Use of routine death and illness surveillance data to provide insight for UK pandemic planning: lessons from COVID-19.
Reporting of COVID-19 cases, deaths and testing has often lacked context for appropriate assessment of disease burden within risk groups. The research considers how routine surveillance data might provide initial insights and identify risk factors, setting COVID-19 deaths early in the pandemic into context. This will facilitate the understanding of wider consequences of a pandemic from the earliest stage, reducing fear, aiding in accurately assessing disease burden and ensuring appropriate disease mitigation. UK, 2020. The study is a secondary analysis of routine, public domain, surveillance data and information from Office for National Statistics (ONS), National Health Service (NHS) 111 and Public Health England (PHE) on deaths and disease. Our principal focus is ONS data on deaths mentioning COVID-19 on the death certificate. We also consider information provided in NHS 111 and PHE data summaries. Deaths with COVID-19 significantly contributed to, yet do not entirely explain, abnormally elevated all-cause mortality in the UK from weeks 12-18 of 2020. Early in the UK epidemic, COVID-19 was the greatest threat to those with underlying illness, rarely endangering people aged under 40 years. COVID-19-related death rates differed by region, possibly reflecting underlying population structure. Risk of COVID-19-related death was greater for healthcare and social care staff and black, Asian and minority ethnic individuals, having allowed for documented risk factors. Early contextualisation of public health data is critical to recognising who gets sick, when and why. Understanding at-risk groups facilitates a targeted response considering indirect consequences of society's reaction to a pandemic alongside disease-related impacts. COVID-19-related deaths mainly mirror historical patterns, and excess non-COVID-19-related deaths partly reflect reduced access to and uptake of healthcare during lockdown. Future outbreak response will improve through better understanding of connectivity between disease monitoring systems to aid interpretation of disease risk patterns, facilitating nuanced mitigation measures.


[An Investigation on a Case of Family-clustered Paragonimiasis in Wenzhou City].
Paragonimus infection was reported in a family of 8 members after consuming wine-preserved raw crabs. Seven members developed symptoms of fever, cough, sputum, fatigueness, chest pain, and abdominal pain during 2-3 months after crab feeding, while one member was normal. Serum samples were collected from the 7 members for anti-paragonimus antibody test, with 5 members showing positive, 1 weakly positive and 1 negative. In view of the epidemiological history, clinical symptoms, imaging and laboratory test results, this case was diagnosed to be a family-clustered paragonimus infection.


A role for p53 in DNA end rejoining by human cell extracts.
The tumor suppressor p53 is a major regulator in the response of human cells to DNA damage. In this study we assessed the role of p53 in the repair of DNA double-strand breaks in plasmid DNA using cell extracts from three human lymphoblastoid cell lines derived from the same donor. TK6, WI-L2-NS and TK6-E6-5e cells express wild-type, mutated and essentially no p53 protein, respectively. Total cellular extracts from TK6, WI-L2-NS and TK6-E6-5e cells were incubated with EcoRI linearized pUC19 DNA. Southern blot analysis of end-rejoined DNA indicated that the major products formed were linear multimers. There was approximately 2-fold greater end rejoining in WI-L2-NS and TK6-E6-5e extracts compared with TK6 extracts. Total DNA from end-rejoining reactions was purified and used to transform bacteria. Using the lacZ reporter gene as a measure of repair fidelity we found that misrepair, as indicated by white colonies, occurred at 4.1% to 6.5% of transformants, with no significant difference between the three cell lines. Gel analysis revealed that misrepair involved only deletions. Sequence analysis of 11 misrepaired products from each cell line showed 12 different deletions from 4 to 48 bp in length, but each cell line yielded similar product types. These results indicate that total cellular extracts from human lymphoblastoid cells lacking p53 or expressing mutated p53 have increased end-rejoining activity as compared with extracts from cells expressing wild-type p53. However, the p53 status does not influence the ratio of misrepair:correct repair, or the type of misrepair events.


Redetermination of brackebuschite, Pb2Mn(3+)(VO4)2(OH).
The crystal structure of brackebuschite, ideally Pb2Mn(3+)(VO4)2(OH) [dilead(II) manganese(III) vanadate(V) hydroxide], was redetermined based on single-crystal X-ray diffraction data of a natural sample from the type locality Sierra de Cordoba, Argentina. Improving on previous results, anisotropic displacement parameters for all non-H atoms were refined and the H atom located, obtaining a significant improvement of accuracy and an unambiguous hydrogen-bonding scheme. Brackebuschite belongs to the brackebuschite group of minerals with general formula A 2 M(T1O4)(T2O4)(OH, H2O), with A = Pb(2+), Ba, Ca, Sr; M = Cu(2+), Zn, Fe(2+), Fe(3+), Mn(3+), Al; T1 = As(5+), P, V(5+); and T2 = As(5+), P, V(5+), S(6+). The crystal structure of brackebuschite is based on a cubic closest-packed array of O and Pb atoms with infinite chains of edge-sharing [Mn(3+)O6] octa-hedra located about inversion centres and decorated by two unique VO4 tetra-hedra (each located on a special position 2e, site symmetry m). One type of VO4 tetra-hedra is linked with the (1) ∞[MnO4/2O2/1] chain by one common vertex, alternating with H atoms along the chain, while the other type of VO4 tetra-hedra link two adjacent octa-hedra by sharing two vertices with them and thereby participating in the formation of a three-membered Mn2V ring between the central atoms. The (1) ∞[Mn(3+)(VO4)2OH] chains run parallel to [010] and are held together by two types of irregular [PbO x ] polyhedra (x = 8, 11), both located on special position 2e (site symmetry m). The magnitude of the libration component of the O atoms of the (1) ∞[Mn(3+)(VO4)2OH] chain increases linearly with the distance from the centerline of the chain, indicating a significant twisting to and fro of the chain along [010]. The hy-droxy group bridges one Pb(2+) cation with two Mn(3+) cations and forms an almost linear hydrogen bond with a vanadate group of a neighbouring chain. The O⋯O distance of this inter-action determined from the structure refinement agrees well with Raman spectroscopic data.


Cognitive load and emotional processing in generalized anxiety disorder: electrocortical evidence for increased distractibility.
Generalized anxiety disorder (GAD) may be characterized by emotion regulation deficits attributable to an imbalance between top-down (i.e., goal-driven) and bottom-up (i.e., stimulus-driven) attention. In prior work, these attentional processes were examined by presenting unpleasant and neutral pictures within a working memory paradigm. The late positive potential (LPP) measured attention toward task-irrelevant pictures. Results from this prior work showed that working memory load reduced the LPP across participants; however, this effect was attenuated for individuals with greater self-reported state anxiety, suggesting reduced top-down control. In the current study, the same paradigm was used with 106 medication-free female participants-71 with GAD and 35 without GAD. Unpleasant pictures elicited larger LPPs, and working memory load reduced the picture-elicited LPP. Compared with healthy controls, participants with GAD showed large LPPs to unpleasant pictures presented under high working memory load. Self-reported symptoms of anhedonic depression were related to a reduced effect of working memory load on the LPP elicited by neutral pictures. These results indicate that individuals with GAD show less flexible modulation of attention when confronted with unpleasant stimuli. Furthermore, among those with GAD, anhedonic depression may broaden attentional deficits to neutral distracters.


Frequency-based multilayer neural network with on-chip learning and enhanced neuron characteristics.
A new digital architecture of the frequency-based multilayer neural network (MNN) with on-chip learning is proposed. As the signal level is expressed by the frequency, the multiplier is replaced by a simple frequency converter, and the neuron unit uses the voting circuit as the nonlinear adder to improve the nonlinear characteristic. In addition, the pulse multiplier is employed to enhance the neuron characteristics. The backpropagation algorithm is modified for the on-chip learning. The proposed MNN architecture is implemented on field programmable gate arrays (FPGA's) and the various experiments are conducted to test the performance of the system. The experimental results show that the proposed neuron has a very good nonlinear function owing to the voting circuit. The learning behavior of the MNN with on-chip learning is also tested by experiments, which show that the proposed MNN has good learning and generalization capabilities. Simple and modular structure of the proposed MNN leads to a massive parallel and flexible network architecture, which is well suited for very large scale integration (VLSI) implementation.


Inducible nonviral gene expression in the treatment of osteochondral defects.
The repair of osteochondral defects with chondrocytes genetically modified to express desired growth factors promises great potential in orthopaedic therapy. Controlled expression of the transgenes is required in many instances. The objective of the present study was to demonstrate the inducibility of tetracycline-responsive transgene expression in osteochondral defects in the knee joint filled with genetically modified chondrocyte implants. An expression plasmid containing the lacZ gene under the control of the minimal CMV promoter fused to the Tet-responsible element (TRE) as well as the reverse transactivator (rtTA2s-M2) was constructed and used to transfect isolated articular chondrocytes from New Zealand white rabbits. rtTA2s-M2 binds to the TRE in the presence of tetracycline and leads to the transcription of the transgene. Different concentrations of DNA and various DNA:lipid ratios were tested to determine best transfection conditions. Transfection efficiency and inducibility were analysed by histochemical analysis and flow-cytometry. To evaluate the system in vivo, collagen-sponges were seeded with transfected autologous chondrocytes and implanted in osteochondral defects in the knees of NZW-rabbits. Gene expression was induced by doxycycline and 3 weeks later, LacZ-expression in isolated knee joints was evaluated in histological sections by X-gal staining. In vitro 13.5% (+/-1.32) of induced primary chondrocytes were LacZ-positive, while non-induced controls showed a background-staining in 0.6% (+/-0.2). In vivo, upon doxycycline treatment, induction of lacZ-gene-expression could be demonstrated in chondrocytes in 3-week-old, well-integrated implants. For the first time, tetracycline-inducible gene expression is demonstrated to work in the treatment of osteochondral defects. This demonstrates the feasibility for a gene therapy-assisted approach using controlled expression of therapeutic growth factors from transplanted genetically modified chondrocytes.


Alternate-form reliability of the Montreal cognitive assessment screening test in a clinical setting.
The Montreal Cognitive Assessment (MoCA) has gained recognition for its validity in detecting cognitive impairment in several clinical populations. For serial assessments, alternate forms are needed to overcome possible practice effects. Our objective was to investigate the reliability of two German MoCA alternate forms for longitudinal assessment applications. The original and one of two alternate forms of the MoCA were administered within a 60-min interval of a clinical interview in a counterbalanced order to 100 healthy elderly controls, 30 patients with mild cognitive impairment (MCI) and 30 patients with Alzheimer's disease (AD). The diagnosis of the majority of patients was supported by in vivo AD pathology biomarkers. There was a strong correlation between the alternate forms and the original MoCA in all groups, but particularly in the clinical samples. Total mean scores did not differ significantly between the MoCA versions, even taking into account the presentation order. As in previous studies, age and education influenced performance in the MoCA. The same pattern of group differences (controls > MCI > AD) was observed for each of the versions. All three forms can be reliably and interchangeably used in serial cognitive assessment, confirming the MoCA's applicability in research and clinical longitudinal approaches.


