Cloning and characterization of a novel Nicotiana tabacum ABC transporter involved in shoot branching.
The ATP-binding cassette (ABC) superfamily is a large protein family with diverse physiological functions in all kingdoms of life. One distinguished subfamily, the pleiotropic drug resistance (PDR) transporters, has only been identified in plants and fungi. Here, we identified a Nicotiana tabacum PDR gene, NtPDR6, which is a homolog of Petunia hybrida PDR1. The full-length cDNA of NtPDR6 had a 4482-bp open reading frame encoding a full-size ABC transporter with 1493 amino acids. Sequence comparison showed that NtPDR6 had high homology with plant PDR proteins. NtPDR6 was strongly induced by phosphate starvation as well as by 1-naphthalene acetic acid. Tissue expression pattern analysis showed that NtPDR6 was detected in all surveyed tissues but preferentially in roots. We cloned the 1.3-kb NtPDR6 promoter and found that there was one phosphate starvation response-related element Pho-like and several root-specific expression-related elements rootmotiftapox1 in the NtPDR6 promoter. A tissue-specific pattern of NtPDR6 promoter-β-glucuronidase expression was dominantly observed in subepidermal cells and the elongation zone of lateral roots. RNA interference technology was used to knock down NtPDR6 expression, and there was a significantly increased branching phenotype in the NtPDR6 knockdown plants. These data suggest that NtPDR6 plays a key role in regulation of shoot branching processes.


Myosin IIA dependent retrograde flow drives 3D cell migration.
Epithelial cell migration is an essential part of embryogenesis and tissue regeneration, yet their migration is least understood. Using our three-dimensional (3D) motility analysis, migrating epithelial cells formed an atypical polarized cell shape with the nucleus leading the cell front and a contractile cell rear. Migrating epithelial cells exerted traction forces to deform both the anterior and posterior extracellular matrix toward the cell body. The cell leading edge exhibited a myosin II-dependent retrograde flow with the magnitude and direction consistent with surrounding network deformation. Interestingly, on a two-dimensional substrate, myosin IIA-deficient cells migrated faster than wild-type cells, but in a 3D gel, these myosin IIA-deficient cells were unpolarized and immobile. In contrast, the migration rates of myosin IIB-deficient cells were similar to wild-type cells. Therefore, myosin IIA, not myosin IIB, is required for 3D epithelial cell migration.


Comparison of cloned Kir2 channels with native inward rectifier K+ channels from guinea-pig cardiomyocytes.
The aim of the study was to compare the properties of cloned Kir2 channels with the properties of native rectifier channels in guinea-pig (gp) cardiac muscle. The cDNAs of gpKir2.1, gpKir2.2, gpKir2.3 and gpKir2.4 were obtained by screening a cDNA library from guinea-pig cardiac ventricle. A partial genomic structure of all gpKir2 genes was deduced by comparison of the cDNAs with the nucleotide sequences derived from a guinea-pig genomic library. The cell-specific expression of Kir2 channel subunits was studied in isolated cardiomyocytes using a multi-cell RT-PCR approach. It was found that gpKir2.1, gpKir2.2 and gpKir2.3, but not gpKir2.4, are expressed in cardiomyocytes. Immunocytochemical analysis with polyclonal antibodies showed that expression of Kir2.4 is restricted to neuronal cells in the heart. After transfection in human embryonic kidney cells (HEK293) the mean single-channel conductance with symmetrical K+ was found to be 30.6 pS for gpKir2.1, 40.0 pS for gpKir2.2 and 14.2 pS for Kir2.3. Cell-attached measurements in isolated guinea-pig cardiomyocytes (n = 351) revealed three populations of inwardly rectifying K+ channels with mean conductances of 34.0, 23.8 and 10.7 pS. Expression of the gpKir2 subunits in Xenopus oocytes showed inwardly rectifying currents. The Ba2+ concentrations required for half-maximum block at -100 mV were 3.24 M for gpKir2.1, 0.51 M for gpKir2.2, 10.26 M for gpKir2.3 and 235 M for gpKir2.4. Ba2+ block of inward rectifier channels of cardiomyocytes was studied in cell-attached recordings. The concentration and voltage dependence of Ba2+ block of the large-conductance inward rectifier channels was virtually identical to that of gpKir2.2 expressed in Xenopus oocytes. Our results suggest that the large-conductance inward rectifier channels found in guinea-pig cardiomyocytes (34.0 pS) correspond to gpKir2.2. The intermediate-conductance (23.8 pS) and low-conductance (10.7 pS) channels described here may correspond to gpKir2.1 and gpKir2.3, respectively.


Hippocampal metabolomics using ultrahigh-resolution mass spectrometry reveals neuroinflammation from Alzheimer's disease in CRND8 mice.
In the wake of genomics, metabolomics characterizes the small molecular metabolites revealing the phenotypes induced by gene mutants. To address the metabolic signatures in the hippocampus of the amyloid-beta (Aβ) peptides produced in transgenic (Tg) CRND8 mice, high-field ion cyclotron resonance-Fourier transform mass spectrometry supported by LC-LTQ-Orbitrap was introduced to profile the extracted metabolites. More than 10,000 ions were detected in the mass profile for each sample. Subsequently, peak alignment and the 80% rule followed by feature selection based on T score computation were performed. The putative identification was also conducted using the highly accurate masses with isotopic distribution by interfacing the MassTRIX database as well as MS/MS fragmentation generated in the LTQ-Orbitrap after chromatographic separation. Consequently, 58 differentiating masses were tentatively identified while up to 44 differentiating elemental compositions could not be biologically annotated in the databases. Nonetheless, of the putatively annotated masses, eicosanoids in arachidonic acid metabolism, fatty acid beta-oxidation disorders as well as disturbed glucose metabolism were highlighted as metabolic traits of Aβ toxicity in Tg CRND8 mice. Furthermore, a web-based bioinformatic tool was used for simulation of the metabolic pathways. As a result of the obtained metabolic signatures, the arachidonic acid metabolism dominates the metabolic perturbation in hippocampal tissues of Tg CRND8 mice compared to non-Tg littermates, indicating that Aβ toxicity functions neuroinflammation in hippocampal tissue and new theranostic opportunities might be offered by characterization of altered arachidonic acid metabolism for Alzheimer's disease.


Disentangling effects of induced plant defenses and food quantity on herbivores by fitting nonlinear models.
Plants can respond to herbivore damage through both broad-scale (systemic) and localized induced responses. While many studies have quantified the impact of systemic responses on herbivores, measuring the impact of localized changes is difficult because plant tissues that have suffered direct damage may represent both a lower quality and a lower quantity of food. This article uses nonlinear models to disentangle the confounding effects of prior herbivory on food quantity and quality. The first (null) model assumes that herbivore performance is determined only by the quantity of food available to an average herbivore. Modified models allow two distinct effects of damage-induced defenses: an increase in the amount of food each herbivore is required to consume in order to achieve maximum performance and a reduction in the maximum performance even when herbivores are fed ad lib. Maximum likelihood methods were used to fit the models to data from field experiments in which Colorado potato beetle (Leptinotarsa decemlineata) larvae were reared on three varieties of potatoes that had been damaged to varying degrees by adult beetles. Prior damage reduced the mean mass of beetles at pupation, and this effect was due to both a decrease in food quantity and induced changes in food quality. In contrast, beetle survival was affected in some cases by reduced food quantity but showed no responses that could be attributed to induced defenses. I discuss this result in the context of previous studies of induced (mostly systemic) responses in the potato-potato beetle system, and I suggest that detailed studies of particular chemical responses and the proposed method of combining bioassays with quantitative models should be used as complementary approaches in future studies of herbivore-induced defenses in plants.


Determining the relationship of heart rate and blood pressure using voluntary cardio-respiratory synchronization (VCRS).
Voluntary cardio-respiratory synchronization (VCRS) was used to investigate heart rate and blood pressure changes in the supine position in 21 subjects. VCRS involves a breathing pattern that is synchronized with the cardiac cycle. The signals to inhale and exhale are derived from the ECG. In this study, the subjects inspired for four heart beats and expired for four heart beats for 35 cycles. This technique is designed to have the heart beat occur at exactly the same phase in the respiratory cycle and lends itself to the study of the influence of the respiration cycle on heart rate and blood pressure changes. The heart rate and blood pressure changed simultaneously in the same direction, with the largest significant positive change occurring on the second heart beat during inspiration. The authors discuss the potential of VCRS for research, and clinical applications as a respiration modulator for hypertension therapy or increased heart rate variability.


[Emotional stress as an illness-related factor].
Emotions represent reactions of living organisms to changes of one's inner sphere, as well as to different factors from our surroundings. Emotions manifest in many ways. Failing to fulfill one's needs on chronic or acute basis leads to conflict situations. These needs may be either biological or social. Conflict situations occur prior to stress itself. Modern technology development provides no friendly environment for workers, with many consequences causing psychosomatic disorders. Researches have been made to evaluate the amount of inflicted stress on patients. One of successful theories regarding this question is the theory of functional systems. The idea was to quantify processes of everyday living. The primary goal was to measure one's adaptation abilities. In order to follow up these parameters a machine was created. This appliance can determine the level of stress, by measuring certain physiological parameters. That is way scientists may apply preventive measures in order to increase stress resistance. We differ hereditary and individual resistance to stress factors. That is why it is necessary to assess each patient individually. This type of individual approach offers an opportunity to create a specific program, in order to increase stress resistance.


