Development and Validation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Glaesserella (Haemophilus) parasuis.
Glaesserella parasuis is a fastidious pathogen that colonizes the respiratory tract of pigs and can lead to considerable economic losses in pig production. Therefore, a rapid detection assay for the pathogen, preferably applicable in the field, is important. In the current study, we developed a new and improved detection method using loop-mediated isothermal amplification (LAMP). This assay, which targets the infB gene, was tested on a collection of 60 field isolates of G. parasuis comprising 14 different serovars. In addition, 63 isolates from seven different closely related species of the family Pasteurellaceae, including A. indolicus, A. porcinus, and A. minor, and a species frequently found in the respiratory tract of pigs were used for exclusivity experiments. This assay showed an analytical specificity of 100% (both inclusivity and exclusivity) and an analytical sensitivity of 10 fg/µL. In further steps, 36 clinical samples were tested with the LAMP assay. An agreement of 77.1 (95% CI: 59.9, 89.6) and 91.4% (95% CI: 75.9, 98.2) to the culture-based and PCR results was achieved. The mean limit of detection for the spiked bronchoalveolar lavage fluid was 2.58 × 102 CFU/mL. A colorimetric assay with visual detection by the naked eye was tested to provide an alternative method in the field and showed the same sensitivity as the fluorescence-based LAMP assay. Overall, the optimized LAMP assay represents a fast and reliable method and is suitable for detecting G. parasuis in the laboratory environment or in the field.


Social Support During COVID-19: Perspectives of Breastfeeding Mothers.
Introduction: Effective social support can have a critical influence on a mother's ability to initiate and continue breastfeeding. Coronavirus disease (COVID-19) has created unprecedented barriers for breastfeeding mothers to obtain various types of support: emotional, instrumental, informational, and appraisal. However, no research has evaluated the influence the pandemic has had on breastfeeding supports. The purpose of this study was to explore perceptions of social support among breastfeeding mothers during the COVID-19 pandemic. Materials and Methods: A cross-sectional phenomenological approach was taken utilizing semistructured interviews (March-June 2020) with currently breastfeeding mothers (n = 29). Data were analyzed through a process of immersion and crystallization. Results: Mothers are still able to obtain each type of support, however, support has been negatively influenced by the pandemic. Mothers reported experiencing increased stress and isolation and had an immense desire to receive in-person support from peers, family, childcare providers, and lactation specialists. Furthermore, mothers of multiple children felt if they did not already have breastfeeding knowledge from previous experiences they would be unsuccessful in breastfeeding due to their current lack of support. Conversely, a majority of mothers felt the pandemic had positively influenced their breastfeeding journeys due to concerns of formula shortages and extended maternity leaves. Finally, mothers were concerned about safely expressing breast milk on their return to work. Conclusion: Mother's ability to obtain breastfeeding support has been negatively impacted by the pandemic due to the inability to engage with individuals in-person and the lack of access to childcare. First-time mothers may be at higher risk of early breastfeeding cessation due to lack of support. However, breastfeeding journeys have also been positively influenced by allowing mothers more time at home with their child. Resources are needed to support expressing breast milk in the workplace during COVID-19.


MID1 catalyzes the ubiquitination of protein phosphatase 2A and mutations within its Bbox1 domain disrupt polyubiquitination of alpha4 but not of PP2Ac.
MID1 is a microtubule-associated protein that belongs to the TRIM family. MID1 functions as an ubiquitin E3 ligase, and recently was shown to catalyze the polyubiquitination of, alpha4, a protein regulator of protein phosphatase 2A (PP2A). It has been hypothesized that MID1 regulates PP2A, requiring the intermediary interaction with alpha4. Here we report that MID1 catalyzes the in vitro ubiquitination of the catalytic subunit of PP2A (PP2Ac) in the absence of alpha4. In the presence of alpha4, the level of PP2Ac ubiquitination is reduced. Using the MID1 RING-Bbox1-Bbox2 (RB1B2) construct containing the E3 ligase domains, we investigate the functional effects of mutations within the Bbox domains that are identified in patients with X-linked Opitz G syndrome (XLOS). The RB1B2 proteins harboring the C142S, C145T, A130V/T mutations within the Bbox1 domain and C195F mutation within the Bbox2 domain maintain auto-polyubiquitination activity. Qualitatively, the RB1B2 proteins containing these mutations are able to catalyze the ubiquitination of PP2Ac. In contrast, the RB1B2 proteins with mutations within the Bbox1 domain are unable to catalyze the polyubiquitination of alpha4. These results suggest that unregulated alpha4 may be the direct consequence of these natural mutations in the Bbox1 domain of MID1, and hence alpha4 could play a greater role to account for the increased amount of PP2A observed in XLOS-derived fibroblasts.


Transmission mode associated with coronavirus disease 2019: a review.
In late December 2019 in Wuhan (China), Health Commission reported a cluster of pneumonia cases of unknown etiology, subsequently isolated and named Severe Acute Respiratory Syndrome (SARS) Coronavirus 2 (CoV-2). In this review, the main transmission routes and causes of mortality associated with COVID-19 were investigated. A review was carried out to recognize relevant research available until 10 April 2020. The main transmission routes of COVID-19 have been the following: animal to human and human-to-human pathways, namely: respiratory transmission; oro-fecal transmission; air, surface-human transmission. Transmission from asymptomatic persons, healthcare transmission, and interfamily transmission have been well documented. SARS-CoV-2 possesses powerful pathogenicity and transmissibility. It is presumed to spread primarily via respiratory droplets and close contact. The most probable transmission pathway is definitely the inter-human one. Asymptomatic patients seem to play a crucial role in spreading the infection. Because of COVID-19 infection pandemic potential, careful surveillance is essential to monitor its future host adaptation, viral evolution, infectivity, transmissibility, and pathogenicity in order to gain an effective vaccine and flock immunity and reduce mortality as soon and as much as it is possible.


Strategies for preventing end stage kidney disease: The impact of kidney stone disease on Chronic Kidney Disease in Pakistan.
Chronic Kidney Disease (CKD) is one of the major noncommunicable diseases that have social and economic impacts along with day-to-day health-related problems. Kidney stone disease is still one of the major causes of CKD in Pakistan. Kidney stone disease is a preventable cause of renal failure, if detected and treated early. Kidney stone is prevalent in the stone belt areas in our country which are located in rural areas. The treatment is very costly, and mostly available only in large cities. The treatment requires tertiary care setup and expertise. Therefore, there is a need to focus on the preventive strategies which are mainly dietary and lifestyle modifications, along with public awareness programmes. Nephrologists and urologists along with the government should take interest and give priority to CKD due to stone disease.


HugeIndex: a database with visualization tools for high-density oligonucleotide array data from normal human tissues.
High-density oligonucleotide arrays are a powerful tool for uncovering changes in global gene expression in various disease states. To this end, it is essential to first characterize the variations of gene expression in normal physiological processes. We established the Human Gene Expression (HuGE) Index database (www.HugeIndex.org) to serve as a public repository for gene expression data on normal human tissues using high-density oligonucleotide arrays. This resource currently contains the results of 59 gene expression experiments on 19 human tissues. We provide interactive tools for researchers to query and visualize our data over the Internet. To facilitate data analysis, we cross-reference each gene on the array with its annotation in the LocusLink database at NCBI.


Recommendations for Pregnancy in Rare Inherited Anemias.
Rare inherited anemias are a subset of anemias caused by a genetic defect along one of the several stages of erythropoiesis or in different cellular components that affect red blood cell integrity, and thus its lifespan. Due to their low prevalence, several complications on growth and development, and multi-organ system damage are not yet well defined. Moreover, during the last decade there has been a lack of proper understanding of the impact of rare anemias on maternal and fetal outcomes. In addition, there are no clear-cut guidelines outlining the pathophysiological trends and management options unique to this special population. Here, we present on behalf of the European Hematology Association, evidence- and consensus-based guidelines, established by an international group of experts in different fields, including hematologists, gynecologists, general practitioners, medical geneticists, and experts in rare inherited anemias from various European countries for standardized and appropriate choice of therapeutic interventions for the management of pregnancy in rare inherited anemias, including Diamond-Blackfan Anemia, Congenital Dyserythropoietic Anemias, Thalassemia, Sickle Cell Disease, Enzyme deficiency and Red cell membrane disorders.


Starvation-induced changes of palmitate metabolism and insulin secretion in isolated rat islets stimulated by glucose.
The influence of 48 h starvation on glucose-induced changes of palmitate metabolism and insulin release in isolated rat islets was investigated. (1) Islet insulin response to 20 mM-glucose was abolished after 48 h starvation, and it was restored by 0.25 mM-2-bromostearate, an inhibitor of fatty acid oxidation. (2) The increase in glucose concentration from 3 to 20 mM was accompanied by a 50% decrease in the oxidation rate of 0.5 mM-[U-14C]palmitate in control (fed) islets, and a concomitant increase (100%) in its incorporation into triacylglycerol and phospholipid fractions. (3) Starvation induced a higher basal (3 mM-glucose) rate of palmitate oxidation, which was resistant to inhibition by 20 mM-glucose. The latter also failed to increase palmitate incorporation into islet triacylglycerols and phospholipids. (4) 2-Bromostearate (0.25 mM) strongly inhibited the high oxidation rate of palmitate in islets of starved rats, and allowed a normal stimulation of its incorporation rate into islet lipids by 20mM-glucose. (5) The results suggest that starvation restricts islet esterification of fatty acids by inducing a higher rate of their oxidative degradation that is insensitive to regulation by glucose.


Tied Pipeline: A Case of Rare Complication.
A 48-year-old female with a symptomatic giant carotid cavernous aneurysm underwent endovascular treatment with multiple Pipeline Flex embolic devices (PED). The delivery catheter had to take a complicated course of multiple turns to cross the aneurysm, and its loop was tied in the aneurysm. When the catheter was to be withdrawn, it was apparent that the tied catheter had made a tight knot that bound the tail of the previous PED together. We successfully retrieved all of the system including the tied PED, and we used telescoping stents with five PEDs in the next operation. Although this is a rare complication, it is worth noting and checking to make sure that there is no knot before deploying the stent.


