In order to evaluate the impacts of volatile organic compounds(VOCs)emissions from building materials on the indoor air quality beyond the standard chamber test conditions and test period,mechanistic emission source m...In order to evaluate the impacts of volatile organic compounds(VOCs)emissions from building materials on the indoor air quality beyond the standard chamber test conditions and test period,mechanistic emission source models have been developed in the past.However,very limited data are available for the required model parameters including the initial concentration(C_(m0)),in-material diffusion coefficient(D_(m)),partition coefficient(Kma),and convective mass transfer coefficient(k_(m)).In this study,a procedure was developed for estimating the model parameters by using VOC emission data from standard small chamber tests.In the procedure,initial values of the model parameters were refined by multivariate regression analysis of the measured emission data.To verify the procedure and estimate its uncertainty,simulated chamber test data were generated by adding 10% experimental uncertainties on the theoretical curve from the analytical solution to a mechanistic emission model.Then the procedure was applied to the generated data to estimate the model parameters.Results indicated that estimates converged to the original parameter values used for the data generation and the error of estimated parameters D_(m1)C_(m0) and K_(ma) were within±10%,±23%,and±25%of the true values,respectively.The procedure was further demonstrated by applying it to estimate the model parameters from real chamber test data.Wide application of the procedure would result in a database of mechanistic source model parameters for assessing the impact of VOC emissions on indoor pollution load,which are essential input data for evaluating the effectiveness of various indoor air quality(IAQ)design and control strategies as well as the energy required for meeting given IAQ requirements.展开更多
The human face is a heritable surface with many complex sensory organs.In recent years,many genetic loci associated with facial features have been reported in different populations,yet there is a lack of studies on th...The human face is a heritable surface with many complex sensory organs.In recent years,many genetic loci associated with facial features have been reported in different populations,yet there is a lack of studies on the Han Chinese population.Here,we report a genome-wide association study of 3D normal human faces of 2,659 Han Chinese with autosegment phenotypes of facial morphology.We identify singlenucleotide polymorphisms(SNPs)encompassing four genomic regions showing significant associations with different facial regions,including SNPs in DENND1 B associated with the chin,SNPs among PISRT1 associated with eyes,SNPs between DCHS2 and SFRP2 associated with the nose,and SNPs in VPS13 B associated with the nose.We replicate 24 SNPs from previously reported genetic loci in different populations,whose candidate genes are DCHS2,SUPT3 H,HOXD1,SOX9,PAX3,and EDAR.These results provide a more comprehensive understanding of the genetic basis of variation in human facial morphology.展开更多
Uncertainty exists in many aspects of building simulation.A deterministic hygrothermal analysis may not sufficiently give a reliable guidance if a number of input variables are subject to uncertainty.In this paper,a p...Uncertainty exists in many aspects of building simulation.A deterministic hygrothermal analysis may not sufficiently give a reliable guidance if a number of input variables are subject to uncertainty.In this paper,a probabilistic-based method was developed to evaluate the hygrothermal performance of building components.The approach accounts for the uncertainties from model inputs and propagates them to the outputs through the simulation model,thus it provides a likelihood of performance risk.Latin hypercube sampling technique,incorporated with correlation structure among the inputs,was applied to generate the random samples that follows the intrinsic relations.The performance of an internally insulated masonry wall was evaluated by applying the proposed approach against different criteria.Thermal performance,condensation and mould growth potential of the renovated wall can overall satisfy the requirements stipulated in multifold standards.The most influential inputs were identified by the standardized regression sensitivity analysis and partial correlation technique.Both methods deliver the same key parameters for the single and time-dependent output variables in the case study.The probabilistic method can provide a comprehensive risk analysis and support the decision-maker and engineer in the design and optimization of building components.展开更多
