mg. These five are known as the fast-acting triptans. Increased dosing can offer increased efficacy but may confer a higher risk of adverse events, which are usually mild to moderate and transient in nature. This paper critically reviews efficacy, safety and tolerability for the different formulations of sumatriptan, zolmitriptan, rizatriptan, naratriptan, almotriptan, eletriptan and frovatriptan.


Sustainable financing for municipal solid waste management in Nepal.
Financing municipal solid waste (MSW) services is one of the key challenges faced by cities in developing countries. This study used plastic waste, a constituent of MSW, to explore the possibility of generating revenue for financing MSW management in the municipalities of Nepal. The results of this study suggest that plastic material recovery could generate revenue, which is equivalent to 1.38 times of the plastic-waste-related management cost when collection efficiency reaches 66.7%. An increase in 1% of recovery rate and collection efficiency could cover an additional 4.64% and 2.06% of the costs of managing plastic waste, respectively. In addition, an increase in tax on imported plastic materials could also motivate recovery of plastic waste for recycle and reuse. An additional 1% tax on plastic imports would be sufficient to cover plastic-related waste management when plastic waste recovery and collection efficiency rates are low. This plastic recovery- revenue exercise could be expanded to other materials such as paper and metal to fully understand the possibility of sustainable financing of MSW management and reducing environmental harm in developing countries like Nepal.


Relay ring-closing metathesis (RRCM): a strategy for directing metal movement throughout olefin metathesis sequences.
The title concept involves the use of structurally modified RCM substrates that contain extender arms, terminating in a remote reactive alkene. Initiation of an RCM sequence at that reactive alkene is followed by rapid intramolecular relay of the metal center to an initially less reactive alkene in the parent substrate. This permits one to control the relative timing (or direction) of a metathesis sequence. For example, one can reverse the inherent tendency of an unsymmetrical alpha,omega-diene substrate to close, say, left-to-right, to that of right-to-left. Four distinct types of application of the RRCM concept are demonstrated. Among other things, they show the preparation of tetrasubstituted electron-deficient alkenes using G1 [(Cy3P)2(Cl2)Ru=CHPh], complementary control of directionality (endedness), auxiliary benefits (enzyme specificity) from the incorporation of additional steric bulk, the activation of otherwise ineffective substrates for RCM closure, the use of unorthodox alkenes as initiation sites for ring closure, and control of product olefin geometry.


Two-photon fluorescence excitation in detection of biomolecules.
Two-photon fluorescence excitation has been found to be a very powerful method for enhancing the sensitivity and resolution in far-field light microscopy. Two-photon fluorescence excitation also provides a substantially background-free detection on the single-molecule level. It allows direct monitoring of formation of labelled biomolecule complexes in solution. Two-photon excitation is created when, by focusing an intensive light source, the density of photons per unit volume and per unit time becomes high enough for two photons to be absorbed into the same chromophore. In this case, the absorbed energy is the sum of the energies of the two photons. In two-photon excitation, dye molecules are excited only when both photons are absorbed simultaneously. The probability of absorption of two photons is equal to the product of probability distributions of absorption of the single photons. The emission of two photons is thus a quadratic process with respect to illumination intensity. Thus in two-photon excitation, only the fluorescence that is formed in the clearly restricted three-dimensional vicinity of the focal point is excited. We have developed an assay concept that is able to distinguish optically between the signal emitted from a microparticle in the focal point of the laser beam, and the signal emitted from the surrounding free labelled reagent. Moreover, the free labels outside the focal volume do not contribute any significant signal. This means that the assay is separation-free. The method based on two-photon fluorescence excitation makes possible fast single-step and separation-free immunoassays, for example, for whole blood samples. Since the method allows a separation-free assay in very small volumes, the method is very useful for high-throughput screening assays. Consequently we believe that two-photon fluorescence excitation will make a remarkable impact as a research tool and a routine method in many fields of analysis.


A personal health information toolkit for health intervention research.
With the emergence of mobile health (mHealth) apps, there is a growing demand for better tools for developing and evaluating mobile health interventions. Recently we developed the Personal Health Intervention Toolkit (PHIT), a software framework which eases app implementation and facilitates scientific evaluation. PHIT integrates self-report and physiological sensor instruments, evidence-based advisor logic, and self-help interventions such as meditation, health education, and cognitive behavior change. PHIT can be used to facilitate research, interventions for chronic diseases, risky behaviors, sleep, medication adherence, environmental monitoring, momentary data collection health screening, and clinical decision support. In a series of usability evaluations, participants reported an overall usability score of 4.5 on a 1-5 Likert scale and an 85 score on the System Usability Scale, indicating a high percentile rank of 95%.


Responding to discontinuities in product-based service supply chains in the COVID-19 pandemic: Towards transilience.
In this study, we identify and characterise how organisations have responded, in ways ranging from restoration to radical change, to discontinuities in their product-based service (PBS) supply chains during the COVID-19 pandemic. Following a theoretical approach that integrates transilience and panarchy theory as a response strategy in PBS supply chains, our qualitative study involved collecting data through 19 semi-structured interviews at six manufacturing firms during the first 6 months of the COVID-19 pandemic (i.e., March to August 2020) and triangulating the findings with the secondary data and that from an industry workshop. Following an inductive approach, we performed thematic data analysis in Nvivo software package. The findings suggest characterising discontinuities in PBS supply chains as unmanageable external supply-side, demand-side or interactional discontinuities or other manageable deliberate or forced organisational discontinuities. Following that characterisation, we developed a conceptual framework combing both resilience and transformation into new service opportunities. We gained insights into the first-response abilities and ways of coping among manufacturing firms during the COVID-19 pandemic. Though our findings capture a contemporary, eye-of-the-storm perspective on future directions, a longitudinal study on the pandemic could further validate and extend the modes of response that complement mitigation with the ability to accelerate change or innovation of internal process or external service offerings. Combining current literature with lessons learned from the firms' immediate responses, this paper's overview and characterisation of discontinuities following the COVID-19 outbreak in PBS supply chains demonstrate how manufacturing firms can foster transilience. As such, it integrates product-based supply chain discontinuities into the domain of service-based supply chains.


Phenotypic features and genetic findings in 2 chinese families with Miyoshi distal myopathy.
Miyoshi distal myopathy (MM) and limb girdle muscular dystrophy type 2B (LGMD2B) were found to map to the same mutant gene encoding for dysferlin on chromosome 2p13. Most reported cases were large inbred kindreds whose members demonstrated both MM and LGMD2B phenotypes. To investigate the clinical, neurophysiological, histopathological, and genetic features in 4 patients with MM from 2 unrelated Chinese families demonstrating linkage to the dysferlin locus. All patients were characterized by early adult onset, preferential atrophy, and weakness of calf muscles, marked elevation of serum creatine kinase levels, and absence of dysferlin staining. Magnetic resonance imaging showed fatty and fibrotic tissue signals in the affected muscles. Genetic analysis revealed novel compound heterozygous mutations, 1310+1G to A and GGG to GTC transition at nucleotide 1650 (G426V ) in one family and another novel compound heterozygous mutation, a deletion of C at nucleotide 477 and a CCG to CTG transition at nucleotide 6576 (P2068L), in the other family. Miyoshi distal myopathy in these 2 Chinese families demonstrated a homogenous phenotype and compound heterozygous mutations. Among the 4 mutations, 3 were novel mutations that, to our knowledge, have not been reported previously.


A Grad-seq View of RNA and Protein Complexes in Pseudomonas aeruginosa under Standard and Bacteriophage Predation Conditions.
The Gram-negative rod-shaped bacterium Pseudomonas aeruginosa is not only a major cause of nosocomial infections but also serves as a model species of bacterial RNA biology. While its transcriptome architecture and posttranscriptional regulation through the RNA-binding proteins Hfq, RsmA, and RsmN have been studied in detail, global information about stable RNA-protein complexes in this human pathogen is currently lacking. Here, we implement gradient profiling by sequencing (Grad-seq) in exponentially growing P. aeruginosa cells to comprehensively predict RNA and protein complexes, based on glycerol gradient sedimentation profiles of >73% of all transcripts and ∼40% of all proteins. As to benchmarking, our global profiles readily reported complexes of stable RNAs of P. aeruginosa, including 6S RNA with RNA polymerase and associated product RNAs (pRNAs). We observe specific clusters of noncoding RNAs, which correlate with Hfq and RsmA/N, and provide a first hint that P. aeruginosa expresses a ProQ-like FinO domain-containing RNA-binding protein. To understand how biological stress may perturb cellular RNA/protein complexes, we performed Grad-seq after infection by the bacteriophage ΦKZ. This model phage, which has a well-defined transcription profile during host takeover, displayed efficient translational utilization of phage mRNAs and tRNAs, as evident from their increased cosedimentation with ribosomal subunits. Additionally, Grad-seq experimentally determines previously overlooked phage-encoded noncoding RNAs. Taken together, the Pseudomonas protein and RNA complex data provided here will pave the way to a better understanding of RNA-protein interactions during viral predation of the bacterial cell.IMPORTANCE Stable complexes by cellular proteins and RNA molecules lie at the heart of gene regulation and physiology in any bacterium of interest. It is therefore crucial to globally determine these complexes in order to identify and characterize new molecular players and regulation mechanisms. Pseudomonads harbor some of the largest genomes known in bacteria, encoding ∼5,500 different proteins. Here, we provide a first glimpse on which proteins and cellular transcripts form stable complexes in the human pathogen Pseudomonas aeruginosa We additionally performed this analysis with bacteria subjected to the important and frequently encountered biological stress of a bacteriophage infection. We identified several molecules with established roles in a variety of cellular pathways, which were affected by the phage and can now be explored for their role during phage infection. Most importantly, we observed strong colocalization of phage transcripts and host ribosomes, indicating the existence of specialized translation mechanisms during phage infection. All data are publicly available in an interactive and easy to use browser.