Improving Food and Drug Administration-Centers for Medicare and Medicaid Services Coordination for Drugs Granted Accelerated Approval.
Policy Points The increasing number of drugs granted accelerated approval by the Food and Drug Administration (FDA) has challenged the Medicare program, which often pays for expensive therapies despite substantial uncertainty about benefits and risks to Medicare beneficiaries. We recommend several administrative and legislative approaches for improving FDA-Centers for Medicare and Medicaid Services (CMS) coordination around accelerated-approval drugs, including promoting earlier discussions among the FDA, the CMS, and drug companies; strengthening Medicare's coverage with evidence development program; linking Medicare payment to evidence generation milestones; and ensuring that the CMS has adequate staffing and resources to evaluate new therapies. These activities can help improve the integrity; transparency; and efficiency of approval, coverage, and payment processes for drugs granted accelerated approval. The Food and Drug Administration (FDA)'s accelerated-approval pathway expedites patient access to promising treatments. However, increasing use of this pathway has challenged the Medicare program, which often pays for expensive therapies despite substantial uncertainty about benefits and risks to Medicare beneficiaries. We examined approaches to improve coordination between the FDA and Centers for Medicare and Medicaid Services (CMS) for drugs granted accelerated approval. We argue that policymakers have focused on expedited pathways at the FDA without sufficient attention to complementary policies at the CMS. Although differences between the FDA and CMS decisions are to be expected given the agencies' different missions and statutory obligations, procedural improvements can ensure that Medicare beneficiaries have timely access to novel therapies that are likely to improve health outcomes. To inform policy options and recommendations, we conducted semistructured interviews with stakeholders to capture diverse perspectives on the topic. We recommend ten areas for consideration: clarifying the FDA's evidentiary standards; strengthening FDA authorities; promoting earlier discussions among the FDA, the CMS, and drug companies; improving Medicare's coverage with evidence development program; tying Medicare payment for accelerated-approval drugs to evidence generation milestones; issuing CMS guidance on real-world evidence; clarifying Medicare's "reasonable and necessary" criteria; adopting lessons from international regulatory-reimbursement harmonization efforts; ensuring that the CMS has adequate staffing and expertise; and emphasizing equity. Better coordination between the FDA and CMS could improve the transparency and predictability of drug approval and coverage around accelerated-approval drugs, with important implications for patient outcomes, health spending, and evidence generation processes. Improved coordination will require reforms at both the FDA and CMS, with special attention to honoring the agencies' distinct authorities. It will require administrative and legislative actions, new resources, and strong leadership at both agencies.


Massively parallel sequencing techniques for forensics: A review.
DNA sequencing, starting with Sanger's chain termination method in 1977 and evolving into the next generation sequencing (NGS) techniques of today that employ massively parallel sequencing (MPS), has become essential in application areas such as biotechnology, virology, and medical diagnostics. Reflected by the growing number of articles published over the last 2-3 years, these techniques have also gained attention in the forensic field. This review contains a brief description of first, second, and third generation sequencing techniques, and focuses on the recent developments in human DNA analysis applicable in the forensic field. Relevance to the forensic analysis is that besides generation of standard STR-profiles, DNA repeats can also be sequenced to look for polymorphisms. Furthermore, additional SNPs can be sequenced to acquire information on ancestry, paternity or phenotype. The current MPS systems are also very helpful in cases where only a limited amount of DNA or highly degraded DNA has been secured from a crime scene. If enough autosomal DNA is not present, mitochondrial DNA can be sequenced for maternal lineage analysis. These developments clearly demonstrate that the use of NGS will grow into an indispensable tool for forensic science.


The role of angiotensin II activation of yes-associated protein/PDZ-binding motif signaling in hypertensive cardiac and vascular remodeling.
Vascular remodeling is the pathogenic basis of hypertension and end organ injury, and the proliferation of vascular smooth muscle cells (VSMCs) is central to vascular remodeling. Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) are key effectors of the Hippo pathway and crucial for controlling cell proliferation, apoptosis and differentiation. The present study investigated the role of YAP/TAZ in cardiac and vascular remodeling of angiotensin II-induced hypertension. Ang II induced YAP/TAZ activation in the heart and aorta, which was prevented by YAP/TAZ inhibitor verteporfin. Treatment with verteporfin significantly reduced Ang II-induced cardiac and vascular hypertrophy with a mild reduction in systolic blood pressure (SBP), verteporfin attenuated Ang II-induced cardiac and aortic fibrosis with the inhibition of transform growth factor (TGF)β/Smad2/3 fibrotic signaling and extracellular matrix collagen I deposition. Ang II induced Rho A, extracellular signal-regulated kinase 1/2 (ERK1/2) and YAP/TAZ activation in VSMCs, either Rho kinase inhibitor fasudil or ERK inhibitor PD98059 suppressed Ang II-induced YAP/TAZ activation, cell proliferation and fibrosis of VSMCs. Verteporfin also inhibited Ang II-induced VSMC proliferation and fibrotic TGFβ1/Smad2/3 pathway. These results demonstrate that Ang II activates YAP/TAZ via Rho kinase/ERK1/2 pathway in VSMCs, which may contribute to cardiac and vascular remodeling in hypertension. Our results suggest that YAP/TAZ plays a critical role in the pathogenesis of hypertension and end organ damage, and targeting the YAP/TAZ pathway may be a new strategy for the prevention and treatment of hypertension and cardiovascular diseases.


Understanding temporal and spatial distribution of intestinal nematodes of horses using faecal egg counts and DNA metabarcoding.
This study reports the spatial and temporal distribution of ascarid and strongylid nematodes in Thoroughbred horses by age category across different climatic zones in Australia over an 18-month period. Faecal samples (n = 2046) from individual horses were analysed using the modified McMaster technique for faecal egg counts (FECs). Strongylids were identified using PCR-directed next-generation sequencing of the second internal transcribed spacer (ITS-2) of the nuclear ribosomal DNA. Yearlings had the highest prevalence (82%) of strongyle eggs followed by weanlings (79%), foals (58%), wet mares (49%) and dry mares (46%). For Parascaris spp., foals had the highest prevalence (35%) followed by weanlings (21%) and yearlings (10%). The highest mean FECs for Parascaris spp. were observed in foals (525 eggs per gram [EPG] of faeces) while those for strongyles were in yearlings (962 EPG). Among horses that were classified as adults at the time of sampling, 77% (860 of 1119) of mares were low (i.e., <250 EPG) strongyle egg-shedders. Mean strongyle FEC counts were highest in the Mediterranean (818 EPG) followed by summer (599 EPG), winter (442 EPG), and non-seasonal (413 EPG) rainfall zones. Twenty-six nematode species were detected, with Cylicostephanus longibursatus (26.5%), Cylicocyclus nassatus (23.7%) and Coronocyclus coronatus (20.5%) being the most frequently detected species. Their richness and relative abundance varied with horse age, season and climatic zone. In addition, Strongylus equinus and Triodontophorus spp. (T. brevicauda and T. serratus) were also detected. This comprehensive study elucidates spatial (climatic zone) and temporal (i.e., seasonal) trends in prevalence and burdens of intestinal nematodes in Australian horses using non-invasive conventional and molecular methods. The information presented in this study is crucial for developing integrated management strategies to control horse parasites in farmed horses.


Activity and Thermostability of GH5 Endoglucanase Chimeras from Mesophilic and Thermophilic Parents.
Cellulases from glycoside hydrolase family 5 (GH5) are key endoglucanase enzymes in the degradation of diverse polysaccharide substrates and are used in industrial enzyme cocktails to break down biomass. The GH5 family shares a canonical (βα)8-barrel structure, where each (βα) module is essential for the enzyme's stability and activity. Despite their shared topology, the thermostability of GH5 endoglucanase enzymes can vary significantly, and highly thermostable variants are often sought for industrial applications. Based on the previously characterized thermophilic GH5 endoglucanase Egl5A from Talaromyces emersonii (TeEgl5A), which has an optimal temperature of 90°C, we created 10 hybrid enzymes with elements of the mesophilic endoglucanase Cel5 from Stegonsporium opalus (SoCel5) to determine which elements are responsible for enhanced thermostability. Five of the expressed hybrid enzymes exhibit enzyme activity. Two of these hybrids exhibited pronounced increases in the temperature optimum (10 and 20°C), the temperature at which the protein lost 50% of its activity (T50) (15 and 19°C), and the melting temperature (Tm ) (16.5 and 22.9°C) and extended half-lives (t1/2) (∼240- and 650-fold at 55°C) relative to the values for the mesophilic parent enzyme and demonstrated improved catalytic efficiency on selected substrates. The successful hybridization strategies were validated experimentally in another GH5 endoglucanase, Cel5 from Aspergillus niger (AnCel5), which demonstrated a similar increase in thermostability. Based on molecular dynamics (MD) simulations of both the SoCel5 and TeEgl5A parent enzymes and their hybrids, we hypothesize that improved hydrophobic packing of the interface between α2 and α3 is the primary mechanism by which the hybrid enzymes increase their thermostability relative to that of the mesophilic parent SoCel5.IMPORTANCE Thermal stability is an essential property of enzymes in many industrial biotechnological applications, as high temperatures improve bioreactor throughput. Many protein engineering approaches, such as rational design and directed evolution, have been employed to improve the thermal properties of mesophilic enzymes. Structure-based recombination has also been used to fuse TIM barrel fragments, and even fragments from unrelated folds, to generate new structures. However, little research has been done on GH5 endoglucanases. In this study, two GH5 endoglucanases exhibiting TIM barrel structure, SoCel5 and TeEgl5A, with different thermal properties, were hybridized to study the roles of different (βα) motifs. This work illustrates the role that structure-guided recombination can play in helping to identify sequence function relationships within GH5 enzymes by supplementing natural diversity with synthetic diversity.


Distinct Core Glycan and O-Glycoform Utilization of SARS-CoV-2 Omicron Variant Spike Protein RBD Revealed by Top-Down Mass Spectrometry.
The SARS-CoV-2 Omicron (B.1.1.529) variant possesses numerous spike (S) mutations particularly in the S receptor-binding domain (S-RBD) that significantly improve transmissibility and evasion of neutralizing antibodies. But exactly how the mutations in the Omicron variant enhance viral escape from immunological protection remains to be understood. The S-RBD remains the principal target for neutralizing antibodies and therapeutics, thus new structural insights into the Omicron S-RBD and characterization of the post-translational glycosylation changes can inform rational design of vaccines and therapeutics. Here we report the molecular variations and O-glycoform changes of the Omicron S-RBD variant as compared to wild-type (WA1/2020) and Delta (B.1.617.2) variants using high-resolution top-down mass spectrometry (MS). A novel O-glycosite (Thr376) unique to the Omicron variant is identified. Moreover, we have directly quantified the Core 1 and Core 2 O-glycan structures and characterized the O-glycoform structural heterogeneity of the three variants. Our findings reveal high resolution detail of Omicron O-glycoforms and their utilization to provide direct molecular evidence of proteoform alterations in the Omicron variant which could shed light on how this variant escapes immunological protection.