Enhancement of suicidal DNA vaccine potency by linking Mycobacterium tuberculosis heat shock protein 70 to an antigen.
Naked DNA vaccines represent an attractive approach for generating antigen-specific immunity because of their stability and simplicity of delivery. There are particular concerns with DNA vaccines however, such as potential integration into the host genome, cell transformation, and limited potency. The usage of DNA-based alphaviral RNA replicons (suicidal DNA vectors) may alleviate the concerns of integration or transformation since suicidal DNA vectors eventually cause lysis of transfected cells. To improve further the potency of suicidal DNA vaccines, we evaluated the effect of linking Mycobacterium tuberculosis heat shock protein 70 (Hsp70) to human papillomavirus type 16 (HPV-16) E7 as a model antigen on antigen-specific immunity generated by a DNA-based Semliki Forest virus (SFV) RNA vector, pSCA1. Our results indicated that this suicidal DNA vaccine containing E7/Hsp70 fusion genes generated significantly higher E7-specific T cell-mediated immune responses than vaccines containing the wild-type E7 gene in vaccinated mice. More importantly, this fusion converted a less effective vaccine into one with significant potency against established E7-expressing metastatic tumors. The antitumor effect was predominantly CD8-dependent. These results indicate that linkage of Hsp70 to the antigen may greatly enhance the potency of suicidal DNA vaccines.


Likelihood of Breastfeeding Within the USDA's Food and Nutrition Service Special Supplemental Nutrition Program for Women, Infants, and Children Population.
Breastfeeding is an important public health initiative. Low-income women benefiting from the U.S. Department of Agriculture's Food and Nutrition Service Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) are a prime population for breastfeeding promotion efforts. Research aim: This study aims to determine factors associated with increased likelihood of breastfeeding for WIC participants. The Preferred Reporting Items for Systematic Reviews and Meta-Analysis statement guided the systematic review of literature. Database searches occurred in September and October 2014 and included studies limited to the previous 10 years. The following search terms were used: low-income; WIC; women, infants, and children; breastfeeding; breast milk; and maternal and child health. The criterion for inclusion was a study sample of women and children enrolled in the WIC program, thereby excluding non-United States-based research. Factors that increased the likelihood of breastfeeding for WIC participants included sociodemographic and health characteristics ( n = 17); environmental and media support ( n = 4); government policy ( n = 2); intention to breastfeed, breastfeeding in hospital, or previous breastfeeding experience ( n = 9); attitudes toward and knowledge of breastfeeding benefits ( n = 6); health care provider or social support; and time exposure to WIC services ( n = 5). The complexity of breastfeeding behaviors within this population is clear. Results provide multisectored insight for future research, policies, and practices in support of increasing breastfeeding rates among WIC participants.


A pattern description and generation method of structural characters.
We report a pattern description and generation method of structural characters, and show some examples of Chinese and Korean character patterns actually generated. We also consider its educational and graphical applications. In this method, any character is regarded as a composite pattern constructed by several simpler subpatterns, and is described in terms of them by introducing three kinds of positional relationships among them. A composite pattern can become a subpattern, too. We call these patterns blocks. Syntactic grammar is defined to encode a pattern expression by two code strings, that is, a string of blocks and a string of production rules. They are used in generating patterns, namely, derivation of pattern expression from the code strings is defined as a process of pattern generation. By this description and generation method, we can encode structural characters without much memory, and generate natural shapes of patterns.


De novo assembly of the pennycress (Thlaspi arvense) transcriptome provides tools for the development of a winter cover crop and biodiesel feedstock.
Field pennycress (Thlaspi arvense L.) has potential as an oilseed crop that may be grown during fall (autumn) and winter months in the Midwestern United States and harvested in the early spring as a biodiesel feedstock. There has been little agronomic improvement in pennycress through traditional breeding. Recent advances in genomic technologies allow for the development of genomic tools to enable rapid improvements to be made through genomic assisted breeding. Here we report an annotated transcriptome assembly for pennycress. RNA was isolated from representative plant tissues, and 203 million unique Illumina RNA-seq reads were produced and used in the transcriptome assembly. The draft transcriptome assembly consists of 33 873 contigs with a mean length of 1242 bp. A global comparison of homology between the pennycress and Arabidopsis transcriptomes, along with four other Brassicaceae species, revealed a high level of global sequence conservation within the family. The final assembly was functionally annotated, allowing for the identification of putative genes controlling important agronomic traits such as flowering and glucosinolate metabolism. Identification of these genes leads to testable hypotheses concerning their conserved function and to rational strategies to improve agronomic properties in pennycress. Future work to characterize isoform variation between diverse pennycress lines and develop a draft genome sequence for pennycress will further direct trait improvement.


Internal Medicine Resident Work Absence During the COVID-19 Pandemic at a Large Academic Medical Center in New York City.
Montefiore Medical Center (MMC) is a large tertiary care center in the Bronx, New York City, with 245 internal medicine residents. Beginning on February 29, 2020, residents became ill with COVID-19-like illness (CLI), which required absence from work. There was initially a shortage of personal protective equipment and delays in SARS-CoV-2 testing, which gradually improved during March and April 2020. We evaluated the relationship between CLI-related work absence rates of internal medicine residents and MMC's COVID-19 hospital census over time. Data on resident work absence between February 29 and May 22 were reviewed along with MMC's COVID-19 hospital census data. To determine the effect of patient exposure on resident CLI incidence, we compared the mean incidence of CLI per patient exposure days (PED = daily hospital census × days pre- or post-peak) before and after peak COVID-19 hospital census. Forty-two percent (103 of 245) of internal medicine residents were absent from work, resulting in 875 missed workdays. At the peak of resident work absence, 16% (38 of 245) were out sick. Residents were absent for a median of 7 days (IQR 6-9.5 days). Mean resident CLI incidence per PED (CLI/PED) was 13.9-fold lower post-peak compared to pre-peak (P = .003). At the beginning of the COVID-19 pandemic in New York City, a large portion of internal medicine residents at this single center became ill. However, the incidence of CLI decreased over time, despite ongoing exposure to patients with COVID-19.


Axonal transport of [3H]serotonin in an identified neuron of Aplysia californica.
A population of characteristic ellipsoidal dense-core vesicles was identified in axons of the giant cerebral neuron of the mollusc Aplysia. We injected [3H]serotonin into the cell body of this identified serotonergic neuron in the isolated central nervous system in order to study the subcellular components associated with the neurotransmitter. Subcellular fractionation by differential centrifugation indicated that injected serotonin was rapidly taken up into particulate form. [3H]Serotonin appeared in the axons within 2 h after injection, and export continued at a constant rate of 6% of the total in the neuron/h thereafter. The dependence of the total amounts of [3H]serotonin which appeared in the axons in 6 h (export from the cell body) on the amounts injected was consistent with the idea that export is a saturable process, possibly depending on the capacity of somatic vesicles or of some unidentified carrier for serotonin. [3H]Serotonin moved into both major branches of the axon, where it was translocated rapidly. The transmitter, which was shown by autoradiography to be restricted to the axons of the injected cell, was distributed along axons in accumulations of radioactivity; in contrast, its precursor, [5-3H]hydroxytryptophan, moved slowly along axons in a smooth, declining curve, its kinetics consistent with diffusion. Quantitative electron microscope autoradiography revealed that the dense-core vesicles and the cytosol of axons fixed with glutaraldehyde were labeled with [3H]serotonin.


Significant increase in astigmatism in children after study at home during the COVID-19 lockdown.
Evaluating changes in refractive astigmatism after 'study at home' during the COVID pandemic may shed light on the aetiology of refractive errors. To investigate whether there has been a change in the proportion of astigmatism among primary school children after the school closure period during the COVID-19 pandemic. This observational study compared cross-sectional (2018: n = 112; 2020: n = 173) and longitudinal data (n = 38) collected from two vision screenings, one in 2018 and the other after the school closure period in 2020, in the same primary school for children aged 8-10 years. Non-cycloplegic refraction and axial length were measured using an open-field auto-refractometer and IOL Master, respectively. A questionnaire focusing on demographic information, near-work time, and outdoor activities was administered to parents of all participants. While there were no significant differences in age, gender, or monthly family income between the two cohorts, astigmatism proportion (Cyl ≥ 0.75 D) in 2020 was 1.5-fold higher than that in 2018 (49.1% vs. 33.9%). The median cylindrical power was significantly higher in 2020 in older children (9 or 10 years old). More importantly, the children participating in both vision screenings had cylindrical power and J0 astigmatism significantly increased by 0.35 ± 0.40 D and 0.21 ± 0.25 D, respectively. A significant increase in astigmatism (both proportion and magnitude) was found after the school closure period. Further studies are needed to investigate the origin of this increased astigmatism.


NMDA-receptor-mediated synaptic currents in guinea pig laterodorsal tegmental neurons in vitro.
1. Whole cell voltage-clamp techniques were used to record glutamate-receptor-mediated synaptic currents from neurons of the laterodorsal tegmental nucleus (LDT). The principal cells of the LDT contain acetylcholine and nitric oxide synthase, and are believed to be involved in the control of sleep-waking behavior via widespread projections to the thalamus and brain stem. LDT cells were recorded from slices of mature guinea pig brain stem with patch pipette solutions containing cesium as the primary cation. 2. Application of N-methyl-D-aspartate (NMDA) elicited currents that were strongly voltage dependent with a mean reversal potential of +16.3 mV. Peak currents occurred near -15 mV, and a region of negative slope conductance was seen at more negative potentials. Application of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid evoked currents that exhibited a nearly linear current-voltage relation with a mean reversal potential of -3.4 mV. 3. Electrical stimulation of local afferents elicited dual-component excitatory postsynaptic currents (EPSCs) with decays that were well fitted by the sum of two exponentials. Mean decay time constants at -60 mV were 8.77 ms for the faster component and 129.4 ms for the slower component. The faster component displayed a linear current-voltage relation and was blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or 6,7-dinitroquinoxaline-2,3-dione, indicating that it was mediated by non-NMDA receptors, whereas the slower component displayed a voltage dependence similar to that for NMDA-evoked currents and was blocked by 2-amino-5-phosphonopentanoic acid (AP-5), indicating its mediation by NMDA receptors. 4. The fractional contribution of NMDA receptors to the EPSC was estimated from double-exponential curve fits to the decay phases. With this method, NMDA receptors were estimated on average to carry 10.1% of the total peak EPSC at -60 mV. Blockade of the non-NMDA-receptor-mediated component with CNQX revealed a residual EPSC whose amplitude was 14.4% of the control value, whereas AP-5 alone reduced the control EPSC peak by 16.1%, both values were comparable with those obtained from curve fit estimates. 5. Previous work has shown that the presence of 4-aminopyridine-sensitive, A-like transient current in LDT cells is correlated with the cholinergic phenotype. The majority of cells in this study exhibited A-like transient currents that were blocked by 4-amino-pyridine, suggesting that the majority of the data were obtained from the cholinergic and NOS-containing neurons of the LDT nucleus. 6. These experiments demonstrate the synaptic activation of functional NMDA and non-NMDA receptors in LDT neurons, and indicate that NMDA receptors contribute to fast excitatory transmission in these cells. The results suggest that afferents releasing excitatory amino acids may play an important role in controlling the state-dependent activity of LDT neurons.