Effect of antidysrhythmic drugs on the heart rate and blood pressure response to treadmill exercise.
The heart rate and blood pressure response to exercise are clinically important parameters evaluated during treadmill testing. These responses may be altered by cardiac drugs. We evaluated the effect of procainamide and disopyramide on the heart rate and blood pressure response to treadmill exercise in 9 healthy volunteers. Each subject performed one Bruce protocol treadmill test while on each of three treatments: placebo, procainamide 1.0 g t.i.d.; disopyramide 150 mg t.i.d. The order of the treatments was randomized in a Latin square design. The study was conducted double-blind. The drugs slightly increased the resting heart rate, but had no significant effect on exercising heart rate or blood pressure. We conclude that the use of procainamide or disopyramide does not interfere with the interpretation of results of treadmill testing.


[Experiences with the psychometric system combitest in the evaluation of the postnarcotic arousal period].
Since the introduction of new German health legislation, operations formerly performed on an in-patient basis have been transferred to the out-patient sector. Due to this development evaluating the postnarcotic arousal period has become increasingly important. Vigilance, reactivity and state of health determine a patient's well-being and time of discharge after an operation and consequently influence economical and legal issues. Former studies investigating the arousal period have mainly focused on patient interviews and assessment of patients by staff. The aim of our study was to objectivize postnarcotic vigilance and reactivity (critical flicker fusion frequency and multiple choice reaction) using neurophysiological monitoring (psychometric system Combitest 2: CFF, EWR, MWR). The applied methods have been defined as valid, reliable and sensitive tools to investigate this question. Inter-individual and gender-specific differences as well as variations due to the type of anaesthetic employed can be detected. The study participants assessed the test battery's practicability and overall described this method as easy to use. Vigilance (CFF), basic and complex sensomotor reactivity (performance in simple and multiple choice tests) were restored after a postanaesthetic phase of 90 minutes (narcotics applied: isofluran and propofol). The results show that the use of modern anaesthetics does not justify the current medical and legal claim to 6 hours of postanaesthetic observation. The computer-supported psychometric system Combitest 2 has recently been used to assess the arousal phase after application of different types of anaesthetical procedures (TIVA, volatile, balanced) and has been found to be superior to parameters traditionally used to assess discharge criteria (time until patient opens eyes, orientation concerning time and location, time spent in postanaesthetic recovery room). Besides employing our study methods, subjective factors such as well-being and pain sensation should also be evaluated when determining the time of a patient's discharge.


Manager Perspectives on Implementation of a Farmers' Market Incentive Program in Maryland.
A growing number of farmers' markets offer incentive programs to increase access to fresh produce in low-income communities and support local farmers, yet program implementation has not been widely studied. This qualitative study explored market manager perspectives on implementing a Maryland statewide farmers' market incentive program. Semistructured interviews with market managers. Participating farmers' markets across Maryland. Managers of participating farmers' markets (n = 19). Barriers and facilitators to program implementation and recommendations for improvement. Interviews were analyzed using a phronetic iterative approach and themes were organized using an adapted version of a multilevel implementation framework. Managers reported overall favorable attitudes toward the incentive program. They identified barriers to implementation such as vendor buy-in and funding uncertainties, facilitators such as private fund-raising and local champions, and opportunities for improvement such as strengthening promotion and increasing the use of technology to reduce the data collection burden. Managers identified implementation barriers (eg, customer confusion) and facilitators (eg, ease of staff training) that may be shared by other incentive programs. Based on these findings, barriers and recommendations may be addressed through collaborative problem-solving with managers, vendors, and customers, and facilitators can be amplified broadly. Future research is warranted to explore incentive program implementation in other settings.


Population structure and growth of two opulations of Aegla Schmitti Hobbs III, 1979 (Anomura, Aeglidae) in the eastern Paraná state, Brazil.
The population structure and growth of two populations (Arraial and Capivari rivers) of Aegla schmitti was evaluated. The following parameters were determined: size, age structure, sex ratio, recruitment period, and sampling method selectivity (traps and handnet). There was variation between males and females. The carapace length size class distribution was unimodal in the Arraial River, and polymodal in the Capivari River. The sampling method influenced the proportion of demographic categories. The recruitment occurred mostly between October and December. The growth curves for the Arraial and Capivari rivers, respectively, were Ct= 26,50[1- e -0,004(t+53,36)] and Ct= 34,79[1- e -0,0024(t+122,34)] for males, and Ct= 23,07[1- e -0.0027(t+23,18)] and Ct= 25.52[1- e -0,0033(t+19,51)] for females. The maximum CL estimated for males and females were 26.50 and 23.07mm for the Arraial River, and 34.79 and 25.52mm for the Capivari River. Longevity was estimated in 2 years for females of both populations, and 2.5 and 3 years for males of the Arraial and Capivari rivers, respectively. The results allow for the characterization of the growth patterns of A. schmitti in two hydrographic basins, providing information for the species' preservation and also demonstrating the selectivity of the different sampling methods used for aeglids.


Regulation of cyclin D1 gene expression.
Cyclin D1 is a key regulator of cell proliferation and its expression is subject to both transcriptional and post-transcriptional regulation. In different cellular contexts, different pathways assume a dominant role in regulating its expression, whereas their disregulation can contribute to overexpression of cyclin D1 in tumorigenesis. Here, we discuss the ability of the NF-kappaB (nuclear factor kappaB)/IKK [IkappaB (inhibitor of NF-kappaB) kinase] pathways to regulate cyclin D1 gene transcription and also consider the newly discovered role of the SNARP (SNIP1/SkIP-associated RNA processing) complex as a co-transcriptional regulator of cyclin D1 RNA stability.


Cree antidiabetic plant extracts display mechanism-based inactivation of CYP3A4.
Seventeen Cree antidiabetic medicinal plants were studied to determine their potential to inhibit cytochrome P450 3A4 (CYP3A4) through mechanism-based inactivation (MBI). The ethanolic extracts of the medicinal plants were studied for their inhibition of CYP3A4 using the substrates testosterone and dibenzylfluorescein (DBF) in high pressure liquid chromatography (HPLC) and microtiter fluorometric assays, respectively. Using testosterone as a substrate, extracts of Alnus incana, Sarracenia purpurea, and Lycopodium clavatum were identified as potent CYP3A4 MBIs, while those from Abies balsamea, Picea mariana, Pinus banksiana, Rhododendron tomentosum, Kalmia angustifolia, and Picea glauca were identified as less potent inactivators. Not unexpectedly, the other substrate, DBF, showed a different profile of inhibition. Only A. balsamea was identified as a CYP3A4 MBI using DBF. Abies balsamea displayed both NADPH- and time-dependence of CYP3A4 inhibition using both substrates. Overall, several of the medicinal plants may markedly deplete CYP3A4 through MBI and, consequently, decrease the metabolism of CYP3A4 substrates including numerous medications used by diabetics.


Mother cells control daughter cell proliferation in intestinal organoids to minimize proliferation fluctuations.
During renewal of the intestine, cells are continuously generated by proliferation. Proliferation and differentiation must be tightly balanced, as any bias toward proliferation results in uncontrolled exponential growth. Yet, the inherently stochastic nature of cells raises the question how such fluctuations are limited. We used time-lapse microscopy to track all cells in crypts of growing mouse intestinal organoids for multiple generations, allowing full reconstruction of the underlying lineage dynamics in space and time. Proliferative behavior was highly symmetric between sister cells, with both sisters either jointly ceasing or continuing proliferation. Simulations revealed that such symmetric proliferative behavior minimizes cell number fluctuations, explaining our observation that proliferating cell number remained constant even as crypts increased in size considerably. Proliferative symmetry did not reflect positional symmetry but rather lineage control through the mother cell. Our results indicate a concrete mechanism to balance proliferation and differentiation with minimal fluctuations that may be broadly relevant for other tissues.


Human biomarker discovery and predictive models for disease progression for idiopathic pneumonia syndrome following allogeneic stem cell transplantation.
Allogeneic hematopoietic stem cell transplantation (SCT) is the only curative therapy for many malignant and nonmalignant conditions. Idiopathic pneumonia syndrome (IPS) is a frequently fatal complication that limits successful outcomes. Preclinical models suggest that IPS represents an immune mediated attack on the lung involving elements of both the adaptive and the innate immune system. However, the etiology of IPS in humans is less well understood. To explore the disease pathway and uncover potential biomarkers of disease, we performed two separate label-free, proteomics experiments defining the plasma protein profiles of allogeneic SCT patients with IPS. Samples obtained from SCT recipients without complications served as controls. The initial discovery study, intended to explore the disease pathway in humans, identified a set of 81 IPS-associated proteins. These data revealed similarities between the known IPS pathways in mice and the condition in humans, in particular in the acute phase response. In addition, pattern recognition pathways were judged to be significant as a function of development of IPS, and from this pathway we chose the lipopolysaccaharide-binding protein (LBP) protein as a candidate molecular diagnostic for IPS, and verified its increase as a function of disease using an ELISA assay. In a separately designed study, we identified protein-based classifiers that could predict, at day 0 of SCT, patients who: 1) progress to IPS and 2) respond to cytokine neutralization therapy. Using cross-validation strategies, we built highly predictive classifier models of both disease progression and therapeutic response. In sum, data generated in this report confirm previous clinical and experimental findings, provide new insights into the pathophysiology of IPS, identify potential molecular classifiers of the condition, and uncover a set of markers potentially of interest for patient stratification as a basis for individualized therapy.


Therapeutic Effects of Diagonal-Transcranial Direct Current Stimulation on Functional Recovery in Acute Stroke: A Pilot Study.
Peak neurologic recovery from acute stroke occurs within the first 3 months, and continues at a slower pace for 6 months. The aim of this pilot study is to clarify the safety and feasibility of multiple diagonal-transcranial direct current stimulation (d-tDCS) sessions up to 3 months with electrodes placed diagonally over the lesional dorsolateral prefrontal cortex and contralesional primary motor cortex for upper limb hemiparesis in acute stroke. Five patients with acute stroke (2 with intracerebral hemorrhage and 3 with cerebral infarction) with upper limb paresis participated. d-tDCS (1 mA, 20 min per day) combined with conventional rehabilitation was given starting 7-21 days after stroke onset. Each session consisted of 10 d-tDCS over 2 weeks and patients received 2 sessions in the acute phase and 2 sessions in the subacute phase for a total 40 treatments. Motor function was assessed using Fugl-Meyer Assessment for upper extremity (FMA-UE) before and after each session, and the period to achieve 70% of maximal potential recovery in FMA-UE was evaluated. All 5 patients completed the intervention and showed no adverse effects throughout the protocol. Of these, 3 (60%) achieved 70% predicted scores within 2 months. Regarding therapeutic gains of FMA-UE in each of the 10 sessions in the acute phase, 4 sessions showed great recovery, 3 sessions showed moderate recovery, and 3 sessions showed poor recovery. These findings suggest that d-tDCS over 3 months may be safe and feasible for acute stroke patients in the acute to subacute phases and have therapeutic potential to promote recovery of upper limb function, although further randomized, double-blind, sham-controlled trial is warranted with larger sample size.


