Neuropathological changes related to the transorbital application of ethyl 2-cyanoacrylate adhesive to the basal cerebral arteries of cats.
The long-term toxic effects of ethyl 2-cyanoacrylate adhesive were evaluated histologically in 25 cats. Fresh medical- or commercial-grade adhesive was introduced transorbitally into the subarachnoid space in the vicinity of the right middle cerebral artery. Three sham-operated animals served as controls. The animals were sacrificed at intervals ranging from 2 days to 6 months. For both medical- and commercial-grade adhesive, neuropathological examination disclosed acute and chronic granulomatous inflammation of the meninges and evidence of severe vascular damage, including vessel wall necrosis, inflammation, thrombosis, and occasionally hemorrhage. Most animals showed cerebral infarcts of variable size in the territories of distribution of the basal arteries which were in contact with adhesive. The results of this study show that ethyl 2-cyanoacrylate is capable of producing severe arterial and parenchymal damage. The risk of its deleterious effects should be weighed against its potential benefits. Clinical experience would suggest that ethyl 2-cyanoacrylate can be used in difficult situations as long as care is taken to protect the brain and local blood vessels.


Observation of Various and Spontaneous Magnetic Skyrmionic Bubbles at Room Temperature in a Frustrated Kagome Magnet with Uniaxial Magnetic Anisotropy.
The quest for materials hosting topologically protected skyrmionic spin textures continues to be fueled by the promise of novel devices. Although many materials have demonstrated the existence of such spin textures, major challenges remain to be addressed before devices based on magnetic skyrmions can be realized. For example, being able to create and manipulate skyrmionic spin textures at room temperature is of great importance for further technological applications because they can adapt to various external stimuli acting as information carriers in spintronic devices. Here, the first observation of skyrmionic magnetic bubbles with variable topological spin textures formed at room temperature in a frustrated kagome Fe3 Sn2 magnet with uniaxial magnetic anisotropy is reported. The magnetization dynamics are investigated using in situ Lorentz transmission electron microscopy, revealing that the transformation between different magnetic bubbles and domains is via the motion of Bloch lines driven by an applied external magnetic field. These results demonstrate that Fe3 Sn2 facilitates a unique magnetic control of topological spin textures at room temperature, making it a promising candidate for further skyrmion-based spintronic devices.


Consistent condom use is associated with lower prevalence of human papillomavirus infection in men.
Reported associations between condom use and human papillomavirus (HPV) infection in men have been inconsistent. We tested 463 men, ages 18-40 years, in 2 cities in the United States for 37 HPV types in samples from 5 anogenital sites. Men answered questionnaires regarding number of partners and frequency of condom use during vaginal sex in the past 3 months (5 categories, from "always" to "never"). Among 393 men who reported > or =1 female partner in the past 3 months, the proportions of men with HPV detected overall and at each anatomic site by frequency of condom use were calculated. Logistic regression was used to examine associations between frequency of condom use and HPV detection. Effect modification by number of recent partners (1 vs >1) was evaluated. The proportion of men positive for HPV ranged from 37.9% among men who reported they "always" used condoms to 53.9% among those who reported they "never" used condoms (P for trend = .008). Always using condoms (vs using them less frequently) was associated with lower odds of HPV detection (adjusted odds ratio, 0.50 [95% confidence interval, 0.30-0.83]). This association was stronger among men with >1 partner than among men with only 1 partner (P for interaction = .05). Consistent condom use was strongly associated with lower HPV prevalence in men.


Detraining reverses positive effects of exercise on the musculoskeletal system in premenopausal women.
We studied the effects of a 6-month withdrawal of exercise after 12 months of progressive impact (jump) plus lower body resistance training on risk factors for hip fracture in premenopausal women (age, 30-45 years). Twenty-nine women completed the 12-month training and detraining programs and were compared with 22 matched controls. Bone mineral density (BMD) at the greater trochanter, femoral neck, lumbar spine, and whole body and body composition (% body fat) were measured by dual energy X-ray absorptiometry (DXA; Hologic QDR-1000/W). Knee extensor and hip abductor strength were assessed via isokinetic dynamometry (Kin-Com 500H); maximum leg power was tested using a Wingate Anaerobic Power test; and dynamic postural stability was measured on a stabilimeter (Biodex). All measurements were conducted at baseline, 12 months and 18 months with an additional midtraining measurement of BMD. Exercisers trained three times per week in a program of 100 jumps and 100 repetitions of resistance exercises at each session. Intensity was increased using weighted vests to final values of 10% and 13% of body weight (BW) for jump and resistance exercises, respectively. Differences between groups from training were analyzed by repeated measures analysis of covariance (ANCOVA), adjusted for baseline values. Detraining effects were analyzed by comparing the changes from training with the changes from detraining using repeated measures analysis of variance (ANOVA). Baseline values were not significantly different between exercisers and controls. Percent change over the training period was significantly greater in the exercise group than in the control group at the greater trochanter (2.7 +/- 2.5% vs. 0.8 +/- 0.8%, respectively; p < 0.01) and approached significance at the femoral neck (1.2 +/- 3.2% vs. -0.3 +/- 1.9%, respectively; p = 0.06). Significant improvements also were observed in exercisers versus controls for strength and power with exercisers increasing 13-15% above controls, whereas stability was not different between groups. After 6 months of detraining, BMD and muscle strength and power decreased significantly toward baseline values, whereas control values did not change. We conclude that the positive benefits of impact plus resistance training on the musculoskeletal system in premenopausal women reverse when training is withdrawn. Therefore, continued training, perhaps at a reduced frequency and intensity, is required to maintain the musculoskeletal benefit from exercise that may lower fracture risk in later life.


Cadmium desorption in sand.
Desorption of cadmium (Cd) from sand was studied by both batch and flow-through methods. Batch experiments were conducted at three pH values (5.5, 6.0 and 6.5). In each case, the amount of Cd desorbed was low compared with the quantity of Cd adsorbed previously. Desorption of Cd in the batch experiments can be described adequately by a Freundlich isotherm. The Freundlich isotherm coefficient, Kf, increased with pH. Hysteresis between the sorption/desorption isotherms was observed in all batch experiments. Flow-through experiments in soil columns were conducted for the same three pH values, with the results used to determine transport and sorption/desorption parameters. Again, the desorption isotherms bore little resemblance to the corresponding adsorption isotherms. The experimental breakthrough curves were well fitted by a nonequilibrium desorption model, however the time scale of the desorption process was much larger than measured in batch experiments. This model was therefore rejected as lacking realism. A simple linear retardation (including hysteresis) model that utilises different isotherms was found to simulate column breakthrough curves well. The Freundlich isotherm coefficients, Kf, in all batch and flow-through desorption experiments were different to values evaluated from the corresponding adsorption experiments. However, in contrast to adsorption, desorption in flow-through experiments was not noticeably affected by changes in pH. The effect of pore-water velocity on desorption was also studied at pH 6.0. No trend was established between flow velocity and the desorption coefficient.


Doctor-patient communication: a review.
Effective doctor-patient communication is a central clinical function in building a therapeutic doctor-patient relationship, which is the heart and art of medicine. This is important in the delivery of high-quality health care. Much patient dissatisfaction and many complaints are due to breakdown in the doctor-patient relationship. However, many doctors tend to overestimate their ability in communication. Over the years, much has been published in the literature on this important topic. We review the literature on doctor-patient communication.


Learning dictionaries of sparse codes of 3D movements of body joints for real-time human activity understanding.
Real-time human activity recognition is essential for human-robot interactions for assisted healthy independent living. Most previous work in this area is performed on traditional two-dimensional (2D) videos and both global and local methods have been used. Since 2D videos are sensitive to changes of lighting condition, view angle, and scale, researchers begun to explore applications of 3D information in human activity understanding in recently years. Unfortunately, features that work well on 2D videos usually don't perform well on 3D videos and there is no consensus on what 3D features should be used. Here we propose a model of human activity recognition based on 3D movements of body joints. Our method has three steps, learning dictionaries of sparse codes of 3D movements of joints, sparse coding, and classification. In the first step, space-time volumes of 3D movements of body joints are obtained via dense sampling and independent component analysis is then performed to construct a dictionary of sparse codes for each activity. In the second step, the space-time volumes are projected to the dictionaries and a set of sparse histograms of the projection coefficients are constructed as feature representations of the activities. Finally, the sparse histograms are used as inputs to a support vector machine to recognize human activities. We tested this model on three databases of human activities and found that it outperforms the state-of-the-art algorithms. Thus, this model can be used for real-time human activity recognition in many applications.


National assessment of business-of-medicine training and its implications for the development of a business-of-medicine curriculum.
The objectives were, first, to determine the current state of business training in otolaryngology residency programs in the United States and, second, to lay the groundwork for development of a business-of-medicine (BOM) curriculum. Cross-sectional survey. A survey concerning methodology and topics for management training of residents was mailed to the chairpersons or program directors of the 102 otolaryngology residency programs. A similar survey was sent to 576 otolaryngology graduates (classes of 2000, 2001, and 2002). An interactive BOM curriculum on CD-ROM was developed based on the results. The response rate among program directors was 74.5% (76 of 102), and among the otolaryngology graduates, 38.2% (220 of 575). Seventy-five percent of graduates rated their BOM training as poor or fair. Only 8% rated their BOM training as excellent. Twenty percent of the graduates responded to having a BOM course during residency. Recent graduates reported that a BOM course can best be taught through lectures and apprenticeship/mentoring, whereas program directors reported that a BOM course can best be taught through lectures and outside consultants. Graduates reported that coding compliance was the topic most neglected in residency, whereas program directors reported that coding compliance was the main topic covered in the business training. Both groups agreed that department attending physicians have the most impact on a resident's business training. Program directors reported that correct coding, planning one's entry into medical practice, risk management, and reimbursement issues are the most important topics for residents to learn, whereas recent graduates stated that the most important topics should be correct coding, office management, risk management, and reimbursement. The present study reflects a perceived necessity for improvement of BOM training in otolaryngology residency programs. Based on this finding, the outcome measures from the survey, and the authors' own experience from business courses given in the first author's department, a BOM curriculum was developed that is general enough to target all otolaryngology residents and intended to provide business skills which result in improved use of resources and, ultimately, higher quality of care.