Coordination of hip and spine in individuals with acute low back pain during unstable sitting.
Trunk postural control differs between individuals with and without chronic low back pain. Whether this corresponds to differences in hip/spine coordination during the early acute phase of LBP (ALBP) is unclear. To compare hip/spine coordination in relation to seat movements between individuals with and without ALBP when balancing on an unstable seat and to identify coordination strategies to maintain balance using cluster analysis. Cross-sectional observational study. ALBP (n=130) and pain-free (n=72) individuals. Frequency domain measures to evaluate hip/spine coordination (amplitude spectrum, phase angle, coherence) and time-series measures to assess overall balance performance (centre of pressure [CoP] reflecting the amount of seat movements, upper thorax motion as a surrogate for head motion). Participants maintained balance while sitting on a seat fixed to a hemisphere. Seat, hip and spine (lower lumbar, lumbar, upper lumbar, thoracic) angular motion and force plate data were recorded. Overall, seat/CoP movements (amplitude spectrum and RMSdisplacement) were greater (in both planes) and sagittal coordination (coherence) between the hip or lower spine and seat movements was lower in ALBP than controls. Cluster analysis using coherence data revealed different coordination strategies to maintain balance. Separate clusters used a "lower lumbar strategy" and "hip strategy" in the sagittal plane, and a "lower and upper lumbar strategy" and "lower lumbar strategy" in the frontal plane. A cluster using a "low coherence strategy" in both planes was also identified. Hip and lower spine coordination was less in individuals with ALBP in conjunction with a lower quality of overall balance performance. However, interpretation of the relationship between coherence and overall balance performance was not straightforward. Clusters in both the ALBP group and the control group adopted a low coherence strategy, and this was not consistently related to poor overall balance performance. This suggests overall balance performance cannot be inferred from coherence alone and requires consideration of interaction of other different features.


Treatment of Obstructive Sleep Apnea-Hypopnea Syndrome With a Mandible Advanced Device Increases Nitric Oxide Release and Ameliorates Pulmonary Hypertension in Rabbits.
To investigate the effects of mandible advanced device (MAD) therapy for obstructive sleep apnea-hypopnea syndrome (OSAHS) on nitric oxide (NO) release and changes in pulmonary artery pressure and structure. Thirty male New Zealand white rabbits were randomly divided into OSAHS, MAD, and control groups (n = 10 per group). The soft palate of rabbits in the OSAHS and MAD groups was injected with hydrophilic polyacrylamide gel to induce OSAHS. The MAD group wore a MAD, and the control group was not treated. Cone-beam computed tomography scans and polysomnography recordings were performed to confirm successful model establishment. All rabbits slept in a supine position for 4 to 6 hours daily and were observed for 8 consecutive weeks. The pulmonary artery pressure was measured by right heart catheterization. Pulmonary artery morphometry was analyzed by hematoxylin and eosin staining. NO levels in plasma and lung homogenate supernatants were detected by Griess reaction assay kits. The OSAHS group exhibited higher pulmonary artery pressure (57.74 ± 1.79 mm Hg) than the MAD (19.99 ± 2.04 mm Hg) and control (14.49 ± 0.54 mm Hg) groups. The media thickness percentage of the pulmonary artery was higher in the OSAHS group (46.89 ± 2.72%) than the control group (15.87 ± 1.18%) and was markedly reduced by MAD (21.64 ± 1.45%). Blood oxygen saturation was positively correlated with the NO concentration in both the lung and plasma, and the NO concentration was negatively correlated with the media thickness percentage and media section percentage. OSAHS induced a decrease in NO and pulmonary hypertension, which was relieved by MAD therapy.


Suppression of in vitro granulocyte-macrophage colony formation by the peripheral mononuclear phagocytic cells of patients with idiopathic aplastic anaemia.
To investigate whether phagocytic cells play a role in the pathogenesis of aplastic anaemia, we studied the effect of the mononuclear phagocytic cells in the peripheral blood from patients with idiopathic aplastic anaemia on granulocyte-macrophage colony (GM-colony) formation. There was no apparent infection in the studied patients. In nine out of 20 cases the mononuclear cells suppressed the formation of GM-colonies by normal bone marrow. Removal of the phagocytic cells by carbonyl iron powder caused a loss of this suppressive effect. These results suggest the requirement for phagocytic cells in this suppressive effect and the possible involvement of phagocyte-mediated immunological mechanism in the pathogenesis of some cases of idiopathic aplastic anaemia.


ergm.userterms: A Template Package for Extending statnet.
Exponential-family random graph models (ERGMs) represent a powerful and flexible class of models for the statistical analysis of networks. statnet is a suite of software packages that implement these models. This paper details how the capabilities for ERGM modeling can be expanded and customized by programming additional network statistics that may be included in ERGMs. We describe a template R package called ergm.userterms that can be modified for this purpose. It is designed to make this process as straightforward as possible. We also explain some of the internal workings of statnet that will help users develop their own network analysis capabilities.


A general system for studying protein-protein interactions in Gram-negative bacteria.
One of the most promising methods for large-scale studies of protein interactions is isolation of an affinity-tagged protein with its in vivo interaction partners, followed by mass spectrometric identification of the copurified proteins. Previous studies have generated affinity-tagged proteins using genetic tools or cloning systems that are specific to a particular organism. To enable protein-protein interaction studies across a wider range of Gram-negative bacteria, we have developed a methodology based on expression of affinity-tagged "bait" proteins from a medium copy-number plasmid. This construct is based on a broad-host-range vector backbone (pBBR1MCS5). The vector has been modified to incorporate the Gateway DEST vector recombination region, to facilitate cloning and expression of fusion proteins bearing a variety of affinity, fluorescent, or other tags. We demonstrate this methodology by characterizing interactions among subunits of the DNA-dependent RNA polymerase complex in two metabolically versatile Gram-negative microbial species of environmental interest, Rhodopseudomonas palustris CGA010 and Shewanella oneidensis MR-1. Results compared favorably with those for both plasmid and chromosomally encoded affinity-tagged fusion proteins expressed in a model organism, Escherichia coli.


The EuroPrevall outpatient clinic study on food allergy: background and methodology.
The EuroPrevall project aimed to develop effective management strategies in food allergy through a suite of interconnected studies and a multidisciplinary integrated approach. To address some of the gaps in food allergy diagnosis, allergen risk management and socio-economic impact and to complement the EuroPrevall population-based surveys, a cross-sectional study in 12 outpatient clinics across Europe was conducted. We describe the study protocol. Patients referred for immediate food adverse reactions underwent a consistent and standardized allergy work-up that comprised collection of medical history; assessment of sensitization to 24 foods, 14 inhalant allergens and 55 allergenic molecules; and confirmation of clinical reactivity and food thresholds by standardized double-blind placebo-controlled food challenges (DBPCFCs) to milk, egg, fish, shrimp, peanut, hazelnut, celeriac, apple and peach. A standardized methodology for a comprehensive evaluation of food allergy was developed and implemented in 12 outpatient clinics across Europe. A total of 2121 patients (22.6% <14 years) reporting 8257 reactions to foods were studied, and 516 DBPCFCs were performed. This is the largest multicentre European case series in food allergy, in which subjects underwent a comprehensive, uniform and standardized evaluation including DBPCFC, by a methodology which is made available for further studies in food allergy. The analysis of this population will provide information on the different phenotypes of food allergy across Europe, will allow to validate novel in vitro diagnostic tests, to establish threshold values for major allergenic foods and to analyse the socio-economic impact of food allergy.


GnRH antagonist-induced down-regulation of the mRNA expression of pituitary receptors: comparisons with GnRH agonist effects.
In order to compare the mechanism for the down regulation of the mRNA expression of pituitary receptors induced by GnRH antagonist (GnRHant) to that by GnRH agonist (GnRHa), we examined the effects of GnRHant (Cetrorelix, 333 mug/kg/day), GnRHa (leuprolide depot, 333 microg/kg), and GnRHant combined with GnRHa on LH response to exogenous GnRH, pituitary LH content, LH beta subunit mRNA, and GnRH receptor (GnRH-R) mRNA levels at 2, 5, 24, 72 hours, and 7 days after the treatment in ovariectomized rats. GnRHant significantly decreased serum LH, the LH response of the pituitary to exogenous GnRH, and the pituitary LH content compared to the control treatment, though GnRHa significantly increased serum LH. GnRHant with GnRHa significantly diminished the GnRHa-induced flare-up phenomenon. GnRHant significantly decreased LH beta mRNA and GnRH-R mRNA levels, but the magnitude of the decrease in these mRNA levels by GnRHant was significantly less than those by GnRHa until 72 hours following treatment. Prolonged treatment of GnRHant caused a marked inhibition of LH beta mRNA and GnRH-R mRNA expression, similar to that caused by GnRHa. Combination treatment with GnRHa and GnRHant was demonstrated to decrease LH beta mRNA and GnRH-R mRNA levels as much as GnRHa alone and GnRHant alone over 7 days of the treatment. The present study showed differences between GnRHant and GnRHa treatment in the reduction of GnRH-R mRNA levels up to 72 hours after the treatment, and indicated that the suppression of GnRH-R mRNA by GnRHant was the maximal by GnRHa 7 days after the treatment because more profound suppression was not observed upon additional treatment with GnRHa. The findings in the present study support the hypothesis that the mechanism by which GnRHant leads to down-regulation of the mRNA expression of pituitary receptors is similar to that of GnRHa.