Elastic and inelastic cross sections for low-energy electron collisions with pyrimidine.
We present theoretical elastic and electronic excitation cross sections and experimental electronic excitation cross sections for electron collisions with pyrimidine. We use the R-matrix method to determine elastic integral and differential cross sections and integral inelastic cross sections for energies up to 15 eV. The experimental inelastic cross sections have been determined in the 15-50 eV impact energy range. Typically, there is quite reasonable agreement between the theoretical and experimental integral inelastic cross sections. Calculated elastic cross sections agree very well with prior results.


Loss of pedunculopontine neurons in progressive supranuclear palsy.
In the present study, the number of neurons (greater than 20 microns in diameter) within the lateral part of the pedunculopontine nucleus pars compacta (PPNc) was determined at six rostrocaudal levels in 3 subjects with progressive supranuclear palsy (PSP), in 9 subjects with Alzheimer's disease, and in 6 age-matched control subjects. At each level examined, significantly fewer neurons were present in patients with PSP than in control subjects (49 to 69% reduction). Significant differences in numbers of neurons were not demonstrated between control subjects and patients with Alzheimer's disease. The extent of pathological changes, particularly neurofibrillary tangles, was examined within the PPNc of subjects in these three groups. The average number of neurofibrillary tangles in a 12-microns-thick midlevel section of the lateral PPNc was 68.7 in subjects with PSP, 18.3 in those with Alzheimer's disease, and 3.0 in aged control subjects. These abnormalities of PPNc neurons in PSP may play important roles in some of the clinical features characteristic of this disease.


[AIDS. The etiologic agent for acquired immunodeficiency syndrome].
In 1981, an unusual case with immunodeficiency was reported and there after an epidemic fo the similar syndrome in population of homosexuals was recognized by Center for Diseases Control(CDC) U. S. A. Later the disease was characterized as an acquired immunodeficiency syndrome(AIDS). In 1983, virologists found the causative agent which is a novel retrovirus named human immunodeficiency virus(HIV). Since then, by use of modern biotechnology, numerous and detailed informations have been accumulated by various researchers. The result, however, demonstrated that there is no a single method of satisfaction to prevent of cure the disease except avoiding from the agent. This problems are discussed by reviewing the results reported up to date.


DTPA Treatment of Wound Contamination in Rats with Americium: Evaluation of Urinary Profiles Using STATBIODIS Shows Importance of Prompt Administration.
In the nuclear industry, wound contamination with americium is expected to increase with decommissioning and waste management. Treatment of workers with diethylenetriaminepentaacetic acid (DTPA) requires optimization to reduce internal contamination and radiation exposure. This work aimed at evaluating and comparing different DTPA protocol efficacies after wound contamination of rats with americium. Wound contamination was simulated in rats by depositing americium nitrate in an incision in the hind limb. Different routes, times, and frequencies of DTPA administration were evaluated. Individual daily urinary americium excretion and tissue retention were analyzed using the statistical tool STATBIODIS. Urinary profiles, urinary enhancement factors, and inhibition percentages of tissue retention were calculated. A single DTPA administration the day of contamination induced a rapid increase in americium urinary excretion that decreased exponentially over 7 d, indicating that the first DTPA administration should be delivered as early as possible. DTPA treatment limited americium uptake in systemic tissues irrespective of the protocol. Liver and skeleton burdens were markedly reduced, which would drive reduction of radiation dose. Local or intravenous injections were equally effective. Inherent difficulties in wound site activity measurements did not allow identification of a significant decorporating effect at the wound site. Repeated intravenous injections of DTPA also increased americium urinary excretion, which supports the use of multiple DTPA administrations shortly after wound contamination. Results from these statistical analyses will contribute to a better understanding of americium behavior in the presence or absence of DTPA and may aid optimization of treatment for workers.


Improved Salinity Tolerance in Carrizo Citrange Rootstock through Overexpression of Glyoxalase System Genes.
Citrus plants are widely cultivated around the world and, however, are one of the most salt stress sensitive crops. To improve salinity tolerance, transgenic Carrizo citrange rootstocks that overexpress glyoxalase I and glyoxalase II genes were obtained and their salt stress tolerance was evaluated. Molecular analysis showed high expression for both glyoxalase genes (BjGlyI and PgGlyII) in 5H03 and 5H04 lines. Under control conditions, transgenic and wild type plants presented normal morphology. In salinity treatments, the transgenic plants showed less yellowing, marginal burn in lower leaves and showed less than 40% of leaf damage compared with wild type plants. The transgenic plants showed a significant increase in the dry weight of shoot but there are no differences in the root and complete plant dry weight. In addition, a higher accumulation of chlorine is observed in the roots in transgenic line 5H03 but in shoot it was lower. Also, the wild type plant accumulated around 20% more chlorine in the shoot compared to roots. These results suggest that heterologous expression of glyoxalase system genes could enhance salt stress tolerance in Carrizo citrange rootstock and could be a good biotechnological approach to improve the abiotic stress tolerance in woody plant species.


Cloud computing applications for biomedical science: A perspective.
Biomedical research has become a digital data-intensive endeavor, relying on secure and scalable computing, storage, and network infrastructure, which has traditionally been purchased, supported, and maintained locally. For certain types of biomedical applications, cloud computing has emerged as an alternative to locally maintained traditional computing approaches. Cloud computing offers users pay-as-you-go access to services such as hardware infrastructure, platforms, and software for solving common biomedical computational problems. Cloud computing services offer secure on-demand storage and analysis and are differentiated from traditional high-performance computing by their rapid availability and scalability of services. As such, cloud services are engineered to address big data problems and enhance the likelihood of data and analytics sharing, reproducibility, and reuse. Here, we provide an introductory perspective on cloud computing to help the reader determine its value to their own research.


The 2-thiouridylase function of the human MTU1 (TRMU) enzyme is dispensable for mitochondrial translation.
MTU1 (TRMU) is a mitochondrial enzyme responsible for the 2-thiolation of the wobble U in tRNA(Lys), tRNA(Glu) and tRNA(Gln), a post-transcriptional modification believed to be important for accurate and efficient synthesis of the 13 respiratory chain subunits encoded by mtDNA. Mutations in MTU1 are associated with acute infantile liver failure, and this has been ascribed to a transient lack of cysteine, the sulfur donor for the thiouridylation reaction, resulting in a mitochondrial translation defect during early development. A mutation in tRNA(Lys) that causes myoclonic epilepsy with ragged-red fibers (MERRF) is also reported to prevent modification of the wobble U. Here we show that mitochondrial translation is unaffected in fibroblasts from an MTU1 patient, in which MTU1 is undetectable by immunoblotting, despite the severe reduction in the 2-thiolation of mitochondrial tRNA(Lys), tRNA(Glu) and tRNA(Gln). The only respiratory chain abnormality that we could observe in these cells was an accumulation of a Complex II assembly intermediate, which, however, did not affect the level of the fully assembled enzyme. The identical phenotype was observed by siRNA-mediated knockdown of MTU1 in HEK 293 cells. Further, the mitochondrial translation deficiencies present in myoblasts from mitochondrial encephalomyopathy, lactic acidosis and stroke-like episode and MERRF patients, which are associated with defects in post-transcriptional modification of mitochondrial tRNAs, did not worsen following knockdown of MTU1 in these cells. This study demonstrates that MTU1 is not required for mitochondrial translation at normal steady-state levels of tRNAs, and that it may possess an as yet uncharacterized function in another sulfur-trafficking pathway.


Cytokine regulation of colony-stimulating factor (CSF) production in cultured human synovial fibroblasts. II. Similarities and differences in the control of interleukin-1 induction of granulocyte-macrophage CSF and granulocyte-CSF production.
Synovial fibroblasts are likely to be a significant source of granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-CSF (G-CSF), which could be crucial to the pathogenesis of rheumatoid arthritis. Using specific enzyme-linked immunosorbent assays (ELISAs) and Northern analysis, GM-CSF and G-CSF expression were followed in human synovial fibroblast-like cells in response to a number of agents, either alone or in the presence of an optimal stimulatory concentration of interleukin-1 (IL-1). For both CSFs, interferon-gamma (100 U/mL) did not increase their levels but dramatically suppressed the stimulatory action of IL-1, while basic fibroblast growth factor (10(-8) mol/L), although nonstimulatory by itself, potentiated IL-1 action. The glucocorticoid, dexamethasone (10(-7) mol/L), inhibited IL-1-stimulated CSF production. However, evidence was obtained for noncoordinated CSF regulation. Cyclooxygenase inhibitors potentiated the action of IL-1 on GM-CSF synthesis but suppressed G-CSF synthesis, suggesting that endogenous cyclooxygenase products can have opposite effects in modulating the levels of each CSF. Also, the lymphokine, IL-4 (250 pmol/L), slightly inhibited GM-CSF formation in the presence of IL-1 but elevated the G-CSF levels in these cultures without having an effect by itself. Transforming growth factor beta (less than or equal to 20 ng/mL) did not modulate levels of either CSF. Mesenchymal cell production of both GM-CSF and G-CSF is generally viewed as being under coordinate control; our findings suggest that their synthesis in IL-1-stimulated human synoviocytes can be modulated by a number of agents, in some cases with divergent actions depending on which CSF is examined.


iBLAST: Incremental BLAST of new sequences via automated e-value correction.
Search results from local alignment search tools use statistical scores that are sensitive to the size of the database to report the quality of the result. For example, NCBI BLAST reports the best matches using similarity scores and expect values (i.e., e-values) calculated against the database size. Given the astronomical growth in genomics data throughout a genomic research investigation, sequence databases grow as new sequences are continuously being added to these databases. As a consequence, the results (e.g., best hits) and associated statistics (e.g., e-values) for a specific set of queries may change over the course of a genomic investigation. Thus, to update the results of a previously conducted BLAST search to find the best matches on an updated database, scientists must currently rerun the BLAST search against the entire updated database, which translates into irrecoverable and, in turn, wasted execution time, money, and computational resources. To address this issue, we devise a novel and efficient method to redeem past BLAST searches by introducing iBLAST. iBLAST leverages previous BLAST search results to conduct the same query search but only on the incremental (i.e., newly added) part of the database, recomputes the associated critical statistics such as e-values, and combines these results to produce updated search results. Our experimental results and fidelity analyses show that iBLAST delivers search results that are identical to NCBI BLAST at a substantially reduced computational cost, i.e., iBLAST performs (1 + δ)/δ times faster than NCBI BLAST, where δ represents the fraction of database growth. We then present three different use cases to demonstrate that iBLAST can enable efficient biological discovery at a much faster speed with a substantially reduced computational cost.