Detection of Kaposi's sarcoma-associated herpesvirus in oral and genital secretions of Zimbabwean women.
Kaposi's sarcoma-associated herpesvirus (KSHV) in oral and genital secretions of women may be involved in horizontal and vertical transmission in endemic regions. Nested polymerase chain reaction assays were used to detect KSHV DNA sequences in one-third of oral, vaginal, and cervical specimens and in 42% of peripheral blood mononuclear cell (PBMC) specimens collected from 41 women infected with human immunodeficiency virus type 1 who had Kaposi's sarcoma (KS). KSHV DNA was not detected in specimens from 100 women without KS, 9 of whom were seropositive for KSHV. A positive association was observed between KSHV DNA detection in oral and genital mucosa, neither of which was associated with KSHV DNA detection in PBMC. These data suggest that KSHV replicates in preferred anatomic sites at levels independent of PBMC viremia. Detection of genital-tract KSHV only among relatively immunosuppressed women may provide an explanation for infrequent perinatal transmission of KSHV.


Accurate visual memory of colors in controlling the pecking behavior of quail chicks.
Animals are predisposed to memorize specific features of objects they encounter, and to link them with behavioral outputs in a selective manner. In this study, we examined whether chicks memorize objects by colors, and how they exploit the memorized color cues for selective pecking in 1- to 2-days-old quail chicks (Coturnix japonica). Ball-shaped beads painted in green (G), yellowish green (YG) and the intermediate color (YGG) were used. Repetitive presentation of a bead (interval: 4.5 min) resulted in gradually fewer pecks (habituation). Subsequent presentation of a different color caused proportionately more pecks (dishabituation); e.g., after habituation to the G bead, the YG bead caused a stronger dishabituation than the YGG bead did. The dishabituation appeared symmetric; e.g., the YG bead caused as strong dishabituation after the G-habituation, as was caused by the G bead after the YG-habituation. Number of pecks could thus reveal the memory-based color perception in chicks. Similar discrimination of beads by memorized color cues was found after one-trial passive avoidance training, where chicks learned to avoid a bitter-tasting object without any differential pre-training experiences. However, proportion of the chicks that discriminated between different colors became progressively smaller at test 15 min, 1 hr, and 24 hr post-training. On the other hand, proportion of chicks that distinguished beads by non-color cues remained unchanged. Chicks may primarily form an accurate memory of colors, but gradually change the link between the color memory and the pecking behavior.


Disentangling the relative effects of bushmeat availability on human nutrition in central Africa.
We studied links between human malnutrition and wild meat availability within the Rainforest Biotic Zone in central Africa. We distinguished two distinct hunted mammalian diversity distributions, one in the rainforest areas (Deep Rainforest Diversity, DRD) containing taxa of lower hunting sustainability, the other in the northern rainforest-savanna mosaic, with species of greater hunting potential (Marginal Rainforest Diversity, MRD). Wild meat availability, assessed by standing crop mammalian biomass, was greater in MRD than in DRD areas. Predicted bushmeat extraction was also higher in MRD areas. Despite this, stunting of children, a measure of human malnutrition, was greater in MRD areas. Structural equation modeling identified that, in MRD areas, mammal diversity fell away from urban areas, but proximity to these positively influenced higher stunting incidence. In DRD areas, remoteness and distance from dense human settlements and infrastructures explained lower stunting levels. Moreover, stunting was higher away from protected areas. Our results suggest that in MRD areas, forest wildlife rational use for better human nutrition is possible. By contrast, the relatively low human populations in DRD areas currently offer abundant opportunities for the continued protection of more vulnerable mammals and allow dietary needs of local populations to be met.


Relationship of Modic Changes, Disk Herniation Morphology, and Axial Location to Outcomes in Symptomatic Cervical Disk Herniation Patients Treated With High-Velocity, Low-Amplitude Spinal Manipulation: A Prospective Study.
The purpose of this study was to evaluate whether cervical disk herniation (CDH) location, morphology, or Modic changes (MCs) are related to treatment outcomes. Magnetic resonance imaging (MRI) and outcome data from 44 patients with CDH treated with spinal manipulative therapy were evaluated. MRI scans were assessed for CDH axial location, morphology, and MCs. Pain (0-10 for neck and arm) and Neck Disability Index (NDI) data were collected at baseline; 2 weeks; 1, 3, and 6 months; and 1 year. The Patient's Global Impression of Change data were collected at all time points and dichotomized into "improved," yes or no. Fischer's exact test compared the proportion improved with MRI abnormalities. Numerical rating scale and NDI scores were compared with MRI abnormalities at baseline and change scores at all time points using the t test or Mann-Whitney U test. Patients who were Modic positive had higher baseline NDI scores (P = .02); 77.8% of patients who were Modic positive and 53.3% of patients who were Modic negative reported improvement at 2 weeks (P = .21). Fifty percent of Modic I and 83.3% of Modic II patients were improved at 2 weeks (P = .07). At 3 months and 1 year, all patients with MCs were improved. Patients who were Modic positive had higher NRS and NDI change scores. Patients with central herniations were more likely to improve only at the 2-week time point (P = .022). Although patients who were Modic positive had higher baseline NDI scores, the proportion of these patients improved was higher for all time points up to 6 months. Patients with Modic I changes did worse than patients with Modic II changes at only 2 weeks.


HCV transmission in industrialized countries and resource-constrained areas.
HCV is a blood-borne virus transmitted by percutaneous exposure to infected blood or blood-derived body fluids. The main routes of transmission are blood transfusions, medical procedures and injection drug use. In industrialized countries, HCV transmission through blood transfusions has been virtually eliminated and iatrogenic transmission occurs only sporadically during local breaches of infection control procedures. As most new cases originate from injection drug use, harm-reduction programmes (including opiate substitution, needle exchange and health education) can greatly reduce HCV transmission. Currently, the main approach to reduce the HCV disease burden is by increasing awareness of both the public and health-care providers to hepatitis C, enhancing screening opportunities and treatment of the infected population. In resource-limited countries, the priority is reducing transmission through blood transfusions and invasive medical procedures. This approach requires training of health-care providers and also structural changes and financial investments in countries where antibody screening, disposable materials and effective sterilization procedures are not routinely available. In these countries, reducing the HCV burden has been hampered by limited access to treatment, largely owing to the cost of drugs. Access to treatment is moving up on the agenda of international and non-governmental organizations in conjunction with the future availability of highly efficacious oral drug regimens.


Is There a Correlation Between Static and Dynamic Postural Balance Among Young Male and Female Dancers?
Aimed to investigate whether young male and female dancers have different patterns of association between static and dynamic postural balance (PB), 60 dancers from the Australian Ballet School (14-19 years old) were tested for static and dynamic PB with head and lumbar accelerometers. Monotonic relationships between static and dynamic PB were found in head movements among young female dancers in all three directions, but were found for young male dancers in the mediolateral (ML) and anteroposterior (AP) directions only. In lumbar movements, monotonic relationships were found for young female dancers in the AP direction only. Comparing head with lumbar movements in static PB, young male dancers demonstrated monotonic relationships between head and lumbar movements in all 3 directions; however, young female dancers demonstrated monotonic relationships in the AP direction only. In the dynamic measurements, both male and female dancers demonstrated monotonic relationships between head and lumbar movements for all parameters measured in the ML and vertical directions (p < .05). In conclusions, among female dancers static PB ability is correlated with their dynamic ability, whereas among male dancers, no relationship between the static and dynamic PB in the AP direction exists. Male dancers showed head and lumbar coordination in the static PB movement, but both genders manifested no head and lumbar coordination in the AP direction measured for dynamic PB.


Human bladder cancer invasion model using rat bladder in vitro and its use to test mechanisms and therapeutic inhibitors of invasion.
As well as being a passive support, the extracellular matrix also regulates key biological processes such as invasion, differentiation and angiogenesis. We have therefore developed an in vitro model of bladder cancer invasion using de-epithelialized rat bladder to allow for tumour cell-extracellular matrix interactions. Onto this we have seeded a panel of human bladder cancer cell lines (RT4, RT112, 253J and EJ28 (T24)) representing progression from well to poorly differentiated phenotypes and used as models of superficial to invasive bladder cancer. The better differentiated cell lines RT4 and RT112 reproducibly grew as stratified epithelium, whereas poorly differentiated EJ28 cells invaded across a broad front. Invasion was not simply related to proliferation rate, measured either as doubling time on plastic (non-invasive 253J and invasive EJ28 having the same doubling time) or by Ki-67 proliferation index within the model. We used the model to test the ability of 4 compounds that interfere with tumour cell-extracellular matrix interactions (suramin, N-acetylcysteine and the urokinase plasminogen activator pathway antagonists A5 compound and monoclonal antibody Mab 3936) to inhibit invasion. At non-toxic concentrations, all significantly inhibited invasion (P< 0.05), although to varying degrees, suramin and A5 almost completely and N-acetylcysteine the least. In conclusion, this model shows the urokinase system is important for bladder invasion and can be used to investigate other mechanisms of bladder cancer invasion and also for the testing of intravesical drugs.


Improving antithrombotic management in patients with atrial fibrillation: current status and perspectives.
Despite overwhelming evidence of the benefits of risk-adjusted oral anticoagulation on stroke reduction in patients with atrial fibrillation (AF), there is still considerable undertreatment. A multidisciplinary expert group was formed to discuss issues surrounding anticoagulant treatment of patients with AF to try and achieve consensus on various aspects of the implementation of guidelines on oral anticoagulation therapy in AF. Panel members were cardiologists, hematologists, and laboratory and primary care physicians with specific expertise from Europe and the United States. One of the most important conclusions of the meeting was to enhance guideline adherence by better communication of the data showing that the benefits of stroke reduction outweigh the risk of bleeding associated with treatment with vitamin K antagonists. Management of oral anticoagulation therapy by dedicated centers, such as anticoagulation clinics, or by patient self-management may improve the quality of anticoagulation and facilitate the management of these patients and thereby further facilitate optimal antithrombotic management in patients with AF.