[Effect of alcohol on interhemispheric functional relations in patients with late sequelae of cranio-cerebral injuries].
Using the backward masking method the time of perception of visual stimuli in the right and the left field of vision was examined. It was found that in patients with craniocerebral injury sequelae, as well as in healthy subjects the time of perception of literal stimuli was shorter in the left field of vision, i.e. the information entering the right hemisphere was processed more rapidly. Under the action of alcohol the asymmetry of the perception time disappeared in the patients, and became inverse in the healthy subjects, i.e. the time of perceiving letters in the left field of vision got longer than in the right field. Under the action the right hemisphere seemed to lose its advantage in the speed of the stimuli perception. It is supposed that the right hemisphere is more sensitive to alcohol.


An investigation of Zr doping in NaBiTi2O6 perovskite by direct hydrothermal synthesis.
The direct crystallisation of perovskites NaBi(Ti(1-x)Zrx)(2)O(6) with x = 0, 0.01, 0.05 and 0.1 at 240 °C is achieved from aqueous alkali (NaOH) solutions of NaBiO(3), TiF(3) and ZrOCl(2). For each material, a single rhombohedral polymorph (R3c a∼ 5.51 Å, c∼ 13.50 Å) can be fitted to powder X-ray diffraction data, with Rietveld refinement showing a linear increase in lattice parameters and unit cell volume with increasing Zr content. Scanning electron microscopy shows micron-sized cube-shaped crystallites for each sample, with energy-dispersive X-ray analysis giving Bi : Ti : Zr ratios consistent with the expected substitution. Raman spectroscopy shows no perturbance of local structure upon Zr doping and the spectrum shows five broad bands, consistent with the literature on similar materials. Attempts to increase the Zr content further (x > 0.1) were unsuccessful by this hydrothermal synthesis method, leading instead to crystalline ZrO(2) by-products. For NaBiTi(2)O(6) and NaBi(Ti(0.99)Zr(0.01))(2)O(6) densified ceramics were prepared (∼95% density of crystallographic value) and their remnant polarisation was found to be reduced upon Zr substitution, along with a higher maximum piezoelectric coefficient, d33, measured and comparable permittivity and dielectric loss to other reported NaBiTi(2)O(6) materials.


Characterization and regulation of the gamma-glutamyl transpeptidase gene from the fission yeast Schizosaccharomyces pombe.
The structural gene for the putative gamma-glutamyl transpeptidase (GGT) was isolated from the chromosomal DNA of the fission yeast Schizosaccharomyces pombe. The determined sequence contained 3324 bp and encoded the predicted 630 amino acid sequence of GGT, which resembles counterparts in Homo sapiens, Rattus norvegicus, Saccharomyces cerevisiae, and Escherichia coli. The S. pombe cells harboring the cloned GGT gene showed about twofold higher GGT activity in the exponential phase than the cells harboring the vector only, indicating that the cloned GGT gene was functional. To monitor the expression of the S. pombe GGT gene, we fused the fragment 1085 bp upstream of the cloned GGT gene into the promoterless beta-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pGT98. The synthesis of beta-galactosidase from the fusion plasmid in S. pombe cells was enhanced by treatments with NO-generating sodium nitroprusside (SN), L-buthionine-(S,R)-sulfoximine (BSO), and glycerol. The GGT mRNA level in the S. pombe cells was increased by SN and BSO. Involvement of Pap1 in the induction of the GGT gene by SN and BSO was observed.


Optimization of Drug Delivery by Drug-Eluting Stents.
Drug-eluting stents (DES), which release anti-proliferative drugs into the arterial wall in a controlled manner, have drastically reduced the rate of in-stent restenosis and revolutionized the treatment of atherosclerosis. However, late stent thrombosis remains a safety concern in DES, mainly due to delayed healing of the endothelial wound inflicted during DES implantation. We present a framework to optimize DES design such that restenosis is inhibited without affecting the endothelial healing process. To this end, we have developed a computational model of fluid flow and drug transport in stented arteries and have used this model to establish a metric for quantifying DES performance. The model takes into account the multi-layered structure of the arterial wall and incorporates a reversible binding model to describe drug interaction with the cells of the arterial wall. The model is coupled to a novel optimization algorithm that allows identification of optimal DES designs. We show that optimizing the period of drug release from DES and the initial drug concentration within the coating has a drastic effect on DES performance. Paclitaxel-eluting stents perform optimally by releasing their drug either very rapidly (within a few hours) or very slowly (over periods of several months up to one year) at concentrations considerably lower than current DES. In contrast, sirolimus-eluting stents perform optimally only when drug release is slow. The results offer explanations for recent trends in the development of DES and demonstrate the potential for large improvements in DES design relative to the current state of commercial devices.


Catalytic asymmetric synthesis of geminal-dicarboxylates.
Stereogenic acetals, spiroacetals and ketals are well-studied stereochemical features that bear two heteroatoms at a common carbon atom. These stereocenters are normally found in cyclic structures while linear (or acyclic) analogues bearing two heteroatoms are rare. Chiral geminal-dicarboxylates are illustrative, there is no current way to access this class of compounds while controlling the stereochemistry at the carbon center bound to two oxygen atoms. Here we report a rhodium-catalysed asymmetric carboxylation of ester-containing allylic bromides to form stereogenic carbon centers bearing two different carboxylates with high yields and enantioselectivities. The products, which are surprisingly stable to a variety of acidic and basic conditions, can be manipulated with no loss of enantiomeric purity as demonstrated by ring closing metathesis reactions to form chiral lactones, which have been extensively used as building blocks in asymmetric synthesis.


The Clinical Efficacy and Safety of Gumiganghwal-Tang in Knee Osteoarthritis: A Phase II Randomized Double Blind Placebo Controlled Study.
Gumiganghwal-tang (GMGHT) is a traditional herbal medicine consisting of nine different herbs. GMGHT inhibits the mRNA expression and production of inflammatory cytokines tumor necrosis factor-α (TNF- α), interleukin-6 (IL-6), and TNF- β on lipopolysaccharide- (LPS-) stimulated peritoneal macrophages in a dose-dependent manner. It is empirically used for the treatment of inflammatory disease, but there are few reports of clinical trials that investigate its efficacy and safety. The current study aimed to investigate the clinical efficacy and safety of GMGHT in patients with knee osteoarthritis (OA). This was a multicenter, two-armed, double-blinded, randomized, placebo controlled study of GMGHT over 6 weeks. Eligible patients who fulfilled the American College of Rheumatology criteria for OA were randomized to receive either GMGHT or the placebo. Clinical assessments included measurement of knee pain and function using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), patient global assessment (PGA), and knee pain scores every 2 weeks. A total of 128 patients were enrolled (91.4% female; mean age, 58.7 ± 8.1 years). At baseline, pain visual analogue score (VAS) was 67.2 ± 1.4, resp. 71.3 ± 1.6 (treatment, resp. placebo group, p=0.84), and total WOMAC score was 55.2 ± 1.6, resp. 55.6 ± 1.5 (p = 0.84). After 6 weeks, the pain VAS was 43.0 ± 2.5, resp. 61.6 ± 2.5 (p < 0.01) and the total WOMAC score was 34.1 ± 2.4, resp. 46.9 ± 1.8 (p < 0.01). No patients withdrew because of treatment emergent adverse events. Expected adverse events including dyspepsia, liver function abnormality, and lower extremity edema were comparable between both groups. Treatment with GMGHT resulted in significant improvement in pain, function, and global assessment, and it was generally safe and well tolerated in patients with OA.


Test Experience, Direct Instruction, and Their Combination Promote Accurate Beliefs about the Testing Effect.
Practice testing is a highly robust learning strategy that promotes long-term retention, especially in comparison to more passive strategies such as restudying-a finding referred to as the testing effect. However, learners do not always appreciate the memorial benefits of practice testing over restudying, which could limit their use of practice testing during self-regulated learning. The current investigation explored the extent to which learners' metacognitive judgments about the testing effect can be improved via test experience, direct instruction, or a combination of both techniques. Prolific participants underwent two learning cycles. In the first cycle, participants were randomly assigned to either (a) experience a testing effect in their own memory performance (i.e., study unrelated word pairs, practice half the pairs through restudying and half through testing with correct-answer feedback, complete a critical test on the pairs, and receive feedback regarding their performance after using each strategy); (b) imagine they had to learn word pairs and read a passage on the purported benefits of practice testing; or (c) undergo both procedures. In the second cycle, all participants learned a novel set of word pairs. Across both learning cycles, participants estimated memory performance for material learned through testing versus restudying. Both test experience and direct instruction-independently and in combination-led to more accurate memory estimates across learning cycles, but no technique was more effective than the other. In summary, people can learn about the memorial benefits of practice testing when they experience a testing effect on their own memory performance and/or when they receive instruction about its benefits.


At last...?: Michelle Obama, Beyoncé, race & history.
In this essay, Griffin brings to the fore two extraordinary black women of our age: First Lady Michelle Obama and entertainment mogul Beyoncé Knowles. Both women signify change in race relations in America, yet both reveal that the history of racial inequality in this country is far from over. As an Ivy League-educated descendent of slaves, Michelle Obama is not just unfamiliar to the mainstream media and the Washington political scene; during the 2008 presidential campaign, she was vilified as angry and unpatriotic. Beyoncé, who controls the direction of her career in a way that pioneering black women entertainers could not, has nonetheless styled herself in ways that recall the distinct racial history of the Creole South. Griffin considers how Michelle Obama's and Beyoncé's use of their respective family histories and ancestry has bolstered or diminished their popular appeal.


Gonadal dysgenesis reveals sexual dimorphism in the embryonic germline of Drosophila.
In hybrid dysgenesis, sterility can occur in both males and females. At 27.5 degrees, however, we found that P element-induced germline death was restricted to females. This sex-specific gonadal dysgenesis (GD) is complete by the first larval instar stage. As such, GD at 27.5 degrees reveals the sexually dimorphic character of the embryonic germline. The only other known dimorphic trait of the embryonic germline is the requirement for ovo. ovo is required for germline development in females only and has been implicated in germline sex determination. Dominant mutations of ovo partially suppressed female GD. Although embryonic germ cells are undifferentiated and morphologically indistinguishable between males and females, the functional dimorphism seen in ovo requirement and GD at 27.5 degrees indicates that sexual identity in Drosophila germ cells is established in embryogenesis.