Associations between cold spells of different time types and coronary heart disease severity.
Limited evidence showed the association between cold spells and the severity of coronary heart disease (CHD). This study was to investigate the association between cold spells and their different time types and CHD severity. We collected data on CHD patients admitted to the Zhongnan Hospital, Wuhan, China from 2016 to 2021. CHD severity was quantified using the SYNTAX score and transformed into a binomial variable. Daily mean, maximum and minimum temperature were collected during the study period. We first used daily mean temperature to find the optimum definition among multiple thresholds and durations. Then daily maximum and minimum temperatures were used to define different types of cold spells (daytime, nighttime and compound) based on the optimum definition. Annual cold spell days were included to assess individual exposure to cold spells. Logistic regression models were performed to fit the association between cold spell days and CHD severity stratified by different tertiles of PM2.5 and NDVI. In this study, 1937 CHD patients were included. The cold spell defined as at least four consecutive days with daily mean temperature below the 5th percentile exhibited the optimum model. We found that a 4-day increase in cold spell days was associated with more severe CHD (OR = 1.170, 95% CI: 1.074, 1.282). And such an association was more pronounced under higher levels of PM2.5 by OR = 1.270 (1.086, 1.494) and lower levels of greenness by OR = 1.240 (1.044, 1.476). Compared with daytime and compound cold spells, nighttime cold spells showed the strongest association with CHD severity by OR = 1.141 (1.026, 1.269). This study showed that exposure to cold spells was positively associated with CHD severity, especially the nighttime cold spells. The association between cold spells and CHD severity was more significant in high levels of PM2.5 and low levels of greenness.


Buserelin as primary therapy in advanced prostatic carcinoma.
The effectiveness of buserelin, a luteinizing hormone-releasing hormone agonist, was tested in 28 patients with Stages C or D adenocarcinoma of the prostate. Of 24 evaluable patients, there were 13 partial responses (54%) by National Prostatic Cancer Project criteria, median duration greater than 6 months. Nine patients had stable disease (38%), median duration greater than 5 months, and only two patients progressed. Performance status improved in 38%, patient-scored pain improved in 46%, and quality of life improved in 57%. Symptoms occurring during treatment consisted of hot flashes, loss of libido, and impotence. A flare of symptoms was observed in only one patient, despite a transient 25% increase in testosterone in 36% of patients. Buserelin is an effective treatment for inducing frequent and meaningful remissions in advanced prostatic cancer.


Erectile Dysfunction in Type-2 Diabetes Mellitus Patients: Predictors of Early Detection and Treatment.
To identify risk factors and potential predictors of erectile dysfunction (ED) in type-2 diabetes mellitus (T2DM) patients for early detection and treatment. A retrospective cohort was used to assess the clinical data of 105 diabetic patients with ED from May 2019 to April 2020 age-matched to 105 diabetic patients without ED. Potential risk factors that could contribute to ED were compared between the groups. Erectile function was evaluated using the International Index of Erectile Function-5 questionnaire. There were higher rates of diabetic peripheral neuropathy (p = 0.036) and retinopathy (p < 0.001), longer duration of diabetes (p < 0.001), lower estimated glomerular filtration rate (p = 0.010) values, and higher uric acid (p < 0.001) and C-reactive protein (p = 0.001) levels in the ED group compared to the non-ED group. Multivariate logistic analysis identified uric acid, diabetic retinopathy, and T2DM course as independent predictors of diabetic ED. Diabetics with retinopathy and T2DM for ≥49 months were 3.028 and 3.860 times more likely to have ED, respectively. Uric acid values ≥392.5 μmol/L were associated with 18.638 times greater risk of having ED, though the values were within normal range. In T2DM patients, higher uric acid (≥392.5 μmol/L), longer diabetes duration (≥49 months), and the presence of diabetic retinopathy were important and reliable predictors for diabetic ED. For patients who have high risk factors for developing ED, diligent screening and early treatment are necessary.


Guillain-Barre syndrome: first description of a snake envenomation aetiology.
Guillain-Barre syndrome (GBS) is considered as an acute, immune-mediated polyradiculoneuropathy with different clinical phenotypes arising after viral or bacterial infections, vaccination or surgery. However, in 40% of GBS patients the aetiology remains unknown. In this manuscript, we report the occurrence of GBS in a patient bitten by a snake (Vipera aspis) for which a cross-reaction was shown between GM2 ganglioside and glycosidic epitopes of venom proteins. The venom of the snake implied in the patient's envenomation was collected. Its composition was characterised by ELISA and SELDI-TOF MS. Cross-reactivities between venom proteins and GM2 gangliosides were identified by Western blot after immunoabsorption of patient's serum with increasing amounts of purified GM2. Enzymatic deglycosylation of the venom was performed to determine the specificity of the patient's serum cross-reaction. We proved the absence of neurotoxicity of the viper venom. The patient's serum presented specific cross-reactions with several glycosylated venom proteins. After deglycolysation of these proteins, the patient's serum cross-reactivity was abolished. Furthermore, we compared the immune response to venom proteins of sera from two groups of patients. The first group showed IgM reactivity against GM2 ganglioside associated with GBS, and cross-reacted with venom proteins. The second group presented an IgM reactivity against CMV, without neurological disorders, and reacted with neither venom proteins nor gangliosides. Our study proved the auto-immunological aetiology of GBS in our patient based on molecular mimicry mechanisms between venom proteins and GM2 ganglioside.


Renal outcomes in adults with HBV, HIV and HBV/HIV coinfection after 3 years of antiviral therapy in urban Tanzania.
An enhanced understanding of renal outcomes in persons with chronic HBV, HIV, and HBV/HIV coinfection is needed to mitigate chronic kidney disease in regions where HBV and HIV are endemic. To investigate changes in estimated glomerular filtration rate (eGFR) in adults with HBV, HIV or HBV/HIV enrolled in a 3 year prospective cohort study of liver outcomes in Dar es Salaam, Tanzania and initiated on antiviral therapy. We compared eGFR between and within groups over time using mixed-effects models. Four hundred and ninety-nine participants were included in the analysis (HBV: 164; HIV: 271; HBV/HIV: 64). Mean baseline eGFRs were 106.88, 106.03 and 107.18 mL/min/1.73 m2, respectively. From baseline to Year 3, mean eGFR declined by 4.3 mL/min/1.73 m2 (95% CI -9.3 to 0.7) and 3.7 (-7.8 to 0.5) in participants with HBV and HIV, respectively, and increased by 5.1 (-4.7 to 14.9) in those with HBV/HIV. In multivariable models, participants with HBV had lower eGFRs compared with those with HIV or HBV/HIV and, after adjusting for HBV DNA level and hepatitis B e antigen (HBeAg) status, significantly lower eGFRs than those with HBV/HIV at all follow-up visits. In this Tanzanian cohort, coinfection with HBV/HIV did not appear to exacerbate renal dysfunction compared with those with either infection alone. Although overall changes in eGFR were small, persons with HBV experienced lower eGFRs throughout follow-up despite their younger age and similar baseline values. Longer-term studies are needed to evaluate continuing changes in eGFR and contributions from infection duration and other comorbidities.


Mutation affecting the 12th amino acid of the c-Ha-ras oncogene product occurs infrequently in human cancer.
A point mutation alters the 12th amino acid of the c-Ha-ras oncogene product p21 in a human bladder cancer cell line. This is, at present, the only mutation known to result in a human transforming gene. This mutation may therefore represent a possible target for mutagenesis leading to carcinogenesis in humans. By means of restriction enzyme analysis, 29 human cancers, including 20 primary tumor tissues, derived from organs commonly exposed to environmental carcinogens, were tested for the presence of this mutation. None of ten primary bladder carcinomas exhibited the mutation; nor did nine colon carcinomas or ten carcinomas of the lung. Thus the point mutation affecting the 12th amino acid of the c-Ha-ras gene product, while a valuable model for carcinogenesis, does not appear to play a role in the development of most human epithelial cancers of the bladder, colon, or lung.


Hemodynamic Markers in the Anterior Circulation as Predictors of Recurrent Stroke in Patients With Intracranial Stenosis.
Background and Purpose- Although aggressive medical therapy was superior to stenting in the SAMMPRIS trial (Stenting and Aggressive Medical Management for Preventing Recurrent Stroke in Intracranial Stenosis), the stroke rate in the medical arm was still high. The aim of this study was to determine the association between hemodynamic markers (borderzone infarct pattern and impaired collateral flow on baseline imaging) and rates of recurrent stroke in patients treated medically in SAMMPRIS. Methods- This was a post hoc analysis of patients whose qualifying event for SAMMPRIS was an infarct in the territory of a stenotic middle cerebral artery or intracranial carotid artery. Infarcts were adjudicated as involving primarily internal or cortical borderzone territories, the core middle cerebral artery territory, or perforator territories, and collateral flow was assessed according to a standard scale (American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology). Log-rank tests and χ2 tests were performed to assess associations of infarct patterns and collateral flow with rates of recurrent stroke. Results- Of 101 patients who qualified, 14 of 53 (26.4%) with borderzone infarcts, 2 of 24 (8.3%) with core middle cerebral artery infarcts, and 3 of 24 (12.5%) with perforator infarcts had a recurrent stroke in the territory (P=0.14 for comparing the 3 groups, P=0.052 for borderzone versus nonborderzone). Of 82 patients with collateral flow assessment, 30 of 43 (70%) with borderzone infarcts, 7 of 19 (37%) with core middle cerebral artery infarcts, and 11 of 20 (55%) with perforator infarcts had impaired collateral flow distal to the stenosis (P=0.049). Patients with borderzone infarcts and impaired collateral flow had the highest risk of recurrent stroke (37%). Conclusions- Borderzone infarcts and impaired collateral flow identify a subgroup of patients with intracranial stenosis who are at particularly high risk of recurrent stroke on medical treatment. Clinical Trial Registration- URL: https://www.clinicaltrials.gov. Unique identifier: NCT00576693.