Long-term noise exposure and the risk for type 2 diabetes: a meta-analysis.
Diabetes mellitus is one of the leading causes for disability and mortality in modern societies. Apart from personal factors its incidence might be influenced by environmental risks such as air pollution and noise. This paper reports a systematic review and meta-analysis on the risk for type 2 diabetes due to long-term noise exposure. Electronic searches in MEDLINE, EMBASE and the Internet yielded 9 relevant studies (5 for residential and 4 for occupational exposure). They were checked against a predefined list of safeguards against bias producing individual quality scores, which were then fed to MetaXL to conduct a quality effects meta-analysis. People exposed at their homes to roughly L(den) > 60 dB had 22% higher risk (95% confidence interval [CI]: 1.09-1.37) for type 2 diabetes in comparison to those exposed to L(den) < 64 dB; when studies reporting contentious exposure categories were excluded, there was still 19% risk (95% CI: 1.05-1.35) for L(den) = 60-70 dB versus L(den) < 60 dB. In occupational environment there was not significant risk (relative risk [RR] = 0.91, 95% CI: 0.78-1.06) for < 85 dB versus >85 dB. There was no heterogeneity in the two groups (I² = 0.00). The results should be interpreted with caution due to methodological discrepancies across the studies; however, they are indicative of the close links that noise pollution might have not only to cardiovascular diseases but to endocrine dysfunction as well.


Unraveling the Molecular Structures of Asphaltenes by Atomic Force Microscopy.
Petroleum is one of the most precious and complex molecular mixtures existing. Because of its chemical complexity, the solid component of crude oil, the asphaltenes, poses an exceptional challenge for structure analysis, with tremendous economic relevance. Here, we combine atomic-resolution imaging using atomic force microscopy and molecular orbital imaging using scanning tunnelling microscopy to study more than 100 asphaltene molecules. The complexity and range of asphaltene polycyclic aromatic hydrocarbons are established in detail. Identifying molecular structures provides a foundation to understand all aspects of petroleum science from colloidal structure and interfacial interactions to petroleum thermodynamics, enabling a first-principles approach to optimize resource utilization. Particularly, the findings contribute to a long-standing debate about asphaltene molecular architecture. Our technique constitutes a paradigm shift for the analysis of complex molecular mixtures, with possible applications in molecular electronics, organic light emitting diodes, and photovoltaic devices.


Countercurrent tangential chromatography for large-scale protein purification.
Recent advances in cell culture technology have created significant pressure on the downstream purification process, leading to a "downstream bottleneck" in the production of recombinant therapeutic proteins for the treatment of cancer, genetic disorders, and cardiovascular disease. Countercurrent tangential chromatography overcomes many of the limitations of conventional column chromatography by having the resin (in the form of a slurry) flow through a series of static mixers and hollow fiber membrane modules. The buffers used in the binding, washing, and elution steps flow countercurrent to the resin, enabling high-resolution separations while reducing the amount of buffer needed for protein purification. The results obtained in this study provide the first experimental demonstration of the feasibility of using countercurrent tangential chromatography for the separation of a model protein mixture containing bovine serum albumin and myoglobin using a commercially available anion exchange resin. Batch uptake/desorption experiments were used in combination with critical flux data for the hollow fiber filters to design the countercurrent tangential chromatography system. A two-stage batch separation yielded the purified target protein at >99% purity with 94% recovery. The results clearly demonstrate the potential of using countercurrent tangential chromatography for the large-scale purification of therapeutic proteins.


Reliability of ultrasound to measure morphology of the toe flexor muscles.
Measuring the strength of individual foot muscles is very challenging; however, measuring muscle morphology has been shown to be associated with strength. A reliable method of assessing foot muscle atrophy and hypertrophy would therefore be beneficial to researchers and clinicians. Thus, the aim of this study was to evaluate the test-retest intra-observer reliability of ultrasound to measure the morphology of the primary toe flexor muscles. The abductor hallucis, flexor hallucis brevis, flexor digitorum brevis, quadratus plantae and abductor digiti minimi muscles in the foot, and the flexor digitorum longus and flexor hallucis longus muscles in the shank were assessed in five males and five females (mean age = 32.1 ± 10.1 years). Muscles were imaged using a GE Venue 40 ultrasound (6-9 or 7.6-10.7 MHz transducer) in a random order, and on two occasions 1-6 days apart. Muscle thickness and cross-sectional area were measured using Image J software with the assessor blinded to muscle and day of scan. Intraclass correlation coefficients (ICC) and limits of agreement were calculated to assess day-to-day repeatability of the measurements. The method was found to have good reliability (ICC = 0.89-0.99) with limits of agreement between 8-28% of the relative muscle size. The protocol described in this paper showed that ultrasound is a reliable method to measure morphology of the toe flexor muscles. The portability and advantages of ultrasound make it a useful tool for clinical and research settings.


Real-time gray-scale photolithography for fabrication of continuous microstructure.
A novel real-time gray-scale photolithography technique for the fabrication of continuous microstructures that uses a LCD panel as a real-time gray-scale mask is presented. The principle of design of the technique is explained, and computer simulation results based on partially coherent imaging theory are given for the patterning of a microlens array and a zigzag grating. An experiment is set up, and a microlens array and a zigzag grating on panchromatic silver halide sensitized gelatin with trypsinase etching are obtained.


A multicopy Y-chromosomal SGNH hydrolase gene expressed in the testis of the platyfish has been captured and mobilized by a Helitron transposon.
Teleost fish present a high diversity of sex determination systems, with possible frequent evolutionary turnover of sex chromosomes and sex-determining genes. In order to identify genes involved in male sex determination and differentiation in the platyfish Xiphophorus maculatus, bacterial artificial chromosome contigs from the sex-determining region differentiating the Y from the X chromosome have been assembled and analyzed. A novel three-copy gene called teximY (for testis-expressed in Xiphophorus maculatus on the Y) was identified on the Y but not on the X chromosome. A highly related sequence called texim1, probably at the origin of the Y-linked genes, as well as three more divergent texim genes were detected in (pseudo)autosomal regions of the platyfish genome. Texim genes, for which no functional data are available so far in any organism, encode predicted esterases/lipases with a SGNH hydrolase domain. Texim proteins are related to proteins from very different origins, including proteins encoded by animal CR1 retrotransposons, animal platelet-activating factor acetylhydrolases (PAFah) and bacterial hydrolases. Texim gene distribution is patchy in animals. Texim sequences were detected in several fish species including killifish, medaka, pufferfish, sea bass, cod and gar, but not in zebrafish. Texim-like genes are also present in Oikopleura (urochordate), Amphioxus (cephalochordate) and sea urchin (echinoderm) but absent from mammals and other tetrapods. Interestingly, texim genes are associated with a Helitron transposon in different fish species but not in urochordates, cephalochordates and echinoderms, suggesting capture and mobilization of an ancestral texim gene in the bony fish lineage. RT-qPCR analyses showed that Y-linked teximY genes are preferentially expressed in testis, with expression at late stages of spermatogenesis (late spermatids and spermatozeugmata). These observations suggest either that TeximY proteins play a role in Helitron transposition in the male germ line in fish, or that texim genes are spermatogenesis genes mobilized and spread by transposable elements in fish genomes.


Dynamic contact angle in rim instability of dewetting holes.
The effects of dynamic contact angle (thetad), between a substrate and the melt of a dewetting polymer thin film, on the evolution of rim instabilities of dewetting holes were reported. Various thetad's were achieved by covering SiOx surfaces with different coverage of octadecyltrichlorosilane. On each surface, the morphology of the dewetting holes was examined in detail as the hole grew to a certain size. Rim instabilities, in terms of undulations in both r and z directions, became more pronounced as thetad increased, under which condition, narrower and higher rims were also observed. Experimentally, atomic force microscopic scans of the rim were used to obtain the rim profile, which was predicted using thetad. The predicted rim profile was used, in combination with the analysis of Rayleigh instability of a cylindrical fluid, to interpret the rim instability. The model captures the basic trend of the rim instability dependency on thetad. The study demonstrates the importance of the substrate properties on the rim instability and the destabilization of polymer thin films during hole growth.


Transcriptome analyses reveal genotype- and developmental stage-specific molecular responses to drought and salinity stresses in chickpea.
Drought and salinity are the major factors that limit chickpea production worldwide. We performed whole transcriptome analyses of chickpea genotypes to investigate the molecular basis of drought and salinity stress response/adaptation. Phenotypic analyses confirmed the contrasting responses of the chickpea genotypes to drought or salinity stress. RNA-seq of the roots of drought and salinity related genotypes was carried out under control and stress conditions at vegetative and/or reproductive stages. Comparative analysis of the transcriptomes revealed divergent gene expression in the chickpea genotypes at different developmental stages. We identified a total of 4954 and 5545 genes exclusively regulated in drought-tolerant and salinity-tolerant genotypes, respectively. A significant fraction (~47%) of the transcription factor encoding genes showed differential expression under stress. The key enzymes involved in metabolic pathways, such as carbohydrate metabolism, photosynthesis, lipid metabolism, generation of precursor metabolites/energy, protein modification, redox homeostasis and cell wall component biogenesis, were affected by drought and/or salinity stresses. Interestingly, transcript isoforms showed expression specificity across the chickpea genotypes and/or developmental stages as illustrated by the AP2-EREBP family members. Our findings provide insights into the transcriptome dynamics and components of regulatory network associated with drought and salinity stress responses in chickpea.


Novel Munc13-4 mutations in children and young adult patients with haemophagocytic lymphohistiocytosis.
Familial haemophagocytic lymphohistiocytosis (FHL) is a genetically heterogeneous disorder characterised by constitutive defects in cellular cytotoxicity resulting in fever, hepatosplenomegaly and cytopenia, and the outcome is fatal unless treated by chemoimmunotherapy followed by haematopoietic stem-cell transplantation. Since 1999, mutations in the perforin gene giving rise to this disease have been identified; however, these account only for 40% of cases. Lack of a genetic marker hampers the diagnosis, suitability for transplantation, selection of familial donors, identification of carriers, genetic counselling and prenatal diagnosis. Mutations in the Munc13-4 gene have recently been described in patients with FHL. We sequenced the Munc13-4 gene in all patients with haemophagocytic lymphohistiocytosis not due to PRF1 mutations. In 15 of the 30 families studied, 12 novel and 4 known Munc13-4 mutations were found, spread throughout the gene. Among novel mutations, 2650C-->T introduced a stop codon; 441del A, 532del C, 3082del C and 3226ins G caused a frameshift, and seven were mis sense mutations. Median age of diagnosis was 4 months, but six patients developed the disease after 5 years of age and one as a young adult of 18 years. Involvement of central nervous system was present in 9 of 15 patients, activity of natural killer cells was markedly reduced or absent in 13 of 13 tested patients. Chemo-immunotherapy was effective in all patients. Munc13-4 mutations were found in 15 of 30 patients with FHL without PRF1 mutations. Because these patients may develop the disease during adolescence or even later, haematologists should include FHL2 and FHL3 in the differential diagnosis of young adults with fever, cytopenia, splenomegaly and hypercytokinaemia.