Intracoronary administration of cardiac progenitor cells alleviates left ventricular dysfunction in rats with a 30-day-old infarction.
Administration of cardiac progenitor cells (CPCs) 4 hours after reperfusion ameliorates left ventricular function in rats with acute myocardial infarction (MI). Clinically, however, this approach is not feasible, because expansion of autologous CPCs after acute MI requires several weeks. Therefore, we sought to determine whether CPCs are beneficial in the more clinically relevant setting of an old MI (scar). One month after coronary occlusion/reperfusion, rats received an intracoronary infusion of vehicle or enhanced green fluorescent protein-labeled CPCs. Thirty-five days later, CPC-treated rats exhibited more viable myocardium in the risk region, less fibrosis in the noninfarcted region, and improved left ventricular function. Cells that stained positive for enhanced green fluorescent protein that expressed cardiomyocyte, endothelial, and vascular smooth muscle cell markers were observed only in 7 of 17 treated rats and occupied only 2.6% and 1.1% of the risk and noninfarcted regions, respectively. Transplantation of CPCs was associated with increased proliferation and expression of cardiac proteins by endogenous CPCs. Intracoronary administration of CPCs in the setting of an old MI produces beneficial structural and functional effects. Although exogenous CPCs can differentiate into new cardiac cells, this mechanism is not sufficient to explain the benefits, which suggests paracrine effects; among these, the present data identify activation of endogenous CPCs. This is the first report that CPCs are beneficial in the setting of an old MI when given by intracoronary infusion, the most widely applicable therapeutic approach in patients. Furthermore, this is the first evidence that exogenous CPC administration activates endogenous CPCs. These results open the door to new therapeutic applications for the use of autologous CPCs in patients with old MI and chronic ischemic cardiomyopathy.


Incomplete ossification of the humeral condyle in spaniels.
An evaluation of 157 dogs with humeral fractures was performed. Cocker spaniels were more likely to have humeral condylar fractures (HCFs) than other breeds (P < .001). Male cocker spaniels were at increased risk (P < .001). Cocker spaniels had more bilateral HCFs than other breeds of dogs (P < .001). Eighteen dogs (17 purebred spaniels and 1 crossbred spaniel) with HCFs of unknown cause or occurring with normal activity were further studied, using radiography of their humeral condyle bilaterally (n = 18), computed tomography (n = 3), biopsy (n = 2), bone scintigraphy (n = 2), and genetic evaluation (n = 8). Fourteen of these 18 dogs had a nonfractured contralateral condyle. Twelve (86%) of the 14 nonfractured humeral condyles had a radiolucent line within the center of the condyle, 13 (93%) had radiographic signs of degenerative joint disease and an abnormal medial coronoid process, and six (43%) had periosteal proliferation involving the lateral epicondyle. Examination of biopsy samples from the fracture sites of two cocker spaniels showed fibrous tissue present at the fracture surfaces. The results of this study suggest an association between incomplete ossification of the humeral condyle in cocker spaniels and Brittany spaniels and a high prevalence of HCFs. Eight affected cocker spaniels with available pedigree information were found to be genetically related, suggesting that incomplete ossification of the humeral condyle may be a genetic disease with a recessive mode of inheritance.


OR Air Quality: Is It Time to Consider Adjunctive Air Cleaning Technology?: 1.3 www.aornjournal.org/content/cme.
Patients undergoing surgery may be at risk for infection from airborne particles such as dust, skin scales, respiratory aerosols, and hair fibers emanating from multiple sources in the OR, including personnel, heater-cooler devices, and surgical smoke. This risk is increased in surgical patients undergoing procedures involving implanted devices. Surgical personnel also are at risk from exposure to surgical smoke, which can contain viable viral particles including human papillomavirus infection. Air quality in the OR is improved by engineering controls (eg, maintaining positive pressure). During the past decade, innovations in the field of adjunctive technology designed to improve OR air quality include using ultraviolet disinfection and mobile ultraviolet disinfection plus high-efficiency particulate air filtration. Some of these technologies additionally provide continuous monitoring of circulating air particle counts. Additional research regarding the benefits of adjunctive air-cleaning technology in the OR is warranted.


Endothelial cell orientation on aortic valve leaflets.
Studies of the structure of blood vessels have commonly found that cells lining the vessels are lengthened in the direction of blood flow, presumably as an effect of shear stress. A study of aortic valves has, however, shown that endothelial cells covering both sides of aortic valve leaflets are arranged across the direction of flow. Aortic valve leaflets taken from seven adult mongrel dogs and fixed either open or closed with glutaraldehyde were studied with a scanning electron microscope after critical point drying. On both open and closed leaflets the endothelial cells tended to be aligned circumferentially with the free edge of the leaflet. The circumferential pattern was particularly unexpected on the leaflet surface facing the left ventricle because this surface could be expected to receive the full effect of shear stress from systolic blood flow. The finding suggested that surface cellular alignment on leaflets is determined by some force other than shear stress. The force apparently responsible for organising the collagenous layers in the leaflet, which are best adapted to bear stress, is diastolic blood pressure. Since the endothelium follows the same pattern of alignment as the layers of collagen it seems reasonable to conclude that endothelial orientation is also a response ultimately to pressure stress. The arrangement of the endothelium then serves as a readily observed indicator of those functional stresses that dominate and organise leaflet structure.


Itraconazole or allopurinol in the treatment of chronic American trypanosomiasis: the results of clinical and parasitological examinations 11 years post-treatment.
Eleven years after they had been given itraconazole or allopurinol for the treatment of chronic American trypanosomiasis, 109 adult patients were checked for electrocardiographic abnormalities and evidence of Trypanosoma cruzi infection. The parasitological investigations included xenodiagnosis, in which the faeces of Triatoma infestans that had fed on the patients were checked under the microscope for flagellates. In addition, a PCR-based assay and a hybridization assay were used to test blood samples from the patients, and faeces from the Tri. infestans that had fed on the patients, for Try. cruzi DNA. For the data analysis, the patients were divided into four groups known as normal/normal, abnormal/normal, normal/abnormal and abnormal/abnormal, according to whether the patients had been found to have normal or abnormal electrocardiograms (ECG) shortly before the first treatment and to have normal or abnormal ECG when checked at the 11-year follow-up. The 51 normal/normal and 24 normal/abnormal patients were assumed to have been in the 'indeterminate' phase of the disease when they were treated, whereas the 16 abnormal/normal and 18 abnormal/abnormal patients all had evidence of chagasic cardiopathy at that time. When checked 11 years post-treatment, 40 (78.4%), 17 (70.8%), 14 (87.5%) and 17 (94.4%) of these patients, respectively, were each found positive for Try. cruzi in at least one of the parasitological tests. The hybridization assay, whether applied to human blood or bug faeces, appeared a significantly more sensitive test than the PCR-based assays or microscopically assessed xenodiagnosis (P<0.05). Only the 21 patients who appeared to be negative for Try. cruzi could be considered parasitologically cured (although all still appeared to have anti-Try. cruzi antibodies in their blood). Only 13 of these parasitologically cured patients (seven of those treated with itraconazole and six of those given allopurinol) had normal ECG at the 11-year follow-up. In Chile at least, itraconazole, which caused fewer adverse effects than the allopurinol while being no less effective at preventing cardiopathy, appears to be the drug of choice to treat chronic American trypanosomiasis in adults.


The bHLH transcription factor MdbHLH3 promotes anthocyanin accumulation and fruit colouration in response to low temperature in apples.
Low environmental temperatures promote anthocyanin accumulation and fruit colouration by up-regulating the expression of genes involved in anthocyanin biosynthesis and regulation in many fruit trees. However, the molecular mechanism by which fruit trees regulate this process in response to low temperature (LT) remains largely unknown. In this study, the cold-induced bHLH transcription factor gene MdbHLH3 was isolated from an apple tree and was found to interact physically and specifically through two regions (amino acids 1-23 and 186-228) at the N terminus with the MYB partner MdMYB1 (allelic to MdMYB10). Subsequently, MdbHLH3 bound to the promoters of the anthocyanin biosynthesis genes MdDFR and MdUFGT and the regulatory gene MdMYB1 to activate their expression. Furthermore, the MdbHLH3 protein was post-translationally modified, possibly involving phosphorylation following exposure to LTs, which enhanced its promoter-binding capacity and transcription activity. Our results demonstrate the molecular mechanism by which MdbHLH3 regulates LT-induced anthocyanin accumulation and fruit colouration in apple.


Cytokinins and Expression of SWEET, SUT, CWINV and AAP Genes Increase as Pea Seeds Germinate.
Transporter genes and cytokinins are key targets for crop improvement. These genes are active during the development of the seed and its establishment as a strong sink. However, during germination, the seed transitions to being a source for the developing root and shoot. To determine if the sucrose transporter (SUT), amino acid permease (AAP), Sugar Will Eventually be Exported Transporter (SWEET), cell wall invertase (CWINV), cytokinin biosynthesis (IPT), activation (LOG) and degradation (CKX) gene family members are involved in both the sink and source activities of seeds, we used RT-qPCR to determine the expression of multiple gene family members, and LC-MS/MS to ascertain endogenous cytokinin levels in germinating Pisum sativum L. We show that genes that are actively expressed when the seed is a strong sink during its development, are also expressed when the seed is in the reverse role of being an active source during germination and early seedling growth. Cytokinins were detected in the imbibing seeds and were actively biosynthesised during germination. We conclude that, when the above gene family members are targeted for seed yield improvement, a downstream effect on subsequent seed germination or seedling vigour must be taken into consideration.