Multifaceted approach to determine the antagonist molecular mechanism and interaction of ibodutant ([1-(2-phenyl-1R-[[1-(tetrahydropyran-4-ylmethyl)-piperidin-4-ylmethyl]-carbamoyl]-ethylcarbamoyl)-cyclopentyl]-amide) at the human tachykinin NK2 receptor.
Ibodutant (MEN15596, [1-(2-phenyl-1R-[[1-(tetrahydropyran-4-ylmethyl)-piperidin-4-ylmethyl]-carbamoyl]-ethylcarbamoyl)-cyclopentyl]-amide) is a tachykinin NK(2) receptor (NK(2)R) antagonist currently under phase II clinical trials for irritable bowel syndrome. This study focuses on the ibodutant pharmacodynamic profile at the human NK(2)R and compares it with two other antagonists, nepadutant (MEN11420, (cyclo-[[Asn(beta-D-GlcNAc)-Asp-Trp-Phe-Dpr-Leu]cyclo(2beta-5beta)]) and saredutant [SR48968, (S)-N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4-dichlorophenyl)butyl]benzamide]. In functional experiments (phosphatidylinositol accumulation) in Chinese hamster ovary cells expressing the human NK(2)R, ibodutant potency measured toward concentration-response curves to neurokinin A as pK(B) was 10.6, and its antagonism mechanism was surmountable and competitive. In the same assay, antagonism equilibration and reversibility experiments of receptor blockade indicated that ibodutant quickly attains equilibrium and that reverts from receptor compartment in a slower manner. Kinetic properties of ibodutant were assessed through competitive binding kinetics experiments performed at [(3)H]nepadutant and [(3)H]saredutant binding sites. Determined K(on) and K(off) values indicated a fast association and slow dissociation rate of ibodutant at the different antagonist binding sites. Last, by radioligand binding experiments at some mutated human tachykinin NK(2)Rs, the amino acidic determinants crucial for the high affinity of ibodutant were identified at the transmembrane (TM) level: Cys167 in TM4; Ile202 and Tyr206 in TM5; Phe270, Tyr266, and Trp263 in TM6; and Tyr289 in TM7. These results indicated an extended antagonist binding pocket in the TM portion of the receptor, which is conceived crucial for TM3 and 6 arrangement and leads to G protein-coupled receptor activation. By combining this information and molecular modeling, the docking mode of ibodutant-human NK(2)R complex is proposed.


Proteomics investigation of human platelets by shotgun nUPLC-MSE and 2DE experimental strategies: a comparative study.
Platelets, the smallest human blood cells component, have a key role in the control of haemostasis and thrombosis but they have also been shown to be implicated in a number of different pathological states because of their involvement also in the process of inflammation end its resolution. Their peculiar anucleated morphology render the proteomics an intriguing approach to understand their biology. Given the high impact of platelet in different diseases we have started a systematic investigation of protein repertoire in controlled platelet preparation. Platelets have been extracted from blood of healthy donors (n=6) collected by venipuncture in Vacutainer. The quality of the preparation was assessed by observation and enumeration in a Bürker chamber with a conventional tissue culture microscope. To characterize human platelets proteome we analysed the pool of purified platelets combining two proteomic approaches: 2-DE separation combined with Mass Spectrometry and nanoscale ultra performances LC-MS(E) shotgun proteomics experiments. The 2D gel analysis leads an average of 1900 protein spots, after the filtering of "noise" and "false positive" spots, over 500 were selected to be eligible for further analysis given their optimal spot quality value. To perform the analysis by ion accounting shotgun proteomic approach, based on nano ultra performance liquid chromatography (nUPLC) coupled to MS(E) processing of continuum LC-MS data, the same pool of samples was subject to liquid phase tryptic digestion and the peptide obtained used for the experiments. All the data obtained were analysed using ProteinLynx GlobalServer v2.3 (PLGS, Waters). Three analytical replicates run were acquire in high/low energy modes and associated to a human protein database returning the identification of 100 distinct genes. Comparative analysis of the Gene Ontology has been performed to evaluate the differential functional representation of the molecular repertoire investigated with these two orthogonal approaches. The overall molecular function classification revealed differences between the two proteomic approaches. In particular, we found significant differences in cytoskeletal proteins (19.65% 2-DE versus 45.60 Shotgun) and receptors (0,92% 2-DE versus 6.90% Shotgun).


Lessons from a community based interdisciplinary learning exposure: benefits for both students and communities in Uganda.
Makerere University implemented a One Health Institute (OHI) in 2016 involving undergraduate students selected from different disciplines. The students were first taken through theoretical principles of One Health followed by a field attachment in communities. The field attachment aimed to expose students to experiential educational opportunities in the communities in a One Health approach. In this paper, we present students' experiences and their contributions to the communities of attachment. This was a cross-sectional study, utilizing qualitative data collection methods. The study involved students who participated in the OHI field attachment and community members in a One Health demonstration site-Western Uganda. Four focus group discussions (FGDs) and four in-depths interviews (IDIs) were conducted among the students, while four FGDs and twelve IDIs were conducted among community members. All interviews were audio-recorded, transcribed and analysed manually. The four themes that emerged are: students' understanding and appreciation of One Health concept, their experiences and gains from the multi-disciplinary field attachment, students' contributions to the community, and challenges faced by the students. Students had good knowledge of One Health. They appreciated that health cannot be achieved by one discipline or sector and thus the need to collaborate across sectors. Regarding experiences and gains during the multi-disciplinary field attachment, the students appreciated that each discipline had a role to play in achieving health in the community. They appreciated the training citing skills gained in communication, team work and collaboration. They also reported a feeling of gratitude and accomplishment because they felt they made a positive change to the community by putting in place interventions to address some of the community challenges. Similarly, the communities appreciated the students' contribution in solving their health challenges, ranging from conducting health education to improving sanitation and hygiene. Through the OHI, students gained One Health competencies including communication, teamwork, and collaboration. Adopting an interdisciplinary model in university teaching system especially during field placement would strengthen skills of collaboration, team work and communication which are critical for a multi-disciplinary approach which is needed among the future workforce in order to solve the current health challenges.


Comparative evaluation of frictional characteristics between nano coated and non coated orthodontic brackets and arch wire configuration-An experimental in vitro study.
To compare and assess the coefficient of frictional resistance between nano-coated orthodontic brackets and orthodontic archwires with conventional orthodontic brackets and archwires. In this experimental study, 128 samples were divided into 4 groups consisting of 32 orthodontic wires and brackets in each group. The samples were randomly allocated into GROUP A- ZNO nanoparticle coated archwires and brackets, GROUP B ZNO nanoparticle coated bracket and conventional archwire, GROUP C-ZNO nanoparticle coated archwire and conventional bracket, and GROUP D- conventional archwire and bracket after positioning them on special jigs frictional resistance was studied and evaluated. Bon - Ferroni test was used for inter group comparison and one way ANOVA was used for intr-group comparison. The lowest mean frictional resistance is seen with Group A (nanocoated archwire with nanocoated bracket) N = 0.3401 ± 0.420; and highest with Group D (conventional brackets with conventional archwires) N = 0.8413 ± 0.60. a significant difference in mean frictional resistance was observed between the groups (P ≤ 0.01). The frictional resistance for the groups was in the following order from lowest to highest: group A < B < C < D. The study showed decreased friction in ZNO nanoparticle coated archwires and brackets than conventional archwires and brackets.


A randomized controlled trial of the GnRH antagonist ganirelix in Chinese normal responders: high efficacy and pregnancy rates.
Gonadotropin-releasing hormone (GnRH) antagonists for controlled ovarian stimulation (COS) were only recently introduced into China. The efficacy and safety of the GnRH antagonist ganirelix was assessed in a multicenter, controlled, open-label study, in which Chinese women were randomized to either ganirelix (n = 113) or a long GnRH agonist protocol of triptorelin (n = 120). The primary end point was the amount of recombinant follicle-stimulating hormone (rFSH) required to meet the human chorionic gonadotropin criterion (three follicles ≥17 mm). The amount of rFSH needed was significantly lower for ganirelix (1272 IU) vs. triptorelin (1416 IU; P< 0.001). Ongoing pregnancy rates per started cycle were 39.8% (ganirelix) and 39.2% (triptorelin). Although both treatments were well tolerated, cancellation due to risk of ovarian hyperstimulation syndrome (OHSS) was less frequent with ganirelix (1.8%) than triptorelin (7.5%) (P = 0.06). Less rFSH was needed in the ganirelix protocol than the long GnRH agonist protocol, with fewer reported cases of OHSS and similar pregnancy rates.


Io's thermal emission from the Galileo photopolarimeter-radiometer.
Galileo's photopolarimeter-radiometer instrument mapped Io's thermal emission during the I24, I25, and I27 flybys with a spatial resolution of 2.2 to 300 kilometers. Mapping of Loki in I24 shows uniform temperatures for most of Loki Patera and high temperatures in the southwest corner, probably resulting from an eruption that began 1 month before the observation. Most of Loki Patera was resurfaced before I27. Pele's caldera floor has a low temperature of 160 kelvin, whereas flows at Pillan and Zamama have temperatures of up to 200 kelvin. Global maps of nighttime temperatures provide a means for estimating global heat flow.


Glucocorticoid withdrawal affects stress-induced changes of urocortin 2 gene expression in rat adrenal medulla and brain.
Corticotropin-releasing factor is well known activator of the hypothalamic-pituitary-adrenocortical axis, that represents crucial system participating on stress response of the organism. Urocortins are members of the corticotropin-releasing factor family of peptides with proposed effects on neuroendocrine and behavioral stress response mechanisms. Urocortin 2, one of three known urocortins, is present in central and peripheral stress response system and its expression can be augmented by glucocorticoids. In the present study we have examined how glucocorticoid withdrawal affects urocortin 2 gene expression after acute immobilization in the adrenal medulla and selected brain areas in rats. We used pharmacological adrenalectomy to block synthesis of corticosterone. Our results show that the immobilization-induced rise in urocortin 2 mRNA levels in rat adrenal medulla was not inhibited by glucocorticoid withdrawal. On the other hand, observed changes in the brain indicate that the effect of stress and pharmacological adrenalectomy on urocortin 2 gene expression is site-specific. While in the paraventricular nucleus and locus coeruleus the immobilization induced rise of urocortin 2 was not inhibited by pharmacological adrenalectomy in the arcuate nucleus and central amygdala it was. Moreover, we have seen a significant depletion of urocortin 2 plasma levels after immobilization. The immobilization induced rise of urocortin 2 gene expression in rat adrenal medulla and brain areas regulating stress response pathways and preservation of its induction after adrenalectomy suggests a role of urocortin 2 in the neuroendocrine stress response of an organism. This article is protected by copyright. All rights reserved.


