Representative Plots Research Articles

Validation of a method for in situ determination of 137Cs soil contamination density in kitchen gardens using the portable spectrometer-dosimeter MKS AT6101D

The collection of representative soil samples in the territory of settlements and subsequent measurements of the content of radionuclides in these samples under laboratory conditions (the so-called “ex situ method”) is a generally accepted technology for determining the density of soil contamination with 137Cs in the populated areas contaminated due to the Chernobyl accident. Recently, as a supplement or alternative to the ex situ method, researchers are developing field (in situ) gamma-spectrometry methods. These methods allow determining the density of soil contamination with 137Cs directly on site, without soil sampling and laboratory analysis. At the same time, the in situ methodology has several limitations, the most important of which is a lack of generally recognized metrological basis for measurements and interpretation of results. Hence, before using a particular technique and measuring device for carrying out large-scale in situ measurements, it is necessary to validate (to assess the suitability) of the selected in situ method using an established ex situ method. The aim of this study was to validate the method for determining the density of 137Cs soil contamination in kitchen gardens using the MKS AT6101D spectrometer-dosimeter in situ. The method was recently presented by a Russian-Swedish-Belarusian group of researchers in an article published in the Journal of Environmental Radioactivity (https://doi.org/10.1016/j.jenvrad.2021.106562). To validate this method, we selected 10 representative kitchen garden plots. The plots were located in six settlements of the Bryansk region in Russia. The territory of the settlements had been heavily contaminated with 137Cs as a result of the Chernobyl accident: the officially established levels of the density of soil contamination by 137Cs ranged from 111 to 511 kBq/m2 in 2017. Field gamma-ray spectra were recorded at a height of 1 m above the ground in the center of kitchen garden plots using the MKS AT6101D device. The measurement duration was in the range of 1207–1801 s (the mean value = 1383 s). Samples of soil in the kitchen gardens were taken layer by layer (with a step of 5 cm) to a depth of 20 cm using a demountable cylindrical sampler. The 137Cs content in each soil layer was determined in the laboratory using a stationary semiconductor gamma spectrometer. The values of the 137Cs contamination density of the sampled soils ranged from 77 to 548 kBq/m2. It was found that the results of the ex situ analyzes of soil samples were in a good agreement with the contamination density values obtained with the in situ method. On average, the difference between two methodologies was 7% (a maximum of 20%). The results of the study confirm that the method proposed by the international group is suitable for determining the density of soil contamination by 137Cs in kitchen gardens in remote period after the Chernobyl accident.

MICAL2PV suppresses the formation of tunneling nanotubes and modulates mitochondrial trafficking.

Tunneling nanotubes (TNTs) are actin‐rich structures that connect two or more cells and mediate cargo exchange between spatially separated cells. TNTs transport signaling molecules, vesicles, organelles, and even pathogens. However, the molecular mechanisms regulating TNT formation remain unclear and little is known about the endogenous mechanisms suppressing TNT formation in lung cancer cells. Here, we report that MICAL2PV, a splicing isoform of the neuronal guidance gene MICAL2, is a novel TNT regulator that suppresses TNT formation and modulates mitochondrial distribution. MICAL2PV interacts with mitochondrial Rho GTPase Miro2 and regulates subcellular mitochondrial trafficking. Moreover, down‐regulation of MICAL2PV enhances survival of cells treated with chemotherapeutical drugs. The monooxygenase (MO) domain of MICAL2PV is required for its activity to inhibit TNT formation by depolymerizing F‐actin. Our data demonstrate a previously unrecognized function of MICAL2 in TNT formation and mitochondrial trafficking. Furthermore, our study uncovers a role of the MICAL2PV‐Miro2 axis in mitochondrial trafficking, providing a mechanistic explanation for MICAL2PV activity in suppressing TNT formation and in modulating mitochondrial subcellular distribution.