Preeclampsia and long-term risk of cardiovascular disease: what do obstetrician-gynecologists know?
Preeclampsia (PE), a hypertensive disorder of pregnancy affects 2-8% of women and is associated with increased cardiovascular disease (CVD) risk later in life. There is little information about the knowledge of obstetrician-gynecologists in German outpatient care setting regarding the future health risk of PE and knowledge of the current guidelines on treatment and counseling patients post PE. This study aimed to assess whether obstetrician-gynecologists are aware of PE's association with maternal long-term adverse outcomes and providing appropriate counseling. A random sample of 500 obstetrician-gynecologists in the federal state of Lower Saxony was mailed a survey and a reminder with a second copy of the survey. The questionnaire elicited both personal information, and knowledge on future disease risks, e.g. cardiovascular disease (CVD) and current guidelines as well as on counseling practice. Descriptive analysis was used to analyze the responses. A total of 212 obstetrician-gynecologists (42.4%) responded to the questionnaire. A large proportion of physicians stated that PE was associated with a higher risk for the development for hypertension (86.6%), stroke (78.5%) and kidney disease (78.0%). Of the participants 75.8% reported that women after PE have a shorter life expectancy. Respondents with knowledge of the current guidelines of the German Association of Obstetrics and Gynecology concerning follow up and risk management of PE (45.2%) were more often aware of the development of CVD and stroke and counseled patients on self -blood-pressure measurement, meaning and long-term-risks of PE and attached importance to family history of PE compared to physicians with no knowledge of the guidelines. Although the majority of obstetrician-gynecologists were aware of higher CVD risk after PE, weaknesses exist in the follow up care and counseling of these patients. These deficiencies would be amendable to directed educational activities to improve the implementation of current guidelines.


[ANMCO/SICP/SIGO Consensus document: Pregnancy and congenital heart disease].
The success of cardiac surgery over the past 50 years has increased numbers and median age of survivors with congenital heart disease (CHD). Adults now represent two-thirds of patients with CHD; in the United States alone the number is estimated to exceed 1 million.In this population many affected women reach reproductive age and wish to have children. While in many CHD patients pregnancy can be accomplished successfully, some special situations with complex anatomy, iatrogenic or residual pathology are associated with an increased risk of severe maternal and fetal complications. Pre-conception counseling allows women to come to truly informed choices. Risk stratification tools can also help high-risk women to eventually renounce to pregnancy and to adopt safe contraception options. Once pregnant, women identified as intermediate or high-risk should receive multidisciplinary care involving a cardiologist, an obstetrician and an anesthesiologist with specific expertise in managing this peculiar medical challenge.This document is intended to provide cardiologists working in hospitals where an Obstetrics and Gynecology Department is available with a streamlined and practical tool, useful for them to select the best management strategies to deal with a woman affected by CHD who desires to plan pregnancy or is already pregnant.


Development and evaluation of a manual for extended brief intervention for alcohol misuse for adults with mild to moderate intellectual disabilities living in the community: The EBI-LD study manual.
Extended brief interventions for alcohol misuse are effective in the general population. The process of manualising the first ever such intervention for people with mild to moderate intellectual disabilities in the UK is the focus of this study. The manual was an adaptation of existing manuals based on Motivational Enhancement and Cognitive Behaviour Therapy and was used in a feasibility randomized controlled trial, the EBI-LD study. The sessions were recorded and scored using an adapted version of the Yale Adherence and Competence Scale (YACS II). Feedback was provided by therapists. The trial is closed. Registered: isrctn.com; ISRCTN58783633. The quality of the sessions provided was rated as good. Therapists were able to cover all topics within each session. Main challenges included session duration and homework task completion. We recommend the duration of the sessions to be extended to 40 min to accommodate carers in the session and to enhance their support in homework task completion.


Complicated Withdrawal Phenomena During Benzodiazepine Cessation in Older Adults.
The cause of prolonged or recurrent symptoms following the cessation of long-term benzodiazepine use is proposed to be related to downregulation and allosteric decoupling of the γ-aminobutyric acid/benzodiazepine receptor complex. This case series describes 2 patients with prolonged (>2 weeks) recurrent complications during attempted tapering of benzodiazepine doses after long-term treatment. Excited catatonia developed in a 90-year-old woman, and prolonged delirium developed in a 69-year-old woman. Both patients showed improvement of symptoms after resumption of higher doses of benzodiazepine treatment and recurrence of symptoms when the dose was again lowered. Caution should be exercised regarding the long-term use of benzodiazepines in older adults (aged ≥65 years). Tapering of benzodiazepines in older patients after long-term treatment may require slow decreases in dose over long periods. Psychotherapeutic interventions, such as brief cognitive therapy with psychoeducation and motivational enhancement, and osteopathic manipulative treatment to decrease paravertebral muscle tension may be beneficial during the tapering process.


Ebbinghaus figures that deceive the eye do not necessarily deceive the hand.
In support of the visual stream dissociation hypothesis, which states that distinct visual streams serve vision-for-perception and vision-for-action, visual size illusions were reported over 20 years ago to 'deceive the eye but not the hand'. Ever since, inconclusive results and contradictory interpretations have accumulated. Therefore, we investigated the effects of the Ebbinghaus figure on repetitive aiming movements with distinct dynamics. Participants performed a Fitts' task in which Ebbinghaus figures served as targets. We systematically varied the three parameters which have been shown to influence the perceived size of the Ebbinghaus figure's target circle, namely the size of the target, its distance to the context circles and the size of the context circles. This paper shows that movement is significantly affected by the context size, but, in contrast to perception, not by the other two parameters. This is especially prominent in the approach phase of the movement towards the target, regardless of the dynamics. To reconcile the findings, we argue that different informational variables are used for size perception and the visual control of movements irrespective of whether certain variables induce (perceptual) illusions.


The nucleus is irreversibly shaped by motion of cell boundaries in cancer and non-cancer cells.
Actomyosin stress fibers impinge on the nucleus and can exert compressive forces on it. These compressive forces have been proposed to elongate nuclei in fibroblasts, and lead to abnormally shaped nuclei in cancer cells. In these models, the elongated or flattened nuclear shape is proposed to store elastic energy. However, we found that deformed shapes of nuclei are unchanged even after removal of the cell with micro-dissection, both for smooth, elongated nuclei in fibroblasts and abnormally shaped nuclei in breast cancer cells. The lack of shape relaxation implies that the nuclear shape in spread cells does not store any elastic energy, and the cellular stresses that deform the nucleus are dissipative, not static. During cell spreading, the deviation of the nucleus from a convex shape increased in MDA-MB-231 cancer cells, but decreased in MCF-10A cells. Tracking changes of nuclear and cellular shape on micropatterned substrata revealed that fibroblast nuclei deform only during deformations in cell shape and only in the direction of nearby moving cell boundaries. We propose that motion of cell boundaries exert a stress on the nucleus, which allows the nucleus to mimic cell shape. The lack of elastic energy in the nuclear shape suggests that nuclear shape changes in cells occur at constant surface area and volume.


Focal High Intensity Focused Ultrasound of Unilateral Localized Prostate Cancer: A Prospective Multicentric Hemiablation Study of 111 Patients.
Up to a third of patients with localized prostate cancer have unilateral disease that may be suitable for partial treatment with hemiablation. To evaluate the ability of high intensity focused ultrasound (HIFU) to achieve local control of the tumor in patients with unilateral localized prostate cancer. The French Urological Association initiated a prospective IDEAL multi-institutional study (2009-2015), to evaluate HIFU-hemiablation as a primary treatment. Multiparametric magnetic resonance imaging and biopsy were used for unilateral cancer diagnosis and control, and HIFU-hemiablation. Primary: absence of clinically significant cancer (CSC) on control biopsy at 1 yr (CSC: Gleason score ≥ 7 or cancer core length>3mm regardless of grade or > 2 positive cores). Secondary: presence of any cancer on biopsy, biochemical response, radical treatment free survival, adverse events, continence (no pad), erectile function (International Index of Erectile Function-5 ≥ 16), and quality of life (European Organization for Research and Treatment of Cancer QLQ-C28) questionnaires. One hundred and eleven patients were treated (mean age: 64.8 yr [standard deviation 6.2]; mean prostate-specific antigen: 6.2 ng/ml [standard deviation 2.6]; 68% low risk, 32% intermediate risk). Of the 101 patients with control biopsy, 96 (95%) and 94 (93%) had no CSC in the treated and contralateral lobes, respectively. Mean prostate-specific antigen at 2 yr was 2.3 ng/ml (standard deviation 1.7). The radical treatment-free survival rate at 2 years was 89% (radical treatments: six radical prostatectomies, three radiotherapies, and two HIFU). Adverse events were Grade 3 in 13%. At 12 mo continence and erectile functions were preserved in 97% and 78%. No significant decrease in quality of life score was observed at 12 mo. One limitation is the number of low-risk patients included in this study. At 1 yr, HIFU-hemiablation was efficient with 95% absence of clinically significant cancer associated with low morbidity and preservation of quality of life. Radical treatment-free survival rate was 89% at 2 yr. This report shows that high intensity focused ultrasound half-gland treatment of unilateral prostate cancer provides promising results with high cancer control and low morbidity.