Antibody response to OspC-I synthetic peptide derived from outer surface protein C of Borrelia burgdorferi in sera from Japanese forestry workers.
The prevalence of antibodies against Lyme disease spirochaetes in serum samples from 80 forestry workers at high occupational risk of Lyme disease was surveyed by enzyme-linked immunosorbent assay (ELISA) with the OspC-I synthetic peptide. The peptide is part of the outer surface protein C (OspC) amino acid sequence located in the region conserved among Borrelia burgdorferi sensu stricto or sensu lato. Positivity for antibodies against OspC-I was observed in 25 (313%) of the forestry workers. Of these positive cases, 12 (15.0%) and 19 (23.8%) were positive for immunoglobulin M (IgM) and IgG antibody, respectively. Among 62 workers who were negative for IgG antibody against B. garinii or B. japonica in our previous study, 9 (14.5%) and 4 (6.5%) were positive for IgM and IgG antibody, respectively, in OspC-I ELISA. These results demonstrate for the first time that Lyme disease in forestry workers can be revealed using OspC-I ELISA. We conclude that forestry workers who show positive results for antibodies against OspC-I have very likely been exposed to Lyme disease spirochaetes, and that those who show positivity for IgM antibody against OspC-I may be in the early stage of Lyme disease.


Factors associated with colorectal cancer in the context of the Mediterranean diet: a case-control study.
Recent evidence demonstrates that increased adherence to the Mediterranean diet (MD) may prevent colorectal cancer (CRC). The aim of this study was to evaluate the association between health-related characteristics and CRC in the context of the MD. This was a case-control study conducted on a sample of 338 consecutive patients with a first diagnosis of CRC recruited in an urban facility in the city of Catania, southern Italy, and matched with 676 apparently healthy subjects without clinical symptoms or signs of any type of cancer. Data regarding sociodemographic, clinical, and lifestyle characteristics were collected, and adherence to the Mediterranean diet pattern was assessed using the MedDietScore. A significant association between a greater adherence to the MD and lower odds of having cancer (odds ratio = 0.46, 95% confidence interval: 0.28-0.75) was found. Also, smoking status, family history of CRC, obesity, diabetes, physical activity, and high intake of alcohol were significantly associated with CRC, but only among subjects less adherent to the MD. MD was associated with a less detrimental effects of several health-related characteristics associated with CRC, suggesting potential benefits of adherence to this dietary pattern with regards to CRC risk factors.


Genomic diversifications of five Gossypium allopolyploid species and their impact on cotton improvement.
Polyploidy is an evolutionary innovation for many animals and all flowering plants, but its impact on selection and domestication remains elusive. Here we analyze genome evolution and diversification for all five allopolyploid cotton species, including economically important Upland and Pima cottons. Although these polyploid genomes are conserved in gene content and synteny, they have diversified by subgenomic transposon exchanges that equilibrate genome size, evolutionary rate heterogeneities and positive selection between homoeologs within and among lineages. These differential evolutionary trajectories are accompanied by gene-family diversification and homoeolog expression divergence among polyploid lineages. Selection and domestication drive parallel gene expression similarities in fibers of two cultivated cottons, involving coexpression networks and N6-methyladenosine RNA modifications. Furthermore, polyploidy induces recombination suppression, which correlates with altered epigenetic landscapes and can be overcome by wild introgression. These genomic insights will empower efforts to manipulate genetic recombination and modify epigenetic landscapes and target genes for crop improvement.


Impacts of a tiered intervention on child internalizing and externalizing behavior in the context of maternal depression.
Greater maternal depressive symptoms are consistently associated with higher levels of behavioral difficulties in children, emerging in early childhood and with long-lasting consequences for children's development. Interventions promoting early relational health have been shown to have benefits for children's behavior; however, these impacts are not always realized in the context of maternal depression. This study examined whether tiered programs could address this limitation by focusing on both parenting, through universal primary prevention, and psychosocial stressors and parent mental health, through tailored secondary prevention. Analysis of a randomized controlled trial (RCT) of the Smart Beginnings (SB) intervention was conducted to determine whether SB attenuated the association between maternal depression and early childhood internalizing and externalizing behaviors. Maternal depression significantly predicted both internalizing and externalizing behaviors in linear regression models. Further, there was a significant interaction between maternal depression and treatment group, such that among mothers with higher depressive symptoms, the SB treatment attenuated the magnitude of the association between depression and child behavior. Findings suggest that while parenting support is important for all families, it may be particularly critical for those with higher levels of depression and underscores the need to consider multidimensional family processes in both research and clinical practice.


Suppression of a VP1 mutant of simian virus 40 by missense mutations in serine codons of the viral agnogene.
We isolated second-site revertants of a partially defective VP1 mutant of simian virus 40. The suppressing mutation in each of these pseudorevertants was mapped to the viral agnogene. Of six independently isolated pseudorevertants, all had a missense mutation in a serine codon, near the beginning of the agnogene, that would cause replacement of serine at position 7, 11, or 17 in the agnoprotein by a hydrophobic amino acid. Our results suggest that the agnoprotein interacts in a specific way with VP1 during the late stages of viral development.


Effect of forage to concentrate ratio and intake level on utilization of early vegetative alfalfa silage by dairy cows.
Two experiments were conducted to measure the effects of intake and forage: grain ratio on utilization of early maturity alfalfa silage in dairy cows. In Experiment 1, diets with three forage: concentrate ratios (percentage of silage, percentage NDF): low (56, 28.3), medium (71, 31.0), or high (86, 33.4) were fed ad libitum to six lactating, ruminally cannulated cows in a replicated 3 x 3 Latin square. The same diets were then fed at 1.3 x maintenance intake to six gestating dry cows. Dairy milk yield and percentage and yield of milk protein and casein were higher for cows fed the low silage diet than for cows receiving other treatments. Fat percentage and yield were not different among diets. Lactating cows consumed more DM on low silage (23.0 kg/d) than on medium or high silage diets (21.4 kg), but NDF intake as percentage of BW was higher for the high silage diet. Digestibility of DM in the lactating (70.7, 69.9, and 67.5% for low, medium, and high) and dry cows (76.7, 73.5, and 69.0%, respectively) decreased as the level of silage increased. Depression in digestibility was greater as dietary concentrate increased. Cows fed the high silage diet had a faster fractional passage rate of solids and higher rumen fill. Digestion of concentrate cell walls appeared to be depressed more than alfalfa cell walls as intake increased.


Dermal contact dermatitis to a disposable electrode plate.
A 50-year-old man developed a dermal type of allergic contact dermatitis after exposure to a disposable electrosurgical electrode. Patch testing did not determine the allergenic component, but the electrode contained none of the substances known to cause dermal contact dermatitis.


Sulfonyl-containing nucleoside phosphotriesters and phosphoramidates as novel anticancer prodrugs of 5-fluoro-2'-deoxyuridine 5'-monophosphate (FdUMP).
A series of sulfonyl-containing 5-fluoro-2'-deoxyuridine (FdU) phosphotriester and phosphoramidate analogues were designed and synthesized as anticancer prodrugs of FdUMP. Stability studies have demonstrated that these compounds underwent pH dependent beta-elimination to liberate the corresponding nucleotide species with half-lives in the range of 0.33-12.23 h under model physiological conditions in 0.1 M phosphate buffer at pH 7.4 and 37 degrees C. Acceleration of the elimination was observed in the presence of human plasma. Compounds with an FdUMP moiety (4-9) were considerably more potent than those without (1-3) as well as 5-fluorouracil (5-FU) against Chinese hamster lung fibroblasts (V-79 cells) in vitro. Addition of thymidine (10 microM) reversed the growth inhibition activities of only 5-FU and the compounds with an FdUMP moiety, but had no effect on those without. These results are consistent with thymidylate synthase as the target of the prodrugs.


What is an artificial muscle? A comparison of soft actuators to biological muscles.
Interest in emulating the properties of biological muscles that allow for fast adaptability and control in unstructured environments has motivated researchers to develop new soft actuators, often referred to as 'artificial muscles'. The field of soft robotics is evolving rapidly as new soft actuator designs are published every year. In parallel, recent studies have also provided new insights for understanding biological muscles as 'active' materials whose tunable properties allow them to adapt rapidly to external perturbations. This work presents a comparative study of biological muscles and soft actuators, focusing on those properties that make biological muscles highly adaptable systems. In doing so, we briefly review the latest soft actuation technologies, their actuation mechanisms, and advantages and disadvantages from an operational perspective. Next, we review the latest advances in understanding biological muscles. This presents insight into muscle architecture, the actuation mechanism, and modeling, but more importantly, it provides an understanding of the properties that contribute to adaptability and control. Finally, we conduct a comparative study of biological muscles and soft actuators. Here, we present the accomplishments of each soft actuation technology, the remaining challenges, and future directions. Additionally, this comparative study contributes to providing further insight on soft robotic terms, such as biomimetic actuators, artificial muscles, and conceptualizing a higher level of performance actuator named artificial supermuscle. In conclusion, while soft actuators often have performance metrics such as specific power, efficiency, response time, and others similar to those in muscles, significant challenges remain when finding suitable substitutes for biological muscles, in terms of other factors such as control strategies, onboard energy integration, and thermoregulation.


What is behind the success of terrestrial bromeliads as microhabitat for the sylvatic triatomine Mepraia spinolai?
Puya alpestris, P. berteroniana and P. chilensis (Bromeliaceae) are terrestrial plants present in north-central Chile, considered important part of the shrub flora due to their abundance and close interaction with animals. A previous study showed that bromeliad cover positively correlates with the abundance of the sylvatic triatomine vector Mepraia spinolai, only when infected by the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Here, we assessed the biological interactions and abiotic conditions provided by these Puya species to M. spinolai. During the austral summers of 2015 and 2016, we sampled 17 sites with presence of M. spinolai colonies. In each site, we estimated the presence of bromeliad and its cover proportion, and placed a camera trap for three months near a M. spinolai colony to record the vertebrates potentially interacting with this triatomine. Three of the camera traps were placed right in front of M. spinolai-colonized bromeliads. At the same time, triatomines present under these bromeliads were collected to assess their blood meal by Next Generation Sequencing. Between July 2017 and January 2018, we examined the abiotic conditions (temperature and humidity) under bromeliad, under rocks and at ground level. We detected 40 vertebrate species associated to Puya spp. (18 birds, 16 mammals, and 6 reptiles). Puya spp. are a resource for keystone species in T. cruzi sylvatic transmission, including small mammals (Octodon degus, Phyllotis darwini) and lizard species (Liolaemus spp.), detecting a positive association between bromeliad presence and availability of reptiles and non-domestic mammals. Native rodents and humans were the most represented vertebrates in the diet of M. spinolai collected under bromeliads. Temperatures were more stable under bromeliad, showing lower amplitude (up to 14.3 °C) compared to under rocks (23.7 °C) or at ground level (49.6 °C). Bromeliads present in the semiarid-Mediterranean ecosystem provide feeding opportunities for triatomines, and act as buffer of abiotic conditions reducing daily thermal amplitude. The presence of bromeliads near human settlements should be carefully monitored, especially because their leaves are consumed by people and browsed by livestock present in endemic areas of Chagas disease.