Trait variation along an urban-rural gradient in Asian dayflower: the contribution of phenotypic plasticity and genetic divergence.
Environmental changes associated with urbanisation can exert pressure that facilitates both adaptations and plastic responses in plants. Field surveys or common garden experiments (CGE) alone cannot differentiate between the observed phenotypic traits resulting from plastic responses versus evolutionary adaptations to urban environments. We conducted a field survey of habitat environmental factors and four vegetative traits in 12 Commelina communis populations along an urban-rural gradient in the Osaka-Kobe megacity area. We collected seedlings from six of the 12 populations, transplanted them into a greenhouse, and measured six vegetative traits. We investigated correlations between pairs of measured traits in both the field survey and CGE. Plant height and leaf area increased significantly in increasingly developed land areas (DLA) in the field survey, whereas no such variations were found in the CGE, suggesting that the observed phenotypic variation was due to a plastic response to urban eutrophication. Leaf number and specific leaf area (SLA) significantly decreased with increasing DLA in the CGE, suggesting the adaptation of these traits to urban environments. Positive correlations between plant height and leaf area were only observed in urban populations in both the field survey and CGE, indicating the evolution of this trait correlation in urban environments. It has been suggested that urban environments promote both plastic response and genetic divergence of a set of traits in native plants. Our findings suggest that low leaf number, SLA, and positive plant height-leaf area correlations have evolved. In addition, larger plant size is achieved via phenotypic plasticity in urban environments.


Prescription stimulant misuse in a military population.
Increased prescription drug misuse has been reported in veterans, yet there has not been a focused look at stimulant misuse in the military community or correlation with deployment injuries and illnesses. Our objective was to identify rates of stimulant misuse and any correlation with deployment in the military population. A prospective, anonymous institutional review board-approved survey in the emergency department waiting room of a military tertiary care hospital using a 12-item questionnaire created with fixed response and multiple-choice questions. Stimulant misuse was defined as taking more than prescribed, obtaining stimulants from others, and taking it for a nonprescribed reason. Proportions were assessed by Chi-square test and Fisher's exact test. 26/498 (5%) of respondents reported misusing stimulants in the last 5 years. Misusers were more likely to have a mental health diagnosis, and they suffered either a deployment-related injury or another injury, as compared to those who used stimulants properly (p<0.05). The stimulant misuse did not correlate with age, gender, active duty status, education, location of deployment, number of times deployed, traumatic brain injury diagnosis, or enlistment status. Stimulant drug misuse in the military community is associated with mental health conditions, deployment-related injuries, or new physical injuries.


An evaluation of germline mutations and reproductive impacts in fathead minnow (Pimephales promelas) exposed to contaminated sediment.
Polycyclic aromatic hydrocarbons (PAHs) have become ubiquitous in the aquatic environment. Some PAHs are mutagenic, potentially causing germline mutations in fish that inhabit PAH contaminated waters. We evaluated the effect of exposure to sediment-borne PAHs on reproduction and germline mutation rates in fathead minnows (Pimephales promelas). Exposure to the contaminated sediment had no significant impact on the reproductive endpoints measured in this study. Germline mutations rates at three microsatellite DNA loci were 1.69 × 10-3 in fish exposed to PAH-contaminated sediment and 0.55 × 10-3 in control fish, with zero mutations being observed in fish exposed to sediment from a reference site. While the difference in mutation rates between treatments was not statistically significant for the sample size used (15-19 families per treatment), the observed mutations rates enabled us to estimate the sample size required to detect a significant effect. To our knowledge, this is the first report of germline mutation rates in fathead minnow exposed to an environmental contaminant, providing baseline data for use in the design of future experiments.


Bone augmentation and autogenous transplantation of premolar to the site of the fissure in a cleft palate patient.
This article presents a case report of autogenous tooth transplantation to the site of the fissure, in addition to bone augmentation with graft of autogenous bone harvested from the iliac crest, performed in a cleft palate patient, who had insufficient bone volume. A non-syndromic 10-year-old girl, with a unilateral cleft lip and palate, incisal transforamen fissures, agenesis of the maxillary left central incisor and both maxillary lateral incisors, was treated with autogenous bone graft in the cleft area. The orthodontic treatment plan was to replace the missing lateral incisors with the maxillary canines and to extract the mandibular first premolars. One of the mandibular premolars was extracted from its site with 2/3 of its root formation completed and transplanted to the maxillary left central incisor area. After orthodontic treatment, the anatomic crowns were characterized with composite resin. Autogenous tooth transplantation can be performed in the area of the fissure in young cleft palate patients, by performing bone graft augmentation before transplantation of the tooth, to gain sufficient recipient alveolar bone volume. A multidisciplinary approach is mandatory for the success of this clinical procedure, especially in cleft palate patients.


Variable leucine-rich repeats of tomato disease resistance genes Cf-2 and Cf-5 determine specificity.
SUMMARY The tomato Cf-2 and Cf-5 genes confer race specific resistance to infection by the leaf mould pathogen Cladosporium fulvum. The encoded proteins induce a defence response upon recognition of the fungal Avr2 and Avr5 determinants, respectively. Each resistance protein is comprised largely of leucine-rich repeats (LRRs) and the specificity of recognition is thought to occur through a particular domain. We have investigated this further using domain swaps between Cf-2 and Cf-5. Engineered chimeric genes containing portions of Cf-2 and Cf-5 were expressed and shown to be functional. The results clearly show that the specificity for the particular avirulence determinant is restricted to a region of each gene that encodes a subset of LRRs containing the highest level of intergenic variability. In addition, two non-functional mutants of Cf-5 were characterized and their significance discussed.


Daily Associations Between Trauma-Related Mental Contamination and Use of Specific Coping Strategies: Results of a Daily Monitoring Study.
Mental contamination (MC)-a sense of dirtiness experienced without contacting an identifiable pollutant-is a distressing and enduring experience among many survivors of sexual trauma. MC has been linked to more frequent use of avoidant coping behaviors (e.g., washing behavior, substance use, binge eating) and approach coping. However, it is unclear if specific approach and avoidant coping strategies are more consistently related to perseverative experiences of trauma-related MC, if the use of certain strategies predicts changes in MC, and if fluctuations in MC predict the use of certain strategies. The present study evaluated contemporaneous and prospective relationships between sexual trauma-related MC and use of 11 specific coping strategies among 41 women with a history of sexual trauma using an experience sampling design. Women completed twice-daily assessments of coping strategy use and MC for 14 days. Between-persons, women reporting more intense MC on average reported more frequent use of distraction, denial, giving up, self-blame, thought suppression, washing behavior, emotional processing, and emotional expression than those experiencing less intense MC. Within-person increases in MC were associated with more frequent concurrent use of all coping strategies except seeking support. Lastly, within-person increases in MC predicted more frequent use of giving up, substance use, and seeking support at the next assessment and within-person increases in substance use predicted less severe MC at the next assessment. Future work should aim to identify factors influencing the selection and/or quality of use of these specific coping strategies among individuals experiencing MC.


Maximizing Production of Human Interferon-γ in HCDC of Recombinant E. coli.
Tuning recombinant protein expression is an approach which can be successfully employed for increasing the yield of recombinant protein production in high cell density cultures. On the other hand, most of the previous results reported the optimization induction conditions during batch and continuous culture of recombinant E. coli, and consequently fed-batch culture have received less attention. Hence, in this research induction conditions for the over-production of recombinant interferon-γ including the amount of inducer, induction time and post-induction duration during chemical induction were optimized. E. coli BL21 (DE3) (pET3a-hifnγ) was used to over-express human interferon-gamma (hIFN-γ) in an exponential fed-batch procedure with a maximum attainable specific growth rate of 0.55 h(-1) at the beginning of feeding and 0.4 h(-1) in induction time. The factors were considered as the amount of inducer (IPTG) in the range of 0.565-22 mg g(-1) L(-1) at seven levels, cell density at induction time as 53, 65 and 75 g (dry cell weight) L(-1), induction duration at different intervals of 3, 4, and 5 h after induction time. The final concentration of biomass and interferon gamma reached to 127 g L(-1) (DCW) and 51 g (hIFN-γ) L(-1) after 17 h, and also the final specific yield and overall productivity were obtained 0.4 g (hIFN-γ) g(-1) DCW and 3 g (hIFN-γ) L(-1) h(-1), respectively, which are the highest amounts of reported specific yield and productivity for recombinant proteins production.


Progression from stenosis to occlusion in the proximal native coronary artery after coronary artery bypass grafting.
Coronary artery bypass grafting (CABG) is an established treatment for multivessel coronary artery disease. However, problematic situations are occasionally encountered after CABG, such as disease progression in the native coronary artery with graft occlusion, which causes difficulty in revascularization. The purpose of this study was to evaluate changes in the native coronary artery after CABG. Between 2009 and 2012 in our institution, 351 patients underwent CABG, and 768 bypass grafts were anastomosed to non-occluded coronary arteries. Of these, 489 bypass grafts had available early postoperative angiographic results (≤6 months) suitable for assessment in this study. We defined malignant graft failure after CABG to be bypass graft occlusion and de novo complete occlusion of the target native coronary artery proximal to the graft anastomosis site. In the early angiographic results, 17 grafts were occluded (17/489; 3.5 %). Two of the grafts displayed malignant graft failure (a saphenous vein graft to the right coronary artery and a saphenous vein graft to the diagonal branch) (2 of 17 occluded grafts, and 2 of 489 studied grafts). Of the patent bypass grafts, 24 involved progression to occlusion in the proximal native coronary artery (19 saphenous vein grafts, 4 left internal thoracic artery grafts, and 1 right internal thoracic artery graft). Malignant graft failure was uncommon during short-term follow-up after CABG. At the same time, disease progression in the proximal native coronary artery from stenosis to occlusion following patent bypass grafting was relatively common, especially for vein grafts.