Magnetic resonance fingerprinting: from evolution to clinical applications.
In 2013, Magnetic Resonance Fingerprinting (MRF) emerged as a method for fast, quantitative Magnetic Resonance Imaging. This paper reviews the current status of MRF up to early 2020 and aims to highlight the advantages MRF can offer medical imaging professionals. By acquiring scan data as pseudorandom samples, MRF elicits a unique signal evolution, or 'fingerprint', from each tissue type. It matches 'randomised' free induction decay acquisitions against pre-computed simulated tissue responses to generate a set of quantitative images of T1 , T2 and proton density (PD) with co-registered voxels, rather than as traditional relative T1 - and T2 -weighted images. MRF numeric pixel values retain accuracy and reproducibility between 2% and 8%. MRF acquisition is robust to strong undersampling of k-space. Scan sequences have been optimised to suppress sub-sampling artefacts, while artificial intelligence and machine learning techniques have been employed to increase matching speed and precision. MRF promises improved patient comfort with reduced scan times and fewer image artefacts. Quantitative MRF data could be used to define population-wide numeric biomarkers that classify normal versus diseased tissue. Certification of clinical centres for MRF scan repeatability would permit numeric comparison of sequential images for any individual patient and the pooling of multiple patient images across large, cross-site imaging studies. MRF has to date shown promising results in early clinical trials, demonstrating reliable differentiation between malignant and benign prostate conditions, and normal and sclerotic hippocampal tissue. MRF is now undergoing small-scale trials at several sites across the world; moving it closer to routine clinical application.


Preventive Effect of Clazosentan against Cerebral Vasospasm after Clipping Surgery for Aneurysmal Subarachnoid Hemorrhage in Japanese and Korean Patients.
Clazosentan has been explored worldwide for the prophylaxis of cerebral vasospasm after aneurysmal subarachnoid hemorrhage (aSAH). In a dose-finding trial (CONSCIOUS-1) conducted in Israel, Europe, and North America, clazosentan (1, 5, and 15 mg/h) significantly reduced the incidence of cerebral vasospasm, but its efficacy in Japanese and Korean patients was unknown. We conducted a double-blind comparative study to evaluate the occurrence of cerebral vasospasm in Japanese and Korean patients with aSAH. The aim of this multicenter, double-blind, randomized, placebo-controlled, dose-finding phase 2 clinical trial, was to evaluate the efficacy, pharmacokinetics, and safety of clazosentan (5 and 10 mg/h) against cerebral vasospasm after clipping surgery in Japanese and Korean patients with aSAH. Patients aged between 20 and 75 years were administered the study drug within 56 h after the aneurysm rupture and up to day 14 post-aSAH. The incidence of vasospasm, defined as an inner artery diameter reduction of major intracranial arteries ≥34% based on catheter angiography, was compared between each treatment group. Cerebral infarction due to vasospasm at 6 weeks and patients' outcome at 3 months was also compared. Among 181 enrolled patients, 158 completed the study and were analyzed. The incidence of vasospasm up to day 14 after aSAH onset was 80.0% in the placebo group (95% CI 67.0-89.6), 38.5% in the 5 mg/h clazosentan group (95% CI 25.3-53.0), and 35.3% in the 10 mg/h clazosentan group (95% CI 22.4-49.9), indicating that the incidence of vasospasm was significantly reduced by clazosentan treatment (placebo vs. 5 mg/h clazosentan, p < 0.0001; placebo vs. 10 mg/h clazosentan, p < 0.0001). The occurrence of cerebral infarction due to vasospasm was 20.8% in the placebo group (95% CI 10.8-34.1), 3.8% in the 5 mg/h clazosentan group (95% CI 0.5-13.2), and 4.2% in the 10 mg/h clazosentan group (95% CI 0.5-14.3), indicating that clazosentan significantly reduced the occurrence of cerebral infarctions caused by vasospasm (placebo vs. 5 mg/h clazosentan, p = 0.0151; placebo vs. 10 mg/h clazosentan, p = 0.0165). The overall incidence of all-cause death and/or vasospasm-related morbidity/mortality was significantly reduced in the 10 mg/h clazosentan group compared with the placebo group (p = 0.0003). These results suggest that clazosentan prevents cerebral vasospasm and subsequent cerebral infarction, and could thereby improve outcomes after performing a clipping surgery for aSAH in Japanese and Korean patients.


Magnetic targeting of adoptively transferred tumour-specific nanoparticle-loaded CD8+ T cells does not improve their tumour infiltration in a mouse model of cancer but promotes the retention of these cells in tumour-draining lymph nodes.
Adoptive T cell-transfer (ATC) therapy is a highly promising cancer-treatment approach. However, in vivo-administered T cells tend to disperse, with only a small proportion reaching the tumour. To remedy this, magnetic targeting of T cells has been recently explored. Magnetic nanoparticles (MNPs) functionalised with antibodies were attached to effector T cells and magnetically recruited to tumour sites under MRI guidance. In this study, we investigated whether 3-aminopropyl-triethoxysilane (APS)-coated MNPs directly attached to CD8+ T cell membranes could also magnetically target and accumulate tumour-specific CD8+ T cells in solid tumours using an external magnetic field (EMF). As it has been shown that T cells associated with APS-coated MNPs are retained in lymph nodes (LNs), and tumour-draining LNs are the most common sites of solid-tumour metastases, we further evaluated whether magnetic targeting of APS-MNP-loaded CD8+ T cells could cause them to accumulate in tumour-draining LNs. First, we show that antigen-specific CD8+ T cells preserve their antitumor activity in vitro when associated with APS-MNPs. Next, we demonstrate that the application of a magnetic field enhanced the retention of APS-MNP-loaded OT-I CD8+ T cells under flow conditions in vitro. Using a syngeneic mouse model, we found similar numbers of APS-MNP-loaded OT-I CD8+ T cells and OT-I CD8+ T cells infiltrating the tumour 14 days after cell transfer. However, when a magnet was placed near the tumour during the transfer of tumour-specific APS-MNP-loaded CD8+ T cells to improve tumour infiltration, a reduced percentage of tumour-specific T cells was found infiltrating the tumour 14 days after cell transfer, which was reflected in a smaller reduction in tumour size compared to tumour-specific CD8+ T cells transferred with or without MNPs in the absence of a magnetic field. Nonetheless, magnet placement near the tumour site during cell transfer induced infiltration of activated tumour-specific CD8+ T cells in tumour-draining LNs, which remained 14 days after cell transfer. The use of an EMF to improve targeting of tumour-specific T cells modified with APS-MNPs reduced the percentage of these cells infiltrating the tumour, but promoted the retention and the persistence of these cells in the tumour-draining LNs.


The use of forensic case data in intelligence-led policing: the example of drug profiling.
To date, forensic science has predominantly focused on generating evidence for judicial proceedings. While many recognise its broader and important contribution to the initial stages of the forensic process, resources do not seem to be employed efficiently. It is often discovered retrospectively that necessary information was previously available in a database or within existing files. Such information could have been proactively used in order to solve a particular case, a number of linked cases or better understand the criminal activity as a whole. This article reviews this broader contribution of forensic science, with a particular emphasis on drug intelligence at the Australian Federal Police (AFP) in Australia. Using the AFP as a model organisation, an overview of the current situation and the contribution of physical and chemical profiling are first discussed. The situation in Europe, and in particular in Switzerland, is also presented. It is argued that a change of attitude towards a more intelligence-led perspective is required in forensic science in general, and in drug profiling in particular.


A comparison of efficacy and safety of vildagliptin and gliclazide in combination with metformin in patients with Type 2 diabetes inadequately controlled with metformin alone: a 52-week, randomized study.
To demonstrate non-inferiority of vildagliptin compared with gliclazide, as an add-on therapy, in patients with Type 2 diabetes inadequately controlled with metformin in a 52-week, randomized, double-blind, active-controlled study. Patients receiving a stable dose of metformin (> or = 1500 mg) were randomized (1 : 1) to receive vildagliptin (50 mg twice daily; n = 513) or gliclazide (up to 320 mg/day; n = 494). Non-inferiority of vildagliptin was demonstrated (95% confidence interval -0.11%, 0.20%) with a mean change (se) from baseline glycated haemoglobin (HbA(1c)) (approximately 8.5% in both groups) to a 52-week endpoint of -0.81% (0.06) with vildagliptin and -0.85% (0.06) with gliclazide. Although a similar proportion of patients reached HbA(1c) < 7.0%, the total number of hypoglycaemic events was lower in the vildagliptin group (6 vs. 11 events). Vildagliptin was non-inferior (margin 0.6 mmol/l) to gliclazide in reducing fasting plasma glucose (1.31 vs. 1.52 mmol/l, P = 0.257). The overall incidence of any adverse events was similar in both groups (approximately 61%), but the number of serious adverse events was higher in the gliclazide group (8.7 vs. 6.7%). The number of patients who discontinued as a result of an unsatisfactory effect was higher in the vildagliptin group (n = 22 vs. 13, respectively) compared with gliclazide, but vildagliptin did not induce weight gain. In patients with Type 2 diabetes inadequately controlled with metformin, addition of vildagliptin provided similar HbA(1c)-lowering efficacy compared with gliclazide after 52 weeks of treatment. Although both treatments were well tolerated, vildagliptin-treated patients had fewer hypoglycaemic events and did not gain weight.


The impact of biosampling procedures on molecular data interpretation.
The separation between biological and technical variation without extensive use of technical replicates is often challenging, particularly in the context of different forms of protein and peptide modifications. Biosampling procedures in the research laboratory are easier to conduct within a shorter time frame and under controlled conditions as compared with clinical sampling, with the latter often having issues of reproducibility. But is the research laboratory biosampling really less variable? Biosampling introduces within minutes rapid tissue-specific changes in the cellular microenvironment, thus inducing a range of different pathways associated with cell survival. Biosampling involves hypoxia and, depending on the circumstances, hypothermia, circumstances for which there are evolutionarily conserved defense strategies in the range of species and also are relevant for the range of biomedical conditions. It remains unclear to what extent such adaptive processes are reflected in different biosampling procedures or how important they are for the definition of sample quality. Lately, an increasing number of comparative studies on different biosampling approaches, post-mortem effects and pre-sampling biological state, have investigated such immediate early biosampling effects. Commonalities between biosampling effects and a range of ischemia/reperfusion- and hypometabolism/anoxia-associated biological phenomena indicate that even small variations in post-sampling time intervals are likely to introduce a set of nonrandom and tissue-specific effects of experimental importance (both in vivo and in vitro). This review integrates the information provided by these comparative studies and discusses how an adaptive biological perspective in biosampling procedures may be relevant for sample quality issues.