[Botox in children with cerebral palsy and triceps syndrome].
Botulinum toxin (botox) therapy was used in the children with cerebral palsy. Nine children, aged 3-6 years, with "triceps-syndrome" domination in a clinical picture, have been examined and treated. Control group included 9 children who have not been treated with botox. A three-week rehabilitation course has been prescribed to all the patients. Children treated by botulinum toxin injections performed better back of the foot bending, some electroneuromyographical indices being also optimized. The results obtained imply a use of botox for treatment of children with spastic forms of cerebral palsy.


Civil Unrest in the Context of Chronic Community Violence: Impact on Maternal Depressive Symptoms.
To examine changes in maternal-child health surrounding the April 2015 civil unrest in Baltimore, Maryland, following Freddie Gray's death while in police custody. We conducted cross-sectional Children's HealthWatch surveys January 2014 through December 2015 in pediatric emergency departments and primary care clinics on maternal-child health and June 2015 through October 2015 on daily and community routines. We used trend analysis and piecewise logistic regression to examine effects of time, residential proximity moderation, and mediation analysis to assess proximity and maternal-child health relations via maternal concerns. Participants comprised 1095 mothers, 93% of whom were African American and 100% of whom had public or no insurance; 73% of participants' children were younger than 24 months. Following the unrest, prevalence of maternal depressive symptoms increased significantly in proximal, but not distal, neighborhoods (b = 0.41; 95% confidence interval [CI] = 0.03, 0.79; P = .03). Maternal concerns were elevated in proximal neighborhoods and associated with depressive symptoms; mediation through maternal concern was not significant. Five months after the unrest, depressive symptoms returned to previous levels. Civil unrest has an acute effect on maternal depressive symptoms in neighborhoods proximal to unrest. Public Health Implications. To mitigate depressive symptoms associated with civil unrest, maintain stability of community routines, screen for maternal depressive symptoms, and provide parent-child nurturing programs.


REFINING THE CENSUS OF THE UPPER SCORPIUS ASSOCIATION WITH GAIA.
We have refined the census of stars and brown dwarfs in the Upper Sco association (~ 10 Myr, ~145 pc) by 1) updating the selection of candidate members from our previous survey to include the high-precision astrometry from the second data release of Gaia, 2) obtaining spectra of a few hundred candidate members to measure their spectral types and verify their youth, and 3) assessing the membership (largely with Gaia astrometry) of 2020 stars toward Upper Sco that show evidence of youth in this work and previous studies. We arrive at a catalog of 1761 objects that are adopted as members of Upper Sco. The distribution of spectral types among the adopted members is similar to those in other nearby star-forming regions, indicating a similar initial mass function. In previous studies, we have compiled mid-infrared photometry from WISE and the Spitzer Space Telescope for members of Upper Sco and used those data to identify the stars that show evidence of circumstellar disks; we present the same analysis for our new catalog of members. As in earlier work, we find that the fraction of members with disks increases with lower stellar masses, ranging from ≲ 10% for > 1 M ⊙ to ~ 22% for 0.01-0.3 M ⊙. Finally, we have estimated the relative ages of Upper Sco and other young associations using their sequences of low-mass stars in M G RP versus G BP - G RP. This comparison indicates that Upper Sco is a factor of two younger than the β Pic association (21-24 Myr) according to both non-magnetic and magnetic evolutionary models.


Influence of the location of the cAMP receptor protein binding site on the geometry of a transcriptional activation complex in Escherichia coli.
The interactions between the cAMP receptor protein (CRP) and RNA polymerase during transcriptional activation at the Escherichia coli malT promoter have been analyzed using a combination of footprinting methods. We show that a closed complex is formed at this promoter in the absence of activator and that CRP merely stabilizes the open complex. The alpha-subunits of the RNA polymerase are involved in this effect as shown by KMnO4 footprinting. The open complex formed in the presence of CRP is structurally identical to the one found at a CRP-independent promoter up-mutant. UV-laser footprinting yields distinct signals for the different protein-DNA interactions within the complex and for interactions between CRP and RNA polymerase. We monitor these signals in promoter variants that place the CRP binding site at different distances upstream of the start site of transcription. Signals within the core promoter region, as well as those located just upstream of the -35 hexamer, are unaffected by the position of the CRP binding site. Contacts of RNA polymerase with the upstream promoter region change in a mutant RNA polymerase containing a truncated alpha-subunit. We conclude that at least one of the alpha-subunits of RNA polymerase binds to DNA upstream of the -35 hexamer and that this interaction is unaffected by the position of the CRP binding site. We discuss models that account for the different activities of CRP in transcriptional activation as a function of promoter geometry.


Soybean (Glycine max L. Merr.) sprouts germinated under red light irradiation induce disease resistance against bacterial rotting disease.
Specific wavelengths of light can exert various physiological changes in plants, including effects on responses to disease incidence. To determine whether specific light wavelength had effects on rotting disease caused by Pseudomonas putida 229, soybean sprouts were germinated under a narrow range of wavelengths from light emitting diodes (LEDs), including red (650-660), far red (720-730) and blue (440-450 nm) or broad range of wavelength from daylight fluorescence bulbs. The controls were composed of soybean sprouts germinated in darkness. After germination under different conditions for 5 days, the soybean sprouts were inoculated with P. putida 229 and the disease incidence was observed for 5 days. The sprouts exposed to red light showed increased resistance against P. putida 229 relative to those grown under other conditions. Soybean sprouts germinated under red light accumulated high levels of salicylic acid (SA) accompanied with up-regulation of the biosynthetic gene ICS and the pathogenesis- related (PR) gene PR-1, indicating that the resistance was induced by the action of SA via de novo synthesis of SA in the soybean sprouts by red light irradiation. Taken together, these data suggest that only the narrow range of red light can induce disease resistance in soybean sprouts, regulated by the SA-dependent pathway via the de novo synthesis of SA and up-regulation of PR genes.


Ring test evaluation of the detection of influenza A virus in swine oral fluids by real-time reverse-transcription polymerase chain reaction and virus isolation.
The probability of detecting influenza A virus (IAV) in oral fluid (OF) specimens was calculated for each of 13 assays based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) and 7 assays based on virus isolation (VI). The OF specimens were inoculated with H1N1 or H3N2 IAV and serially diluted 10-fold (10(-1) to 10(-8)). Eight participating laboratories received 180 randomized OF samples (10 replicates × 8 dilutions × 2 IAV subtypes plus 20 IAV-negative samples) and performed the rRT-PCR and VI procedure(s) of their choice. Analysis of the results with a mixed-effect logistic-regression model identified dilution and assay as variables significant (P < 0.0001) for IAV detection in OF by rRT-PCR or VI. Virus subtype was not significant for IAV detection by either rRT-PCR (P = 0.457) or VI (P = 0.101). For rRT-PCR the cycle threshold (Ct) values increased consistently with dilution but varied widely. Therefore, it was not possible to predict VI success on the basis of Ct values. The success of VI was inversely related to the dilution of the sample; the assay was generally unsuccessful at lower virus concentrations. Successful swine health monitoring and disease surveillance require assays with consistent performance, but significant differences in reproducibility were observed among the assays evaluated.


Diabetes Induces Aberrant DNA Methylation in the Proximal Tubules of the Kidney.
Epigenetic mechanisms may underlie the progression of diabetic kidney disease. Because the kidney is a heterogeneous organ with different cell types, we investigated DNA methylation status of the kidney in a cell type-specific manner. We first identified genes specifically demethylated in the normal proximal tubules obtained from control db/m mice, and next delineated the candidate disease-modifying genes bearing aberrant DNA methylation induced by diabetes using db/db mice. Genes involved in glucose metabolism, including Sglt2, Pck1, and G6pc, were selectively hypomethylated in the proximal tubules in control mice. Hnf4a, a transcription factor regulating transporters for reabsorption, was also selectively demethylated. In diabetic mice, aberrant hypomethylation of Agt, Abcc4, Cyp4a10, Glut5, and Met and hypermethylation of Kif20b, Cldn18, and Slco1a1 were observed. Time-dependent demethylation of Agt, a marker of diabetic kidney disease, was accompanied by histone modification changes. Furthermore, inhibition of DNA methyltransferase or histone deacetylase increased Agt mRNA in cultured human proximal tubular cells. Aberrant DNA methylation and concomitant changes in histone modifications and mRNA expression in the diabetic kidney were resistant to antidiabetic treatment with pioglitazone. These results suggest that an epigenetic switch involving aberrant DNA methylation causes persistent mRNA expression of select genes that may lead to phenotype changes of the proximal tubules in diabetic kidney disease.


Immediate response of mammalian target of rapamycin (mTOR)-mediated signalling following acute resistance exercise in rat skeletal muscle.
The purpose of the present investigation was to determine whether mammalian target of rapamycin (mTOR)-mediated signalling and some key regulatory proteins of translation initiation are altered in skeletal muscle during the immediate phase of recovery following acute resistance exercise. Rats were operantly conditioned to reach an illuminated bar located high on a Plexiglass cage, such that the animals completed concentric and eccentric contractions involving the hindlimb musculature. Gastrocnemius muscle was extracted immediately after acute exercise and 5, 10, 15, 30 and 60 min of recovery. Phosphorylation of protein kinase B (PKB) on Ser-473 peaked at 10 min of recovery (282% of control, P < 0.05) with no significant changes noted for mTOR phosphorylation on Ser-2448. Eukaryotic initiation factor (eIF) 4E-binding protein-1 (4E-BP1) and S6 kinase-1 (S6K1), both downstream effectors of mTOR, were altered during recovery as well. 4E-BP1 phosphorylation was significantly elevated at 10 min (292%, P < 0.01) of recovery. S6K1 phosphorylation on Thr-389 demonstrated a trend for peak activation at 10 min following exercise (336%, P = 0.06) with ribosomal protein S6 phosphorylation being maximally activated at 15 min of recovery (647%, P < 0.05). Components of the eIF4F complex were enhanced during recovery as eIF4E association with eIF4G peaked at 10 min (292%, P < 0.05). Events regulating the binding of initiator methionyl-tRNA to the 40S ribosomal subunit were assessed through eIF2B activity and eIF2 alpha phosphorylation on Ser-51. No differences were noted with either eIF2B or eIF2 alpha. Collectively, these results provide strong evidence that mTOR-mediating signalling is transiently upregulated during the immediate period following resistance exercise and this response may constitute the most proximal growth response of the cell.