In vitro toxicity of different-sized ZnO nanoparticles in Caco-2 cells.
There has been rapid growth in nanotechnology in both the public and private sectors worldwide, but concern about nanosafety exists. To assess size-dependent cytotoxicity on human cancer cells, we studied the cytotoxic effect of three kinds of zinc oxide nanoparticles (ZnO NPs) on human epithelial colorectal adenocarcinoma (Caco-2) cells. Nanoparticles were first characterized by size, distribution, and intensity. Multiple assays have been adopted to measure the cell activity and oxidative stress. The cytotoxicity of ZnO NPs was time dependent and dose dependent. The 24-h exposure was chosen to confirm the viability and accessibility of the cells and taken as the appropriate time for the following test system. The IC50 value was found at a low concentration. The oxidative stress elicited a significant reduction in glutathione with increase in reactive oxygen species and lactate dehydrogenase. The toxicity resulted in a deletion of cells in the G1 phase and an accumulation of cells in the S and G2/M phases. One type of metallic oxide (ZnO) exerted different cytotoxic effects according to different particle sizes. Data from the previous experiments showed that 26-nm ZnO NPs appeared to have the highest toxicity to Caco-2 cells. The study demonstrated the toxicity of ZnO NPs to Caco-2 cells and the impact of particle size, which could be useful in the medical applications.


Falciform ligament aperture causing intestinal strangulation.
In rare cases intestinal obstruction may be caused by herniation of small bowel through a defect in the falciform ligament. Such a case was that of a 28-year-old woman who had a 26-week twin pregnancy. She had acute upper abdominal pain. The diagnosis was acute appendicitis, but at operation a loop of ileum was found to have passed through an aperture in the falciform ligament. The loop was gangrenous and required resection. Her course was complicated by delivery of two infants on the day after operation; one died within a few hours and the other 4 months later. The patient recovered after a second operation 2 weeks later for small bowel obstruction due to adhesions. Only six other patients with this anomaly have been reported. Two were neonates. Three patients died. This complication of pregnancy has not been described before. Dislocation of the small intestine by the enlarged uterus may have contributed to the herniation. Prompt operation is mandatory in this condition as in any case of closed-loop bowel obstruction.


Ventilator-associated pneumonia, liver disease and oral chlorhexldine.
As part of the ventilator care bundle, the Department of Health (DH) in the U.K. recommends the use of chlorhexidine (CHX) for oral care to prevent the occurrence of ventilator-associated pneumonia (VAP) in all mechanically ventilated patients. Due to the heterogenous nature of this population, however, it is important to consider whether such recommendations are also relevant to specific critical care patient population groups. This article reviews the available scientific evidence on the use of CHX in the prevention of VAP, with a focus on critically ill mechanically ventilated patients who have liver dysfunction. Findings will be discussed with reference to the wider research literature in order to make recommendations for future practice.


P450 aromatase expression in the temperature-sensitive sexual differentiation of salamander (Hynobius retardatus) gonads.
Sex differentiation of gonads in amphibians is believed to be controlled genetically, but altered epigenetically or environmentally. When larvae of the salamander Hynobius retardatus were reared at defined temperatures from hatching to metamorphic stages, a high temperature (28 degrees C) induced exclusively female gonads (ovaries), whereas intermediate (20 and 23 degrees C) or lower (16 degrees C) temperatures produced a 1:1 sex ratio of the morphological gonads. The thermosensitive period was determined to be restricted from 15 to 30 days after hatching, just before or when sexual differentiation occurred. Hynobius P450 aromatase (P450arom) cDNA was isolated from adult gonads and the partial nucleotide or deduced amino acid sequences were determined, showing a high level of identity with various vertebrate species. The P450arom gene was expressed predominantly in the adult ovary and brain, weakly in testis, but not in other somatic organs. A typical sexual dimorphism in P450arom expression was detected in normally developing larvae by a quantitative competitive RT-PCR; strong expression in the female gonads but very weak in male gonads. The dimorphism was detected much earlier than the morphological sexual differentiation of the gonads. When larvae were reared at the female-producing temperature (28 degrees C), strong expression was detected in all the temperature-treated larvae, suggesting that P450arom was up-regulated, even in genetic males. Our results confirm the importance of the P450arom regulation in the sexual differentiation of gonads and demonstrate that an up-regulation of P450arom is involved in the process of temperature-sensitive sex reversal in this species.


Can the vicious cycle of obscure or intractable gastrointestinal bleeding be broken in patients with atrial fibrillation subject to anticoagulant therapy?
Gastrointestinal bleeding of obscure origin or with an intractable cause is particularly common in patients with atrial fibrillation subject to oral anticoagulant therapy. This condition is highly recurrent and therefore gives rise to high morbidity and mortality rates, thus entailing a vicious cycle that is difficult to solve.Percutaneous left atrial appendage closure has become a therapeutic alternative for patients with atrial fibrillation and a contraindication for oral anticoagulation. This technique would allow the discontinuation of oral anticoagulants, thus helping to reduce the risk for gastrointestinal bleeding, and would also be protective against embolic events in this group of patients, thereby eventually breaking this vicious cycle.We report our experience with percutaneous left atrial appendage closure in the management of patients with atrial fibrillation who are subject to oral anticoagulation therapy and suffer from obscure or intractable gastrointestinal bleeding.


Prepartum nutritional strategy affects reproductive performance in dairy cows.
Negative energy balance during early postpartum is associated with reduced reproductive performance in dairy cows. A pooled statistical analysis of 7 studies completed in our group from 1993 to 2010 was conducted to investigate the association between prepartum energy feeding regimen and reproductive performance. The interval from calving to pregnancy (days to pregnancy, DTP) was the dependent variable to assess reproductive performance. Individual data for 408 cows (354 multiparous and 54 primiparous) were included in the analysis. The net energy for lactation (NEL) intake was determined from each cow's average dry matter intake and calculated dietary NEL density. Treatments applied prepartum were classified as either controlled-energy (CE; limited NEL intake to ≤100% of requirement) or high-energy (HE; cows were allowed to consume >100%) diets fed during the far-off (FO) or close-up (CU) dry periods. Cow was the experimental unit. The Cox proportional hazard model revealed that days to pregnancy was shorter for CE (median=157 d) than HE (median=167 d) diets during the CU period [hazard ratio (HR)=0.70]. Cows fed HE diets during the last 4 wk prepartum lost more body condition score in the first 6 wk postpartum than those fed CE diets (-0.43 and -0.30, respectively). Cows fed CE diets during the FO period had lower nonesterified fatty acids concentrations in wk 1, 2, and 3 of lactation than cows fed HE diets. Higher nonesterified fatty acids concentration in wk 1 postpartum was associated with a greater probability of disease (n=251; odds ratio=1.18). Cows on the CE regimen during the FO period had greater plasma glucose concentrations during wk 1 and 3 after calving than cows fed the HE regimen. Higher plasma glucose (HG) concentration compared with lower glucose (LG) in wk 3 (HG: n=154; LG: n=206) and wk 4 (HG: n=71; LG: n=254) after calving was associated with shorter days to pregnancy (wk 3: median=151 and 171 d for HG and LG, respectively, and HR=1.3; wk 4: median=148 and 167 d, respectively, and HR=1.4). In the first 2 wk after calving, cows that received HE diets in the FO period had higher concentrations of total lipids and triglyceride and greater ratio of triglyceride to glycogen in liver than cows fed CE diets. In conclusion, cows fed CE diets during the CU period had a shorter interval between parturition and conception, which may be explained by increased NEL intake during the first 4 wk postpartum and lower incidence of peripartal diseases. Lower body condition score loss during the first 6 wk postpartum and slightly higher glucose concentration at wk 3 likely contributed to improved reproductive performance.


Additive Manufacturing of Biomaterials-Design Principles and Their Implementation.
Additive manufacturing (AM, also known as 3D printing) is an advanced manufacturing technique that has enabled progress in the design and fabrication of customised or patient-specific (meta-)biomaterials and biomedical devices (e.g., implants, prosthetics, and orthotics) with complex internal microstructures and tuneable properties. In the past few decades, several design guidelines have been proposed for creating porous lattice structures, particularly for biomedical applications. Meanwhile, the capabilities of AM to fabricate a wide range of biomaterials, including metals and their alloys, polymers, and ceramics, have been exploited, offering unprecedented benefits to medical professionals and patients alike. In this review article, we provide an overview of the design principles that have been developed and used for the AM of biomaterials as well as those dealing with three major categories of biomaterials, i.e., metals (and their alloys), polymers, and ceramics. The design strategies can be categorised as: library-based design, topology optimisation, bio-inspired design, and meta-biomaterials. Recent developments related to the biomedical applications and fabrication methods of AM aimed at enhancing the quality of final 3D-printed biomaterials and improving their physical, mechanical, and biological characteristics are also highlighted. Finally, examples of 3D-printed biomaterials with tuned properties and functionalities are presented.


Physiological regulation of isocitrate dehydrogenase and the role of 2-oxoglutarate in Prochlorococcus sp. strain PCC 9511.
The enzyme isocitrate dehydrogenase (ICDH; EC 188.8.131.52) catalyzes the oxidative decarboxylation of isocitrate, to produce 2-oxoglutarate. The incompleteness of the tricarboxylic acids cycle in marine cyanobacteria confers a special importance to isocitrate dehydrogenase in the C/N balance, since 2-oxoglutarate can only be metabolized through the glutamine synthetase/glutamate synthase pathway. The physiological regulation of isocitrate dehydrogenase was studied in cultures of Prochlorococcus sp. strain PCC 9511, by measuring enzyme activity and concentration using the NADPH production assay and Western blotting, respectively. The enzyme activity showed little changes under nitrogen or phosphorus starvation, or upon addition of the inhibitors DCMU, DBMIB and MSX. Azaserine, an inhibitor of glutamate synthase, induced clear increases in the isocitrate dehydrogenase activity and icd gene expression after 24 h, and also in the 2-oxoglutarate concentration. Iron starvation had the most significant effect, inducing a complete loss of isocitrate dehydrogenase activity, possibly mediated by a process of oxidative inactivation, while its concentration was unaffected. Our results suggest that isocitrate dehydrogenase responds to changes in the intracellular concentration of 2-oxoglutarate and to the redox status of the cells in Prochlorococcus.