Hemodynamic, autonomic and neurohormonal behaviour of familial amyloidotic polyneuropathy and neurally mediated syncope patients during supine and orthostatic stress.
Orthostatic intolerance (OI) syndromes are frequent and share symptoms like dizziness and orthostatic syncope. Their pathophysiology however seems to be different. The aim of our work was to evaluate autonomic and hemodynamic behaviour in patients with familial amyloidotic polyneuropathy and neurally mediated syncope in supine position and after acute orthostatic passive stress. We studied 12 patients with autonomic failure (group A), 12 patients with neurally mediated syncope (group B) and 16 aged matched normal controls (group C), in supine position and during the first 10 min of head-up tilt test (HUTT). Beat-by-beat blood pressure and heart rate were continuously monitored and digitised at 500 Hz. The baroreceptor alfa-index gain (vagal reflex-BRG), high frequency of RR variability (HFRR, vagal tonus) and low frequency of systolic arterial pressure variability (LFSAP, sympathetic tone) were calculated. Catecholamines, plasma brain (BNP) and atrial natriuretic (ANP) peptides were also measured. Hemodynamic data were derived and calculated by the non-invasive modelflow method. During supine position, cardiac output (CO) and stroke volume (SV) were similar in all groups. Mean arterial pressure (MAP) and BNP were higher in group A. Noradrenaline (NOR), BRG, HFRR and LFSAP were extremely low in this group. BRG and adrenaline (ADR) were higher in group B than in controls. Within the first 10 min of HUTT, there was a huge drop of CO, SV and MAP in group A, maintenance of very low levels of neurohormones and lack of autonomic function. HR, LFSAP and ADR had a higher rise at HUTT in group B compared with controls (p<0.01) but a significant decrease of BRG was noted (p<0.05). ANP or BNP did not change with tilt in any group. Different orthostatic intolerance syndromes may show important hormonal, autonomic and hemodynamic differences during supine rest and enhanced after passive orthostatism.


[Study of molecular mobility in biological membranes by 2-mm band ESR spectroscopy].
Temperature relationship was measured of ESR spectra of spin probes--derivatives of fatty acids whose nitroxyl fragments are incorporated into surface and inner layers of phosphatidylcholine liposomes (T = 180-260 K), as well as of the probe in model ethanol solution (T = 110-220 K). Data on the nature and rotation frequency of probe nitroxyl fragments were obtained from the analysis of the spectra. It was shown that the frequency of the probes movement in the systems under study corresponds to the slow movement region, i.e. it is not above 10(7) s-1.


4-Iodine N-Methylpyridinium-Mediated Peptide Synthesis.
Through systematic optimization of halopyridinium compounds, we established a peptide coupling protocol utilizing 4-iodine N-methylpyridinium (4IMP) for solid-phase peptide synthesis (SPPS). The 4IMP coupling reagent is easily prepared, bench stable, and cost-effective. Employing 4IMP in the SPPS process has showcased remarkable chemoselectivity and efficiency, effectively eliminating racemization and epimerization. This achievement has been substantiated through the successful synthesis of a range of peptides via the direct utilization of commercially available amino acid substrates for SPPS.


Longitudinal study of dogs living in an area of Spain highly endemic for leishmaniasis by serologic analysis and the leishmanin skin test.
The literature contains few longitudinal studies that have assessed areas endemic for canine leishmaniasis and over the same time interval Leishmania-specific cellular and humoral immunity in healthy dogs. Fourteen dogs, three mixed breed and 11 Ibizian hounds, living in an area of Spain that was highly endemic for leishmaniasis were followed-up over a three-year period by serologic analysis and the leishmanin skin test (LST). All but one of these dogs remained clinically healthy during the study period. Seroconversion was observed in four dogs. The three mixed breed dogs had a negative reaction in the LST in the first and third years. The general trend in the Ibizian hounds was an increase in the diameter of the LST reaction at both the 48- and 72-hour readings in the third year. This study demonstrates that in addition to an increase in Leishmania-specific humoral immune response in Ibizian hounds, a parallel increase in cellular immune response was observed.


Studies of the glomerular mesangium and the juxtaglomerular apparatus in the genetically diabetic mouse.
Intact and uninephrectomized genetically diabetic (db/db) mice (C57BL/KsJ) and their nondiabetic littermates (dm/m) had renal biopsies performed at 6 months of age. Renal tissues were studied by regular light microscopy and by a variety of immunohistochemical techniques. Intact db/db mice had peripheral mesangial thickening as compared to db/m mice. This thickening, predominantly due to increased mesangial matrix material, extended to the glomerular hilum and the extraglomerular mesangium of the juxtaglomerular apparatus. This abnormality was markedly increased an uninephrectomized db/db mice (db/db-UN) compared to intact db/db mice. Db/m-UN animals had slightly greater mesangial thickness than intact db/mice but less than that of db/db mice. Intact db/db mice had increased mesangial IgM staining compared to db/m mice and these differences were magnified by uninephrectomy. The IgM staining, especially in the diabetic mice, involved the peripheral mesangium and the glomerular hilum extending into extraglomerular mesangium and the distal tubule at the level of the macula densa. The tubular staining in the region of the juxtaglomerular apparatus was between the tubular basement membrane and the epithelial cells and between epithelial cells. The distal tubular epithelial cell cytoplasm also showed increased staining for IgM as the tubule coursed away from the glomerulus. These studies amplify the argument that alterations in glomerular hemodynamics influence the rate of development of diabetic glomerular lesions. Further, these diabetic mice appear to represent an important model for the study of mesangial macromolecular processing mechanisms.


Serum resistance associated with virulence in Yersinia enterocolitica.
Yersinia enterocolitica strains that exhibited a calcium requirement for growth and autoagglutination at 37 degrees C were invariably virulent in rabbits, causing diarrhea and a high degree of lethality, and were capable of colonizing the intestinal lumen and establishing foci of infection on the Peyer's patches of mice. Strains that had lost the properties of calcium dependency and autoagglutinability were totally avirulent in rabbits and were quickly eliminated from the intestinal lumen and tissues of mice. Virulent and avirulent strains were shown to be equally invasive to HeLa cells. However, the virulent strains were resistant to the bactericidal action of normal serum, and this serum resistance was lost with the loss of virulence. Furthermore, the serum resistance of virulent strains was expressed, as were other properties, when strains were grown at 37 degrees C, but not at 27 degrees C. These results suggest that a virulence factor associated with serum resistance plays an essential role in the pathogenicity of Y. enterocolitica.


High-resolution profiling of histone h3 lysine 36 trimethylation in metastatic renal cell carcinoma.
Mutations in SETD2, a histone H3 lysine trimethyltransferase, have been identified in clear cell renal cell carcinoma (ccRCC); however it is unclear if loss of SETD2 function alters the genomic distribution of histone 3 lysine 36 trimethylation (H3K36me3) in ccRCC. Furthermore, published epigenomic profiles are not specific to H3K36me3 or metastatic tumors. To determine if progressive SETD2 and H3K36me3 dysregulation occurs in metastatic tumors, H3K36me3, SETD2 copy number (CN) or SETD2 mRNA abundance was assessed in two independent cohorts: metastatic ccRCC (n=71) and the Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma data set (n=413). Although SETD2 CN loss occurs with high frequency (>90%), H3K36me3 is not significantly impacted by monoallelic loss of SETD2. H3K36me3-positive nuclei were reduced an average of ~20% in primary ccRCC (90% positive nuclei in uninvolved vs 70% positive nuclei in ccRCC) and reduced by ~60% in metastases (90% positive in uninvolved kidney vs 30% positive in metastases) (P<0.001). To define a kidney-specific H3K36me3 profile, we generated genome-wide H3K36me3 profiles from four cytoreductive nephrectomies and SETD2 isogenic renal cell carcinoma (RCC) cell lines using chromatin immunoprecipitation coupled with high-throughput DNA sequencing and RNA sequencing. SETD2 loss of methyltransferase activity leads to regional alterations of H3K36me3 associated with aberrant RNA splicing in a SETD2 mutant RCC and SETD2 knockout cell line. These data suggest that during progression of ccRCC, a decline in H3K36me3 is observed in distant metastases, and regional H3K36me3 alterations influence alternative splicing in ccRCC.


Maternal Lifetime Trauma Exposure, Prenatal Cortisol, and Infant Negative Affectivity.
Little research has examined the impact of maternal lifetime trauma exposure on infant temperament. We examined associations between maternal trauma history and infant negative affectivity and modification by prenatal cortisol exposure in a sociodemographically diverse sample of mother-infant dyads. During pregnancy, mothers completed measures of lifetime trauma exposure and current stressors. Third-trimester cortisol output was assessed from maternal hair. When infants were 6 months old, mothers completed the Infant Behavior Questionnaire-Revised. In analyses that controlled for infant sex and maternal age, education, race/ethnicity, and stress during pregnancy, greater maternal trauma exposure was associated with increased infant distress to limitations and sadness. Higher and lower prenatal cortisol exposure modified the magnitude and direction of association between maternal trauma history and infant rate of recovery from arousal. The association between maternal trauma history and infant distress to limitations was somewhat stronger among infants exposed to higher levels of prenatal cortisol. The analyses suggested that maternal lifetime trauma exposure is associated with several domains of infant negative affectivity independently of maternal stress exposures during pregnancy and that some of these associations may be modified by prenatal cortisol exposure. The findings have implications for understanding the intergenerational impact of trauma exposure on child developmental outcomes.


Assessment of Weight Loss With the Intragastric Balloon in Patients With Different Degrees of Obesity.
The intragastric balloon (IGB) is an endoscopic device for the treatment of obesity. Best results are observed in patients who follow a dietary program but few studies have assessed the results of this treatment in patients with different degrees of obesity. The aim of this study is to compare the efficacy of IGB in patients with different degrees of obesity. A total of 2002 patients with IGB were retrospectively evaluated and were divided into groups according to initial body mass index (BMI) range, as follows: group 1, 27 to 29.9 kg/m; group 2, 30 to 34.9 kg/m; group 3, 35 to 39.9 kg/m; group 4, 40 to 44.9 kg/m; group 5, ≥45 kg/m. Weight was assessed in 3 different times: before (T0), 1 month (T1), 6 months (T2), and 6 months after removal of the IGB (T3). A total of 946 patients lost follow-up. Overall, 40 (3.78%) removed the device before programmed by intolerance, and 1016 patients completed the 6-month treatment. The mean weight loss was 18.9%, excess weight loss 60.1% and an BMI reduction of 6.76 points. 6 months after removal of the balloon 842 patients had continued follow-up (82.8%). At this time, weight loss was 19.84%, excess weight loss was 59.49%, and BMI reduction of 7.06 points. In all groups there was statistical difference between the times T0 and T1 and between T1 and T2 (P<0.001). There was no statistical difference between T2 and T3, in any group. IGB provided sustained weight loss in patients who remained in dietary follow-up for 1 year.