Simultaneous imaging using Si-PM-based PET and MRI for development of an integrated PET/MRI system.
The silicon photomultiplier (Si-PM) is a promising photo-detector for PET for use in magnetic resonance imaging (MRI) systems because it has high gain and is insensitive to static magnetic fields. Recently we developed a Si-PM-based depth-of-interaction PET system for small animals and performed simultaneous measurements by combining the Si-PM-based PET and the 0.15 T permanent MRI to test the interferences between the Si-PM-based PET and an MRI. When the Si-PM was inside the MRI and installed around the radio frequency (RF) coil of the MRI, significant noise from the RF sequence of the MRI was observed in the analog signals of the PET detectors. However, we did not observe any artifacts in the PET images; fluctuation increased in the count rate of the Si-PM-based PET system. On the MRI side, there was significant degradation of the signal-to-noise ratio (S/N) in the MRI images compared with those without PET. By applying noise reduction procedures, the degradation of the S/N was reduced. With this condition, simultaneous measurements of a rat brain using a Si-PM-based PET and an MRI were made with some degradation in the MRI images. We conclude that simultaneous measurements are possible using Si-PM-based PET and MRI.


Myosin heavy chain IIX overshoot in human skeletal muscle.
The distribution of myosin heavy chain (MHC) isoforms, fiber type composition, and fiber size of the vastus lateralis muscle were analyzed by sodium dodecylsulfate polymerase gel electrophoresis (SDS-PAGE), ATPase histochemistry, and immunocytochemistry in a group of adult sedentary men before and after 3 months of heavy-load resistance training and, subsequently, after 3 months of detraining. Following the period of resistance training, MHC IIX content decreased from 9.3 +/- 2.1% to 2.0 +/- 0.8% (P < 0.01), with a corresponding increase in MHC IIA (42.4 +/- 3.9% vs. 49.6 +/- 4.0% [P < 0.05]). Following detraining the amount of MHC IIX reached values that were higher than before and after resistance training (17.2 +/- 3.2% [P < 0.01]). Changes in fiber type composition resembled the changes observed in MHC isoform content. Significant hypertrophy was observed for the type II fibers after resistance training. Maximal isometric quadriceps strength increased after resistance training, but returned to pretraining levels after detraining. The present results suggest that heavy-load resistance training decreases the amount of MHC IIX while reciprocally increasing MHC IIA content. Furthermore, detraining following heavy-load resistance training seems to evoke an overshoot in the amount of MHC IIX to values markedly higher than those observed prior to resistance training.


Radio-ulnar joint supinates around ball release during baseball fastball pitching.
This study was conducted to determine whether a supination phase of the forearm exists around ball release (BRL), and, if present, to determine whether this supination is explained by a reaction force or by the body configuration required for this task. A 16-camera motion analysis system with a sampling frequency of 1,000 Hz recorded 20 healthy male semi-professional pitchers pitching from an indoor pitching mound. A short supination phase around BRL was confirmed for all participants in the current study. Correlation analyses revealed that the supination angle at BRL had significant relationships with several measurements of shoulder movement kinematics. Mechanical work analysis of the forearm's longitudinal axis revealed several variations in joint power curve and various patterns of mechanical work among the participants, suggesting that a reaction force originating from accelerating a ball might not be the main cause of supination. The raw data also were down-sampled to a sampling frequency of 250 Hz, to match previous studies and to investigate the discrepancy among previous studies concerning the existence of the supination phase. The experience of participants and methodological differences, such as the definition of BRL and the time-normalisation technique, influenced whether the supination phase was observed.


Feasibility and Acceptability of a Multidisciplinary Academic Telemedicine System for Memory Care in Response to COVID-19.
In response to the restrictions imposed by the COVID-19 pandemic, the University of California San Francisco Memory and Aging Center (UCSF MAC) has deployed a comprehensive telemedicine model for the diagnosis and management of Alzheimer disease and related dementias. This review summarizes a large academic behavioral neurology clinic's experience transitioning to telemedicine services, including the impact on clinic care indicators, access metrics, and provider's experience. We compared these outcomes from 3 years before COVID-19 to 12 months after the transition to video teleconferencing (VTC) encounters. Patient demographics and appointment data (dates, visit types, and departments) were extracted from our institution's electronic health record database from January 1, 2017, to May 1, 2021. We present data as descriptive statistics and comparisons using Wilcoxon rank-sum tests and Fisher exact tests. The results of anonymous surveys conducted among the clinic's providers are reported as descriptive findings. After the implementation of telemedicine services, the proportion of clinic encounters completed via VTC increased from 1.9% to 86.4%. There was a statistically significant decline in both the percentage of scheduled appointments that were canceled (32.9% vs 27.9%; p < 0.01) and total cancelations per month (mean 240.3 vs 179.4/mo; p < 0.01). There was an increase in the percentage of completed scheduled appointments (60.2% vs 64.8%; p < 0.01) and an increase in the average estimated commuting distance patients would need to drive for follow-up appointments (mean 49.8 vs 54.7 miles; p < 0.01). The transition to telemedicine services did not significantly affect the clinic's patient population as measured by age, gender, estimated income, area deprivation index, or self-reported racial/ethnic identity. The results of the provider survey revealed that physicians reported a more positive experience relative to neuropsychologists. Both types of providers reported telemedicine services as a reasonable equivalent and acceptable alternative to in-person evaluations with notable caveats. UCSF MAC's comprehensive integration of telemedicine services maintained critical ambulatory care to patients living with dementia during the COVID-19 pandemic. The recognized benefits of our care model suggest dementia telemedicine may be used as a feasible and equivalent alternative to in-person ambulatory care in the after COVID-19 era.


Drugs associated with teratogenic mechanisms. Part II: a literature review of the evidence on human risks.
What is the current state of knowledge on the human risks of drugs suspected to be associated with teratogenic mechanisms? Evidence for the presence or absence of human risks of birth defects is scarce or non-existent for the majority of drugs associated with teratogenic mechanisms. Medical drugs suspected to be associated with teratogenic mechanisms are dispensed to a significant proportion of women in the first trimester of pregnancy. However, an overview of the current state of knowledge on the human teratogenic effects of these drugs is lacking. We performed an extensive literature review of studies in the English language which examined the associations between selected drugs and specific birth defects. The literature was identified from MEDLINE and EMBASE from database inception (January 1946 and January 1974, respectively) through December 2012 using 287 terms for the drugs of interest. We only included studies if they specified birth defect subtypes and, specifically for cohort studies, involved live born infants. Of 14 406 potentially relevant articles, 556 full-text articles were assessed for eligibility and 250 met the inclusion criteria. The studies included were divided into four categories according to their design to increase the validity of our study. Epidemiologic studies assessing teratogenic risks were identified for less than half of the drugs included in the review. A substantial variation in study design and data collection methods was observed. When the data collection method is of questionable validity, study quality may be affected considerably. For only 15 drugs of interest, birth defects were assessed in at least 1000 infants in cohort studies, and 13 of these were associated with one or more specific birth defects. The majority of associations observed in case-control studies are as yet unconfirmed. For most drugs and drug groups, however, the numbers of exposed infants studied were too small to draw any conclusions regarding their human teratogenic risks. The validity of our review is limited by the validity and reporting of the studies from which the data were extracted. Some relevant studies might have been missed owing to the exclusion of articles not in the English language and publication bias. It is a cause of concern that the drugs most often dispensed in the first trimester of pregnancy are not necessarily the drugs for which teratogenic risks have been studied. Future studies should focus on those drugs that are most commonly used during pregnancy and for which the teratogenic risks are unknown, such as iron preparations, serotonin receptor agonists or antagonists, drugs used in fertility treatment, dihydrofolate reductase inhibitors. Marleen van Gelder was supported by the Netherlands Organisation for Scientific Research/NWO (grant no. 021.001.008). No competing interests are declared. N/A.


Abscopal effect following nivolumab induction in a patient with metastatic renal cell carcinoma-unique pathological features of the primary specimen: A case report.
The case of a patient with metastatic renal cell carcinoma who exhibited the abscopal effect following treatment by anti-programmed death-1 (PD-1) antibody is presented. A 40-year-old woman was diagnosed with an 8.2-cm renal tumor without distant metastases, and radical nephrectomy was subsequently performed. Pathological examination revealed a clear cell renal cell carcinoma. At 3 months after surgery, the patient developed one lung metastasis. Following treatment with interferon and three types of tyrosine kinase inhibitors, anti-PD1 antibody (nivolumab) was started. During the treatment, para-aortic/supraclavicular lymph nodes and several lung lesions remained, although other lesions decreased markedly. The patient was subsequently treated by palliative radiotherapy to the para-aortic and supraclavicular lymph nodes for pain control. After the radiotherapy, the lung lesions previously refractory to nivolumab started to decrease, probably due to an abscopal effect. Additionally, the laboratory data and Karnofsky Performance Status improved. Histological re-examination of the primary lesion revealed heterogeneity of the immunological microenvironment, which may be associated with the heterogeneity of treatment sensitivity.


Short-term and long-term attentional biases to frequently encountered target features.
It has long been known that frequently occurring targets are attended better than infrequent ones in visual search. But does this frequency-based attentional prioritization reflect momentary or durable changes in attention? Here we observed both short-term and long-term attentional biases for visual features as a function of different types of statistical associations between the targets, distractors, and features. Participants searched for a target, a line oriented horizontally or vertically among diagonal distractors, and reported its length. In one set of experiments we manipulated the target's color probability: Targets were more often in Color 1 than in Color 2. The distractors were in other colors. Participants found Color 1 targets more quickly than Color 2 targets, but this preference disappeared immediately when the target's color became random in the subsequent testing phase. In the other set of experiments, we manipulated the diagnostic values of the two colors: Color 1 was more often a target than a distractor; Color 2 was more often a distractor than a target. Participants found Color 1 targets more quickly than Color 2 targets. Importantly, and in contrast to the first set of experiments, the featural preference was sustained in the testing phase. These results suggest that short-term and long-term attentional biases are products of different statistical information. Finding a target momentarily activates its features, inducing short-term repetition priming. Long-term changes in attention, on the other hand, may rely on learning diagnostic features of the targets.