The effects of meclofenamate, captopril and phentolamine on organ blood flow in the conscious rabbit.
1. Systemic and regional vascular changes were measured in conscious rabbits after intravenous sodium meclofenamate, captopril and phentolamine. These drugs were used respectively to inhibit prostaglandin synthesis and angiotensin-converting enzyme, and to block alpha-adrenoceptors. 2. Meclofenamate reduced renal and adrenal blood flow by 11 and 28% respectively, and doubled hepatic artery flow. The effect on renal and adrenal flow persisted in the presence of phentolamine. 3. Captopril decreased estimated peripheral resistance and increased cardiac output without changing arterial pressure. Kidney and adrenal flow increased by 70 and 21% respectively. 4. Phentolamine reduced arterial pressure and doubled flow to skeletal muscle and increased hepatic artery flow to the liver. 5. Splenic blood flow was unaffected by meclofenamate, captopril or phentolamine alone. Meclofenamate given after captopril caused a halving of splenic flow and a rise in arterial pressure; these effects were prevented by phentolamine. 6. The results point to a continuing effect of prostaglandin synthesis in maintaining blood flow to the kidney and adrenal gland independent of alpha-adrenoceptor activation in resting conscious rabbits. An important modulating effect of prostaglandins on sympathetic vascular tone in the spleen is suggested.


Nonsteroidal anti-inflammatory drugs (NSAIDs) cause male-biased sex differentiation in zebrafish.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used pharmaceuticals to treat pain, fever and inflammation. NSAIDs are also known to have many side effects including adverse effects on reproduction in both humans and animals. As NSAIDs usage is not regulated they are frequently detected at high concentrations in the environment. In order to understand the effect of NSAIDs on zebrafish sex differentiation, we used seven different NSAIDs which were either Cox-1 selective, Cox-1 biased, non-selective or COX-2 selective. We show that at higher concentration, NSAIDs are toxic to zebrafish embryo as they lead to mortality and hatching delay. Gene expression analysis following short term exposure of NSAIDs led to downregulation of female specific genes including zp2, vtg2 foxl2 and wnt4. Long term exposure of larvae to environmentally relevant concentrations of Cox-2 selective and non-selective NSAIDs resulted in male-biased sex ratio which confirmed the qRT-PCR analysis. However, the Cox-1 selective acetylsalicylic acid and the Cox-1 biased ketoprofen did not alter sex ratio. The observed male-biased sex ratio could also be due to induction of apoptosis process as the genes including p21 and casp8 were significantly upregulated following exposure to the Cox-2 selective and the non-selective NSAIDs. The present study indicates that NSAIDs alter sex differentiation in zebrafish, primarily through inhibition of Cox-2. This study clearly demonstrates that the use of NSAIDs and their release into the aquatic environment should be carefully monitored to avoid adverse effects to the aquatic organisms.


Do BDNF and NT-4/5 exert synergistic or occlusive effects on corticostriatal transmission in a male mouse model of Huntington's disease?
Brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT-4/5) are trophic factors belonging to the neurotrophin family; in addition to their trophic role, both neurotrophins play an important role in modulating corticostriatal synaptic transmission. Failures in BDNF supply and mitochondrial dysfunction are among the factors involved in the striatal degeneration that occurs in Huntington's disease (HD). While the effects of BDNF have been widely studied in striatal degeneration, the role of NT-4/5 has been less addressed. NT-4/5 does not appear to exert effects similar to those of BDNF in HD. The physiological roles of these molecules in corticostriatal transmission have been evaluated separately, and we have demonstrated that sequential exposure to both neurotrophins results in different modulatory effects on corticostriatal transmission depending on the exposure order. In the present study, we evaluated the effects of BDNF followed by NT-4/5 or NT-4/5 followed by BDNF on corticostriatal synaptic transmission with field recordings in a male mouse model of HD produced by in vivo treatment with the mitochondrial toxin 3-nitropropionic acid. Here, we show that these neurotrophins elicit an antagonistic or synergistic effect that depends on the activation of the truncated isoform or the stimulation of the full-length isoform of the tropomyosin receptor kinase B.


Characteristics, service use and mortality of clusters of multimorbid patients in England: a population-based study.
Multimorbidity is associated with mortality and service use, with specific types of multimorbidity having differential effects. Additionally, multimorbidity is often negatively associated with participation in research cohorts. Therefore, we set out to identify clusters of multimorbidity patients and how they are differentially associated with mortality and service use across age groups in a population-representative sample. Linked primary and secondary care electronic health records contributed by 382 general practices in England to the Clinical Practice Research Datalink (CPRD) were used. The study included a representative set of multimorbid adults (18 years old or more, N = 113,211) with two or more long-term conditions (a total of 38 conditions were included). A random set of 80% of the multimorbid patients (N = 90,571) were stratified by age groups and clustered using latent class analysis. Consistency between obtained multimorbidity phenotypes, classification quality and associations with demographic characteristics and primary outcomes (GP consultations, hospitalisations, regular medications and mortality) was validated in the remaining 20% of multimorbid patients (N = 22,640). We identified 20 patient clusters across four age strata. The clusters with the highest mortality comprised psychoactive substance and alcohol misuse (aged 18-64); coronary heart disease, depression and pain (aged 65-84); and coronary heart disease, heart failure and atrial fibrillation (aged 85+). The clusters with the highest service use coincided with those with the highest mortality for people aged over 65. For people aged 18-64, the cluster with the highest service use comprised depression, anxiety and pain. The majority of 85+-year-old multimorbid patients belonged to the cluster with the lowest service use and mortality for that age range. Pain featured in 13 clusters. This work has highlighted patterns of multimorbidity that have implications for health services. These include the importance of psychoactive substance and alcohol misuse in people under the age of 65, of co-morbid depression and coronary heart disease in people aged 65-84 and of cardiovascular disease in people aged 85+.


Genetic dissection of retinoid dehydrogenases.
Biochemical studies indicate that alcohol dehydrogenase (ADH) metabolizes retinol to retinal, and that aldehyde dehydrogenase (ALDH) metabolizes retinal to retinoic acid, a molecule essential for growth and development. Summarized herein are several genetic studies supporting in vivo functions for ADH and ALDH in retinoic acid synthesis. Gene targeting was used to create knockout mice for either Adh1 or Adh4. Both knockout mice were viable and fertile without obvious defects. However, when wild-type and Adh4 knockout mice were subjected to vitamin A deficiency during gestation, the survival rate at birth was 3.3-fold lower for Adh4 knockout mice. When adult mice were examined for production of retinoic acid following retinol administration, Adh1 knockout mice exhibited 10-fold lower retinoic acid levels in liver compared with wild-type, whereas Adh4 knockout mice differed from wild-type by less than 2-fold. Thus, Adh1 plays a major role in the metabolism of a large dose of retinol to retinoic acid in adults, whereas Adh4 plays a role in maintaining sufficient retinol metabolism for development during retinol deficiency. ALDHs were examined by overexpression studies in frog embryos. Injection of mRNAs for either mouse Raldh1 or Raldh2 stimulated retinoic acid synthesis in frog embryos at the blastula stage when retinoic acid is normally undetectable. Overexpression of human ALDH2, human ALDH3, and mouse Aldh-pb did not stimulate retinoic acid production. In addition, Raldh2 knockout mice exhibit embryonic lethality with defects in retinoid-dependent tissues. Overall, these studies provide genetic evidence that Adh1, Adh4, Raldh1, and Raldh2 encode retinoid dehydrogenases involved in retinoic acid synthesis in vivo.


Long-term intrathecal infusion of outer surface protein C from Borrelia burgdorferi causes axonal damage.
Lyme neuroborreliosis (LNB) is the most frequent tick-borne infectious disease of the central nervous system. In acute LNB and the rare chronic state of infection, patients can experience cognitive deficits such as attention and memory disturbances. During LNB, single compounds of Borrelia burgdorferi sensu lato are released into the subarachnoid space.To investigate the pathogenesis of neurologic dysfunction in LNB, we determined that the outer surface protein C (OspC), a major virulence factor of B. burgdorferi, stimulated mouse microglial cells in a dose-dependent manner to release nitric oxide (EC50 = 0.24 mg/L) in vitro. To mimic pathophysiologic conditions of long-term release of this bacterial component in vivo, we treated C57BL/6 mice with recombinant OspC from Borrelia garinii or buffer by intraventricular infusion and tested them for behavioral deficits. After 4weeks, brains were examined by routine histology and immunohistochemistry. Assessment of spatial learning and memory of treated mice during OspC exposure did not reveal significant differences from controls. Continuous exposure to intrathecal B. burgdorferi OspC led to activation of microglia and axonal damage without demonstrable cognitive impairment in experimental mice. These results suggest that long-term intrathecal exposure to OspC resulted in axonal damage that may underlie the neurologic manifestations in chronic LNB.


Primary blast survival and injury risk assessment for repeated blast exposures.
The widespread use of explosives by modern insurgents and terrorists has increased the potential frequency of blast exposure in soldiers and civilians. This growing threat highlights the importance of understanding and evaluating blast injury risk and the increase of injury risk from exposure to repeated blast effects. Data from more than 3,250 large animal experiments were collected from studies focusing on the effects of blast exposure. The current study uses 2,349 experiments from the data collection for analysis of the primary blast injury and survival risk for both long- and short-duration blasts, including the effects from repeated exposures. A piecewise linear logistic regression was performed on the data to develop survival and injury risk assessment curves. New injury risk assessment curves uniting long- and short-duration blasts were developed for incident and reflected pressure measures and were used to evaluate the risk of injury based on blast over pressure, positive-phase duration, and the number of repeated exposures. The risk assessments were derived for three levels of injury severity: nonauditory, pulmonary, and fatality. The analysis showed a marked initial decrease in injury tolerance with each subsequent blast exposure. This effect decreases with increasing number of blast exposures. The new injury risk functions showed good agreement with the existing experimental data and provided a simplified model for primary blast injury risk. This model can be used to predict blast injury or fatality risk for single exposure and repeated exposure cases and has application in modern combat scenarios or in setting occupational health limits.