Prediction of venous thromboembolism using clinical and serum biomarker data from a military cohort of trauma patients.
Venous thromboembolism (VTE) is a frequent complication of trauma associated with high mortality and morbidity. Clinicians lack appropriate tools for stratifying trauma patients for VTE, thus have yet to be able to predict when to intervene. We aimed to compare random forest (RF) and logistic regression (LR) predictive modelling for VTE using (1) clinical measures alone, (2) serum biomarkers alone and (3) clinical measures plus serum biomarkers. Data were collected from 73 military casualties with at least one extremity wound and prospectively enrolled in an observational study between 2007 and 2012. Clinical and serum cytokine data were collected. Modelling was performed with RF and LR based on the presence or absence of deep vein thrombosis (DVT) and/or pulmonary embolism (PE). For comparison, LR was also performed on the final variables from the RF model. Sensitivity/specificity and area under the curve (AUC) were reported. Of the 73 patients (median Injury Severity Score=16), nine (12.3%) developed VTE, four (5.5%) with DVT, four (5.5%) with PE, and one (1.4%) with both DVT and PE. In all sets of predictive models, RF outperformed LR. The best RF model generated with clinical and serum biomarkers included five variables (interleukin-15, monokine induced by gamma, vascular endothelial growth factor, total blood products at resuscitation and presence of soft tissue injury) and had an AUC of 0.946, sensitivity of 0.992 and specificity of 0.838. VTE may be predicted by clinical and molecular biomarkers in trauma patients. This will allow the development of clinical decision support tools which can help inform the management of high-risk patients for VTE.


Stearidonic acid-enhanced soybean oil: a plant-based source of (n-3) fatty acids for foods.
Omega-3 (n-3) fatty acids have been reported to have a variety of cardiovascular and neuropsychiatric benefits. Although obtaining the preformed fatty acids EPA and DHA from their traditional source (fish) is optimal, such an approach may not be realistic for meeting the world's growing demand for (n-3) fatty acids; therefore, a more sustainable and dependable source is needed. Stearidonic acid (SDA) is a metabolic precursor of EPA that can be provided by SDA-enhanced soybean oil. Such a product can provide a sustainable source of (n-3) fatty acids that does not endanger fish stocks. Two clinical trials have demonstrated that SDA-enhanced soybean oil can significantly improve an emerging marker of cardiovascular health, the omega-3 index (RBC EPA+DHA). The increase in the Index seen in these trials was used to estimate the potential clinical benefit of SDA consumption based on prior prospective cohort studies. In this analysis, risk for sudden cardiac death and the rate of cellular aging would both theoretically be reduced. The lower risk for major cardiac events seen in the Japan EPA Lipid Intervention Study (which used EPA supplementation) suggests that raising EPA tissue levels, independent of changes in DHA, can have clinical benefit. Thus, the consumption of foods containing SDA-enhanced soybean oil may be both a practical and sustainable approach to enriching tissues with beneficial (n-3) fatty acids.


Predictors of independent living status in adult survivors of childhood cancer: a report from the Childhood Cancer Survivor Study.
Adult survivors of childhood cancer and their siblings are compared on one of the most salient developmental milestones of adulthood, the ability to live independently. Adult survivors of childhood cancers (n = 6,047) and siblings (n = 2,326), all 25 years of age and older, completed a long-term follow-up questionnaire that assessed adaptive, neurocognitive, and psychological functioning, as well as demographic and health status. Multivariable logistic regression analyses and structural equation modeling (SEM) were used to identify predictors of independent living. Compared to siblings (n = 206, 8.7%), survivors (n = 1063; 17.7%) were more than twice as likely to live dependently (OR 2.07; 95% confidence interval [CI] 1.77-2.42). Survivors diagnosed with CNS tumors (OR 0.13, 95% CI 0.10-0.18) or leukemia (OR 0.29, 95% CI 0.23-0.27) were significantly less likely to live independently compared to those diagnosed with Hodgkin lymphoma. Other risk factors for reduced independent living included cranial radiation (≤ 24 Gy OR 0.76, 95% CI 0.62-0.93; >24 Gy OR 0.31, 95% CI 0.24-0.41), use of neuroleptic, anticonvulsant, or psychostimulant medication (OR 0.32, 95% CI 0.24-0.43), attention and processing speed problems (OR 0.58, 95% CI 0.47-0.71), poor physical functioning (OR 0.49, 95% CI 0.38-0.63), depression (OR 0.68, 95% CI 0.53-0.88), and racial/ethnic minority status (OR 0.39, 95% CI 0.30-0.51). SEM demonstrated that neurocognitive functioning had both direct effects on independent living status, and indirect effects through use of neurologically directed medication, depression, and poor mental health. Adult survivors of childhood cancer who experience neurocognitive, psychological, or physical late effects are less likely to live independently as adults.


In-silico prediction of novel genes responsive to drought and salinity stress tolerance in bread wheat (Triticum aestivum).
Common wheat (Triticum aestivum) is the most widely grown cereal crop and is cultivated extensively in dry regions. Water shortage, resulting from either drought or salinity, leads to slow growth and loss of wheat yield. In order to predict new genes responsive to the drought and salt stresses in wheat, 6,717 expressed sequence tags (ESTs), expressed in drought and salinity stress conditions were collected from the National Center for Biotechnology Information (NCBI). The downloaded ESTs were clustered and assembled into 354 contigs; 14 transcription factor families in 29 contigs were identified. In addition, 119 contigs were organized in five enzyme classes. Biological functions were obtained for only 324 of the 354 contigs using gene ontology. In addition, using Kyoto Encyclopedia of Genes and Genomes database, 191 metabolic pathways were identified. The remaining contigs were used for further analysis and the search for new genes responsive to drought and salt stresses. These contigs were mapped on the International Wheat Genome Sequencing Consortium RefSeq v1.0 assembly, the most complete version of the reference sequence of the bread wheat variety Chinese Spring. They were found to have from one to three locations on the subgenomes A, B, and D. Full-length gene sequences were designed for these contigs, which were further validated using promoter analysis. These predicted genes may have applications in molecular breeding programs and wheat drought and salinity research.


Economic and epidemiological impact of early antiretroviral therapy initiation in India.
Recent WHO guidance advocates for early antiretroviral therapy (ART) initiation at higher CD4 counts to improve survival and reduce HIV transmission. We sought to quantify how the cost-effectiveness and epidemiological impact of early ART strategies in India are affected by attrition throughout the HIV care continuum. We constructed a dynamic compartmental model replicating HIV transmission, disease progression and health system engagement among Indian adults. Our model of the Indian HIV epidemic compared implementation of early ART initiation (i.e. initiation above CD4 ≥350 cells/mm(3)) with delayed initiation at CD4 ≤350 cells/mm(3); primary outcomes were incident cases, deaths, quality-adjusted-life-years (QALYs) and costs over 20 years. We assessed how costs and effects of early ART initiation were impacted by suboptimal engagement at each stage in the HIV care continuum. Assuming "idealistic" engagement in HIV care, early ART initiation is highly cost-effective ($442/QALY-gained) compared to delayed initiation at CD4 ≤350 cells/mm(3) and could reduce new HIV infections to <15,000 per year within 20 years. However, when accounting for realistic gaps in care, early ART initiation loses nearly half of potential epidemiological benefits and is less cost-effective ($530/QALY-gained). We project 1,285,000 new HIV infections and 973,000 AIDS-related deaths with deferred ART initiation with current levels of care-engagement in India. Early ART initiation in this continuum resulted in 1,050,000 new HIV infections and 883,000 AIDS-related deaths, or 18% and 9% reductions (respectively), compared to current guidelines. Strengthening HIV screening increases benefits of earlier treatment modestly (1,001,000 new infections; 22% reduction), while improving retention in care has a larger modulatory impact (676,000 new infections; 47% reduction). Early ART initiation is highly cost-effective in India but only has modest epidemiological benefits at current levels of care-engagement. Improved retention in care is needed to realize the full potential of earlier treatment.


A whole-genome screen identifies Salmonella enterica serovar Typhi genes involved in fluoroquinolone susceptibility.
A whole-genome screen at sub-gene resolution was performed to identify candidate loci that contribute to enhanced or diminished ciprofloxacin susceptibility in Salmonella enterica serovar Typhi. A pool of over 1 million transposon insertion mutants of an S. Typhi Ty2 derivative were grown in a sub-MIC concentration of ciprofloxacin, or without ciprofloxacin. Transposon-directed insertion site sequencing (TraDIS) identified relative differences between the mutants that grew following the ciprofloxacin treatment compared with the untreated mutant pool, thereby indicating which mutations contribute to gain or loss of ciprofloxacin susceptibility. Approximately 88% of the S. Typhi strain's 4895 annotated genes were assayed, and at least 116 were identified as contributing to gain or loss of ciprofloxacin susceptibility. Many of the identified genes are known to influence susceptibility to ciprofloxacin, thereby providing method validation. Genes were identified that were not known previously to be involved in susceptibility, and some of these had no previously known phenotype. Susceptibility to ciprofloxacin was enhanced by insertion mutations in genes coding for efflux, other surface-associated functions, DNA repair and expression regulation, including phoP, barA and marA. Insertion mutations that diminished susceptibility were predominantly in genes coding for surface polysaccharide biosynthesis and regulatory genes, including slyA, emrR, envZ and cpxR. A genomics approach has identified novel contributors to gain or loss of ciprofloxacin susceptibility in S. Typhi, expanding our understanding of the impact of fluoroquinolones on bacteria and of mechanisms that may contribute to resistance. The data also demonstrate the power of the TraDIS technology for antibacterial research.


Peripartum and Postpartum Analgesia and Pain in Women Prescribed Buprenorphine for Opioid Use Disorder Who Deliver by Cesarean Section.
Little is known about whether pain can be effectively managed in pregnant women with opioid use disorder (OUD) during delivery hospitalization, particularly those undergoing surgery and taking buprenorphine as medication for OUD (MOUD). To address this question, we compared pain scores and opioid analgesic utilization during delivery hospitalization in women taking their pre-hospital dose of buprenorphine who delivered by cesarean section to matched controls. To inform future research efforts, we also began to explore opioid analgesic utilization and pain scores by type of anesthesia as this variable is often not included in related literature. Retrospective matched cohort study of 46 women prescribed buprenorphine during pregnancy who delivered by cesarean section during a 7-year period. When compared to matched controls, women taking their pre-hospital dose of buprenorphine undergoing cesarean section utilized more opioid analgesics as measured by morphine milligram equivalents (MME) (mean MME first 48 hours 153.0 mg vs 175.1 mg, respectively, P < .01) but had similar pain scores during delivery hospitalization. There was no difference in MME utilization by maternal dose of buprenorphine though sample sizes were small. Women on buprenorphine who received spinal anesthesia with morphine had mean pain scores that were 1.4 points lower (P = .01) during the first 48 hours than women on buprenorphine receiving other methods of anesthesia. Pregnant women taking their pre-hospital dose of buprenorphine throughout their surgical delivery hospitalization were able to achieve pain relief similar to women not on MOUD but had higher MME requirements. Our results add to the emerging body of evidence suggesting that individuals on MOUD can achieve adequate post-surgical pain management without adjusting their pre-hospital dose of buprenorphine. Further research is required to fully understand the optimal buprenorphine dosing regimen during surgical hospitalizations. Our results also provide important preliminary evidence that spinal anesthesia containing opioids can be used effectively in individuals with OUD requiring surgical intervention.