Perspectives of a mobile application for people with communication disabilities in the community.
Purpose To determine the perceptions of people with complex communication needs (CCN) and business staff regarding the uses and functionality of a mobile application to aid communication access. Method A qualitative study using thematic analysis of transcripts and field notes from focus groups and interviews of 19 people with CCN and nine business staff. Results Four themes and 10 subthemes were drawn from the data. Themes highlighted the desire for: increased communication strategies to support customer interactions, increased access to information, functionality of a mobile application to increase its utility, and preferred technical and visual features of mobile applications. Conclusion People with CCN and business staff perceived a mobile application as a useful tool to aid communication access. This research highlighted the importance of facilitating strategies to communicative interactions and information in the community as the fundamental goal of a mobile application developed to support communication access. Implications for Rehabilitation Mobile applications are widely accepted and used in modern customer service industries and have been identified as tools to increase communication access for people with complex communication needs (CCN). People with CCN identified accessibility, presentation, and customisation as important features of mobile applications for communication access. The diversity of user preferences and needs, and the rapid development of new technologies limit the applicability of a single design for mobile applications for people with CCN. People with CCN should be involved in application design and development. A mobile application for communication access would support customer-business interactions as well as enable more accessible information sharing about disability needs and services.


Effective protective agents against organ toxicity of deoxynivalenol and their detoxification mechanisms: A review.
Deoxynivalenol (DON) is one of the most prevalent mycotoxins in feed, which causes organ toxicity in animals. Therefore, reducing DON-induced organ toxicity can now be accomplished effectively using protective agents. This review provides an overview of multiple studies on a wide range of protective agents and their molecular mechanisms against DON organ toxicity. Protective agents include plant extracts, yeast products, bacteria, peptides, enzymes, H2, oligosaccharides, amino acids, adsorbents, vitamins and selenium. Among these, biological detoxification of DON using microorganisms to reduce the toxicity of DON without affecting the growth performance of pigs may be the most promising detoxification strategy. This paper also evaluates future developments related to DON detoxification and discusses the detoxification role and application potential of protective agents. This paper provides new perspectives for future research and development of safe and effective feed additives.


[The physician's humanizing role. Philosophical view of socio-anthropological work in medicine].
The paper presents a critical analysis of current society making emphasis on the human needs to generate domain among equals, as well as the sense of competition and division that this creates. It addresses the process of dehumanization that the same civilization has generated focusing on laboral paradigm of our days and the weak social justice that unequal wealth distribution has generated. It has been seen that health professionals involved in such context, have not escaped socio-cultural pressure and postmodern neo-liberal position. The paper discusses about the humanizing role health professionals have in society, and the relevance of the philosophical exercise in their acts above human frailty. Finally, it is suggested an alternative for repositioning to himself and society.


Enzymatic oxidation of 2-phenylethylamine to phenylacetic acid and 2-phenylethanol with special reference to the metabolism of its intermediate phenylacetaldehyde.
2-phenylethylamine is an endogenous constituent of the human brain and is implicated in cerebral transmission. This bioactive amine is also present in certain foodstuffs such as chocolate, cheese and wine and may cause undesirable side effects in susceptible individuals. Metabolism of 2-phenylethylamine to phenylacetaldehyde is catalysed by monoamine oxidase B but the oxidation to its acid is usually ascribed to aldehyde dehydrogenase and the contribution of aldehyde oxidase and xanthine oxidase, if any, is ignored. The objective of this study was to elucidate the role of the molybdenum hydroxylases, aldehyde oxidase and xanthine oxidase, in the metabolism of phenylacetaldehyde derived from its parent biogenic amine. Treatments of 2-phenylethylamine with monoamine oxidase were carried out for the production of phenylacetaldehyde, as well as treatments of synthetic or enzymatic-generated phenylacetaldehyde with aldehyde oxidase, xanthine oxidase and aldehyde dehydrogenase. The results indicated that phenylacetaldehyde is metabolised mainly to phenylacetic acid with lower concentrations of 2-phenylethanol by all three oxidising enzymes. Aldehyde dehydrogenase was the predominant enzyme involved in phenylacetaldehyde oxidation and thus it has a major role in 2-phenylethylamine metabolism with aldehyde oxidase playing a less prominent role. Xanthine oxidase does not contribute to the oxidation of phenylacetaldehyde due to low amounts being present in guinea pig. Thus aldehyde dehydrogenase is not the only enzyme oxidising xenobiotic and endobiotic aldehydes and the role of aldehyde oxidase in such reactions should not be ignored.


Delayed gastric emptying after palliative gastrojejunostomy for carcinoma of the pancreas.
Fifty-seven patients with carcinoma of the pancreas underwent gastrojejunostomy (GJ) alone or in conjunction with biliary bypass. The mortality rate for GJ alone was 18%; for the combined biliary and duodenal bypass operation it was 5%. Fifteen patients (26%) had delayed gastric emptying (DGE) postoperatively for periods extending from nine to 31 days (average, 16 days); five patients (33%) died. Eight (57%) of 14 patients with preoperative duodenal obstruction and five (42%) of 12 patients with retrocolic GJ experienced DGE postoperatively. Stomal diameter in the patients with DGE averaged 6.5 cm, and 8.4 cm was the average in those without DGE. We conclude that DGE is a frequent and serious problem after GJ for patients with unresectable pancreatic cancer.


Nitrogen-corrected True Metabolizable Energy and Amino Acid Digestibility of Chinese Corn Distillers Dried Grains with Solubles in Adult Cecectomized Roosters.
This study was conducted to evaluate chemical composition, nitrogen-corrected true metabolizable energy (TMEn) and true amino acids digestibility of corn distillers dried grains with solubles (DDGS) produced in China. Twenty five sources of corn DDGS was collected from 8 provinces of China. A precision-fed rooster assay was used to determine TMEn and amino acids digestibility with 35 adult cecectomized roosters, in which each DDGS sample was tube fed (30 g). The average content of ash, crude protein, total amino acid, ether extract, crude fiber and neutral detergent fiber were 4.81, 27.91, 22.51, 15.22, 6.35 and 37.58%, respectively. TMEn of DDGS ranged from 1,779 to 3,071 kcal/kg and averaged 2,517 kcal/kg. Coefficient of variation for non-amino acid crude protein, ether extract, crude fiber and TMEn were 55.0, 15.7, 15.9 and 17.1%, respectively. The average true amino acid digestibility was 77.32%. Stepwise regression analysis obtained the following equation: TMEn, kcal/kg = -2,995.6+0.88×gross energy+49.63×a* (BIC = 248.8; RMSE = 190.8; p<0.01). Removing gross energy from the model obtained the following equation: TMEn, kcal/kg = 57.88×ether extracts+87.62×a* (BIC = 254.3, RMSE = 223.5; p<0.01). No correlation was found between color scores and lysine true digestibility (p>0.05). These results suggest that corn DDGS produced in China has a large variation in chemical composition, and gross energy and a* value can be used to generate TMEn predict equation.


Recent approaches of lipid-based delivery system for lymphatic targeting via oral route.
Lymphatic system is a key target in research field due to its distinctive makeup and huge contributing functions within the body. Intestinal lymphatic drug transport (chylomicron pathway) is intensely described in research field till date because it is considered to be the best for improving oral drug delivery by avoiding first pass metabolism. The lymphatic imaging techniques and potential therapeutic candidates are engaged for evaluating disease states and overcoming these conditions. The novel drug delivery systems such as self-microemulsifying drug delivery system, nanoparticles, liposomes, nano-lipid carriers, solid lipid carriers are employed for delivering drugs through lymphatic system via various routes such as subcutaneous route, intraperitoneal route, pulmonary route, gastric sub-mucosal injection, intrapleural and intradermal. Among these colloidal particles, lipid-based delivery system is considered to be the best for lymphatic delivery. From the last few decades, mesenteric lymph duct cannulation and thoracic lymph duct cannulation are followed to assess lymphatic uptake of drugs. Due to their limitations, chylomicrons inhibitors and in-vitro models are employed, i.e. lipolysis model and permeability model. Currently, research on this topic still continues and drainage system used to deliver the drugs against lymphatic disease as well as targeting other organs by modulating the chylomicron pathway.


Necessity of Bumped Kinase Inhibitor Gastrointestinal Exposure in Treating Cryptosporidium Infection.
There is a substantial need for novel therapeutics to combat the widespread impact caused by Crytosporidium infection. However, there is a lack of knowledge as to which drug pharmacokinetic (PK) characteristics are key to generate an in vivo response, specifically whether systemic drug exposure is crucial for in vivo efficacy. To identify which PK properties are correlated with in vivo efficacy, we generated physiologically based PK models to simulate systemic and gastrointestinal drug concentrations for a series of bumped kinase inhibitors (BKIs) that have nearly identical in vitro potency against Cryptosporidium but display divergent PK properties. When BKI concentrations were used to predict in vivo efficacy with a neonatal model of Cryptosporidium infection, these concentrations in the large intestine were the sole predictors of the observed in vivo efficacy. The significance of large intestinal BKI exposure for predicting in vivo efficacy was further supported with an adult mouse model of Cryptosporidium infection. This study suggests that drug exposure in the large intestine is essential for generating a superior in vivo response, and that physiologically based PK models can assist in the prioritization of leading preclinical drug candidates for in vivo testing.


Pb deposition on I-coated Au(111). UHV-EC and EC-STM studies.
This article concerns the growth of an atomic layer of Pb on the Au(111)( radical3 x radical3)R30 degrees -I structure. The importance of this study lies in the use of Pb underpotential deposition (UPD) as a sacrificial layer in surface-limited redox replacement (SLRR). SLRR reactions are being applied in the formation of metal nanofilms via electrochemical atomic layer deposition (ALD). Pb UPD is a surface-limited reaction, and if it is placed in a solution of ions of a more noble metal, redox replacement can occur, but limited by the amount of Pb present. Pb UPD is a candidate for use as a sacrificial layer for replacement by any more noble element. It has been used by this group for both Cu and Pt nanofilm formation using electrochemical ALD. The I atom layer was intended to facilitate electrochemical annealing during nanofilm growth. Two distinctly different Pb atomic layer structures are reported, studied using in situ scanning tunneling microscopy (STM) with an electrochemical flow cell and ultrahigh vacuum surface analysis combined directly with electrochemical reactions (UHV-EC). Starting with the initial Au(111)( radical3 x radical3)R30 degrees -I, 1/3 monolayer of I on the Au(111) surface, Pb deposition began at approximately 0.1 V. The first Pb UPD structure was observed just below -0.2 V and displayed a (2 x radical3)-rect unit cell, for a structure composed of 1/4 monolayer each of Pb and I. The I atoms fit in Pb 4-fold sites, on the Au(111) surface. The structure was present in domains rotated by 120 degrees. Deposition to -0.4 V resulted in complete loss of the I atoms and formation of a Pb monolayer on the Au(111), which produced a Moiré pattern, due to the Pb and Au lattice mismatch. These structures represent two well-defined starting points for the growth of nanofilms of other more noble elements. It is apparent from these studies that the adsorption of I- on Pb is weak, and it will rinse away. If Pb is used as a sacrificial metal in an electrochemical ALD cycle and adsorbed I atoms are employed for electrochemical annealing, I atoms will need to be applied each cycle.