Recombinant Human Proteoglycan‐4 Regulates Phagocytic Activation of Monocytes and Reduces IL‐1b Secretion by Urate Crystal Stimulated PBMCs

Introduction Gout is a chronic inflammatory disease due to monosodium urate (MSU) crystal deposits in articular joints. MSU phagocytosis by monocytes/macrophages results in interleukin-1 beta (IL-1β) secretion. Pharmacologic treatments of gout are inadequate and are associated with significant side effects. Interleukin-1 receptor antagonist (IL-1RA) is an endogenous inhibitor of IL-1b signaling and is efficacious in acute gout management where other treatments are contraindicated. Proteoglycan-4 (PRG4) is an anti-inflammatory glycoprotein that has shown efficacy in reducing MSU induced IL-1b release from macrophages. Our objective was to compare the activation of monocytes, of normal subjects and gout patients, by MSU crystals and examine the comparative efficacy of rhPRG4 and rhIL-1RA in regulating phagocytic activation of monocytes and IL-1b secretion. Methods Collection of peripheral blood mononuclear cells (PBMCs) and sera from patients with gout was approved by the IRB at RIH. PBMCs and sera of normal subjects were obtained from ATCC. IL-1b and IL-1RA levels were determined by ELISA. PBMCs were seeded (1 million cells/well) in 6 well plates and treated with Pam3CSK4 (1mg/mL; TLR2 agonist) ± rhIL-1RA or rhPRG4 for 24 h followed by MSU crystals (200mg/mL) for 6 h. Monocytes in PBMCs were identified by flow cytometry using anti-CD14-APC-Cy7 and anti-CD45-PE-A antibodies (BioLegend). Monocytes’ phagocytic activity was determined by co-incubation of PBMCs with FITC-labeled rabbit IgG-coated latex beads (Cayman Chemicals) for 2 h followed by determination of percent bead-positive monocytes. Statistical analyses included Student's t-test and one-way ANOVA followed by post-hoc Tukey's test. Results Sera from patients with gout had lower IL-1RA levels compared to sera from normal subjects (p<0.01; fig. 1A). The gating of monocytes and corresponding bead phagocytosis is shown in fig. 1B. Representative plots showing increased bead phagocytosis by gout PBMCs compared to normal PBMCs are shown in fig. 1C. Basal phagocytic activities of gout PBMCs were higher than corresponding activities of normal PBMCs (p<0.0001; fig. 1D). IL-1b secretion in MSU crystal stimulated gout PBMCs was higher than corresponding secretion in normal PBMCs (p<0.01; fig. 1E). rhIL-1A and rhPRG4 dose-dependently reduced IL-1b secretion by normal PBMCs (p<0.001; fig. 2A & 2B). rhPRG4 was efficacious in reducing phagocytic activities of monocytes from normal and goutsubjects (p<0.001; fig. 2C & 2E). rhPRG4 exhibited a non-significant trend towards reducing IL-1b secretion by gout PBMCs (p=0.13) and the efficacy of rhPRG4 was not different from IL-1RA (p>0.05; fig. 2D). Conclusion Gout PBMCs secreted more IL-1b associated with higher phagocytic activation of monocytes. Uniquely, rhPRG4 reduced phagocytic activation of monocytes, from normal and gout patients. rhIL-1RA and rhPRG4 showed comparable anti-inflammatory efficacy. rhPRG4 is a promising new systemic therapy in gout.

Neurological Manifestations of COVID-19 Associated Multi-system Inflammatory Syndrome in Children: A Systematic Review and Meta-analysis.

Children comprise only 1-5% of COVID-19 cases. Recent studies have shown that COVID-19 associated multisystem inflammatory syndrome in children (MIS-C) can present with neurological signs and symptoms. In this systematic review and meta-analysis, we have reviewed neurological involvement in these patients. A comprehensive electronic literature search was done on PubMed, Google Scholar, Embase, Cochrane database, and SCOPUS for the published English language articles from December 1, 2019, to February 28, 2021. A meta-analysis of the proportion was expressed as a pooled proportion with a 95% confidence interval (CI). Representative forest plots showing individual studies and the combined effect size were generated to provide an overview of the results. This systematic review and meta-analysis analyzed 15 published MIS-C studies with a total of 785 patients. Neurological manifestations in patients with MIS-C was found in 27.1%. We found that 27% developed headaches, 17.1% developed meningism/meningitis and 7.6 % developed encephalopathy. Other uncommon neurological manifestations of MIS-C includes anosmia, seizures, cerebellar ataxia, global proximal muscle weakness and bulbar palsy. In MIS-C patients with neurological feature, neuroimaging showed signal changes in the splenium of the corpus callosum. Electroencephalography showed slow wave pattern and nerve conduction studies and electromyography showed mild myopathic and neuropathic changes. Our study revealed that neurological manifestations are not uncommon in patients with MIS-C. Further large prospective studies are needed to better explore the disease spectrum and to unravel the underlying pathophysiology.