Role of copper in regression of cardiac hypertrophy.
Pressure overload causes an accumulation of homocysteine in the heart, which is accompanied by copper depletion through the formation of copper-homocysteine complexes and the excretion of the complexes. Copper supplementation recovers cytochrome c oxidase (CCO) activity and promotes myocardial angiogenesis, along with the regression of cardiac hypertrophy and the recovery of cardiac contractile function. Increased copper availability is responsible for the recovery of CCO activity. Copper promoted expression of angiogenesis factors including vascular endothelial growth factor (VEGF) in endothelial cells is responsible for angiogenesis. VEGF receptor-2 (VEGFR-2) is critical for hypertrophic growth of cardiomyocytes and VEGFR-1 is essential for the regression of cardiomyocyte hypertrophy. Copper, through promoting VEGF production and suppressing VEGFR-2, switches the VEGF signaling pathway from VEGFR-2-dependent to VEGFR-1-dependent, leading to the regression of cardiomyocyte hypertrophy. Copper is also required for hypoxia-inducible factor-1 (HIF-1) transcriptional activity, acting on the interaction between HIF-1 and the hypoxia responsible element and the formation of HIF-1 transcriptional complex by inhibiting the factor inhibiting HIF-1. Therefore, therapeutic targets for copper supplementation-induced regression of cardiac hypertrophy include: (1) the recovery of copper availability for CCO and other critical cellular events; (2) the activation of HIF-1 transcriptional complex leading to the promotion of angiogenesis in the endothelial cells by VEGF and other factors; (3) the activation of VEGFR-1-dependent regression signaling pathway in the cardiomyocytes; and (4) the inhibition of VEGFR-2 through post-translational regulation in the hypertrophic cardiomyocytes. Future studies should focus on target-specific delivery of copper for the development of clinical application.


A novel function for UDP glycosyltransferase 8: galactosidation of bile acids.
The human UDP glycosyltransferase (UGT) superfamily comprises four families of enzymes that catalyze the addition of sugar residues to small lipophilic chemicals. The UGT1 and UGT2 enzymes use UDP-glucuronic acid, and UGT3 enzymes use UDP-N-acetylglucosamine, UDP-glucose, and UDP-xylose to conjugate xenobiotics, including drugs and endobiotics such as metabolic byproducts, hormones, and signaling molecules. This metabolism renders the substrate more polar and more readily excreted from the body and/or functionally inactive. The fourth UGT family, called UGT8, contains only one member that, unlike other UGTs, is considered biosynthetic. UGT8 uses UDP galactose to galactosidate ceramide, a key step in the synthesis of brain sphingolipids. To date other substrates for this UGT have not been identified and there has been no suggestion that UGT8 is involved in metabolism of endo- or xenobiotics. We re-examined the functions of UGT8 and discovered that it efficiently galactosidates bile acids and drug-like bile acid analogs. UGT8 conjugates bile acids ∼60-fold more efficiently than ceramide based on in vitro assays with substrate preference deoxycholic acid > chenodeoxycholic acid > cholic acid > hyodeoxycholic acid > ursodeoxycholic acid. Activities of human and mouse UGT8 are qualitatively similar. UGT8 is expressed at significant levels in kidney and gastrointestinal tract (intestine, colon) where conjugation of bile acids is likely to be metabolically significant. We also investigate the structural determinants of UDP-galactose selectivity. Our novel findings suggest a new role for UGT8 as a modulator of bile acid homeostasis and signaling.


How should we grow cities to minimize their biodiversity impacts?
Urbanization causes severe environmental degradation and continues to increase in scale and intensity around the world, but little is known about how we should design cities to minimize their ecological impact. With a sprawling style of urban development, low intensity impact is spread across a wide area, and with a compact form of development intense impact is concentrated over a small area; it remains unclear which of these development styles has a lower overall ecological impact. Here, we compare the consequences of compact and sprawling urban growth patterns on bird distributions across the city of Brisbane, Australia. We predicted the impact on bird populations of adding 84,642 houses to the city in either a compact or sprawling design using statistical models of bird distributions. We show that urban growth of any type reduces bird distributions overall, but compact development substantially slows these reductions at the city scale. Urban-sensitive species particularly benefited from compact development at the city scale because large green spaces were left intact, whereas the distributions of nonnative species expanded as a result of sprawling development. As well as minimizing ecological disruption, compact urban development maintains human access to public green spaces. However, backyards are smaller, which impacts opportunities for people to experience nature close to home. Our results suggest that cities built to minimize per capita ecological impact are characterized by high residential density, with large interstitial green spaces and small backyards, and that there are important trade-offs between maintaining city-wide species diversity and people's access to biodiversity in their own backyard.


Terminal change across facets of affective experience and domain satisfaction: Commonalities, differences, and bittersweet emotions at the end of life.
General well-being is known to deteriorate sharply at the end of life. However, it is an open question how rates of terminal change differ across affective and evaluative facets of well-being and if individual difference correlates operate in facet-specific ways. We examined how discrete affective states (happy, angry, fearful, sad) and satisfaction with key life domains (health, leisure, family) change as people approach death and how differences in end-of-life trajectories are related to sociodemographic (age, gender, education), physical health (disability, body mass index, physician visits), and psychosocial characteristics (perceived control, social orientation, living with a partner). We applied growth models to 9-year annual longitudinal data of 864 participants (age at death: M = 75 years, 41% women) from the nationwide German Socio-Economic Panel (SOEP). Findings revealed commonalities and specificities in terminal change: Six of seven facets became increasingly fragile late in life (6 to 35 times steeper terminal change than age change), but at vastly different rates of change (e.g., steep declines in happiness and satisfaction with health vs. stability in anger) and at different levels at which changes occurred. Commonalities and differences also emerged for the correlates: Those who perceived more control over their lives experienced generally more favorable late-life affect and satisfaction trajectories, whereas other correlates operated in more facet-specific ways. For example, participants living with a partner were happier and more satisfied with family life throughout their last years, but also reported more fear and steeper increases in sadness, a picture of bittersweet emotions at the end of life. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Bioinformatic Atlas of Radical SAM Enzyme-Modified RiPP Natural Products Reveals an Isoleucine-Tryptophan Crosslink.
Ribosomally synthesized and post-translationally modified peptides (RiPPs) are a growing family of natural products with diverse activities and structures. RiPP classes are defined by the tailoring enzyme, which can introduce a narrow range of modifications or a diverse set of alterations. In the latter category, RiPPs synthesized by radical S-adenosylmethionine (SAM) enzymes, known as RaS-RiPPs, have emerged as especially divergent. A map of all RaS-RiPP gene clusters does not yet exist. Moreover, precursor peptides remain difficult to predict using computational methods. Herein, we have addressed these challenges and report a bioinformatic atlas of RaS-RiPP gene clusters in available microbial genome sequences. Using co-occurrence of RaS enzymes and transporters from varied families as a bioinformatic hook in conjunction with an in-house code to identify precursor peptides, we generated a map of ∼15,500 RaS-RiPP gene clusters, which reveal a remarkable diversity of syntenies pointing to a tremendous range of enzymatic and natural product chemistries that remain to be explored. To assess its utility, we examined one family of gene clusters encoding a YcaO enzyme and a RaS enzyme. We find the former is noncanonical, contains an iron-sulfur cluster, and installs a novel modification, a backbone amidine into the precursor peptide. The RaS enzyme was also found to install a new modification, a C-C crosslink between the unactivated terminal δ-methyl group of Ile and a Trp side chain. The co-occurrence search can be applied to other families of RiPPs, as we demonstrate with the emerging DUF692 di-iron enzyme superfamily.


Ethane ocean on titan.
It is proposed that Saturn's satellite Titan is covered by an ocean one to several kilometers deep consisting mainly of ethane. If the ocean is in thermodynamic equilibrium with an atmosphere of 3 percent (mole fraction) methane, then its composition is roughly 70 percent ethane, 25 percent methane, and 5 percent nitrogen. Photochemical models predict that ethane is the dominant end product of methane photolysis so that the evolving ocean is both the source and sink for continuing photolysis. The coexisting atmosphere is compatible with Voyager data.


Photorefractive damage thresholds in Ti:LiNbO(3) channel waveguides.
The photorefractive effect in titanium-in-diffused LiNbO(3) waveguides is studied both experimentally and theoretically. Measurements of mode size and transmitted optical power as a function of input optical power are presented. The diffusion constants in the diffusion model and the unknown parameters in Kukhtarev's model are determined from measurements of the near-field intensity profiles at low and at high intensity levels. These parameters can then be used to predict waveguide behavior as a function of the input power level. The simulated behavior closely resembles that observed experimentally. Calculated corrections for thermally induced index perturbation show that the thermal effects are higher-order corrections to the dominant photorefractive effect.