[A pharmacokinetic and clinical evaluation of ceftazidime in neonates and premature infants. A study of ceftazidime in the perinatal co-research group].
Ceftazidime (CAZ) was evaluated for its pharmacokinetics and clinical usefulness in neonates and premature infants. The results obtained were summarized below. Following intravenous injection of CAZ 10 or 20 mg/kg to neonates and premature infants, dose response was observed in serum concentrations ranging from 5.1 to 21.9 micrograms/ml at 6 hours after the injection. The serum half-life tended to be longer in premature infants than in neonates; the half-life being longer for an infant with lower day-age. Urinary recovery rates during the first 6 hours after single administrations of 10 mg/kg of CAZ tended to be higher in neonates than in premature infants, and higher rates were observed in older infants. However, no noticeable difference was observed after the administration of CAZ 20 mg/kg. Clinical efficacy was evaluated in 99 neonates and 55 premature infants (156 infections), daily doses ranging from 21.1 to 246.4 mg/kg. Out of 105 cases of common infections, mainly 44 cases with causative organisms identified (including 17 of sepsis, 7 of pneumonia, 4 of purulent meningitis, 11 of urinary tract infections) were examined for the clinical efficacy. The efficacy of CAZ was excellent in 21, good in 18, fair in 1 and poor in 4, with the efficacy rate of 88.6%. In the remaining 61 cases, i.e., 37 with causative organisms unknown and 24 with signs of intrauterine infections, the efficacy rate was 95.1%. Other than these cases, additional 51 cases were given CAZ solely for prophylaxis of infections, and the results were found satisfactory. On the whole, clinical efficacy rate of CAZ was 94.9% in 156 cases. Out of the 44 cases examined for bacteriological responses, 38 were evaluated as 'eradicated', 3 'persisted' and 3 'unknown' with eradication rate of 92.7%. Replacement of organisms (superinfection) was observed in 3 cases. Out of 179 cases in which adverse effects were assessable, adverse effects were observed in a total of 4 cases (2.2%), i.e., 3 cases of diarrhea (1.7%) and 1 case of rash (0.6%), and abnormal laboratory findings were observed in a total of 14 cases (7.8%), i.e., increase in eosinophiles count in 8 (4.5%), elevation of GOT in 3 (1.7%), increase in platelet, elevation of GOT . GPT, and elevation of GOT . GPT . BUN in 1 case each (0.6%). None of them were severe and they were transient. Elevations of bilirubin and cases of positive PIVKA II associated with CAZ were not observed.(ABSTRACT TRUNCATED AT 400 WORDS)


Evaluation of three rapid RNA extraction reagents: relevance for use in RT-PCR's and measurement of low level gene expression in clinical samples.
Although peripheral blood and bone marrow are usually readily available from patients, present techniques of RNA extraction are tedious, require millilitres of starting material and removal of red blood cells before RNA purification. Further, successful reverse transcriptase polymerase chain reaction (RT-PCR) amplification requires the removal of haemoglobin derivatives which interfere with the PCR process. Recently, one step rapid use reagents have become available, claiming to be useful for obtaining high quality RNA from microlitre quantities of whole blood drawn directly from the patient. Their use to date in clinical samples appears limited with little information in the literature documented. In an attempt to overcome this, we tested the Trizol-LS, RNA-STAT-50 and Ultraspec-3 reagents upon a statistically significant number of clinical isolates of fresh and cryopreserved peripheral blood, bone marrow, blood apheresis products and a breast cancer cell line (MCF7) in order to evaluate whether these methods could be applied to routine laboratory use in an RT-PCR method capable of detecting rare gene expression. Our findings showed that there was some variation in the quality of RNA extracted which was indicated by absorbance spectrophotometry at 260 and 280 nm. 1% agarose gel electrophoresis showed that each of these methods could yield total RNA capable of generating the signature 18S and 28S rRNA bands. Using the Kruskal-Wallis non-parametric anova test combined with Dunn's multiple comparison test, the only statistically significant difference (p<0.05) indicated that Trizol-LS was more reliable than RNA-STAT-50-LS and Ultraspec-3 at extracting RNA from fresh peripheral blood. RNA extracted with the Trizol-LS and RNA STAT-50 reagents was successfully amplified in a multiplex RT-PCR reaction for detection of the multi-drug resistance related genes MDR1, the multi-drug resistance related protein (MRP) and topoisomerase IIalpha. Low level MDR1 gene expression could be detected in frozen whole blood. However, PCR products were only seen when the anti-coagulant heparin was removed from all samples prior to cDNA production. RT-PCR amplification was not 100% successful with RNA extracted with Ultraspec-3 reagent. In conclusion, we found that the RNA extracted from whole blood with the Trizol-LS and the RNA-STAT-50 are suitable for use in clinically relevant molecular biology protocols that analyze rare event genes without further purification. Our results indicated that the Trizol-LS reagent was generally more consistent in obtaining a pure and sufficient quantity of RNA from patient material as shown by the mean result of purity and quantity in comparison to either Ultraspec-3 or RNA-STAT-50-LS reagents. Ultraspec-3 is not easily suited for direct use with whole blood products.


Stoppa Approach for Anterior Plate Fixation in Unstable Pelvic Ring Injury.
The Stoppa (intrapelvic) approach has been introduced for the treatment of pelvic-acetabular fractures; it allows easy exposure of the pelvic brim, where the bone quality is optimal for screw fixation. The purpose of our study was to investigate the surgical outcomes of unstable pelvic ring injuries treated using the Stoppa approach for stable anterior ring fixation. We analyzed 22 cases of unstable pelvic ring injury treated with plate fixation of the anterior ring with the Stoppa approach. We excluded cases of nondisplaced rami fracture, simple symphyseal diastasis, and parasymphyseal fractures, which can be easily treated with other techniques. The average age of the study patients was 41 years (range, 23 to 61 years). There were 10 males and 12 females. According to the Young and Burgess classification, there were 12 lateral compression, 4 anteroposterior compression, and 6 vertical shear fracture patterns. The fracture location on the anterior ring was near the iliopectineal eminence in all cases and exposure of the pelvic brim was required for plate fixation. All patients were placed in the supine position. For anterior plate fixation, all screws were applied to the anterior ramus distally and directed above the hip joint proximally. Radiologic outcomes were assessed by union time and quality of reduction by Matta method. The Merle d'Aubigne-Postel score was used to evaluate the functional results. The average radiologic follow-up period was 16 months (range, 10 to 51 months). All fractures united at an average of 3.5 months (range, 3 to 5 months). According to the Matta method, the quality of reduction was classified as follows: 16 anatomical (73%) and 6 nearly anatomical (27%) reductions. There were no cases of screw or implant loosening before bone healing. The functional results were classified as 7 excellent (32%), 12 good (55%), and 3 fair (13%) by the Merle d'Aubigne-Postel score. There were no wound complications, neurovascular injuries, or other complications related to the surgical approach. Stable anterior ring fixation placed via the Stoppa approach can result in excellent reduction and stable screw fixation with a low complication rate.


Association of Concomitant Use of Cholinesterase Inhibitors or Memantine With Cognitive Decline in Alzheimer Clinical Trials: A Meta-analysis.
Clinical trials in Alzheimer disease (AD) generally allow participants to continue receiving concomitant medications, including cholinesterase inhibitors (ChEIs) and memantine, if the dose is stable. Previous analysis of observational studies indicates such individuals experience greater rate of decline on cognitive testing than those not receiving such medications. To investigate whether concomitant use of ChEIs or memantine is associated with cognitive outcomes in AD clinical trials. Meta-database of 18 studies from the Alzheimer Disease Cooperative Study and Alzheimer Disease Neuroimaging Initiative. All studies with data on ChEI and memantine use that included assessment of specified outcome measures. The analysis estimated annual rate of decline on the Alzheimer Disease Assessment Scale-cognitive subscale (ADAS-cog) using linear mixed-effects models, and compared rates for participants receiving ChEIs and memantine, alone and combined, with participants not receiving either medication using random-effects meta-analysis. Annual rate of change on the ADAS-cog. Across 10 studies, of 2714 participants, the mean (SD) age was 75.0 (8.2) years, 58% were female, and 9% were racial/ethnic minorities. There were 906 participants (33.4%) receiving ChEIs, 143 (5.3%) receiving memantine, 923 (34.0%) receiving both, and 742 (27.3%) receiving neither. Meta-analysis showed those receiving ChEIs or memantine were associated with significantly greater annual rate of decline on the ADAS-cog than those receiving neither medication (1.4 points/y; 95% CI, 0.1-2.7). Similar to observational studies, many participants in AD clinical trials receiving ChEIs or memantine experience greater cognitive decline. This difference is nearly as large as the hypothesized effect sizes of the treatments investigated in the trials. Concomitant use of ChEIs or memantine may be confounded with outcomes on the ADAS-cog and should be considered in design of clinical trials of potential therapeutic agents for AD. Post hoc analyses stratifying by ChEIs or memantine must be interpreted cautiously given the potential for confounding.