There is a fundamental body of evidence suggesting that activated apoptosis signaling in ejaculated human sperm negatively influences their fertilization potential. However, it is still controversial whether this apop...There is a fundamental body of evidence suggesting that activated apoptosis signaling in ejaculated human sperm negatively influences their fertilization potential. However, it is still controversial whether this apoptotic signaling is a relic of an abortive apoptosis related to spermatogenesis or if it should be regarded as a functional preformed pathway in mature sperm leading to stereotypical morphological changes reflecting nuclear disassembly. To address this question, apoptosis was induced using betulinic acid in mature and immature ejaculated human sperm enriched by density gradient centrifugation. Execution of apoptosis was monitored by observing ultra-morphological changes via transmission electron microscopy. Typical morphological signs of apoptosis in somatic cells include plasma membrane blebbing with the formation of apoptotic bodies, impaired mitochondrial integrity, defects of the nuclear envelope, and nuclear fragmentation; these morphologies have also been observed in human sperm. In addition, these apoptotic characteristics were more frequent in immature sperm compared to mature sperm. Following betulinic acid treatment, apoptosis-related morphological changes were induced in mature sperm from healthy donors. This effect was much less pronounced in immature sperm. Moreover, in both fractions, the betulinic acid treatment increased the percentage of acrosome-reacted sperm. The results of our ultra-morphological study prove the functional competence of apoptosis in mature ejaculated human sperm. The theory of a sole abortive process may be valid only for immature sperm. The induction of the acrosome reaction by stimulating apoptosis might shed light on the biological relevance of sperm apoptosis.展开更多
The Mesta-Nestos river basin in Bulgaria and Greece is a case study for transboundary decision-making support in south-eastern Europe and a show-case for the development of methodologies and information-gathering for ...The Mesta-Nestos river basin in Bulgaria and Greece is a case study for transboundary decision-making support in south-eastern Europe and a show-case for the development of methodologies and information-gathering for the integrated regional planning of water resources. Land-use conflicts in this water-scarce region cover a wide spectrum of activities like agricultural irrigation, drinking water production, diversions for industrial water, and risk of pollution from mining, to name a few examples. Measurements of the water quality were carried out in the upper basin. Results will be illustrated by the example of the environmental situation in the alpine region of the Pirin National Park as well as in the Razlog Basin with a stronger anthropogenic impact and pollution around a former uranium mine near the village of Elesnica. The social and economic development of this transboundary region is a recently established priority for mean an increase in water usage the water resources if regional the future. It will and more stress for impacts of global climate change are verified. Problem-focused management of the catchment area as a whole on the basis of proved geo-data sets is needed for the future.展开更多
Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the eff...Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.展开更多
Formation of claudin-10 based tight junctions(TJs)is paramount to paracellular Na+transport in multiple epithelia.Sequence variants in CLDN10 have been linked to HELIX syndrome,a salt-losing tubulopathy with altered h...Formation of claudin-10 based tight junctions(TJs)is paramount to paracellular Na+transport in multiple epithelia.Sequence variants in CLDN10 have been linked to HELIX syndrome,a salt-losing tubulopathy with altered handling of divalent cations accompanied by dysfunctional salivary,sweat,and lacrimal glands.Here,we investigate molecular basis and phenotypic consequences of a newly identified homozygous CLDN10 variant that translates into a single amino acid substitution within the fourth transmembrane helix of claudin-10.In addition to hypohidrosis(H),electrolyte(E)imbalance with impaired urine concentrating ability,and hypolacrimia(L),phenotypic findings include altered salivary electrolyte composition and amelogenesis imperfecta but neither ichthyosis(I)nor xerostomia(X).Employing cellular TJ reconstitution assays,we demonstrate perturbation of cis-and trans-interactions between mutant claudin-10 proteins.Ultrastructures of reconstituted TJ strands show disturbed continuity and reduced abundance in the mutant case.Throughout,both major isoforms,claudin-10a and claudin-10b,are differentially affected with claudin-10b showing more severe molecular alterations.However,expression of the mutant in renal epithelial cells with endogenous TJs results in wild-type-like ion selectivity and conductivity,indicating that aberrant claudin-10 is generally capable of forming functional paracellular channels.Thus,mutant proteins prove pathogenic by compromising claudin-10 TJ strand assembly.Additional ex vivo investigations indicate their insertion into TJs to occur in a tissue-specific manner.展开更多