Osteoporosis, osteonecrosis, and orthodontics.
Osteoporosis, whether senile, postmenopausal, or corticosteroid-induced, has been shown to affect orthodontic tooth movement and the periodontium. There have also been reports of osteonecrosis related to oral bisphosphonate therapy during treatment for osteoporotic bone diseases. This article summarizes the current knowledge of the relationship between osteoporosis and osteonecrosis and their effects on orthodontic tooth movements. The information will be important in orthodontic treatment planning and realization.


Best Practice Guidelines for Pre-Launch Characterization and Calibration of Instruments for Passive Optical Remote Sensing.
The pre-launch characterization and calibration of remote sensing instruments should be planned and carried out in conjunction with their design and development to meet the mission requirements. The onboard calibrators such as blackbodies and the sensors such as spectral radiometers should be characterized and calibrated using SI traceable standards. In the case of earth remote sensing, this allows inter-comparison and intercalibration of different sensors in space to create global time series of climate records of high accuracy where some inevitable data gaps can be easily bridged. The recommended best practice guidelines for this pre-launch effort is presented based on experience gained at National Institute of Standards and Technology (NIST), National Aeronautics and Space Administration (NASA) and National Oceanic and Atmospheric Administration (NOAA) programs over the past two decades. The currently available radiometric standards and calibration facilities at NIST serving the remote sensing community are described. Examples of best practice calibrations and intercomparisons to build SI (international System of Units) traceable uncertainty budget in the instrumentation used for preflight satellite sensor calibration and validation are presented.


Shoulder vibratory exercises improves shoulder external rotation muscle strength and shoulder function: Randomized comparison trial.
Based on electromyography measurements, shoulder vibratory exercises efficiently stimulate shoulder muscles activity. Yet very few studies have supported that shoulder vibratory exercises increased shoulder muscles strength and function, and the noninferiority compared with conventional elastic resistance exercises remains unknown. This study investigated the effect of vibratory exercises versus conventional elastic resistance exercises with elastic bands on shoulder external rotation muscles strength and functional performance in young adults. 26 young adults (7 males and 19 females, with age 23.89 ± 3.02) were recruited and randomly allocated to shoulder vibratory exercises with FLEXI-BAR (FLEXI-BAR group) or conventional resistance exercises with elastic band (TheraBand group) for 3 times/week, 4 weeks totally. Shoulder external rotator muscles strength test and Underkofler softball distance throw test (USDTT) were performed before and after the training period. After 4 weeks training, shoulder external rotator muscles strength increased 22.25 ± 15.06 N (P= 0.004, effect size = 1.48) within FLEXI-BAR group and 22.81 ± 14.94 N (P= 0.007, effect size = 1.53) within TheraBand group. There were no statistically significant differences between groups in the three muscle strength tests (P> 0.65). Regarding shoulder function, FLEXI-BAR exercises increased the throw distance 0.81 ± 0.92 meters in USDTT (P= 0.041, effect size = 0.88) while TheraBand exercise did not (P= 0.284), yet there was no statistically significant between group effects (P= 0.608). These findings suggest that shoulder vibratory exercises can improve shoulder muscles strength in young adults and can be a useful alternative to the conventional elastic resistance exercises to improve the shoulder muscles strength and function. This provides therapists with more options in terms of choosing training equipment for rehabilitation programs.


Fano-ADC(2,2) method for electronic decay rates.
Fano-ADC is a family of ab initio methods for the prediction of electronic decay widths in excited, singly and doubly ionized systems. It has been particularly successful in elucidating the geometry dependence of the inter-atomic decay widths in clusters and facilitated the prediction of new electronic decay phenomena. However, the available Fano-ADC schemes are limited to the second-order treatment of the initial state and the first-order treatment of the final states of the decay. This confines the applicability of the Fano-ADC approach to first-order decay processes, e.g., normal but not double Auger decay (DAD), and compromises the numerical accuracy of the schemes through the unbalanced treatment of electronic correlation. Here, we introduce the ADC(2,2) approximation for singly ionized states, which describes both initial and final states of the decay up to second order. We use the new scheme to construct the Fano-ADC(2,2) approximation for the decay widths and show that it provides superior accuracy for the decay widths of a series of processes. Moreover, the Fano-ADC(2,2) method provides access to second-order decay processes, such as DAD, which are qualitatively beyond the reach of the previously available Fano-ADC implementations.


Fellowship Training in Nepal: Current Prospects.
The need of subspecialty training has been acutely felt in Nepal now with the rapid advancement in the field of medical sciences, the changing burden of diseases, the acute shortage of highly specialized health professionals, the evolving concept of learning and medical education and the ever-increasing focus on right and safety of patients.Fellowship Training is obtained in a specific specialty or subspecialty. Fully academic 3-year Fellowship programsin Pulmonary and Critical Care Medicine, Hepatopancreatobiliary Surgery and Cardiologyhavebeen initiated for certification of the training for the first time in Nepal at Chitwan Medical College (CMC) and all three are recognized by the Nepal Medical Council.The three Fellowship training programs, run in affiliation with Lincoln University, at the CMC have all the training criteria of DM/MCh included along with extra requirement of one more paper publication and the trainees are paid adequate stipend without any tuition fee or bond. The high volume of cases and procedures in CMC, exposes the FCMC Fellows to an increased chance of appropriate skill development along with appropriate postings in various other institutions for adequate experience of hands on training.Fellowship programs are the cornerstone of providing super specialized patient care.The need today is to focus on the basic training concepts and criteria, rather than on various names like diploma, MD, MS, DM/MCh of training.Nepal should focus onthe prerequisites, infrastructure and human resource for expanding the quality Fellowship trainingprograms in the country and thereby enhancing the standard of patient care. Keywords: Cardiology; DM; FCMC; fellowship; hepatopancreatobiliary surgery; MCh; PCCM.


The acquisition of Streptococcus mutans by infant monkeys (Macaca fascicularis) and its relationship to the initiation of dental caries.
The acquisition and transmission of Streptococcus mutans by 16 consecutively born infant monkeys (Macaca fascicularis) was studied. The 16 infant monkeys were weaned in four groups and caged together to form a commune. Transmission from mother to infant occurred infrequently. Streptococcus mutans was isolated from nine of the mothers but only from the dental plaque of two infant monkeys at weaning at which time the predominant streptococci were S. sanguis and S. mitior. One further animal was colonized by S. mutants during the formation of the commune, but only after it was caged with an infant harbouring the same organism. For 46 d after the completion of the commune, the monkeys were fed a starch-based diet during which time S. mutans of serotypes c, e or h were isolated from the faeces of all 16 animals and from the dental plaque taken from the developmental groove of the first deciduous molar of 11 animals. Faecal transmission appeared to be an important factor in the spread of S. mutans between monkeys in the commune. The monkeys were then fed a caries-promoting high sucrose diet resulting in a rapid increase in the proportion of S. mutans in the plaque and in the faeces. Streptococcus mutans serotype e was more frequently isolated from both plaque and faeces and its predominance may in part be due to the production of a bacteriocin active in vitro against S. mutans serotype h and other species of oral streptococci isolated from monkey dental plaque. The proportion of S. mutans in the developmental groove 8 d after the introduction of the high sucrose diet was significantly related to both the caries status of the groove and the total caries score 6 months later. The results suggest that, in this model of human dental caries, S. mutans is the major bacterial factor in the initiation of tooth decay.


Public health and ambient air pollution in arctic and subarctic cities of Russia.
In arctic and subarctic regions of Russia, there are 34 industrial cities where the largest metallurgic, paper, and pulp production in the world takes place. This article reviews investigations in environmental epidemiology, which are just beginning. The investigations are hindered by the absence of computer databases and by the migration of the population into other regions. Marked changes in the status of public health can be seen in Norilsk and Montchegorsk, the centers of metallurgy, as well as in cities near paper and pulp production. Improvement of epidemiologic investigations will require better analytical methods and the consideration of information from international sources, for example, the foundation of a cancer register and a register for congenital abnormalities in newborns.


Assessing the Evidence of Micronutrients on Depression among Children and Adolescents: An Evidence Gap Map.
There is some evidence indicating that nutrition may have the ability to prevent, treat, and/or influence the severity of depression. The aims of this evidence gap map (EGM) are to provide an overview and to determine evidence gaps in the existing research on micronutrients and their impact on depression among children and adolescents. We conducted a comprehensive search in multiple databases of primary and secondary literature assessing the impact of micronutrients on depression-related outcomes such as unipolar depression, major depressive disorders, dysthymia, acute depression, and mood disorders. Abstracts and full-text articles were dual-screened based on predefined eligibility criteria. A total of 30 primary research publications were included in the EGM. About 47% of included studies focused on late adolescents (15-19 y), ∼40% on early adolescents (10-14 y), and ∼13% on children aged 6-9 y. Among the included studies, 8 studies examined a single micronutrient intervention and 22 studies examined micronutrient concentrations (either intake or serum), and their impact on depression. The most frequently studied micronutrients were vitamin D (n = 8), zinc (n = 8), iron (n = 6), folate (n = 7), and vitamin B-12 (n = 5). More longitudinal studies and trials are needed to determine the role of micronutrients in the etiology and treatment of depression among children and adolescents.


Assessing the evolution of redundancy/synergy of spontaneous variability regulation with age.
We exploited a model-based Wiener-Granger causality method in the information domain for the evaluation of the transfer entropy (TE) and interaction TE (ITE), the latter taken as a measure of the net balance between redundancy and synergy, to describe the interactions between the spontaneous variability of heart period (HP) and systolic arterial pressure (SAP) and the effect of respiration (R) on both variables. Cardiac control was typified via the genuine TE from SAP to HP, that from R to HP, and the ITE from SAP and R to HP, while vascular control was characterized via the genuine TE from HP to SAP, that from R to SAP, and the ITE from HP and R to SAP. The approach was applied to study age-related modifications of cardiac and vascular controls in a cohort of 100 healthy humans (age from 21 to 70 years, 54 males) recorded at supine rest (REST) and during active standing (STAND). A surrogate approach was exploited to test the significance of the computed quantities. Trends of the genuine information transfer with age, already present in literature, were here confirmed. We originally found that: (i) at REST redundancy was predominant over synergy in both vascular and cardiac controls; (ii) the predominance of redundancy of the cardiac control was not affected by postural challenge, while STAND reduced redundancy of vascular control; (iii) the net redundancy of the cardiac control at REST gradually decreased with age, while that of vascular control remained stable; (iv) during STAND net redundancy of both cardiac and vascular controls was stable with age. The study confirms the relevance of computing genuine information transfer in cardiovascular control analysis and stresses the importance of evaluating the ITE to quantify the degree of redundancy of physiological mechanisms operating to maintain cardiovascular homeostasis.