Adoptive transfer of CD8(+) T cells from transforming growth factor beta receptor type II (dominant negative form) induces autoimmune cholangitis in mice.
We recently reported that mice with a T cell-restricted expression of a dominant negative form of transforming growth factor beta receptor type II (dnTGFbetaRII) spontaneously develop autoimmune cholangitis that resembles human primary biliary cirrhosis (PBC), including antimitochondrial antibodies (AMAs) and extensive portal CD4(+) and CD8(+) lymphocytic infiltrates. On the basis of these data, we performed a series of experiments to determine whether the pathology was secondary to direct dnTGFbetaRII disruption of the liver and/or alternatively the appearance of autoreactive T cells. First, using dnTGFbetaRIIRag1(-/-) mice, we noted a normal hepatic and biliary structure. Hence, we performed a rigorous series of adoptive transfer studies, transferring Ly5.1(+) unfractionated spleen cell CD4(+) or CD8(+) T cells from dnTGFbetaRII mice into B6/Rag(-/-) (Ly 5.2) recipients. In unmanipulated dnTGFbetaRII mice, there was a marked increase in CD4(+) and CD8(+) T cell biliary infiltrates with AMA. Indeed, B6/Rag(-/-) recipients of dnTGFbetaRII unfractionated cells develop features of liver disease similar to PBC, suggesting that splenic loss of self-tolerance alone is sufficient to cause disease in this model and therefore that there is no specific abnormality in the biliary targets required for appearance of disease. More importantly, adoptive transfer of CD8(+) but not CD4(+) T cells into B6/Rag(-/-) mice led to liver histopathology remarkably similar to PBC, emphasizing a prominent role for CD8 T cell-mediated pathogenesis. In contrast, B6/Rag(-/-) recipients of CD4(+) T cells from dnTGFbetaRII mice predominantly developed inflammatory bowel disease associated with higher levels of serum interferon gamma and tumor necrosis factor alpha. These data suggest that in this model of PBC, autoreactive CD8(+) cells destroy bile ducts.


Prevention of contrast-induced nephropathy with Na/K citrate.
Contrast-induced nephropathy (CIN) is a frequent complication of many radiological procedures involving the application of contrast media. It represents a significant health problem that causes the increase in mortality, morbidity, and medical costs. For the prevention of CIN, a number of methods have been proposed to be effective. Among them, alkalinization of urine takes an important place. Although the Na/K citrate is a well-known agent for urine alkalinization, it has not been studied in the prevention of CIN. Two hundred and two patients who underwent coronary angiography were included in the study. They were randomized into groups receiving the drug Na/K citrate per os and to the control group. Serum creatinine and glomerular filtration rate were determined in all patients immediately before coronary angiography, and 48 h after the procedure. CIN criteria were a creatinine increase of >25%, reduction in the glomerular filtration rate by >25%, or an increase in serum creatinine of >44 μmol/L. The incidence of CIN in the group receiving Na/K citrate was significantly lower when compared with the control group (4% compared with 20%, P = 0.0001). Patients who had a urine pH <6 had a more than ten-fold higher incidence of contrast nephropathy compared with patients whose urine pH was >6. Alkalinization of urine using the Na/K citrate may reduce the incidence of CIN.


Molecular strategies for interrupting arthropod-borne virus transmission by mosquitoes.
Arthropod-borne virus (arbovirus) infections cause a number of emerging and resurgent human and veterinary infectious diseases. Traditional means of controlling arbovirus diseases include vaccination of susceptible vertebrates and mosquito control, but in many cases these have been unavailable or ineffective, and so novel strategies for disease control are needed. One possibility is genetic manipulation of mosquito vectors to render them unable to transmit arboviruses. This review describes recent work to test the concept of pathogen-derived resistance in arthropods by expression of viral genes in mosquito cell cultures and mosquitoes. Sense and antisense genome sequences from La Crosse virus (LAC) (a member of the Bunyaviridae) and dengue viruses serotypes 1 to 4 (DEN-1 to DEN-4) (members of the Flaviviridae) were expressed in mosquito cells from double-subgenomic and replicon vectors based on Sindbis virus (a member of the Togaviridae). The cells were then challenged with homologous or related viruses. For LAC, expression of antisense sequences from the small (S) genome segment, particularly full-length antisense S RNA, effectively interfered with replication of challenge virus, whereas expression of either antisense or sense RNA from the medium (M) segment was completely ineffective in LAC inhibition. Expression of sense and antisense RNA derived from certain regions of the DEN genome also blocked homologous virus replication more effectively than did RNA from other regions. Other parameters of RNA-mediated interference have been defined, such as the time when replication is blocked and the minimum size of effector RNA. The mechanism of RNA inhibition has not been determined, although it resembles double-stranded RNA interference in other nonvertebrate systems. Prospects for application of molecular strategies to control arbovirus diseases are briefly reviewed.


Cryopreserved homograft and autologous deep vein replacement for infrarenal aorto and iliaco-femoral graft infection: early and late results.
The aim of the study was to evaluate the early and late results of aortic replacement using cryopreserved homografts and autologous deep veins for infected infrarenal prosthetic reconstructions and the influence of type of bacteria on the mortality. Thirty-three patients were treated from 30 March 1994 to 01 September 2008 for aorto-femoral or iliaco-femoral graft infections with homografts (HG:19) or autologous deep veins (DV:14). The diagnosis was based on physical signs, bacteriological tests and computed tomography(CT) scans. We obtain cryopreserved homografts from our non-profit vessel bank, deep veins were harvested before the arterial reconstruction. Patients were followed by clinical examination and ultrasound. The infections occurred 47.78 ± 20 months after the primary operations; 45% of the infections were caused by Gram-negative bacteria. Treatment indications included seven aortoduodenal fistula (21.2%) and six septic para-anastomotic bleedings (18.2%). There were six in-hospital deaths (18.2%). All the deceased patients had Gram-negative bacteria in cultures and pluribacterial infections. No patient died with single staphylococcus, streptococcus or MRSA infection. At three years freedom from reinfection was 100% in DV group and 82% (CI:0.56-0.92) in the homograft group. Survival after three years was 71% (CI:0.48-0.88) in HG group and 79% (CI:0.49-0.94) in DV group. The difference is not significant. For infrarenal graft infection homograft replacement or deep vein implantation is durable method and eradicate late reinfection. The in-hospital mortality is significantly higher in patients with aortoduodenal fistulas with Gram-negative organisms regardless of the conduit used for aortic replacement.


Acute stress response and posttraumatic stress disorder in traffic accident victims: a one-year prospective, follow-up study.
This study was designed to assess the natural course of posttraumatic symptoms formation, as well as the degree to which acute stress reactions predict later posttraumatic stress disorder (PTSD) in injured traffic accident victims. A prospective, 1-year follow-up study was carried out on 74 injured traffic accident victims and a comparison group of 19 patients who were hospitalized for elective orthopedic surgery. Participants were interviewed within the first week following the accident, and follow-up interviews were performed 1, 3, 6, and 12 months after the accident. At 12 months, a structured clinical interview was administered to determine a formal DSM-III-R diagnosis of PTSD. Twenty-four (32%) of the 74 traffic accident victims, but none of the 19 comparison subjects, met DSM-III-R criteria for PTSD at 1 year. Traffic accident victims who developed PTSD had higher levels of premorbid and comorbid psychopathology. Levels of posttraumatic symptoms were significantly higher from the outset in the subjects who developed PTSD and worsened progressively over the first 3 months, in contrast to subjects without PTSD, who manifested gradual amelioration of symptoms during this time. Existence of posttraumatic symptoms immediately after the accident was a better predictor of later PTSD than was accident or injury severity. In this study, a significant portion of injured traffic accident victims manifested PTSD 1 year after the event. The development of PTSD at 1 year can be predicted as early as 1 week after the accident on the basis of the existence and severity of early PTSD-related symptoms. However, the first 3 months following the accident appear to be the critical period for the development of PTSD.


In vivo imaging of Hedgehog pathway activation with a nuclear fluorescent reporter.
The Hedgehog (Hh) pathway is essential for embryonic development and tissue regeneration, and its dysregulation can lead to birth defects and tumorigenesis. Understanding how this signaling mechanism contributes to these processes would benefit from an ability to visualize Hedgehog pathway activity in live organisms, in real time, and with single-cell resolution. We report here the generation of transgenic zebrafish lines that express nuclear-localized mCherry fluorescent protein in a Gli transcription factor-dependent manner. As demonstrated by chemical and genetic perturbations, these lines faithfully report Hedgehog pathway state in individual cells and with high detection sensitivity. They will be valuable tools for studying dynamic Gli-dependent processes in vertebrates and for identifying new chemical and genetic regulators of the Hh pathway.


Cometary Dust.
This review presents our understanding of cometary dust at the end of 2017. For decades, insight about the dust ejected by nuclei of comets had stemmed from remote observations from Earth or Earth's orbit, and from flybys, including the samples of dust returned to Earth for laboratory studies by the Stardust return capsule. The long-duration Rosetta mission has recently provided a huge and unique amount of data, obtained using numerous instruments, including innovative dust instruments, over a wide range of distances from the Sun and from the nucleus. The diverse approaches available to study dust in comets, together with the related theoretical and experimental studies, provide evidence of the composition and physical properties of dust particles, e.g., the presence of a large fraction of carbon in macromolecules, and of aggregates on a wide range of scales. The results have opened vivid discussions on the variety of dust-release processes and on the diversity of dust properties in comets, as well as on the formation of cometary dust, and on its presence in the near-Earth interplanetary medium. These discussions stress the significance of future explorations as a way to decipher the formation and evolution of our Solar System.


NADH-glutamate synthase in alfalfa root nodules. Genetic regulation and cellular expression.
NADH-dependent glutamate synthase (NADH-GOGAT; EC 188.8.131.52) is a key enzyme in primary nitrogen assimilation in alfalfa (Medicago sativa L.) root nodules. Here we report that in alfalfa, a single gene, probably with multiple alleles, encodes for NADH-GOGAT. In situ hybridizations were performed to assess the location of NADH-GOGAT transcript in alfalfa root nodules. In wild-type cv Saranac nodules the NADH-GOGAT gene is predominantly expressed in infected cells. Nodules devoid of bacteroids (empty) induced by Sinorhizobium meliloti 7154 had no NADH-GOGAT transcript detectable by in situ hybridization, suggesting that the presence of the bacteroid may be important for NADH-GOGAT expression. The pattern of expression of NADH-GOGAT shifted during root nodule development. Until d 9 after planting, all infected cells appeared to express NADH-GOGAT. By d 19, a gradient of expression from high in the early symbiotic zone to low in the late symbiotic zone was observed. In 33-d-old nodules expression was seen in only a few cell layers in the early symbiotic zone. This pattern of expression was also observed for the nifH transcript but not for leghemoglobin. The promoter of NADH-GOGAT was evaluated in transgenic alfalfa plants carrying chimeric beta-glucuronidase promoter fusions. The results suggest that there are at least four regulatory elements. The region responsible for expression in the infected cell zone contains an 88-bp direct repeat.