This research reveals relationships between climate variables and inter-annual dynamics in the area of the glacieret located in the cirque Golemiya Kazan in the Pirin Mountains. The study period is 1993–2017. The cor...This research reveals relationships between climate variables and inter-annual dynamics in the area of the glacieret located in the cirque Golemiya Kazan in the Pirin Mountains. The study period is 1993–2017. The correlations are identified using statistical methods. Also, a statistical model is constructed, including some climate variables as predictors. Despite the evident decrease of the glacieret's size in the period from the 1950 s onwards, the long-term trends for the last decades have been insignificant. The main climatic factors influencing the inter-annual dynamics in the area of the glacieret are air temperature, precipitation, zonal and meridional winds and relative humidity. With respect to the dynamics in the area of the glacieret, the important trends in the different climate variables are those of the warm period air temperatures and zonal(u) wind. They also determine to a great extent its future development by acting in two opposite directions – rising temperatures in the warm period will lead to a rapid decrease of its area by the end of the melting season, while the change of wind directionfrom west to east in the warm period will increase its area. The influence of the zonal wind in the warm period is explained mainly by the location of the glacieret in the cirque. Generally, the glacieret is tilted downwards from west to east. Thus, westerly winds facilitate blowing away the snow from the surface of the glacieret, assisting its melting in the warm period. Easterly winds do not have such an effect. The combination of the opposite effects of these two most important climate variables leads to the most likely scenario for the future development of the glacieret, according to which by the middle of this century it is expected to turn into a semi-permanent snow patch, which disappears after some summers, and by the end of the century to completely melt every year before the end of the melting season.展开更多
New 4^(th)-generation synchrotron sources,with their increased bilince,promise to greatly improve the performances of coherent X-ray microscopy.This perspective is of major interest for crystal microscopy,which aims a...New 4^(th)-generation synchrotron sources,with their increased bilince,promise to greatly improve the performances of coherent X-ray microscopy.This perspective is of major interest for crystal microscopy,which aims at revealing the 3D crystalline structure of matter at the nanoscale,an approach strongly limited by the available coherent flux.Our results,based on Bragg ptychography experiments performed at the frst 4-generation synchrotron source,demonstrate the possibility of retrieving a high-quality image of the crystalline sample,with unprecedented quality.Importantly,the larger available coherent flux produces datasets with enough information to overcome experimental limitations,such as strongly deteriorated scanning conditions.We show this achievement would not be posible with 30-generation sources,a limit that has inhibited the development of this otherwise powerful microscopy method,so far.Hence,the advent of next-generation synchrotron sources not only makes Bragg ptychography suitable for high throughput studies but also strongly relaxes the associated experimental constraints,making it compatible with a wider range of experimental set-ups at the new synchrotrons.展开更多
Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used t...Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays： （i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP） （n = 17）; （iii） externalization of phosphatidylserine （EPS） （n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.展开更多
Soil types, humus types and vegetation as well as their hypsometric variation were analysed in terms of sequences in the northern part of the high mountains of the Pirin National Park at altitudes between looo and 240...Soil types, humus types and vegetation as well as their hypsometric variation were analysed in terms of sequences in the northern part of the high mountains of the Pirin National Park at altitudes between looo and 2400 m a.s.1. The study area is characterised by a large variety of natural parameters like petrology （mainly marble and granite）, morphology （different slope deposits, exposition） and the orographic climate gradient. Statistical analyses using these parameters provided a basis for the soil group classification of the sites. Based on a Digital Terrain