Mind the gaps: Comparison of representative vs opportunistic assessment of tree regeneration in Central European beech forests

Tree regeneration (TR) in canopy gaps is a key process to understand how forest ecosystems might adapt to future environmental changes. Since successful TR is the result of a complex interplay of several stochastic events such as gap formation, seed production, ungulate pressure and diseases, some of the processes have been neglected or strongly reduced for model applications. Most empirical data on TR originate from (national) forest inventories based on statistically optimized designs for timber resource estimations and use geographically fixed permanent plots. Consequently, these representative samples record TR both in gaps as well as under closed canopy conditions. In this study, we compared TR in 63 representative plots of the Swiss National Forest Inventory (‘NFI sampling’), located in beech and silver fir-beech forests along an elevation gradient in the Jura Mts., with an opportunistic ‘gap sampling’ strategy targeting TR-clusters. We analyzed quantitative (e.g., number of individuals) and qualitative (e.g., species diversity, similarity to surrounding canopy) differences between the ‘NFI sampling’ and ‘gap sampling,’ as well as the environmental factors (e.g., gap size, cluster age, elevation) influencing the composition of TR in small canopy gaps. The ‘gap sampling’ recorded higher median TR densities (+62%, stems ha−1) and species richness (4.0 ± 2.1, mean ± sd) than the ‘NFI sampling’. More importantly, the ‘gap sampling’ provided much more consistent results among sites (i.e., 4x lower variance). Consequently, the ‘gap sampling’ revealed not only general but also species-specific patterns across TR size classes and along elevational gradients in contrast to the lack of patterns in the ‘NFI sampling’. Species richness decreased as TR grew taller, and the tallest TR size class (>130 cm) was almost solely comprised of the dominant species in the surrounding canopy (direct ingrowth). The high amount of direct ingrowth in TR indicates that small canopy gaps might offer only limited management potential to alter forests and that extensive disturbances or management at early TR stages (sapling) would be necessary to promote species better suited to the expected future site conditions. We raise the question of whether a forest monitoring that is statistically optimized for timber resource estimation is best suited to detect adaptation effects to quickly changing environments best visible at the regeneration stage, or whether forest inventory tools should be extended by methods that focus on disturbed parts of forests where changes and adaptations take place. We propose the implementation of an opportunistic gap-based monitoring of TR that essentially serves to identify the best forest management strategies (e.g. gap size, intervention intervals) in forests adapting to environmental change.

Evaluation of Ranzoni etal.: Integrative Single-Cell RNA-Seq and ATAC-Seq Analysis of Human Developmental Hematopoiesis.

This article shows an example of the peer review process for "Integrative Single-Cell RNA-Seq and ATAC-Seq Analysis of Human Developmental Hematopoiesis" (Ranzoni etal., 2021).