Zoledronate reverses mandibular bone loss in osteoprotegerin-deficient mice.
To characterize the changes in osteoprotegerin-deficient (OPG-/-) mice mandibles and the possible mandibular bone loss prevention by zoledronate. This preventive effect in the mandible differed from that in the proximal tibia and was independent of the OPG pathway. The study aimed to characterize both the changes in the mandible in osteoprotegerin-deficient (OPG-/-) mice and possible mandibular bone loss prevention by zoledronate. Twenty-eight 6-week-old female mice (C57BL/6J), including OPG-/- (n = 21) and wild-type (WT) (n = 7) mice, were assigned to four groups after 2 weeks of acclimatization to local vivarium conditions: wild mice with vehicle (WT group); OPG-/- mice with vehicle (OPG-/- group); and OPG-/- mice that were subcutaneously injected with either 50 or 150 microg/kg zoledronate (Zol-50 and Zol-150 groups, respectively). Mice were sacrificed at 4 weeks after these treatments and after fasting for 12 h. Sera were harvested for biochemical analyses. The right mandible and tibia of each mouse were selected for microCT analysis. Student's t-test was performed for comparisons of bone parameters at different sites in the WT group. Analysis of variance (ANOVA) was used to compare the biomarkers and bone parameters in the different treatment groups. Serum bone-specific alkaline phosphatase (B-ALP) and tartrate-resistant acid phosphatase 5b (TRACP-5b) were significantly decreased in WT mice as compared to the levels in the OPG-/- mice (P < 0.05). Zoledronate treatment decreased the high serum B-ALP activity observed in OPG-/- mice to the levels seen in WT mice, while serum TRACP-5b concentrations were decreased to levels even lower than those in WT mice. There were substantial variations in BMD and microstructure of the mandibular and proximal tibial trabeculae. Mandibular bone loss was less affected by OPG gene deprivation than the proximal tibia was. Both zoledronate groups showed greater BMD, trabecular BV/TV, Tb.Th, Tb.N, and Conn.D and a significant decrease in Tb.Sp and SMI as compared to the findings in OPG-/- mice (P < 0.05). However, higher apparent BMD and more compact plate-like trabeculae were observed in the mandible after treatment with zoledronate as compared to the findings in the proximal tibia. No significant differences were found in any parameter in both zoledronate groups. The present study showed that zoledronate could reverse the significant bone loss in mice mandibles that was induced by OPG gene deficiency. This preventive effect, which was accompanied with considerable inhibition of bone turnover, differed in the mandible and in the proximal tibia and was independent of the OPG pathway.


A new flap design: neural-island flap.
This report introduces the "neural-island flap" concept, which represents a consistent and reliable skin flap design supplied only by the intrinsic vasculature of a cutaneous nerve. In this study, the lateral femoral cutaneous nerve was selected as the pedicle of the neural-island flap, and a standard skin flap, which is the territory of the accompanying vessels (i.e., iliac branches of the iliolumbar artery and vein), was elevated on the lower dorsal region of the rats. In a total of 92 Wistar rats, three experiments were performed. In part I (n = 24), the vascular anatomy of the lateral femoral cutaneous nerve was established by the methods of dissection, microangiography, nerve mapping, perfusion with colored latex and India ink, and histologic analysis. In part II (n = 46), the role of the cutaneous nerve in supporting an acutely elevated skin flap was explored by creating five flap groups as follows: group 1, conventional flap (artery, vein, and nerve intact); group 2, neural island flap (only the nerve intact); group 3, neurocutaneous flap (vein and nerve intact); group 4, denervated flap (artery and vein intact); and group 5, skin graft. In part III (n = 22), the role of a preliminary surgical delay procedure to augment the survival of the neural island flap was investigated. Results of the anatomic studies indicated a consistent perineural vasculature by the accompanying iliolumbar artery. Skin flaps survived totally in groups where the artery and vein were intact, whereas mean survival rates for the neural island flap and the neurocutaneous flap were 38.2 +/- 3.1 percent and 44.5 +/- 3.8 percent, respectively (p > 0.05). Results of part III of the experiment demonstrated a significantly higher survival for the delayed neural island flap (94.5 +/- 5.5 percent) compared with the acutely elevated neural island flap (p < 0.05). The perineural and intraneural vessels were found to be greatly dilated after a delay procedure, demonstrated by direct observation, microangiography, histologic analysis, dye injection study, and scanning electron microscopy. On the basis of this promising series of experiments, a clinical technique was developed using the sural neural-island flap. The flap was used to reconstruct lower extremity defects in four cases. A delay procedure was accomplished in the first stage by elevating a fasciocutaneous flap from the midcalf region based on a posterior skin bridge and the sural nerve. After a 2-week delay period, a sural neural-island flap was created based on the nerve and transposed to the defect. Flap survival was complete in all cases, with a satisfactory result. The authors conclude that this report proves for the first time that a robust and reliable skin flap can be created pedicled only by the intrinsic vasculature of a cutaneous nerve, after a proper surgical delay. The so-created neural-island flap design offers two novel advantages: (1) a very narrow pedicle and (2) a pedicle without any restriction to a specific pivot point, in addition to the previously described unique advantages of preservation of a major artery and avoidance of microvascular anastomoses.


The electron-transfer reaction between azurin and the cytochrome c oxidase from Pseudomonas aeruginosa.
A stopped-flow investigation of the electron-transfer reaction between oxidized azurin and reduced Pseudomonas aeruginosa cytochrome c-551 oxidase and between reduced azurin and oxidized Ps. aeruginosa cytochrome c-551 oxidase was performed. Electrons leave and enter the oxidase molecule via its haem c component, with the oxidation and reduction of the haem d1 occurring by internal electron transfer. The reaction mechanism in both directions is complex. In the direction of oxidase oxidation, two phases assigned on the basis of difference spectra to haem c proceed with rate constants of 3.2 X 10(5)M-1-S-1 and 2.0 X 10(4)M-1-S-1, whereas the haem d1 oxidation occurs at 0.35 +/- 0.1S-1. Addition of CO to the reduced enzyme profoundly modifies the rate of haem c oxidation, with the faster process tending towards a rate limit of 200S-1. Reduction of the oxidase was similarly complex, with a fast haem c phase tending to a rate limit of 120S-1, and a slower phase with a second-order rate of 1.5 X 10(4)M-1-S-1; the internal transfer rate in this direction was o.25 +/- 0.1S-1. These results have been applied to a kinetic model originally developed from temperature-jump studies.


Basal hypothalamo-pituitary-adrenal axis activity and corticotropin feedback in young and older men: relationships to magnetic resonance imaging-derived hippocampus and cingulate gyrus volumes.
Alterations in basal cortisol secretion and feedback sensitivity are reported in aging. However, it is not known whether these hypothalamus-pituitary-adrenal (HPA) axis alterations are related to structural brain changes. This study was designed to investigate these relationships in the human. Nine young (24.0 +/- 1.2 years; mean +/- SE; range: 19-30) and 11 older (69.0 +/- 1.8 years; range: 59-76) men, in addition to having standardized magnetic resonance imaging of their brains, were given 0.5 mg/kg cortisol or placebo intravenously in a double-blind, crossover study. As expected, older men had significantly smaller volumes for all brain regions. Although the groups did not differ in baseline HPA axis activity, there were significant and specific relationships between the brain volumes and the baseline measures of HPA activity. Namely, for young and older subjects combined and after controlling for age and cerebral vault size, hippocampal volumes were inversely associated with 24-hour urinary cortisol and basal corticotropin (ACTH) levels, and the anterior cingulate gyrus volume was negatively correlated with baseline ACTH. Elderly subjects had a slower decrease in ACTH levels (percent of baseline level) during the first 30 min after cortisol administration. However, no associations were observed between the ACTH feedback indices and any brain measure. This report, although based on a small number of subjects, supports previous studies showing a blunted ACTH fast feedback during normal aging. Hippocampal atrophy appears to be related to increased basal measures of HPA axis activity, but not to fast ACTH feedback. It remains possible that age-associated changes in fast feedback may be related to changes to other brain sites, such as hypothalamus or pituitary.


Role of perigastric arcade removal in serous epithelial ovarian cancers.
Omentectomy is an essential part of cytoreductive surgery (CRS). However, removal of perigastric arcade (PGA) of the omentum is a controversial aspect of omentectomy in view of the fear of injury, vascular compromise and gastroparesis. Hence, we conducted a study to evaluate the necessity and effect of removal of PGA during omentectomy. The nature of the study was a prospective observational study. The study period was for 1 year between 1.3.2019 and 29.2.2020. Patients with stage III to IV serous epithelial ovarian cancers - chemo naive/post neoadjuvant chemotherapy, without macroscopic involvement of the PGA were included in the study. Patients were divided into two groups - those who had PGA removed (group 1) and those whose PGA was preserved (group 2). Pre, intra and postoperative factors between the two groups were compared using standard statistical methods. Micrometastasis to PGA was present in 36.4% of the patients in group 1. The predictors for this involvement included gross involvement and microscopic involvement of the mobile part of the omentum (p < 0.001), pre surgery Meyer's score (p < 0.05) and requirement of peritonectomy (p < 0.05) during the CRS implying that higher the peritoneal carcinomatosis, more are the chances of microscopic involvement of PGA. On comparing postoperative outcomes between the two groups, we noted a statistically significant difference in intra-operative time (p < 0.01), prolonged recovery time with increased intensive care unit and hospital stay (p < 0.001) in group 1, although all with small absolute difference. However, there was no significant difference in major post-operative complications or time taken to tolerate soft diet. Micrometastasis to PGA was noted in significant number of cases. Its removal is also a safe procedure with minimal morbidity and good postoperative outcomes especially in cases with significant peritoneal carcinomatosis. Hence, it should be considered, provided we are achieving a complete cytoreduction otherwise.