Influenza and oseltamivir phosphate (Tamiflu) in infants: what you need to know.
Influenza is a highly contagious virus that causes an average of 20,000 hospitalizations a year in children under five years of age. As of March 30, 2013, the 2012-2013 flu season had seen 111 pediatric deaths, with 21 deaths in the 0- to 23-month-old range. Rates of influenza-associated hospitalization are substantially higher among infants and young children than among older children, and those under six months old are at the highest risk. Research shows that influenza vaccine is not as effective in children two years of age relative to adults, and the vaccine is not approved in infants younger than six months. Thus, preventing influenza and proper treatment are important for keeping this high-risk group safe from complications. With infants being highest at risk for complications and the extrapolation of efficacy and safety from the older population, the U.S. Food and Drug Administration (FDA) recently approved the use of the antiviral oseltamivir phosphate (Tamiflu) for treatment of uncomplicated influenza in patients two weeks and older. This young population is susceptible to the benefits as well as the risks of the drug. Health care providers must be aware of dosing, adverse reactions, and monitoring parameters to better treat and educate their patients.


Risk Prediction Models and Novel Prognostic Factors for Heart Failure with Preserved Ejection Fraction: A Systematic and Comprehensive Review.
Patients with heart failure with preserved ejection fraction (HFpEF) have large individual differences, unclear risk stratification, and imperfect treatment plans. Risk prediction models are helpful for the dynamic assessment of patients' prognostic risk and early intensive therapy of high-risk patients. The purpose of this study is to systematically summarize the existing risk prediction models and novel prognostic factors for HFpEF, to provide a reference for the construction of convenient and efficient HFpEF risk prediction models. Studies on risk prediction models and prognostic factors for HFpEF were systematically searched in relevant databases including PubMed and Embase. The retrieval time was from inception to February 1, 2023. The Quality in Prognosis Studies (QUIPS) tool was used to assess the risk of bias in included studies. The predictive value of risk prediction models for end outcomes was evaluated by sensitivity, specificity, the area under the curve, C-statistic, C-index, etc. In the literature screening process, potential novel prognostic factors with high value were explored. A total of 21 eligible HFpEF risk prediction models and 22 relevant studies were included. Except for 2 studies with a high risk of bias and 2 studies with a moderate risk of bias, other studies that proposed risk prediction models had a low risk of bias overall. Potential novel prognostic factors for HFpEF were classified and described in terms of demographic characteristics (age, sex, and race), lifestyle (physical activity, body mass index, weight change, and smoking history), laboratory tests (biomarkers), physical inspection (blood pressure, electrocardiogram, imaging examination), and comorbidities. It is of great significance to explore the potential novel prognostic factors of HFpEF and build a more convenient and efficient risk prediction model for improving the overall prognosis of patients. This review can provide a substantial reference for further research.


MVA85A vaccine to enhance BCG for preventing tuberculosis.
Tuberculosis causes more deaths than any other infectious disease globally. Bacillus Calmette-Guérin (BCG) is the only available vaccine, but protection is incomplete and variable. The modified Vaccinia Ankara virus expressing antigen 85A (MVA85A) is a viral vector vaccine produced to prevent tuberculosis. To assess and summarize the effects of the MVA85A vaccine boosting BCG in humans. We searched the Cochrane Infectious Diseases Group Specialized Register; Central Register of Controlled Trials (CENTRAL); MEDLINE (PubMed); Embase (Ovid); and four other databases. We searched the WHO ICTRP and ClinicalTrials.gov. All searches were run up to 10 May 2018. We evaluated randomized controlled trials of MVA85A vaccine given with BCG in people regardless of age or HIV status. Two review authors independently assessed the eligibility and risk of bias of trials, and extracted and analyzed data. The primary outcome was active tuberculosis disease. We summarized dichotomous outcomes using risk ratios (RR) and risk differences (RD), with 95% confidence intervals (CI). Where appropriate, we combined data in meta-analyses. Where meta-analysis was inappropriate, we summarized results narratively. The search identified six studies relating to four Phase 2 randomized controlled trials enrolling 3838 participants. Funding was by government bodies, charities, and philanthropic donors. Five studies included infants, one of them infants born to HIV-positive mothers. One study included adults living with HIV. All trials included authors from Oxford University who led the laboratory development of the vaccine. Participants received intradermal MVA85A after BCG in some studies, and before selective deferred BCG in HIV-exposed infants.The largest trial in 2797 African children was well conducted with low risk of bias for most parameters. Risk of bias was uncertain for selective reporting because there were no precise case definition endpoints for active tuberculosis published prior to the trial analysis.MVA85A added to BCG compared to BCG alone probably has no effect on the risk of developing microbiologically confirmed tuberculosis (RR 0.97, 95% CI 0.58 to 1.62; 3439 participants, 2 trials; moderate-certainty evidence), or the risk of starting on tuberculosis treatment (RR 1.10, 95% CI 0.92 to 1.33; 3687 participants, 3 trials; moderate-certainty evidence). MVA85A probably has no effect on the risk of developing latent tuberculosis (RR 1.01, 95% CI 0.85 to 1.21; 3831 participants, 4 trials; moderate-certainty evidence). Vaccinating people with MVA85A in addition to BCG did not cause life-threatening serious adverse effects (RD 0.00, 95% CI -0.00 to 0.00; 3692 participants, 3 trials; high-certainty evidence). Vaccination with MVA85A is probably associated with an increased risk of local skin adverse effects (3187 participants, 3 trials; moderate-certainty evidence), but not systemic adverse effect related to vaccination (144 participants, 1 trial; low-certainty evidence). This safety profile is consistent with Phase 1 studies which outlined a transient, superficial reaction local to the injection site and mild short-lived symptoms such as malaise and fever. MVA85A delivered by intradermal injection in addition to BCG is safe but not effective in reducing the risk of developing tuberculosis.


Strength of social tie predicts cooperative investment in a human social network.
Social networks--diagrams which reflect the social structure of animal groups--are increasingly viewed as useful tools in behavioural ecology and evolutionary biology. Network structure may be especially relevant to the study of cooperation, because the action of mechanisms which affect the cost:benefit ratio of cooperating (e.g. reciprocity, punishment, image scoring) is likely to be mediated by the relative position of actor and recipient in the network. Social proximity could thus affect cooperation in a similar manner to biological relatedness. To test this hypothesis, we recruited members of a real-world social group and used a questionnaire to reveal their network. Participants were asked to endure physical discomfort in order to earn money for themselves and other group members, allowing us to explore relationships between willingness to suffer a cost on another's behalf and the relative social position of donor and recipient. Cost endured was positively correlated with the strength of the social tie between donor and recipient. Further, donors suffered greater costs when a relationship was reciprocated. Interestingly, participants regularly suffered greater discomfort for very close peers than for themselves. Our results provide new insight into the effect of social structure on the direct benefits of cooperation.


Assessment of the Portable C-320 Electronic Nose for Discrimination of Nine Insectivorous Bat Species: Implications for Monitoring White-Nose Syndrome.
The development of new C-320 electronic-nose (e-nose) methods for pre-symptomatic detection of White-Nose Syndrome (WNS) in bats has required efficacy studies of instrument capabilities to discriminate between major sources of volatile organic compounds (VOCs) derived from clinical samples. In this phase-2 study, we further tested this e-nose for capabilities to distinguish between bat species based on differences in whole-body VOC emissions. Live healthy individuals of nine bat species were temporarily captured outside of caves in Arkansas and Louisiana. VOC emissions from bats were collected using newly developed portable air collection and sampling-chamber devices in tandem. Sensor-array output responses to bat VOC emissions were compared to those of 22 pure VOC analytical standards from five chemical classes. Distinct smellprint signatures were produced from e-nose analyses of VOC metabolites derived from individual bat species. Smellprint patterns were analyzed using 2-dimensional and 3-dimensional Principal Component Analysis (PCA) to produce aroma map plots showing effective discrimination between bat species with high statistical significance. These results demonstrate potential instrument efficacy for distinguishing between species-specific, bat-derived VOC metabolite emissions as major components of clinical samples collected from bats in caves for disease detection prior to symptom development. This study provided additional information required to fully test the efficacy of a portable e-nose instrument for diagnostic applications in subsequent phase-3 testing of noninvasive, early WNS disease detection in intra-cave hibernating bats.


The type I interferon system in idiopathic inflammatory myopathies.
Polymyositis (PM), dermatomyositis (DM) and inclusion body myositis (IBM) are chronic inflammatory diseases that are characterized by muscle weakness and inflammatory cells in muscle tissue. Autoantibodies are common, some of them are specific for myositis, the most frequent being the anti-Jo-1 antibody which is associated not only with myositis but also with interstitial lung disease and arthritis. A role of type I interferons in disease mechanisms of myositis was first supported by the reported onset of PM and DM during treatment with type I interferon. More recently an interferon signature has been reported in muscle tissue of DM and PM patients both as gene and protein expression, and type I IFN expression in peripheral blood cells seems to correlate with disease activity. Different mechanisms could induce type I interferon in PM and DM like viral infections or endogenous factors as suggested by the observation that sera from myositis patients with anti-Jo-1 antibodies as well as anti-SSA and anti-SSB antibodies have an interferon inducible capacity. Accumulating data indicate a role of the type I interferon in myositis, particularly in juvenile and adult DM and in anti-Jo-1 or anti-SSA positive PM.