冀北山地天然次生林土壤生态化学计量特征及影响因素

为了探究天然次生林植被及立地因子对土壤养分含量的影响，量化植被与立地因子对土壤养分的影响比例，为合理抚育冀北山区次生林，提高次生林生产力提供依据，以冀北山地分布的天然次生林为研究对象，选取代表性的地段并设置面积为20 m×20 m的样地，以均匀布点与多点混合为原则，按照0-20、20-40、40-60 cm土层取样。测定土壤pH值、土壤有机碳（Soil organic carbon，SOC）、全氮（Total nitrogen，TN）、全磷（Total phosphorus，TP）、全钾（Total potassium，TK）、速效磷（Available phosphorus，AP）、速效钾（Available potassium，AK）含量，并计算土壤碳、氮、磷计量比。结果表明：不同深度土层土壤TN与SOC之间保持显著相关（P<0.001），SOC、C∶N与AK之间呈显著正相关（P<0.01）；研究区域冀北山地次生林3层不同深度土壤C∶N平均值（27.74、29.96、29.44）与C∶P平均值（158.86、128.63、119.20）明显高于全国土壤C∶N（11.90）与C∶P（60.00）平均值，N∶P平均值（6.45、4.59、4.70）与全国平均水平（3.56）基本相当，对各深度层土壤N∶P值与土壤TN、TP值进行相关性分析发现，TN值与N∶P的相关性更加显著，说明影响N∶P值的主要因素为氮素量的限制。环境因子对3层不同深度土壤的养分指标分别做出48.64%、53.45%及49.23%的解释比例（P<0.001），因子交互后的解释能力相较因子自身的解释能力更强，地形因子与其他因子相交互后，解释比率分别为其单独解释比例的6.45、8.06、3.21倍。从立地条件角度讲，地形因子能够对土壤养分空间分布产生一定的影响，抚育过程中对地形因子的作用应当予以重视。;In order to explore the effects of natural secondary forest vegetation and site factors on soil nutrient concentration, quantify the ratio of vegetation and site factors on soil nutrition, provide a basis for rationally tending secondary forests in the mountainous area of northern Hebei and improving secondary forest productivity, the natural secondary forests distributed in the mountains of northern Hebei Province were selected as the research object and a representative plot was selected and a sample plot with an area of 20m×20m was selected. The principle of uniform distribution and multi-site mixing was adopted when 0-20 cm, 20-40 cm and 40-60 cm soil layer sampling. Soil pH value (Soil pH value), soil organic carbon (SOC), total nitrogen (TN), total phosphorus (TP), total potassium (TK), and available phosphorus (AP), available potassium (AK) content were measured in experiment. Based on the results of experiment, soil carbon, nitrogen, and phosphorus stoichiometric ratio were calculated. The results showed that the soil TN and SOC of different depths maintained an extremely significant correlation(P<0.001)among different soil layers, there was also a significant positive correlation between SOC, C∶N and AK(P<0.01).The average values of C∶N ratio of three layers of soils at different depths in the secondary forest in northern Hebei mountainous area (27.74,29.96 and 29.44) and C∶P ratio (158.86, 128.63 and 119.20) were significantly higher than the average level of national soil C∶N (11.90) and C∶P (60.00).Besides, N∶P averages (6.45, 4.59 and 4.70) was basically equivalent to the national average (3.56). The correlation analysis of the soil N∶P value and the soil TN and TP values of each depth layer showed that the correlation between the TN value and the N∶P value was more significant, indicating that the factor why N∶P value was affected was the limitation of nitrogen. Environmental factors accounted for 48.64%, 53.45%, and 49.23% of soil nutrient index of the three layers of soil at different depths (P<0.001). The explanatory power after the factor interaction was stronger than that of the factor itself. After interacting with other factors, the explanatory ratios of terrain factors were 6.45, 8.06, and 3.21 times of the separate ones. From the perspective of site conditions, topographical factors did have a certain impact on the spatial distribution of soil nutrition, and the role of topographical factors in the process of tending should be paid attention to.

Evaluation of University Campus Landscape Based on AHP: A Case Study of Southwest University of Science and Technology

In order to quantitatively evaluate the qualitative problem of campus landscape comfort of Southwest University of Science and Technology, based on the principles of landscape design, the Analytic Hierarchy Process (AHP method) and comprehensive index method are used to establish a reasonable overall landscape evaluation model for the university campus. Firstly, professional teachers are invited to compare the importance of various factors of the landscape to determine the weight of each evaluation factor, and then conduct a field survey on various factors of the landscape of Southwest University of Science and Technology, and select representative sample plots for questionnaire survey and statistics. Finally, the comfort score evaluation and comprehensive evaluation grade analysis of the overall campus landscape are carried out. The results show that the campus landscape in the Southwest University of Science and Technology is generally better, and there is still much room for improvement. By summarizing and analyzing the existing problems of the campus landscape and putting forward suggestions for optimization, it aims to provide a certain reference for the future campus landscape construction.

Regeneration Characteristics and Spatial Pattern of &amp;lt;i&amp;gt;Platycladus orientalis&amp;lt;/i&amp;gt; in Mount Tai, China

Platycladus orientalis is one of the main tree species in Mount Tai, and its sustainable development is of great significance to the protection and development of the forest ecological environment of Mount Tai. In this study, a representative sample plot of Platycladus orientalis with a size of 30 m * 20 m was selected on Mount Tai. The growth and distribution of the parent trees and seedlings of Platycladus orientalis were investigated in the sample plots. The results showed that both the mother cypress trees and seedlings inside the plot present an aggregated distribution pattern within a certain scale, which conforms to the distribution of most natural communities; and the mother cypress can promote seedlings within a certain scale. Although Platycladus orientalis has good natural regeneration ability, it is difficult to succeed in understory regeneration solely relying on natural fertility, and it is not conducive to the long-term stable development of forest stands. Therefore, the thinning of Platycladus orientalis and the adjustment of the forest stand structure and artificial promotion of natural regeneration can not only reduce operating costs, but also conform to the idea of forest near-natural management and maintain sustainable forest development.