Blue eyes in lemurs and humans: same phenotype, different genetic mechanism.
Almost all mammals have brown or darkly-pigmented eyes (irises), but among primates, there are some prominent blue-eyed exceptions. The blue eyes of some humans and lemurs are a striking example of convergent evolution of a rare phenotype on distant branches of the primate tree. Recent work on humans indicates that blue eye color is associated with, and likely caused by, a single nucleotide polymorphism (rs12913832) in an intron of the gene HERC2, which likely regulates expression of the neighboring pigmentation gene OCA2. This raises the immediate question of whether blue eyes in lemurs might have a similar genetic basis. We addressed this by sequencing the homologous genetic region in the blue-eyed black lemur (Eulemur macaco flavifrons; N = 4) and the closely-related black lemur (Eulemur macaco macaco; N = 4), which has brown eyes. We then compared a 166-bp segment corresponding to and flanking the human eye-color-associated region in these lemurs, as well as other primates (human, chimpanzee, orangutan, macaque, ring-tailed lemur, mouse lemur). Aligned sequences indicated that this region is strongly conserved in both Eulemur macaco subspecies as well as the other primates (except blue-eyed humans). Therefore, it is unlikely that this regulatory segment plays a major role in eye color differences among lemurs as it does in humans. Although convergent phenotypes can sometimes come about via the same or similar genetic changes occurring independently, this does not seem to be the case here, as we have shown that the genetic basis of blue eyes in lemurs differs from that of humans.


The endocannabinoid system controls food intake via olfactory processes.
Hunger arouses sensory perception, eventually leading to an increase in food intake, but the underlying mechanisms remain poorly understood. We found that cannabinoid type-1 (CB1) receptors promote food intake in fasted mice by increasing odor detection. CB1 receptors were abundantly expressed on axon terminals of centrifugal cortical glutamatergic neurons that project to inhibitory granule cells of the main olfactory bulb (MOB). Local pharmacological and genetic manipulations revealed that endocannabinoids and exogenous cannabinoids increased odor detection and food intake in fasted mice by decreasing excitatory drive from olfactory cortex areas to the MOB. Consistently, cannabinoid agonists dampened in vivo optogenetically stimulated excitatory transmission in the same circuit. Our data indicate that cortical feedback projections to the MOB crucially regulate food intake via CB1 receptor signaling, linking the feeling of hunger to stronger odor processing. Thus, CB1 receptor-dependent control of cortical feedback projections in olfactory circuits couples internal states to perception and behavior.


Large buoyant LDL-like particles in hepatic lipase deficiency.
Hepatic lipase (HL) is thought to play a role in processing very low density lipoprotein to low density lipoprotein (LDL). To analyze the relationship between HL and LDL, the density, size, and chemical composition of LDL isolated from 18 normal subjects and from three subjects with reduced or absent levels of HL activity were compared. In an HL-deficient subject, the major peak of apoprotein (apo) B-containing lipoproteins ('LDL') had a density of 1.023 g/ml and a diameter of 26.4 nm compared to male control subjects (1.044 +/- 0.006 g/ml and 25.3 +/- 0.3 nm). Two half-sisters of the HL-deficient subject with half the normal levels of HL activity had LDL that also were more buoyant and slightly larger than the LDL isolated from female control subjects. The peak density and average diameter of LDL were correlated with HL activity, consistent with the hypothesis that HL influenced formation and physical characteristics of typical LDL. Apo B-100 was the major apoprotein in the 'LDL' isolated from the HL-deficient subject and contained a greater proportion of triglyceride compared to the control subjects' LDL. The absence of HL appears to prevent the production of classical LDL. Our data support the hypothesis that HL helps determine normal LDL characteristics.


Hepatocellular carcinoma in patients with nonalcoholic steatohepatitis.
There have been few reports on hepatocellular carcinoma (HCC) in nonalcoholic steatohepatitis (NASH) and the natural history of NASH. Accordingly, we assessed the clinical features of HCC in NASH, the risk factors for HCC, and natural history of NASH with advanced fibrosis. There were 34 NASH patients with HCC and 348 NASH patients without HCC. To clarify the clinical features of NASH patients with HCC and to determine the risk factors for HCC, we compared NASH patients with HCC with those without HCC. Univariate and multivariate logistic regression models were used for statistical analysis. A prospective cohort study of the outcomes of 137 NASH with advanced fibrosis was started in 1990. The impact of baseline risk factors on the development of HCC and survival was evaluated by Cox regression analysis. In total, 88% of patients with HCC had advanced fibrosis, with a median age of 70 years. Older age, low level of AST, low grade of histological activity, and advanced stage of fibrosis were risk factors for HCC. A prospective cohort study showed that the 5-year cumulative incidence of HCC was 7.6%, and the 5-year survival rate was 82.8%. HCC was the leading cause of death. The present study confirmed that older age and advanced fibrosis were important risk factors for HCC, and that HCC was the major cause of mortality in NASH patients with advanced fibrosis. Regular screening for HCC is thus extremely important for NASH patients with advanced fibrosis.


Feasibility study on the use of visible-near-infrared spectroscopy for the screening of individual and total glucosinolate contents in broccoli.
The potential of visible-near-infrared spectroscopy to determine selected individual and total glucosinolates in broccoli has been evaluated. Modified partial least-squares regression was used to develop quantitative models to predict glucosinolate contents. Both the whole spectrum and different spectral regions were separately evaluated to develop the quantitative models; in all cases the best results were obtained using the near-infrared zone between 2000 and 2498 nm. These models have been externally validated for the screening of glucoraphanin, glucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin, and total glucosinolates contents. In addition, discriminant partial least-squares was used to distinguish between two possible broccoli cultivars and showed a high degree of accuracy. In the case of the qualitative analysis, best results were obtained using the whole spectrum (i.e., 400-2498 nm) with a correct classification rate of 100% in external validation being obtained.


Msh2 acts in medium-spiny striatal neurons as an enhancer of CAG instability and mutant huntingtin phenotypes in Huntington's disease knock-in mice.
The CAG trinucleotide repeat mutation in the Huntington's disease gene (HTT) exhibits age-dependent tissue-specific expansion that correlates with disease onset in patients, implicating somatic expansion as a disease modifier and potential therapeutic target. Somatic HTT CAG expansion is critically dependent on proteins in the mismatch repair (MMR) pathway. To gain further insight into mechanisms of somatic expansion and the relationship of somatic expansion to the disease process in selectively vulnerable MSNs we have crossed HTT CAG knock-in mice (HdhQ111) with mice carrying a conditional (floxed) Msh2 allele and D9-Cre transgenic mice, in which Cre recombinase is expressed specifically in MSNs within the striatum. Deletion of Msh2 in MSNs eliminated Msh2 protein in those neurons. We demonstrate that MSN-specific deletion of Msh2 was sufficient to eliminate the vast majority of striatal HTT CAG expansions in HdhQ111 mice. Furthermore, MSN-specific deletion of Msh2 modified two mutant huntingtin phenotypes: the early nuclear localization of diffusely immunostaining mutant huntingtin was slowed; and the later development of intranuclear huntingtin inclusions was dramatically inhibited. Therefore, Msh2 acts within MSNs as a genetic enhancer both of somatic HTT CAG expansions and of HTT CAG-dependent phenotypes in mice. These data suggest that the selective vulnerability of MSNs may be at least in part contributed by the propensity for somatic expansion in these neurons, and imply that intervening in the expansion process is likely to have therapeutic benefit.


Three-dimensional reconstruction of human cystic fibrosis transmembrane conductance regulator chloride channel revealed an ellipsoidal structure with orifices beneath the putative transmembrane domain.
The cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is a membrane-integral protein that belongs to an ATP-binding cassette superfamily. Mutations in the CFTR gene cause cystic fibrosis in which salt, water, and protein transports are defective in various tissues. Here we expressed wild-type human CFTR as a FLAG-fused protein in HEK293 cells heterologously and purified it in three steps: anti-FLAG and wheat germ agglutinin affinity chromatographies and size exclusion chromatography. The stoichiometry of the protein was analyzed using various biochemical approaches, including chemical cross-linking, blue-native PAGE, size exclusion chromatography, and electron microscopy (EM) observation of antibody-decorated CFTR. All these data support a dimeric assembly of CFTR. Using 5,039 automatically selected particles from negatively stained EM images, the three-dimensional structure of CFTR was reconstructed at 2-nm resolution assuming a 2-fold symmetry. CFTR, presumably in a closed state, was shown to be an ellipsoidal particle with dimensions of 120 x 106 x 162 A. It comprises a small dome-shaped extracellular and membrane-spanning domain and a large cytoplasmic domain with orifices beneath the putative transmembrane domain. EM observation of CFTR.anti-regulatory domain antibody complex confirmed that two regulatory domains are located around the bottom end of the larger oval cytoplasmic domain.


Characterization of local chicken production systems and their potential under different levels of management practice in Jordan.
This study aimed to characterize the local chickens and their production systems and to investigate the opportunities for improvement. The study was carried out in 18 villages in northern Jordan with the participation of 120 households. Data were collected by individual interviews and surveys supported with structured questionnaires. A scoring system was developed to study the effect of different levels of management on chicken performance. The main reasons for keep local chickens were egg production and generation of income. The main features of the production systems were improper housing and inadequate hygiene. Newcastle disease, predators, and parasites were the main causes of flock losses. Purchasing rate was controlled by the chickens' phenotype, sex and age, and by season of year. The average flock size was 41.6 (+/-32.9) chickens per household, with a hen:cock ratio of 6.4:1. The average effective population size was 15.35, which indicated a high rate of inbreeding (5.52%). The main selection criterion adopted by farmers was egg production. There were positive significant correlations (p<0.001) between management level and chickens' performance. Hatchability, survivability, flock size, number of clutches, egg weight and egg mass were the major parameters that improved significantly with improvement in management level. Local chickens fulfil significant functions in the livelihood of rural smallholders; however, many constraints affected the efficiency of the production systems. Solutions should start with improving the management practices and establishing an effective breeding system.


Ancient DNA provides new insights into the evolutionary history of New Zealand's extinct giant eagle.
Prior to human settlement 700 years ago New Zealand had no terrestrial mammals--apart from three species of bats--instead, approximately 250 avian species dominated the ecosystem. At the top of the food chain was the extinct Haast's eagle, Harpagornis moorei. H. moorei (10-15 kg; 2-3 m wingspan) was 30%-40% heavier than the largest extant eagle (the harpy eagle, Harpia harpyja), and hunted moa up to 15 times its weight. In a dramatic example of morphological plasticity and rapid size increase, we show that the H. moorei was very closely related to one of the world's smallest extant eagles, which is one-tenth its mass. This spectacular evolutionary change illustrates the potential speed of size alteration within lineages of vertebrates, especially in island ecosystems.