On-demand generation of singlet oxygen from a smart graphene complex for the photodynamic treatment of cancer cells.
Graphene oxide (GO) has been proven to be a highly efficient long-range quencher for various fluorescence processes, which intrinsically work through a photophysical mechanism similar to that of singlet oxygen generation (SOG). Under our hypothesis that GO may be capable of quenching the SOG process, here we design and synthesize a novel nanocomplex consisting of GO, a photosensitizer and an aptamer. We demonstrate that GO is an ideal SOG controller, which can reversibly quench and recover SOG depending on the interaction intensity between GO and a photosensitizer. Additionally, it can simultaneously act as a carrier for the efficient loading and delivery of the photosensitizers to cancer cells. Thus, during the delivery process, SOG of the nanocomplex can be completely inhibited by the quenching capacity of the GO even though there is light present; however, when the nanocomplex enters into cancer cells where target molecules are present, SOG is triggered by a target binding event and singlet oxygen is reversibly released from the nanocomplex, ultimately inducing significant cell death in the presence of light. This proof-of-concept study provides a new chemical strategy for creating a highly selective photodynamic therapy with low toxicity, using hydrophilic GO-based systems.


Topical and peripheral ketamine as an analgesic.
Ketamine, in subanesthetic doses, produces systemic analgesia in chronic pain settings, an action largely attributed to block of N-methyl-D-aspartate receptors in the spinal cord and inhibition of central sensitization processes. N-methyl-D-aspartate receptors also are located peripherally on sensory afferent nerve endings, and this provided the initial impetus for exploring peripheral applications of ketamine. Ketamine also produces several other pharmacological actions (block of ion channels and receptors, modulation of transporters, anti-inflammatory effects), and while these may require higher concentrations, after topical (e.g., as gels, creams) and peripheral application (e.g., localized injections), local tissue concentrations are higher than those after systemic administration and can engage lower affinity mechanisms. Peripheral administration of ketamine by localized injection produced some alterations in sensory thresholds in experimental trials in volunteers and in complex regional pain syndrome subjects in experimental settings, but many variables were unaltered. There are several case reports of analgesia after topical application of ketamine given alone in neuropathic pain, but controlled trials have not confirmed such effects. A combination of topical ketamine with several other agents produced pain relief in case, and case series, reports with response rates of 40% to 75% in retrospective analyses. In controlled trials of neuropathic pain with topical ketamine combinations, there were improvements in some outcomes, but optimal dosing and drug combinations were not clear. Given orally (as a gargle, throat swab, localized peritonsillar injections), ketamine produced significant oral/throat analgesia in controlled trials in postoperative settings. Topical analgesics are likely more effective in particular conditions (patient factors, disease factors), and future trials of topical ketamine should include a consideration of factors that could predispose to favorable outcomes.


Rapid method of determining cholera vibrio biotype.
The characteristic motility of cholera vibrios, as viewed through a dark-field microscope, and the adhesiveness of chicken cell-positive vibrios provide a means for rapidly identifying and biotyping cholera vibrios. Dilute suspensions of vibrios, such as one might find in a fresh rectal swab specimen from a cholera patient, when mixed with a 0.25% suspension of chicken erythrocytes in saline, can be used to biotype the cholera vibrios without prior isolation in pure culture. This is accomplished by using a dark-field microscope through which the chicken cell-positive cholera vibrios are observed to attach to the scattered erythrocytes and to propel them with a characteristic flipping motion.


Development of tip-splitting and side-branching patterns in elastic fingering.
Elastic fingering supplements the already interesting features of the traditional viscous fingering phenomena in Hele-Shaw cells with the consideration that the two-fluid separating boundary behaves like an elastic membrane. Sophisticated numerical simulations have shown that under maximum viscosity contrast the resulting patterned shapes can exhibit either finger tip-splitting or side-branching events. In this work, we employ a perturbative mode-coupling scheme to get important insights into the onset of these pattern formation processes. This is done at lowest nonlinear order and by considering the interplay of just three specific Fourier modes: a fundamental mode n and its harmonics 2n and 3n. Our approach further allows the construction of a morphology diagram for the system in a wide range of the parameter space without requiring expensive numerical simulations. The emerging interfacial patterns are conveniently described in terms of only two dimensionless controlling quantities: the rigidity fraction C and a parameter Γ that measures the relative strength between elastic and viscous effects. Visualization of the rigidity field for the various pattern-forming structures supports the idea of an elastic weakening mechanism that facilitates finger growth in regions of reduced interfacial bending rigidity.


Assessing polyphenols content and antioxidant activity in coffee beans according to origin and the degree of roasting
The roasting stage constitutes a key component in the manufacturing process of natural coffee because temperature elicits changes in bioactive compounds such as polyphenols and that Maillard-reaction compounds appear, thus affecting the product's sensory and antioxidant properties. Actual contents of these compounds may depend on which region the coffee is cultivated as well as the extent to which the beans are roasted To determine polyphenols content and antioxidant activity in the 'Arabica' coffee type coming from various world regions of its cultivation and which have undergone industrial roasting. Also to establish which coffee, taking into account the degree of roasting (ie. light, medium and strong), is nutritionally the most beneficial The study material was natural coffee beans (100% Arabica) roasted to various degrees, as aforementioned, that had been cultivated in Brazil, Ethiopia, Columbia and India. Polyphenols were measured in the coffee beans by spectrophotometric means based on the Folin-Ciocalteu reaction, whereas antioxidant activity was measured colourimetrically using ABTS+ cat-ionic radicals Polyphenol content and antioxidant activity were found to depend both on the coffee's origin and degree of roasting. Longer roasting times resulted in greater polyphenol degradation. The highest polyphenol concentrations were found in lightly roasted coffee, ranging 39.27 to 43.0 mg/g, whereas levels in medium and strongly roasted coffee respectively ranged 34.06 to 38.43 mg/g and 29.21 to 36.89 mg/g. Antioxidant activity however significantly rose with the degree of roasting, where strongly roasted coffee had higher such activity than lightly roasted coffee. This can be explained by the formation of Maillard-reaction compounds during roasting, leading then to the formation of antioxidant melanoidin compounds which, to a large extent, compensate for the decrease in polyphenols during roasting Polyphenols levels and antioxidant activities in the studied Arabica coffee beans that had undergone roasting depended on the cultivation region of the world. Longer roasting caused a significant decline in polyphenols compound levels (from 7.3% to 32.1%) in the coffee beans. Antioxidant activities of coffee increased with roasting, despite reduced levels of natural antioxidants. From a nutritional standpoint, the most favoured coffees are those lightly or medium roasted


Phylogenetic analysis of the structure of RNase MRP RNA in yeasts.
RNase MRP is a ribonucleoprotein enzyme involved in processing precursor rRNA in eukaryotes. To facilitate our structure-function analysis of RNase MRP from Saccharomyces cerevisiae, we have determined the likely secondary structure of the RNA component by a phylogenetic approach in which we sequenced all or part of the RNase MRP RNAs from 17 additional species of the Saccharomycetaceae family. The structure deduced from these sequences contains the helices previously suggested to be common to the RNA subunit of RNase MRP and the related RNA subunit of RNase P, an enzyme cleaving tRNA precursors. However, outside this common region, the structure of RNase MRP RNA determined here differs from a previously proposed universal structure for RNase MRPs. Chemical and enzymatic structure probing analyses were consistent with our revised secondary structure. Comparison of all known RNase MRP RNA sequences revealed three regions with highly conserved nucleotides. Two of these regions are part of a helix implicated in RNA catalysis in RNase P, suggesting that RNase MRP may cleave rRNA using a similar catalytic mechanism.


Alpha-adrenoceptors and facilitation at a sympathetic neuroeffector junction.
Excitatory junction potentials were recorded from the mouse vas deferens following 5 stimuli at 1Hz and a single stimulus at times from 100 msec to 9 sec later. E.j.ps facilitate and facilitation decayed over a biexponential time course, with time constants of 120 msec and 4.3 sec. Clonidine (10(-8)-10(-7) M) depressed e.j.p. amplitude, and also accelerated both phases of the decay of facilitation. Yohimbine (10(-7) M) and piperoxan (10(-7)-10(-6) M) increased the amplitude of all but the first e.j.p. in a train, and slowed both phases of the decay of facilitation. One way in which prejunctional modulation by alpha-adrenoceptors may occur is therefore by modifying the carry-over of facilitation from one stimulus to the next.


Diminishing morbidity and mortality of bone marrow transplantation.
The reasons for the operational failure of bone marrow transplantation (BMT) are familiar and varied. They are the following: nonmarrow toxicity of the preparative regimen, marrow toxicity of the preparative regimen, acute graft-versus-host disease (GVHD), opportunistic infections, especially viral infections and recurrence of hematologic malignancy in the case of transplants for leukemia. This review examines operational failure of BMT and presents measures for its prevention.


Repetitive Transcranial Magnetic Stimulation For The Treatment of Chronic Tinnitus: A Preliminary Study of The Influence of Traumatic Brain Injury on Treatment Response.
This study aims to test whether the efficacy of repetitive Transcranial Magnetic Stimulation (rTMS) differs between patients who developed tinnitus following a traumatic brain injury (TBI), and those without a history of TBI. This was a parallel pilot, open-label, non-randomized, clinical trial to compare the efficacy of low frequency rTMS on tinnitus symptoms in patients with and without a TBI history. Patients with moderate to severe tinnitus symptoms based on the Tinnitus Handicap Inventory (THI) and the Tinnitus Functional Index (TFI) were enrolled in the study. Validated questionnaires (THI and TFI) were used to quantify the severity of tinnitus symptoms and hearing impairment (Hearing Handicap Index - HHI) before and after ten sessions of rTMS of the left primary auditory cortex. Hearing threshold levels as well as speech reception and speech discrimination thresholds were also compared. The number of patients who experienced a reduction in their subjective tinnitus symptoms was greater and sustained longer in patients without a history of TBI. The same was seen with subjective symptoms of hearing impairment. In conclusion, our preliminary results suggest tinnitus patients without a history of TBI respond better to low frequency rTMS than those with a history of TBI, suggesting that treatments could be more effective if tailored to tinnitus etiology.