An Investigation on the Impact of Different Vegetation Types on Microenvironment in Winter

The carefully designed greening system can significantly improve the thermal comfort of a microenvironment. A greening system is usually composed of different vegetation types, which have different impacts on their surrounding microenvironment. In order to evaluate the impacts of different vegetation types on campus microenvironment, four representative plots with different vegetation types (arbor + lawn, shrub + lawn, lawn, bamboo forest + lawn) in a university campus in northwest China were selected for investigation. HOBO temperature and humidity recorder were used to monitor the temperature and humidity of four plots at a level of 1.5m high in for 24 consecutive hours in 5 days in winter of 2019. The results show that, during the observation period, all the four kinds of vegetation types have a certain influence on the microenvironment, and the degree of influence shows a trend of bamboo forest + lawn> arbor + lawn> shrub + lawn> lawn. The results of this research are expected to provide a reference in the selection of vegetation types for the design of campus greening system.

Clinical Relevance of Polymorphonuclear Myeloid-Derived Suppressor Cells in Autoimmune-Blistering Disorders Pemphigus Vulgaris and Bullous Pemphigoid

Clinical Relevance of Polymorphonuclear Myeloid-Derived Suppressor Cells in Autoimmune-Blistering Disorders Pemphigus Vulgaris and Bullous Pemphigoid

Bacterial diversity on an abandoned, industrial wasteland contaminated by polychlorinated biphenyls, dioxins, furans and trace metals

Most former industrial sites are contaminated by mixtures of trace elements and organic pollutants. Levels of pollutants do not provide information regarding their biological impact, bioavailability and possible interactions between substances. There is genuine interest in combining chemical analyses with biological investigations. We studied a brownfield where several industrial activities were carried out starting in the 1970s, (incineration of pyralene transformers, recovery of copper by burning cables in the open air). Four representative plots showing different levels of polychlorobiphenyls were selected. Organic and trace metal levels were measured together with soil pedological characteristics. The bacterial community structure and functional diversity were assessed by 16S metagenomics with deep sequencing and community-level physiological profiling. Additionally, a vegetation survey was performed. Polychlorobiphenyls (8 mg.kg−1 to 1500 mg.kg−1) were from 2.4 × 103-fold to 6 × 105-fold higher than the European background level of 2.5 μg.kg−1. Polychlorinated dibenzo-p-dioxins and dibenzofurans ranged from 0.5 to 8.0 μg.kg−1. The soil was also contaminated with trace metals, i.e., Cu > 187, Zn > 217 and Pb > 372 mg.kg−1. Location within the study area, trace metal content and soil humidity were stronger determinants than organic pollutants of bacterial community structures and activities. Thus, the highest biological activity and the greatest bacteriological richness were observed in the plot that was less contaminated with trace metals, despite the high level of organic pollutants in the plot. Moreover, trace element pollution was associated with a relatively low presence of Actinobacteria and Rhizobia. The plot with the highest metal contamination was rich in metal-resistant bacteria such as Sphingomonadales, Geodermatophilaceae and KD4–96 (Chloroflexi phylum). Acidobacteria and Sphingomonadales, capable of resisting trace metals and degrading persistent organic pollutants, were dominant in the plots that had accumulated metal and organic contamination, but bacterial activity was lower in these plots than in the other plots.

Soil Cover Diversity and Its Spatial Organization at Different Map Scales

We summarized the results of studies of pedodiversity and contrast and variability of soil properties based on the analyses of soil maps of different scales for the republics of Karelia and Dagestan and the Moscow region. We used fragments of the Soil Map of the Russian Federation on a scale of 1 : 2 500 000 as a small-scale map and three different medium-scale maps (from 1 : 300 000 to 1 : 500 000). Large-scale maps (1 : 10 000) were compiled for representative plots within the studied territories specifically for the study of pedodiversity. All the maps were digitized, map legends were converted to the new Russian soil classification system and to the World Reference Base for Soil Resources (WRB), and Shannon–Wiener diversity indices were calculated. Comparison of indicators revealed the dependence of the values of the diversity indices on the map scale. The indicators for the large-scale map were relatively low and did not differ much when using Russian and international classification. We concluded that for the successful assessment of soil diversity at different scales, it is important to use several large-scale maps that reflect the heterogeneity of regional soilscapes. Natural boundaries between soilscapes and soil regions can be found using the pedodiversity index map. In almost all cases, maps with a WRB-based legend had lower indicators of pedodiversity than maps with a legend based on the Russian soil classification. This confirms the fact that the international classification is less detailed than the Russian national classification, which was originally intended for large-scale mapping of soils.