Model （DTM） and a geological map of the Pirin National Park, the results of these statistical analyses were used to generate a ＂map of potential soil groups＂ （regionalisation using GIS）. Six potential soil groups could be determined. The resulting map exhibits a confidence level of 68 % on 74.4 % of the covered area. Rendzic Leptosols, in combination with Folic Histosols and Histi-lithic Leptosols occur in the alpine and subalpine regions on calcareous substrates. With decreasing altitude they are replaced by a mosaic of Rendzic Leptosols, Phaeozems and an increasing occurrence of Cambic Umbrisols. Umbrisols found on silicatic substrates in the alpine region are replaced by Cambic Umbrisols with decreasing altitude as well. Hence, pedogenesis is characterised by increasing browning and depth of the soil profiles with decreasing altitude. The pH-level is slightly acidic to neutral in lower zones and on calcareous rocky bases. Acidification increases in the subalpine zone. Soil pH decreases down to 4 on silicate subtrates. Typical humic values in mineral topsoils are 10 to 12 %, and in organic layers of the soils above 2000 m a.s.1, they are even more. The C：N ratio closely ranges around 20 （median）.展开更多
L.G.and S.M.also thank Valentina Shumakova,Elizaveta Gangrskaia,Alessandra Bellissimo,Audrius Pugzlys,and Andrius Baltu?ka for fruitful discussions and computational support by the Vienna Scientific Cluster(VSC 4).The...L.G.and S.M.also thank Valentina Shumakova,Elizaveta Gangrskaia,Alessandra Bellissimo,Audrius Pugzlys,and Andrius Baltu?ka for fruitful discussions and computational support by the Vienna Scientific Cluster(VSC 4).The authors thankfully acknowledge the computer resources at MareNostrum and SCAYLE,and the technical support provided by Barcelona Supercomputing Center(RES-FI-2022-3-0041).R.M.-H.,L.S.-T.,L.P.,E.C.J.,and C.H.-G.acknowledge funding from the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program(Grant Agreement No.851201)and support from Ministerio de Ciencia e Innovación(PID2022-142340NB-I00,PID2019-106910GB-I00).This research was funded in whole or in part by the Austrian Science Fund(FWF)(10.55776/ZK91).For open access purposes,the author has applied a CC BY public copyright license to any author accepted manuscript version arising from this submission.展开更多
血管胰岛素抵抗是肥胖的一个特点,可导致血管异常与疾病的发生。然而,目前对肥胖相关的血管胰岛素抵抗及功能异常的潜在分子机制仍了解很少。该文假设TRAF3交互蛋白2(TRAF3 interacting protein 2,TRAF3IP2),一个已知在心血管病激活病...血管胰岛素抵抗是肥胖的一个特点,可导致血管异常与疾病的发生。然而,目前对肥胖相关的血管胰岛素抵抗及功能异常的潜在分子机制仍了解很少。该文假设TRAF3交互蛋白2(TRAF3 interacting protein 2,TRAF3IP2),一个已知在心血管病激活病理应激通路的促炎症接头分子,与肥胖相关的血管胰岛素抵抗与功能异常有因果联系。研究人员通过体外在内皮细胞、离体动脉中,体内在喂食诱发肥胖的TRAF3IP2敲除小鼠模型中进行基因调控.展开更多
In on-shell Zγ production, deviations from the Standard Model for the Zγ＂ and ZγZ＊ couplings may be described in terms of eight parameters, hV （i = 1, 4; V = 7, Z） [1]. The parameters hiγ describe the Zγγ＊ ...In on-shell Zγ production, deviations from the Standard Model for the Zγ＂ and ZγZ＊ couplings may be described in terms of eight parameters, hV （i = 1, 4; V = 7, Z） [1]. The parameters hiγ describe the Zγγ＊ couplings and the parameters hz the Ziγ,ZγZ＊couplings.展开更多
文摘In order to evaluate the impacts of volatile organic compounds(VOCs)emissions from building materials on the indoor air quality beyond the standard chamber test conditions and test period,mechanistic emission source models have been developed in the past.However,very limited data are available for the required model parameters including the initial concentration(C_(m0)),in-material diffusion coefficient(D_(m)),partition coefficient(Kma),and convective mass transfer coefficient(k_(m)).In this study,a procedure was developed for estimating the model parameters by using VOC emission data from standard small chamber tests.In the procedure,initial values of the model parameters were refined by multivariate regression analysis of the measured emission data.To verify the procedure and estimate its uncertainty,simulated chamber test data were generated by adding 10% experimental uncertainties on the theoretical curve from the analytical solution to a mechanistic emission model.Then the procedure was applied to the generated data to estimate the model parameters.Results indicated that estimates converged to the original parameter values used for the data generation and the error of estimated parameters D_(m1)C_(m0) and K_(ma) were within±10%,±23%,and±25%of the true values,respectively.The procedure was further demonstrated by applying it to estimate the model parameters from real chamber test data.Wide application of the procedure would result in a database of mechanistic source model parameters for assessing the impact of VOC emissions on indoor pollution load,which are essential input data for evaluating the effectiveness of various indoor air quality(IAQ)design and control strategies as well as the energy required for meeting given IAQ requirements.