[Classification of color/race in children from indigenous households in Brazil].
Studies on racial classification systems in Brazil reveal the influence of socioeconomic factors in the expression of color/race categories, especially for whites and blacks. The aim of this study was to analyze specific family arrangements between fathers, mothers, and children, at least one of whom was indigenous. Based on the sample from the 2010 Population Census, we selected households with at least three residents (father, mother, and children), at least one of whom was indigenous. Children were characterized according to color/race (white, brown, and indigenous), sex, age, per capita household income, maternal schooling, and number of urban and rural household residents. Descriptive and multinomial logistic regression analyses were performed. We estimated a total of 290.247 children (of whom 77.1% were classified as indigenous, 13.8% brown, and 9.1% white), 74.3% living in rural households and 41.3% in the North region of Brazil; children classified as white and brown were located mostly in urban areas. The odds of children of indigenous fathers or mothers being classified as white were higher in the Southeast and South. The odds of children being classified as white or brown increased proportionally with monthly income and maternal schooling. The findings show that socioeconomic status is significantly associated with color/race classification in Brazil, including in indigenous households.


Syntaxin 1B suppresses macropinocytosis and semaphorin 3A-induced growth cone collapse.
Growth cone collapse is a crucial process for repulsive axon guidance and is accompanied by a reduction in growth cone surface area. This process of reduction may be regulated by endocytosis; however, its molecular mechanism is unclear. Macropinocytosis is a clathrin-independent form of endocytosis in which large areas of plasma membrane can be engulfed. We have reported previously that macropinocytosis is induced in growth cones of chick dorsal root ganglion neurons by semaphorin 3A (Sema3A), a repulsive axon guidance cue, and that Sema3A-induced reduction in growth cone surface area and macropinocytic vacuole area were correlated, suggesting a positive role for macropinocytosis in Sema3A-induced growth cone collapse. In the present study, we found that syntaxin 1B (Syx1B), a membrane trafficking protein, is a negative regulator of macropinocytosis, and its expression is downregulated by Sema3A signaling. Macropinocytosis inhibitor ethylisopropylamiloride or Syx1B overexpression suppressed Sema3A-induced macropinocytosis and growth cone collapse. These results indicate that Syx1B couples macropinocytosis-mediated massive internalization of the plasma membrane to Sema3A-induced growth cone collapse.


Effort-Reward Imbalance Is Associated With Alcohol-Related Problems. WIRUS-Screening Study.
There is ample evidence of associations between a perceived stressful working environment and several health-related outcomes. To better understand potential mechanisms behind these observations some studies have focused on the relationship between effort-reward imbalance at work and alcohol consumption. So far, the findings have been inconsistent. One reason for this inconsistency might come from the focus on alcohol consumption per se, while disregarding other aspects such as adverse consequences related to the consumption of alcohol. The aim of the present study was to explore associations between perceived effort and reward, effort-reward imbalance and overcommitment, and alcohol-related problems. Using data from the alcohol screening component in the Norwegian WIRUS-project (N = 5,080), we ascertained the perceived effort, reward, effort-reward imbalance (ERI) and overcommitment using the effort-reward imbalance questionnaire. Alcohol-related problems was determined using a cut-off ≥8 on the Alcohol Use Disorder Identification Test (AUDIT). Associations were estimated using crude and adjusted logistic regression models. Covariates were age, gender and education. We found associations between different aspects of ERI and overcommitment, and alcohol-related problems. Specifically, the main analysis indicated that there was an increased odds for alcohol-related problems among those who reported high levels of ERI in conjunction with high overcommitment [adjusted OR: 1.40 (CI 95% 1.10-1.78)] compared to those with low levels of ERI and low overcommitment. Our findings suggest that ERI and overcommitment is associated with increased likelihood of alcohol-related problems. These findings indicate that individual and work-related factors should be taken into account collectively when aiming to determine the impact of psychosocial work environment on alcohol-related problems. Due to the cross-sectional nature of the present study, we are not able to determine the direction of the associations, and future studies should aim to investigate this.


Mutual attachment, personality, and drug use: pathways from childhood to young adulthood.
Young adult drug use stemming from childhood aggression, the parent-child mutual attachment relationship, and the effect of unconventionality were studied. Youngsters and their mothers were interviewed when the former were early adolescents, late adolescents, and young adults. Additional data were collected from the mothers when their youngsters were children. The analysis was conducted on youngsters who had complete data at all 4 points in time. The findings were in accord with the family interactional model; that is, the parent-child mutual attachment relationship affects unconventionality in the youngster, which, in turn, affects young adult drug use. The results indicate that the parent-child mutual attachment relationship does so through (a) the stability of the attachment relationship from childhood to young adulthood, (b) the stability of unconventional personality and behavioral attributes from early adolescence to young adulthood, and (c) the stability of drug use from early adolescence to young adulthood. The findings imply that (a) early intervention with respect to aggression, (b) interventions that focus on strengthening the parent-child bond and conventional behavior, and (c) interventions aimed at early drug use should be most effective in reducing young adult drug use.


Synthesis of Semliki-forest virus in polyamine-depleted baby-hamster kidney cells.
The role of polyamines in macromolecular synthesis has been studied using the synthesis of Semliki-Forest virus (SF virus) in normal and alpha-difluoromethylornithine-treated baby-hamster kidney (BHK21) cells as a model system. The activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase, the rate-limiting enzymes in polyamine biosynthesis, decreased rapidly in mock- and SF-virus-infected cells, indicating that virus production in BHK21 cells was not dependent on polyamines formed after infection. A prolonged treatment of BHK21 cells with alpha-difluoro-methylornithine, a specific inhibitor of polyamine synthesis, resulted in a marked inhibition of the initial rate of virus production, which appeared 72 h after the beginning of the treatment. This inhibition was reversed by putrescine, spermidine and spermine, and at last partially by several other diamines and polyamine homologues. Polyamine-depletion also markedly reduced viral RNA polymerase activity in SF-virus infected cells. Addition of spermidine to the culture medium rapidly increased viral RNA polymerase activity in the inhibitor-treated cells but had no effect on the enzyme activity when added directly to the assay mixture. The results indicated that polyamines are needed for maximum initial rate of SF-virus replication and suggest that the inhibition of virus production in polyamine-depleted cells is at least partly due to malfunction of the protein-synthetic machinery of the host cell.


Crosslinking of trimethylene carbonate and D, L-lactide (co-) polymers by gamma irradiation in the presence of pentaerythritol triacrylate.
High-molecular-weight (co)polymers of trimethylene carbonate and D,L-lactide are efficiently crosslinked using PETA during gamma irradiation. Form-stable networks with gel contents of 86 ± 5 to 96 ± 1 are obtained from non-crystalline (co)polymers. Glass transition temperatures and elastic moduli of the networks can be varied by adjusting the copolymer composition. The PETA-containing (co)polymer networks are not cytotoxic. Upon incubation in macrophage cultures for 14 d, all (co)polymer films and PETA-containing network films erode to varying extents, showing that these materials can be degraded by cell-mediated erosion processes. This method is very useful for the facile preparation of TMC- and DLLA-containing form-stable networks from high-molecular-weight polymers.


A new protein A assay based on Raman reporter labeled immunogold nanoparticles.
A unique, sensitive, highly specific, and photobleaching-resistant immunoassay system utilizing gold nanoparticles and surface-enhanced Raman scattering (SERS) is described. This new system, featuring a capability of bifunctional analysis, is manufactured by chemisorption of antibody immunoglobulin G (IgG) on gold nanoparticles (AuNP), followed by coupling the Raman-active reporter molecule, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) to the surface of IgG-AuNP. The adsorbed DTNB molecules exhibit strong Raman signals via both electromagnetic and chemical enhancement. The narrow spectral widths and high photostability assure the system to be an excellent detection label. This SERS-based immunoassay is applied to the detection of protein A, which is a specific surface antigen of Staphylococcus aureus. A working curve is obtained by plotting the intensity of the SERS signal of symmetric NO(2) stretching of DTNB at 1,333 cm(-1) versus the concentration of the analyte (antigen). A dynamic range of two to three orders of magnitude and a detection limit of 1 pg/mL of protein A are achieved.


Directly diode-pumped Yb3+:SrY4(SiO4)3O regenerative amplifier.
We report a regenerative amplifier based on an Yb-doped apatite crystal: Yb3+:SrY4(SiO4)3O (Yb:SYS). We obtained 420-fs pulses at a central wavelength of 1066 nm with an energy of 100 microJ at 300 Hz after compression. To the best of our knowledge, this system is the first regenerative amplifier based on an Yb:SYS crystal and provides duration among the shortest ones generated by a directly diode-pumped regenerative amplifier.


A national study of the clinical management of HIV-positive women in Australia: what are the successes and where are the gaps?
Background Women comprise ~10% of people living with HIV in Australia, so are often underrepresented in research. This study invited clinicians providing care to women living with HIV to complete an anonymous survey containing questions related to four key areas: HIV (including diagnosis, treatment and virological outcomes), reproductive health (including sexual activity, contraception, pregnancy and outcomes) and linkage and retention in care. In total, 484 surveys were received, with responses from all states and territories. Most women living with HIV in Australia are on treatment (>90%) and virologically suppressed (>90% have a viral load <50 copies mL-1). Almost 75% of women have had at least one switch in treatment (with toxicity almost as common as simplification as the indication). Treatment interruption is also relatively common, but is more likely the longer a woman has been diagnosed, if she is on benefits (P = 0.007) and is the primary carer of children without a partner (P = 0.001). In Australia, women living with HIV are a diverse heterogeneous group, with over 70 different countries of birth and almost half speaking a language other than English at home. Mental health diagnosis was the most common co-morbid condition identified. A total of 21% of women were post-menopausal, with 42% reporting symptoms to their healthcare provider, but only 17% were receiving treatment for symptoms attributed to menopause. As well as strategies to support women vulnerable to treatment interruption, important areas for future investment in research and clinical care include co-morbid mental health and menopause symptoms and treatment.