Efficacy and safety of rituximab in autoimmune encephalitis: A meta-analysis.

Autoimmune encephalitis (AE) is a rare but debilitating neurological disease where the body develops antibodies against neuronal cell surface/synaptic proteins. Rituximab is an anti-CD20 chimeric monoclonal antibody which shows promise in AE treatment observational studies. To our knowledge, there has been no previous meta-analysis providing robust evidence on the effectiveness and safety of rituximab as second-line therapy for the treatment for AE. This study was conducted according to the PRISMA (Preferred Reporting Items for Systematic review and Meta-Analysis) statement. Investigators independently searched PubMed, Web of Science, Google Scholar, WANFANG, CNKI, and J-STAGE for studies. Meta-analysis via representative forest plots was conducted for good functional outcome (mRS≤2), proportion of relapse, and mRS score change pre- and post-treatment. Good functional outcome at last follow-up following rituximab therapy occurred in 72.2% of patients (95% CI: 66.3%-77.4%). Mean mRS score decreased by 2.67 (95% CI: 2.04-3.3; P<.001). Relapses following the rituximab therapy occurred in only 14.2% of patients (95% CI: 9.5%-20.8%). Infusion related reactions, pneumonia, and severe sepsis were seen in 29 (15.7%), 11 (6.0%), and two patients (1.1%), respectively. The efficacy and side effect profile of rituximab are comparable to outcomes seen in rituximab use in other autoimmune and inflammatory CNS disease. Our meta-analysis showed that rituximab is an effective second-line agent for AE with an acceptable toxicity profile.

Dominant species' dominant role and spatial stability are enhanced with increasing stocking rate

Stipa breviflora Grisb. (S. breviflora) is a dominant species in the desert steppe of northern China. Its function and role at the plant community level increases with increasing stocking rate. However, the response of spatial stability remains unclear. We selected treatment areas representing no grazing (CK), light grazing (LG), moderate grazing (MG) and heavy grazing (HG) in a long-term grazing experiment (2004–2017) in a S. breviflora desert steppe in Inner Mongolia, northern China. Using a mechanical sampling method, 40 m × 40 m representative sample plots were selected to obtain the height, coverage and density of the S. breviflora population and community, and we computed the standing crop of mechanical sampling quadrats based on a random sample of cutting quadrats. Analysis of standing crop, density of S. breviflora population and its ratio in the plant community showed that the dominant role of S. breviflora population in the plant community increased with increasing grazing intensity, while the spatial stability of S. breviflora population not only had many dimensions, but also many states. The dimension or combination of dimensions of its stability performance and its adaptive state varied under different disturbance intensities and frequencies.

ВЫДЕЛЕНИЕ РЕПРЕЗЕНТАТИВНЫХ УЧАСТКОВ ЛЕСНЫХ ЭКОСИСТЕМ В РАМКАХ FSC СЕРТИФИКАЦИИ

Voluntary forest certification is a widespread mechanism of a market economy aimed at balancing economic, environmental and social interests. The article provides a methodology for identifying representative areas of forest ecosystems when forming a network of protected areas that form the ecological framework of the leased area. As an example, we took a logging company that leases two forest plots in Chebulinsky forestry of the Kemerovo region with different characteristics of the forest fund and the history of forest use. Based on the basic requirements of the current FSC forest management standard, a methodology for identifying representative sites has been developed. As a result of its application, it was found that in the area with more intensive forest management, 41% of the forest types present in the entire forestry area are found, despite its smaller area (3837 hectares). It is 21% on a larger, but less transformed and fragmented plot (5224 ha). It was revealed that the share of the frequent forest types (with the occurrence above 1%), which is 18%, is common for both leased plots. The area of representative plots is also approximately the same - 4.16 and 3.79%, respectively. It is noteworthy that frequent types of forests are already present in sufficient quantities in the network of protective forests and especially protective areas in production forests. But rare forest types, as a rule, are slightly represented in them. It enables us to conclude that it is important to form a network of representative sites in forest certification. It has been established that both coniferous and deciduous stands fall into rare forest ecosystems. The average area under allotment in the older cultivated leased plot is 8.8 hectares (with a variation from 0.8 to 55.0 hectares), and in less cultivated - 13.3 hectares (with a variation from 1.8 to 28.7 hectares), which is quite logical. However, in the first leased area, only 9%, and in the second - 6% of representative areas have an area of over 20 ha, which is accepted as a threshold in several countries