Intramuscular deoxygenation during exercise in patients who have chronic anterior compartment syndrome of the leg.
Currently, the definitive diagnosis of chronic compartment syndrome is based on invasive measurements of intracompartmental pressure. We measured the intramuscular pressure and the relative oxygenation in the anterior compartment of the leg in eighteen patients who were suspected of having chronic compartment syndrome as well as in ten control subjects before, during, and after exercise. Chronic compartment syndrome was considered to be present if the intramuscular pressure was at least fifteen millimeters of mercury (2.00 kilopascals) before exercise, at least thirty millimeters of mercury (4.00 kilopascals) one minute after exercise, or at least twenty millimeters of mercury (2.67 kilopascals) five minutes after exercise. Changes in relative oxygenation were measured with use of the non-invasive method of near-infrared spectroscopy. In all patients and subjects, there was rapid relative deoxygenation after the initiation of exercise, the level of oxygenation remained relatively stable during continued exercise, and there was reoxygenation to a level that exceeded the pre-exercise resting level after the cessation of exercise. During exercise, maximum relative deoxygenation in the patients who had chronic compartment syndrome (mean relative deoxygenation [and standard error], -290 +/- 39 millivolts) was significantly greater than that in the patients who did not have chronic compartment syndrome (-190 +/- 10 millivolts) and that in the control subjects (-179 +/- 14 millivolts) (p < 0.05 for both comparisons). In addition, the interval between the cessation of exercise and the recovery of the pre-exercise resting level of oxygenation was significantly longer for the patients who had chronic compartment syndrome (184 +/- 54 seconds) than for the patients who did not have chronic compartment syndrome (39 +/- 19 seconds) and the control subjects (33 +/- 10 seconds) (p < 0.05 for both comparisons).


Transcriptional silencing and promoter methylation triggered by double-stranded RNA.
Double-stranded RNA induces a post-transcriptional gene silencing process, termed RNAi, in diverse organisms. It is shown here that transcriptional gene silencing accompanied by de novo methylation of a target promoter in plants can be triggered by a double-stranded RNA containing promoter sequences. Similar to the double-stranded RNA involved in RNAi, this promoter double-stranded RNA, which is synthesized in the nucleus, is partially cleaved into small RNAs approximately 23 nucleotides in length. Both transcriptional and post-transcriptional gene silencing can thus be initiated by double-stranded RNAs that enter the same degradation pathway. The results also implicate double-stranded RNA in directing DNA methylation. Different constructs designed to produce double-stranded promoter RNA in various ways were evaluated for their ability to induce gene silencing in tobacco and Arabidopsis. RNA hairpins transcribed from inverted DNA repeats were the most effective trans-acting silencing signals. This strategy could be useful for transcriptionally downregulating genes in a variety of plants.


Venous thrombosis in neonates.
The incidence of thrombotic disorders in neonates and children is increasing with advances in diagnostic modalities, supportive care, and management of many health conditions. The developing coagulation system, need for intensive care, including catheterization, and co-morbid conditions are responsible for the relatively high risk of thrombosis in neonates compared to older children. This review addresses the advances over the last 3 years in neonatal thrombosis, with a focus on epidemiology, cerebral sinovenous thrombosis (CSVT), renal vein thrombosis (RVT), and portal vein thrombosis (PVT). The incidence of neonatal thrombosis in the contemporary era is reported to be 6.9-15 per 1,000 neonatal intensive care unit (NICU) admissions, compared to 2.4 per 1,000 NICU admissions reported in older registry data. The majority of recently published studies are small, retrospective, and from single centers, albeit they emphasize the need for definitive data to support the efficacy and safety of anticoagulation therapy (ACT) in the management of CSVT, RVT, and PVT. We highlight two important international initiatives geared towards improving the evidence base for these conditions. The International Pediatric Thrombosis Network (IPTN) is a collaboration of 74 centers across 27 countries (as of January 2021) which has started important projects like the international neonatal RVT registry, while the International Pediatric Stroke Study (IPSS) group is in the planning stages of a randomized controlled trial to evaluate the utility of ACT in the management of neonatal CSVT.


Use of oily contrast medium for selective drug targeting to tumor: enhanced therapeutic effect and X-ray image.
Highly malignant rabbit tumor (VX-2) was implanted at the periphery of the liver in 63 rabbits. Selective delivery of the anticancer agent copoly(styrenemaleic acid) conjugated to neocarzinostatin (SMANCS), which was dissolved in an oil contrast medium (Lipiodol), was performed by injection via the proper hepatic artery. The anticancer effect was also evaluated by various parameters. By using low-kVp X-ray examination of the resected rabbit liver, Lipiodol was found to distribute throughout the entire liver arterial lumina and was retained there for about 24 hr, but disappeared from the normal liver arterial lumina gradually. However, Lipiodol was retained in the tumor tissue and vessels for at least 7 days, whereas it was undetectable in any other organs. A radioactive analogue of Lipiodol, a chloroiodinated fatty acid, was prepared by using [14C]linoleic acid. This analogue was used in the study of the distribution by low-kVp X-ray examination, Sudan III staining, and autoradiography. Lipiodol remained in the tumor vessels as well as the tumor cells. The use of the radioisotope yielded a quantitative profile of Lipiodol accumulation in tumor tissues; approximately 1000 times more at 15 min and 100 times more at 3 days after the injection than that of most other organs or plasma. Its major excretion route appeared to be through the bile and then the feces. The biological activity of SMANCS was also determined and was found to be significant in both tumor and liver even 7 days after injection. No activity was found in any other organ or tissue. The relatively high biological activity of SMANCS in the nontumorous liver adjacent to the tumor may be the result of continuous drug release from SMANCS-Lipiodol in the tumor tissue. By histological examination, massive tumor necrosis and infiltration of the inflammatory cells were found in the rabbits treated with SMANCS-Lipiodol. In the rabbits treated with Lipiodol alone, necrosis of the tumor was only minimal, and no infiltration of inflammatory cells was observed. Survival periods of the treated rabbits (n = 14) were significantly longer than those of controls (n = 10); 23.1 +/- 5.5 (S.D.) days versus 16.1 +/- 2.9 days (p less than 0.005), respectively, even though only one injection was used for this highly malignant tumor. Mean tumor size for both groups at laparotomy was 163.3 +/- 83.0 sq mm and 160.5 +/- 76.5 sq mm, respectively (not significant).(ABSTRACT TRUNCATED AT 400 WORDS)


Effects of in vivo metabolic acidosis on midcortical bone ion composition.
Chronic metabolic acidosis increases urine calcium excretion without altering intestinal calcium absorption, suggesting that bone mineral is the source of the additional urinary calcium. During metabolic acidosis there appears to be an influx of protons into bone mineral, lessening the magnitude of the decrement in pH. Although in vitro studies strongly support a marked effect of metabolic acidosis on the ion composition of bone, there are few in vivo observations. We utilized a high-resolution scanning ion microprobe with secondary ion mass spectroscopy to determine whether in vivo metabolic acidosis would alter bone mineral in a manner consistent with its purported role in buffering the increased proton concentration. Postweanling mice were provided distilled drinking water with or without 1.5% NH(4)Cl for 7 days; arterial blood gas was then determined. The addition of NH(4)Cl led to a fall in blood pH and HCO(-)(3) concentration. The animals were killed on day 7, and the femurs were dissected and split longitudinally. The bulk cortical ratios Na/Ca, K/Ca, total phosphate/carbon-nitrogen bonds [(PO(2) + PO(3))/CN], and HCO(-)(3)/CN each fell after 1 wk of metabolic acidosis. Because metabolic acidosis induces bone Ca loss, the fall in Na/Ca and K/Ca indicates a greater efflux of bone Na and K than Ca, suggesting H substitution for Na and K on the mineral. The fall in (PO(2) + PO(3))/CN indicates release of mineral phosphates, and the fall in HCO(-)(3)/CN indicates release of mineral HCO(-)(3). Each of these mechanisms would result in buffering of the excess protons and returning the systemic pH toward normal.


Characterization of Nicotine Catabolism through a Novel Pyrrolidine Pathway in Pseudomonas sp. S-1.
Nicotine is a major toxic alkaloid in wastes generated from tobacco production and cigarette manufacturing. In the present work, a nicotine-degrading bacterial strain was isolated from tobacco powdery waste. The isolate was identified as Pseudomonas sp. S-1 based on morphology, physiology, and 16S rRNA gene sequence. Suitable conditions of isolate S-1 for nicotine degradation were pH 7.0 and 30 °C. Catabolic intermediates of nicotine were isolated with preparative-HPLC and characterized with LC-HRMS and NMR. The catabolic pathways of nicotine were involved in dehydrogenation, oxidation, hydrolysis, and hydroxylation. Interestingly, nicotine catabolism in strain S-1 undergoes a new pyrrolidine pathway that differs from the other three catabolic pathways in bacterial species. This work sheds light on catabolic diversity of nicotine and heteroaromatics.


The Digenea parasite Opisthorchis felineus: a target for the discovery and development of novel drugs.
Opisthorchosis is a helminthiasis affecting mainly the hepatobiliary system and pancreas; its most dramatic complication is malignization of the organs infected by the parasites. The causative agents of opisthorchosis are two species of liver flukes, the trematodes belonging to the family Opisthorchiidae--Opisthorchis felineus and O. viverrini. The Chinese liver fluke, Clonorchis sinensis, also member of the family Opisthorchiidae, causes clonorchosis, a disease very close in symptomatology. According to different estimations, up to 40 million people are currently infected with these liver flukes and up to 600-750 million people in Eurasian countries constitute the risk group. These parasites colonize ever-increasing new areas in Eurasia where this disease has never been previously reported. Opisthorchiases are gradually transforming from a local problem of individual geographic regions to a widespead problem; in particular, O. viverrini is now referred to as "an underestimated parasite." As we see it, O. felineus has all the reasons to share this status. First and foremost, the observed expansion is likely to be connected with the ever-increasing intensity of traffic flows and migration of the infection carriers between cities, regions, and countries. This review briefs the characteristics of O. felineus and the other liver flukes persisting in various countries of Eurasia, clinical manifestations of opisthorchosis, the drugs for chemotherapy of trematodiasis, and the strategy for discovery of new antihelminthic drugs.


Histological Changes of the Middle Ear Ossicles Harvested during Cholesteatoma Surgery.
In the cholesteatoma surgery ossicles can be replaced to reconstruct middle ear function. It is important that these ossicles are free of squamous epithelium, to prevent residual disease. This study focuses on the histological findings of the malleus and incus harvested during cholesteatoma surgery. Eighty middle ears ossicles were examined in vivo and histologically to consider the relationship of cholesteatoma to ossicles, grade of bone destruction and invasion of cholesteatoma to deeper layers of bone. Serious ossicular destruction was observed more frequently in incus compared to malleus (p=0.0065). Difference of ossicles destruction between children and adults was not significant (p=0.3032). Deep invasion of cholesteatoma into the vascular spaces or inner core of the bone was not observed. Autograft ossicles from cholesteatomatous ears should not necessarily be rejected for reconstruction of the ossicular chain. Regarding the histological finding, the authors suggest mechanical cleaning of the ossicle surface to eliminate residual disease.