Combined multivariate and pathway analyses show that allergen-induced gene expression changes in CD4+ T cells are reversed by glucocorticoids.
Glucocorticoids (GCs) play a key role in the treatment of allergy. However, the genome-wide effects of GCs on gene expression in allergen-challenged CD4(+) T cells have not been described. The aim of this study was to perform a genome-wide analysis to investigate whether allergen-induced gene expression changes in CD4(+) T cells could be reversed by GCs. Gene expression microarray analysis was performed to profile gene expression in diluent- (D), allergen- (A), and allergen + hydrocortisone- (T) challenged CD4(+) T cells from patients with seasonal allergic rhinitis. Principal component analysis (PCA) showed good separation of the three groups. To identify the correlation between changes in gene expression in allergen-challenged CD4(+) T cells before and after GC treatment, we performed orthogonal partial least squares discriminant analysis (OPLS-DA) followed by Pearson correlation analysis. This revealed that allergen-induced genes were widely reversed by GC treatment (r = -0.77, P<0.0001). We extracted 547 genes reversed by GC treatment from OPLS-DA models based on their high contribution to the discrimination and found that those genes belonged to several different inflammatory pathways including TNFR2 Signalling, Interferon Signalling, Glucocorticoid Receptor Signalling and T Helper Cell Differentiation. The results were supported by gene expression microarray analyses of two independent materials. Allergen-induced gene expression changes in CD4(+) T cells were reversed by treatment with glucocorticoids. The top allergen-induced genes that reversed by GC treatment belonged to several inflammatory pathways and genes of known or potential relevance for allergy.


Safety and efficacy of pharmacological cardioversion of atrial fibrillation using intravenous vernakalant, a new antiarrhythmic drug with atrial selectivity.
Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia that due to its frequent hospitalizations and increased complication rates imposes a significant health economic burden. Many patients with recurrent AF are admitted to the hospital for cardioversion to restore sinus rhythm. Given this knowledge, it is clearly important to identify a feasible and effective approach for cardioversion of these patients. Cardioversion always requires careful assessment of potential complications, which apart from thromboembolic risks, include proarrhythmias and those related to the deep sedation required for electrical cardioversion. Even though electrical cardioversion is proven to be safe and effective, the need for anesthesia makes alternative strategies more attractive. The research discussed is the alternative strategies for cardioversion, including electrical cardioversion and the new relatively atrial-selective antiarrhythmic drug, vernakalant. The literature search methodology undertaken included search in PubMed (cardioversion, vernakalant, conversion as key words). Vernakalant is shown to have good conversion rates, an apparently safe antiarrhythmic profile and is well tolerated in patients with a history of ischemic heart disease. In most cases of recent-onset AF, pharmacological cardioversion can provide a probably more cost-effective and safer alternative to electrical cardioversion, which can then be used as a second option for those who failed the first attempt of cardioversion.


N and P constrain C in ecosystems under climate change: Role of nutrient redistribution, accumulation, and stoichiometry.
We use the Multiple Element Limitation (MEL) model to examine responses of 12 ecosystems to elevated carbon dioxide (CO2 ), warming, and 20% decreases or increases in precipitation. Ecosystems respond synergistically to elevated CO2 , warming, and decreased precipitation combined because higher water-use efficiency with elevated CO2 and higher fertility with warming compensate for responses to drought. Response to elevated CO2 , warming, and increased precipitation combined is additive. We analyze changes in ecosystem carbon (C) based on four nitrogen (N) and four phosphorus (P) attribution factors: (1) changes in total ecosystem N and P, (2) changes in N and P distribution between vegetation and soil, (3) changes in vegetation C:N and C:P ratios, and (4) changes in soil C:N and C:P ratios. In the combined CO2 and climate change simulations, all ecosystems gain C. The contributions of these four attribution factors to changes in ecosystem C storage varies among ecosystems because of differences in the initial distributions of N and P between vegetation and soil and the openness of the ecosystem N and P cycles. The net transfer of N and P from soil to vegetation dominates the C response of forests. For tundra and grasslands, the C gain is also associated with increased soil C:N and C:P. In ecosystems with symbiotic N fixation, C gains resulted from N accumulation. Because of differences in N versus P cycle openness and the distribution of organic matter between vegetation and soil, changes in the N and P attribution factors do not always parallel one another. Differences among ecosystems in C-nutrient interactions and the amount of woody biomass interact to shape ecosystem C sequestration under simulated global change. We suggest that future studies quantify the openness of the N and P cycles and changes in the distribution of C, N, and P among ecosystem components, which currently limit understanding of nutrient effects on C sequestration and responses to elevated CO2 and climate change.


Visualizing double-stranded RNA distribution and dynamics in living cells by dsRNA binding-dependent fluorescence complementation.
Double-stranded RNA (dsRNA) is an important type of RNA that plays essential roles in diverse cellular processes in eukaryotic organisms and a hallmark in infections by positive-sense RNA viruses. Currently, no in vivo technology has been developed for visualizing dsRNA in living cells. Here, we report a dsRNA binding-dependent fluorescence complementation (dRBFC) assay that can be used to efficiently monitor dsRNA distribution and dynamics in vivo. The system consists of two dsRNA-binding proteins, which are fused to the N- and C-terminal halves of the yellow fluorescent protein (YFP). Binding of the two fusion proteins to a common dsRNA brings the split YFP halves in close proximity, leading to the reconstitution of the fluorescence-competent structure and restoration of fluorescence. Using this technique, we were able to visualize the distribution and trafficking of the replicative RNA intermediates of positive-sense RNA viruses in living cells.


Computational Studies of SARS-CoV-2 3CLpro: Insights from MD Simulations.
Given the enormous social and health impact of the pandemic triggered by severe acute respiratory syndrome 2 (SARS-CoV-2), the scientific community made a huge effort to provide an immediate response to the challenges posed by Coronavirus disease 2019 (COVID-19). One of the most important proteins of the virus is an enzyme, called 3CLpro or main protease, already identified as an important pharmacological target also in SARS and Middle East respiratory syndrome virus (MERS) viruses. This protein triggers the production of a whole series of enzymes necessary for the virus to carry out its replicating and infectious activities. Therefore, it is crucial to gain a deeper understanding of 3CLpro structure and function in order to effectively target this enzyme. All-atoms molecular dynamics (MD) simulations were performed to examine the different conformational behaviors of the monomeric and dimeric form of SARS-CoV-2 3CLpro apo structure, as revealed by microsecond time scale MD simulations. Our results also shed light on the conformational dynamics of the loop regions at the entry of the catalytic site. Studying, at atomic level, the characteristics of the active site and obtaining information on how the protein can interact with its substrates will allow the design of molecules able to block the enzymatic function crucial for the virus.


Increased incidence of unsulphated and 4-sulphated residues in the chondroitin sulphate linkage region observed by high-pH anion-exchange chromatography.
We report the isolation, characterization and quantification of five octasaccharides, four hexasaccharides and two tetrasaccharides, derived from the chondroitin sulphate (CS) linkage region of 6-8-year-old bovine articular cartilage aggrecan, following digestion with chondroitin ABC endolyase. Using a novel high-pH anion-exchange chromatography (HPAEC) method, in conjunction with one- and two-dimensional (1)H-NMR spectroscopy, we have identified the following basic structure for the CS linkage region of aggrecan: DeltaUA(beta1-3)GalNAc[0S/4S/6S](beta1-4)GlcA(beta1-3)GalNAc[0S/4S/6S](beta1-4)GlcA(beta1-3)Gal[0S/6S](beta1-3)Gal(beta1-4)Xyl, where DeltaUA represents 4,5-unsaturated hexuronic acid, and 4S and 6S represent an O-ester sulphate group on C-4 and C-6 respectively. The octa-, hexa- and tetra-saccharide linkage region fragments were used to develop a HPAEC fingerprinting method, with detection at A(232 nm), and a linear response to approx. 0.1 nmol of substance. The sulphation patterns of CS linkage regions, of up to octasaccharide in size, from articular and tracheal cartilage aggrecan were examined. The results show that in articular cartilage, for the majority (53%) of octasaccharides the 2-deoxy-2-N-acetyl amino-D-galactose (GalNAc) residues closest to the linkage region are both 6-sulphated; however, in a significant portion (34%), one or more of these GalNAc residues are unsulphated, and in 8% both are unsulphated. Approximately 10-18% of the chains have a 4-sulphated GalNAc in the first disaccharide, and 12% have a sulphated linkage region Gal residue. No evidence was found for uronic acid sulphation. These data show that there is a significant increase in the incidence of unsulphated and 4-sulphated GalNAc residues adjacent to the linkage region compared with the rest of the chain. Bovine tracheal cartilage linkage regions displayed very similar sulphation profiles to those from articular cartilage, despite the presence of a higher level of GalNAc 4-sulphation within the repeat region of the main CS chain.


Understanding the Effects of Growing Seasons, Genotypes, and Their Interactions on the Anthesis Date of Wheat Sown in North China.
Quantitative studies on the effects of growing season, genotype (including photoperiod genes and vernalization genes), and their interaction (GGI) on the anthesis date of winter wheat (Triticum aestivum L.) are helpful to provide a scientific reference for selecting or developing adaptive varieties in target environments. In this study, we collected 100 winter wheat varieties with ecological adaptability in North China and identified the anthesis date under field conditions for three consecutive years from 2016 to 2019 with mapped photoperiod and vernalization alleles. Our results showed that the number of the photoperiod-insensitive Ppd-D1a allele increased with variety replacement, while the haplotype Ppd-A1b + Ppd-D1b + vrn-D1 (A4B2) decreased from the 1940s to 2000s. The anthesis date of A4B2 was significantly delayed due to the photoperiod-insensitive alleles Ppd-A1b and Ppd-D1b. The additive main effect and multiplicative interaction (AMMI) model and GGI biplot analysis were used for data analysis. A large portion of the total variation was explained by growing seasons (66.3%), while genotypes and GGIs explained 21.9% and 10.1% of the anthesis dates, respectively. The varieties from the 1940s and before had a great influence on the anthesis date, suggesting these germplasms tend to avoid premature anthesis and could facilitate the development of phenological resilient varieties.