Risk factors for invasive reptile-associated salmonellosis in children.
Reptile-associated salmonellosis (RAS) in children has been reported primarily due to direct contact with turtles, but recently also due to indirect contact with more exotic reptiles, causing disease in infants. To evaluate risk factors for RAS, we reviewed the RAS cases published in the literature since 1965. A case was defined as a child ≤18 years of age with an epidemiological link by identification of Salmonella enterica in cultures from both the affected child and the exposed reptile. We identified a total of 177 otherwise healthy children (median age 1.0 years, range 2 days to 17.0 years). RAS manifested mainly with gastrointestinal disease, but 15% presented with invasive RAS, including septicemia, meningitis, and bone and joint infection. The children with invasive RAS were significantly younger than children with noninvasive disease (median age 0.17 and 2.0 years, p<0.0001). RAS is most frequently seen after exposure to turtles (42%). However, children with invasive RAS had been exposed more often (p≤0.001) to reptiles other than turtles, including iguanas, bearded dragons, snakes, chameleons, and geckos. Children exposed to those latter reptiles usually kept indoors were younger than children exposed to turtles mostly kept outdoors (p<0.0001). RAS in children is significantly associated with invasive disease at young age, in particular infants <6 months of age. Exposure to reptiles, other than turtles, kept indoors is associated with RAS at younger age and more invasive disease. This finding is helpful for recognizing or even preventing invasive RAS in young infants that are at highest risk.


Novel technical strategies to optimize cryopreservation of goat semen using cholesterol-loaded cyclodextrin.
Artificial insemination is well-established in dairy cattle, with sires housed in commercial studs for processing. In some species, however, sires located on-farm are used for artificial insemination by shipping their semen to an off-site laboratory for processing within 24 h of collection. To expedite semen transport from the farm to laboratory, protocols must be uncomplicated. For goat semen, an obstacle is the seminal plasma, which must be removed because it contains proteins that impede sperm quality. Our objective is to develop a simple strategy to transiently store goat semen for 24 h prior to freezing. Cholesterol-loaded cyclodextrin (CLC) has been demonstrated to improve sperm tolerance to cryopreservation. Therefore, we hypothesized that CLC improves goat sperm resistance to seminal plasma damage, over 24 h prior to cryopreservation. We first evaluated the ability of CLC to protect goat sperm against seminal plasma damage by treating fresh semen with or without seminal plasma prior to cryopreservation. Second, fresh goat semen with seminal plasma was extended in skim milk-based extender ± CLC and held for 24 h at 5 °C prior to freezing. Our results indicate that CLC treatment improves goat sperm resistance to seminal plasma-mediated injury and protects sperm quality over 24 h prior to freezing (P < 0.05). Although the in vivo fertility of semen must first be assessed, it is possible that protocols for goat semen cryopreservation can be simplified by including CLC and eliminating seminal plasma removal. Processing and distribution of goat semen for AI would thereby be facilitated.


Predominantly Pro-Inflammatory Phenotype with Mixed M1/M2 Polarization of Peripheral Blood Classical Monocytes and Monocyte-Derived Macrophages among Patients with Excessive Ethanol Intake.
Excessive alcohol consumption impairs the immune system, induces oxidative stress, and triggers the activation of peripheral blood (PB) monocytes, thereby contributing to alcoholic liver disease (ALD). We analyzed the M1/M2 phenotypes of circulating classical monocytes and macrophage-derived monocytes (MDMs) in excessive alcohol drinkers (EADs). PB samples from 20 EADs and 22 healthy controls were collected for isolation of CD14+ monocytes and short-term culture with LPS/IFNγ, IL4/IL13, or without stimulation. These conditions were also used to polarize MDMs into M1, M2, or M0 phenotypes. Cytokine production was assessed in the blood and culture supernatants. M1/M2-related markers were analyzed using mRNA expression and surface marker detection. Additionally, the miRNA profile of CD14+ monocytes was analyzed. PB samples from EADs exhibited increased levels of pro-inflammatory cytokines. Following short-term culture, unstimulated blood samples from EADs showed higher levels of soluble TNF-α and IL-8, whereas monocytes expressed increased levels of surface TNF-α and elevated mRNA expression of pro-inflammatory cytokines and inducible nitric oxide synthase. MDMs from EADs showed higher levels of TNF-α and CD206 surface markers and increased IL-10 production. LPS/IFNγ induced higher mRNA expression of Nrf2 only in the controls. miRNA analysis revealed a distinctive miRNA profile that is potentially associated with liver carcinogenesis and ALD through inflammation and oxidative stress. This study confirms the predominantly pro-inflammatory profile of PB monocytes among EADs and suggests immune exhaustion features in MDMs.


Single-molecule DNA sequencing of a viral genome.
The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing.


Dried-leaf Artemisia annua: A practical malaria therapeutic for developing countries?
Artemisinin from the plant Artemisia annua (A. annua) L, and used as artemisinin combination therapy (ACT), is the current best therapeutic for treating malaria, a disease that hits children and adults especially in developing countries. Traditionally, A. annua was used by the Chinese as a tea to treat "fever". More recently, investigators have shown that tea infusions and oral consumption of the dried leaves of the plant have prophylactic and therapeutic efficacy. The presence of a complex matrix of chemicals within the leaves seems to enhance both the bioavailability and efficacy of artemisinin. Although about 1000-fold less potent than artemisinin in their antiplasmodial activity, these plant chemicals are mainly small molecules that include other artemisinic compounds, terpenes (mainly mono and sesqui), flavonoids, and polyphenolic acids. In addition, polysaccharide constituents of A. annua may enhance bioavailability of artemisinin. Rodent pharmacokinetics showed longer T1/2 and Tmax and greater Cmax and AUC in Plasmodium chabaudi-infected mice treated with A. annua dried leaves than in healthy mice. Pharmacokinetics of deoxyartemisinin, a liver metabolite of artemisinin, was more inhibited in infected than in healthy mice. In healthy mice, artemisinin serum levels were > 40-fold greater in dried leaf fed mice than those fed with pure artemisinin. Human trial data showed that when delivered as dried leaves, 40-fold less artemisinin was required to obtain a therapeutic response compared to pure artemisinin. ACTs are still unaffordable for many malaria patients, and cost estimates for A. annua dried leaf tablet production are orders of magnitude less than for ACT, despite improvements in the production capacity. Considering that for > 2000 years this plant was used in traditional Chinese medicine for treatment of fever with no apparent appearance of artemisinin drug resistance, the evidence argues for inclusion of affordable A. annua dried leaf tablets into the arsenal of drugs to combat malaria and other artemisinin-susceptible diseases.


Azelnidipine, a dihydropyridine-based calcium antagonist, inhibits angiotensin II-induced oxidative stress generation and downregulation of pigment epithelium-derived factor mRNA levels in microvascular endothelial cells.
We have previously shown that azelnidipine, a long-acting dihydropyridine-based calcium antagonist (DHP), inhibited tumor necrosis factor-alpha-induced endothelial cell (EC) activation through its antioxidative properties. However whether azelnidipine could also block the angiotensin II (Ang II)-signaling in ECs remains to be elucidated. Since Ang II-type 1 receptor interaction could contribute to exacerbation of diabetic retinopathy by downregulating pigment epithelium-derived factor (PEDF) gene expression in ECs, we examined here whether azelnidipine inhibited the Ang II-induced reactive oxygen species (ROS) generation and subsequent PEDF gene suppression in microvascular ECs. Azelnidipine, but not nitrendipine, the other popular DHFP completely inhibited the Ang II-induced ROS generation in ECs. Furthermore, azelnidipine, but not nitrendipine, was found to partially restore decreased PEDF mRNA levels in Ang II-exposed ECs. The present study suggests that azelnidipine could inhibit the Ang II-induced decrease in PEDF mRNA levels in ECs through its antioxidative properties. Upregulation of PEDF by azelnidipine may become a therapeutic target for the treatment of diabetic retinopathy associated with hypertension.


Astaxanthin: a potential therapeutic agent in cardiovascular disease.
Astaxanthin is a xanthophyll carotenoid present in microalgae, fungi, complex plants, seafood, flamingos and quail. It is an antioxidant with anti-inflammatory properties and as such has potential as a therapeutic agent in atherosclerotic cardiovascular disease. Synthetic forms of astaxanthin have been manufactured. The safety, bioavailability and effects of astaxanthin on oxidative stress and inflammation that have relevance to the pathophysiology of atherosclerotic cardiovascular disease, have been assessed in a small number of clinical studies. No adverse events have been reported and there is evidence of a reduction in biomarkers of oxidative stress and inflammation with astaxanthin administration. Experimental studies in several species using an ischaemia-reperfusion myocardial model demonstrated that astaxanthin protects the myocardium when administered both orally or intravenously prior to the induction of the ischaemic event. At this stage we do not know whether astaxanthin is of benefit when administered after a cardiovascular event and no clinical cardiovascular studies in humans have been completed and/or reported. Cardiovascular clinical trials are warranted based on the physicochemical and antioxidant properties, the safety profile and preliminary experimental cardiovascular studies of astaxanthin.


Sequencing of the entire coding region of the receptor associated protein (RAP) in patients with primary hypothyroidism of unknown origin.
The LDL receptor-associated protein (RAP) is involved in secretion of thyroglobulin (Tg) from the thyrocyte to the colloid. Disruption of the RAP gene in mice results in a reduced Tg content within the colloid, leading to subclinical hypothyroidism and histological alterations resembling early goiter. Here we studied the entire coding sequence of RAP in genomic DNA samples from 18 patients with primary hypothyroidism not due to thyroid autoimmunity or dysgenesis. The control group included 21 subjects with no evidence of thyroid alterations. Eleven different polymorphisms with amino-acid substitution and 4 different missense polymorphisms without amino-acid substitution were found in various regions of the RAP gene. Only one polymorphism in exone 7 (V311M) was observed exclusively in patients, but it had been previously reported in normal subjects as well. The remaining polymorphisms were found either both in patients and controls or only in controls and had not been previously reported. The frequency of the various polymorphisms did not differ significantly between patients and controls. Based on these findings, we conclude that alterations of the RAP gene are not a common cause of hypothyroidism, although it cannot be excluded that other, rarer alterations with a pathogenic effect exist, and that they should be investigated in a larger number of patients.