Multiconfigurable logic gates based on fluorescence switching in adaptive coordination polymer nanoparticles.
Fluorescence switching of guest molecules confined in coordination polymer nanoparticles (CPNs) generated from nucleotides and lanthanide ions are used to construct multiconfigurable logic gates. Moreover, the potential of the material as fluorescent probe with large Stokes shift is demonstrated for cellular imaging. In this work the logic gate is integrated into the therapeutic agent and this will be highly beneficial in future molecular computing.


Glial Cell Line-Derived Neurotrophic Factor and Chondroitinase Promote Axonal Regeneration in a Chronic Denervation Animal Model.
Functional recovery following nerve injury declines when target re-innervation is delayed. Currently, no intervention exists to improve outcomes after prolonged denervation. We explored the neuroregenerative effects of glial cell line-derived neurotrophic factor (GDNF) and chondroitinase (CDN) in a chronic denervation animal model. A fibrin-based sustained delivery method for growth factors was optimized in vitro and in vivo, and then tested in our animal model. GDNF, CDN, and GDNF+CDN were injected into the denervated stump at the time of nerve repair. Histomorphometry and retrograde labeling were used to assess axonal regeneration. The mechanisms promoting such regeneration were explored with immunofluorescence. Five weeks after repair, the GDNF+CDN group had the highest number and maturity of axons. GDNF was noted to preferentially promote axonal maturity, whereas CDN predominantly increased the number of axons. GDNF favored motor neuron regeneration, and upregulated Ki67 in Schwann cells. CDN did not favor motor versus sensory regeneration and was noted to cleave inhibitory endoneurial proteoglycans. Early measures of nerve regeneration after delayed repair are improved by activating Schwann cells and breaking down the inhibitory proteoglycans in the distal nerve segment, suggesting a role for GDNF+CDN to be translated for human nerve repairs.


Feasibility of telaprevir-based triple therapy in liver transplant patients with hepatitis C virus: SVR 24 results.
Management of recurrent Hepatitis C virus (HCV) infection following liver transplantation remains a major challenge. In non-transplanted HCV genotype 1 patients, the introduction of protease inhibitor-based regimens has significantly increased the rate of sustained virological response. In this follow-up study, on the first published cohort of post-liver transplant patients treated with telaprevir-based triple therapy, we investigated both efficacy and safety data in follow-up to 24 weeks (SVR 24) after end of treatment (EOT). SVR 24 efficacy and safety data from 9 liver transplant HCV patients being treated with telaprevir, pegylated interferon, and ribavirin, showed 5 of the transplanted patients accomplished the full duration of the 48 week triple therapy. Notable were the 4 patients found to be HCV RNA-negative at week 4, and 8 patients at week 12. Upon EOT, at week 48, 6 patients were HCV RNA-negative. Importantly, at follow-up (24 weeks after EOT), a favorable sustained virological response rate was observed in 5 of these patients with HCV RNA remaining negative, including in one patient who discontinued treatment prematurely. Due to side effects, 2 patients discontinued, 2 suffered from virological breakthrough after the telaprevir treatment phase, and 1 patient had a relapse after EOT. Two thirds of patients exhibited hematological side effects requiring ribavirin dose reductions, administration of erythropoetin, or even blood transfusions. This retrospective analysis provides evidence that--with respect to SVR 24--liver transplant patients suffering from HCV genotype 1 recurrence may benefit from a telaprevir-based triple therapy as this new regimen showed acceptable antiviral efficacy in this small cohort of mostly pre-treated patients. Management of drug-drug interactions is challenging, but feasible. In part severe side effects are frequent during treatment and require therapeutic interventions.


Airport risk of importation and exportation of the COVID-19 pandemic.
On March 11, 2020, the Director-General of the World Health Organization (WHO) characterized the spread of the coronavirus disease, COVID-19, as a pandemic on the basis of "alarming levels of spread and severity, and by the alarming levels of inaction." Hence, it is urgent and imperative to evaluate the risk of COVID-19 for as many global locations as possible. This study calculates the relative risk of the importation and exportation of the COVID-19 virus. The study's most important contribution is the calculation of the overall relative risk of the importation and exportation of COVID-19 from every airport in local municipalities around the world, based on global spatial and mapping information. Three scenarios of air travel restriction are considered, and the change in the risk of importation and exportation of COVID-19 is calculated. The relative risk of importation and exportation of COVID-19 clearly shows that not only China, Europe, Middle East, and East Asia, but also the U.S., Australia, and countries in northeast Asia and Latin America are subject to risk. Further, a larger reduction in air travel through airports in a large part of the cumulative incidence area would lead to a gradual decrease in the risk flow. Importantly, the exportation risk of the disease from some airports in China, Iran, and European countries has a higher global spread than the importation risk during the pandemic stage. Therefore, every airport, or government with airports in their jurisdiction, should implement strict countermeasures. It is also indispensable for these countries to undertake countermeasures for COVID-19, such as home quarantine within each country and restricting infected or suspected individuals from flying on airplanes.


Expression of myelin basic protein isoforms in nonglial cells.
The myelin basic proteins (MBPs) mediate the cytoplasmic apposition of the oligodendrocyte plasma membrane to form the major dense line of central nervous system myelin. Four major isoforms of murine MBP, obtained by alternative splicing of seven exons from a single primary transcript, display distinct developmental profiles. We expressed these major MBPs individually in HeLa cells and mapped their distributions by immunofluorescence and confocal microscopy. The 14- and 18.5-kD MBPs that are the predominant forms in compact myelin distributed primarily in the perinuclear regions of the cell in configurations highly suggestive of close association with membranes. We infer that these MBP isoforms possess strong, nonspecific membrane-binding properties that have been adapted by the oligodendrocyte to mediate compaction of the sheaths of plasma membrane that form myelin. In contrast, the 17- and 21.5-kD isoforms distributed diffusely in both the cytoplasm and the nucleoplasm and often accumulated within the nucleus. This distribution can be correlated with the presence of the peptide segment encoded by exon II, which is unique to these isoforms. The physiological significance of the nuclear targeting displayed by the 17- and 21.5-kD MBP isoforms in HeLa cells remains to be determined.


Simultaneous assessment of iodine, iron, vitamin A, malarial antigenemia, and inflammation status biomarkers via a multiplex immunoassay method on a population of pregnant women from Niger.
Deficiencies of vitamin A, iron, and iodine are major public health concerns in many low- and middle-income countries, but information on their status in populations is often lacking due to high costs and logistical challenges associated with assessing micronutrient status. Accurate, user-friendly, and low-cost analytical tools are needed to allow large-scale population surveys on micronutrient status. We present the expansion of a 7-plex protein microarray tool for the simultaneous measurement of up to seven biomarkers with relevance to the assessment of the key micronutrients iron, iodine, and vitamin A, and inflammation and malaria biomarkers: α-1-acid glycoprotein, C-reactive protein, ferritin, retinol binding protein 4, soluble transferrin receptor, thyroglobulin, and histidine-rich protein II. Assay performance was assessed using international reference standards and then verified by comparing the multiplexed and conventional immunoassay results on a training panel of plasma samples collected from US adults. These data were used to assign nominal concentrations to the calibrators of the assay to further improve performance which was then assessed by interrogating plasma samples from a cohort of pregnant women from Niger. The correlation between assays for each biomarker measured from this cohort was typically good, with the exception of thyroglobulin, and the sensitivity ranged from 74% to 93%, and specificity from 81% to 98%. The 7-Plex micronutrient assay has the potential for use as an affordable tool for population surveillance of vitamin A, iron, and iodine deficiencies as well as falciparum malarial parasitemia infectivity and inflammation. The assay is easy-to-use, requires minimal sample volume, and is scalable, rapid, and accurate-needing only a low-cost reader and basic equipment present in most reference laboratory settings and so may be employed by low and middle income countries for micronutrient surveillance to inform on status in key populations. Micronutrient deficiencies including iron, iodine, and vitamin A affect a significant portion of the world's population. Efforts to assess the prevalence of these deficiencies in vulnerable populations are challenging, partly due to measurement tools that are inadequate for assessing multiple micronutrients in large-scale population surveys. We have developed a 7-plex immunoassay for the simultaneous measurement of seven biomarkers relevant to assessing iodine, iron, and vitamin A status, inflammation and Plasmodium falciparum parasitemia by measuring levels of thyroglobulin, ferritin, soluble transferrin receptor, retinol binding protein 4, α-1-acid glycoprotein, C-reactive protein, and histidine-rich protein II. This 7-plex immunoassay technique has potential as a rapid and effective tool for use in large-scale surveys and assessments of nutrition intervention programs in low- and middle-income countries.


Effects of low-intensity resistance exercise with vascular occlusion on physical function in healthy elderly people.
Some successful fall prevention programs include resistance or balance training, but less is known about the effects of low-intensity resistance exercise with moderate vascular occlusion (LIO) on physical function in healthy elderly people. In LIO, appropriate pressure is applied to the proximal parts of the upper and lower extremities with a specially designed belt. The reduction of muscle blood flow is considered likely to induce the secretion of growth hormone. The aim of this study was to compare the effects of two training programs, LIO versus dynamic balance exercise (DBE) in elderly people in a community. Fifty-one healthy subjects aged 65 and older were randomly assigned to the LIO program (n = 24) or the DBE program (n = 27). Performance, balance, muscle strength were measured in both groups before and after the 8-week programs. In addition, blood was sampled from LIO participants (n = 11) and analyzed for growth hormone and lactate. Overall improvements, but no group differences, were found in performance and balance after the programs. Muscle strength in the lower extremities was significantly increased in the LIO group, but not in the DBE group. Growth hormone was significantly increased immediately after LIO. The 8-week LIO program improved physical function, especially muscle strength, which may be associated with the exercise-induced secretion of growth hormone. Further studies are needed to determine the contents and duration of an LIO program for elderly people.