基金funded by the Max-Planck-Gesellschaft Partner Group Grant(K.T.)the National Natural Science Foundation of China(31371267,31322030,and 91331108,K.T.+10 种基金91631307,S.W.30890034,31271338,L.J.and 31525014,91731303,31771388,31961130380,and 32041008,S.X.)supported by the National Basic Research Program(2015FY111700,L.J.)Shanghai Municipal Science and Technology Major Project(2017SHZDZX01,L.J.,S.X.,and S.W.)the Ministry of Education(311016,L.J.)Strategic Priority Research Program of the Chinese Academy of Sciences(CAS)(XDB13040000,S.X.and S.W.)the UK Royal Society-Newton Advanced Fellowship(NAFn R1n191094)Key Research Program of Frontier Sciences(QYZDJ-SSW-SYS009)of the Chinese Academy of Sciencesthe support of a National Thousand Young Talents Award and a Max Planck-CAS Paul Gerson Unna Independent Research Group Leadership Award(S.W.)the Science and Technology Commission of Shanghai Municipality(16JC1400504,S.W.)。
文摘The human face is a heritable surface with many complex sensory organs.In recent years,many genetic loci associated with facial features have been reported in different populations,yet there is a lack of studies on the Han Chinese population.Here,we report a genome-wide association study of 3D normal human faces of 2,659 Han Chinese with autosegment phenotypes of facial morphology.We identify singlenucleotide polymorphisms(SNPs)encompassing four genomic regions showing significant associations with different facial regions,including SNPs in DENND1 B associated with the chin,SNPs among PISRT1 associated with eyes,SNPs between DCHS2 and SFRP2 associated with the nose,and SNPs in VPS13 B associated with the nose.We replicate 24 SNPs from previously reported genetic loci in different populations,whose candidate genes are DCHS2,SUPT3 H,HOXD1,SOX9,PAX3,and EDAR.These results provide a more comprehensive understanding of the genetic basis of variation in human facial morphology.
基金This study is financially supported by the Programme of Introducing Talents of Discipline to Universities,project No.B13011.
文摘Uncertainty exists in many aspects of building simulation.A deterministic hygrothermal analysis may not sufficiently give a reliable guidance if a number of input variables are subject to uncertainty.In this paper,a probabilistic-based method was developed to evaluate the hygrothermal performance of building components.The approach accounts for the uncertainties from model inputs and propagates them to the outputs through the simulation model,thus it provides a likelihood of performance risk.Latin hypercube sampling technique,incorporated with correlation structure among the inputs,was applied to generate the random samples that follows the intrinsic relations.The performance of an internally insulated masonry wall was evaluated by applying the proposed approach against different criteria.Thermal performance,condensation and mould growth potential of the renovated wall can overall satisfy the requirements stipulated in multifold standards.The most influential inputs were identified by the standardized regression sensitivity analysis and partial correlation technique.Both methods deliver the same key parameters for the single and time-dependent output variables in the case study.The probabilistic method can provide a comprehensive risk analysis and support the decision-maker and engineer in the design and optimization of building components.