CD44‐Targeted Monoclonal Antibody is Anti‐Inflammatory in an In‐Vivo Model of Monosodium Urate Crystal Induced Peritonitis

CD44‐Targeted Monoclonal Antibody is Anti‐Inflammatory in an <i>In‐Vivo</i> Model of Monosodium Urate Crystal Induced Peritonitis

Fingolimod (FTY720) Activates Protein Phosphatase‐2A in Human Monocytes and Inhibits Uric Acid Crystal‐Induced Interleukin‐1 Beta Production

IntroductionGout is a chronic inflammatory disease due to monosodium urate (MSU) crystal deposits in articular joints. Phagocytosis of MSU crystals by monocytes/macrophages results in downstream expression and production of interleukin‐1 beta (IL‐1β). The two‐step process of gout inflammation involves 1) priming of monocytes with danger signals e.g. lipopolysaccharide (LPS) or soluble uric acid (UA) and 2) MSU phagocytosis and activation of the NLRP3 inflammasome in monocytes. Protein‐phosphatase‐2A (PP2A) is a serine/threonine phosphatase, and a reduction in intracellular macrophage PP2A activity was shown to aggravate LPS‐induced proinflammatory cytokine production. The prodrug Fingolimod (FTY720) and its phosphorylated active metabolite (p‐FTY720) are efficacious in multiple sclerosis and their mechanism of action involves activation of intracellular PP2A. The objective of this study is to investigate the role of PP2A in regulating monocyte priming and activation by MSU crystals and whether intracellular PP2A activation exerts an anti‐inflammatory effect in MSU‐stimulated monocytes.MethodsHuman THP‐1 monocytes (ATCC; 500,000 cells per well) were treated with UA (50 mg/dL) and LPS (10ng/mL) for 24 hours. Subsequently, cells were treated with MSU crystals (100ug/mL) and MSU phagocytosis was determined at 4 hours using flow cytometry and the percentage of cells displaying side‐scattering above a threshold value, indicative of crystal phagocytosis, were determined. Intracellular PP2A activity was determined at 6 hours following MSU stimulation using the PP2A Immunoprecipitation Phosphatase Assay Kit (Fisher Scientific). IL‐1β expression and protein levels at 6 hours were determined using quantitative polymerase chain reaction and ELISA, respectively. THP‐1 monocytes were treated with FTY720 or p‐FTY720 (2.5mM for both treatments; Cayman Chemicals) and PP2A activity was determined at 1, 3, 6 and 24 hours. THP‐1 monocytes were treated with UA+LPS followed by MSU ± p‐FTY720 (2.5mM for 6 hours) and IL‐1β expression and protein levels were determined. Statistical analyses were performed using analysis of variance (ANOVA) with Tukey’s post‐hoc test.ResultsUA+LPS pre‐treatment increased MSU phagocytosis by THP‐1 monocytes (fig. 1A and 1B; p&lt;0.001) and IL‐1β expression (fig. 1C; p&lt;0.001) and production (fig. 1D; p&lt;0.001). This effect was associated with a reduction in intracellular PP2A activity (fig. 1E; p&lt;0.001). FTY720 and p‐FTY720 increased intracellular PP2A activity in monocytes (fig. 2A; p&lt;0.001) with a higher peak effect achieved at 1 hour for p‐FTY720 (p&lt;0.01). p‐FTY720 treatment reduced IL‐1β expression (fig. 2B; p&lt;0.001) and mature IL‐1β production (fig. 2C; p&lt;0.001) in UA+LPS pre‐treated human monocytes following MSU stimulation.ConclusionUA and LPS enhanced MSU phagocytosis, expression and production of IL‐1β, potentially via a reduction in intracellular PP2A activity. FTY720 and p‐FTY720 activated PP2A in monocytes with an onset of effect of 1 hour and correspondingly reduced IL‐1β expression and production. PP2A is a novel therapeutic target for gout treatment.Impact of soluble uric acid (UA) and lipopolysaccharide (LPS) pre‐treatments on THP‐1 monocyte phagocytic activity towards monosodium urate (MSU) crystals, interleukin‐1 beta (IL‐1β) expression and production and association with intracellular protein phosphatase‐2A (PP2A) activity. A) Representative flow cytometry scatter plots showing enhanced MSU crystal phagocytosis (as shown by more cells with increased side‐scatter in P2 region of interest) by THP‐1 monocytes following UA+LPS pre‐treatment. B) UA and UA+LPS pre‐treatment increased the percentage of THP‐1 monocytes positive for MSU phagocytosis. C) UA+LPS pre‐treatment increased IL‐1β expression by THP‐1 monocytes following MSU challenge. D) UA+LPS pre‐treatment increased IL‐1β production by THP‐1 monocytes following MSU challenge. E) Intracellular PP2A activity was reduced in UA or UA+LPS pre‐treated THP‐1 monocytes. *p&lt;0.001; **p&lt;0.01; n.s.: non significant.Figure 1Impact of FTY720 (Fingolimod; prodrug) and phosphorylated FTY720 (p‐FTY‐720; active metabolite) treatments on protein phosphatase‐2A (PP2A) activity in human THP‐1 monocytes and impact of p‐FTY720 treatment on interleukin‐1 beta (IL‐1β) expression and production by monosodium urate (MSU) crystal stimulated THP‐1 monocytes ± soluble uric acid (UA) and lipopolysaccharide (LPS) pre‐treatment. Treatments were conducted using 2.5μM concentrations. A) FTY720 and p‐FTY720 increased intracellular PP2A activity in THP‐1 monocytes. B) p‐ FTY720 reduced IL‐1β expression in THP‐1 monocytes. C) p‐FTY720 reduced IL‐1β production in THP‐1 monocytes. *p&lt;0.001; **p&lt;0.01; n.s.: non significant.Figure 2

The distribution of pine mistletoe (Viscum album ssp. austriacum) in Scots pine (Pinus sylvestris) forests: from stand to tree level

ABSTRACTThis paper reports and discusses the distribution of pine mistletoe in Scots pine forests at stand and tree level in relation to some site, stand, and tree characteristics. Population characteristics (age, number and biomass) and distribution of pine mistletoe in the canopy of Scots pine trees were also evaluated. A total of 1408 trees were measured in 114 representative sample plots in the Eastern Black Sea Region of Turkey. Of the measured trees, 648 (46%) were mistletoe infected and 760 (54%) were uninfected. The 6-class dwarf mistletoe rating system (DMRS) was used for infection severity rating. Correlation analysis results indicated significant relationships between mistletoe infection severity and stand and tree characteristics. The t-test results showed that there was a significant difference between mean stand and tree characteristics in mistletoe infected and uninfected stands. The results of the binary logistic regression analysis indicated that altitude and stand volume were the most significant predictor of the distribution of pine mistletoe at stand level. As for the population characteristics of pine mistletoe, the results indicated that the life span of pine mistletoe was about 24 years and that biomass distribution gradually increased from lower to upper part of the crowns.

Mitochondrial transfer from MSCs to T cells induces Treg differentiation and restricts inflammatory response.

Mesenchymal stem cells (MSCs) have fueled ample translation for the treatment of immune-mediated diseases. They exert immunoregulatory and tissue-restoring effects. MSC-mediated transfer of mitochondria (MitoT) has been demonstrated to rescue target organs from tissue damage, yet the mechanism remains to be fully resolved. Therefore, we explored the effect of MitoT on lymphoid cells. Here, we describe dose-dependent MitoT from mitochondria-labeled MSCs mainly to CD4+ T cells, rather than CD8+ T cells or CD19+ B cells. Artificial transfer of isolated MSC-derived mitochondria increases the expression of mRNA transcripts involved in T-cell activation and T regulatory cell differentiation including FOXP3, IL2RA, CTLA4, and TGFβ1, leading to an increase in a highly suppressive CD25+ FoxP3+ population. In a GVHD mouse model, transplantation of MitoT-induced human T cells leads to significant improvement in survival and reduction in tissue damage and organ T CD4+ , CD8+ , and IFN-γ+ expressing cell infiltration. These findings point to a unique CD4+ T-cell reprogramming mechanism with pre-clinical proof-of-concept data that pave the way for the exploration of organelle-based therapies in immune diseases.