文摘There is a fundamental body of evidence suggesting that activated apoptosis signaling in ejaculated human sperm negatively influences their fertilization potential. However, it is still controversial whether this apoptotic signaling is a relic of an abortive apoptosis related to spermatogenesis or if it should be regarded as a functional preformed pathway in mature sperm leading to stereotypical morphological changes reflecting nuclear disassembly. To address this question, apoptosis was induced using betulinic acid in mature and immature ejaculated human sperm enriched by density gradient centrifugation. Execution of apoptosis was monitored by observing ultra-morphological changes via transmission electron microscopy. Typical morphological signs of apoptosis in somatic cells include plasma membrane blebbing with the formation of apoptotic bodies, impaired mitochondrial integrity, defects of the nuclear envelope, and nuclear fragmentation; these morphologies have also been observed in human sperm. In addition, these apoptotic characteristics were more frequent in immature sperm compared to mature sperm. Following betulinic acid treatment, apoptosis-related morphological changes were induced in mature sperm from healthy donors. This effect was much less pronounced in immature sperm. Moreover, in both fractions, the betulinic acid treatment increased the percentage of acrosome-reacted sperm. The results of our ultra-morphological study prove the functional competence of apoptosis in mature ejaculated human sperm. The theory of a sole abortive process may be valid only for immature sperm. The induction of the acrosome reaction by stimulating apoptosis might shed light on the biological relevance of sperm apoptosis.
文摘The Mesta-Nestos river basin in Bulgaria and Greece is a case study for transboundary decision-making support in south-eastern Europe and a show-case for the development of methodologies and information-gathering for the integrated regional planning of water resources. Land-use conflicts in this water-scarce region cover a wide spectrum of activities like agricultural irrigation, drinking water production, diversions for industrial water, and risk of pollution from mining, to name a few examples. Measurements of the water quality were carried out in the upper basin. Results will be illustrated by the example of the environmental situation in the alpine region of the Pirin National Park as well as in the Razlog Basin with a stronger anthropogenic impact and pollution around a former uranium mine near the village of Elesnica. The social and economic development of this transboundary region is a recently established priority for mean an increase in water usage the water resources if regional the future. It will and more stress for impacts of global climate change are verified. Problem-focused management of the catchment area as a whole on the basis of proved geo-data sets is needed for the future.
文摘Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMR,Δψ_m) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.
基金supported by Deutsche Forschungsgemeinschaft(DFG)grants(No.GU 447/14-1,14-2 to DG,PI 837/4-1,4-2 to JP,and(No.HA 6908/2-1 to JH,respectively)by Else Kroner-Fresenius-Stiftung grant(No.2016_A52 to JH)+3 种基金JH receives additional funding from the DFG(No.6908/3-1)CB is an employee of Limbach and holds a part-time faculty appointment at the University of FreiburgHis research laboratory receives support from the DFG(No.BE 3910/8-1 and BE 3910/9-1)from the Federal Ministry of Education and Research(BMBF,No.01GM1903I and 01GM1903G).
文摘Formation of claudin-10 based tight junctions(TJs)is paramount to paracellular Na+transport in multiple epithelia.Sequence variants in CLDN10 have been linked to HELIX syndrome,a salt-losing tubulopathy with altered handling of divalent cations accompanied by dysfunctional salivary,sweat,and lacrimal glands.Here,we investigate molecular basis and phenotypic consequences of a newly identified homozygous CLDN10 variant that translates into a single amino acid substitution within the fourth transmembrane helix of claudin-10.In addition to hypohidrosis(H),electrolyte(E)imbalance with impaired urine concentrating ability,and hypolacrimia(L),phenotypic findings include altered salivary electrolyte composition and amelogenesis imperfecta but neither ichthyosis(I)nor xerostomia(X).Employing cellular TJ reconstitution assays,we demonstrate perturbation of cis-and trans-interactions between mutant claudin-10 proteins.Ultrastructures of reconstituted TJ strands show disturbed continuity and reduced abundance in the mutant case.Throughout,both major isoforms,claudin-10a and claudin-10b,are differentially affected with claudin-10b showing more severe molecular alterations.However,expression of the mutant in renal epithelial cells with endogenous TJs results in wild-type-like ion selectivity and conductivity,indicating that aberrant claudin-10 is generally capable of forming functional paracellular channels.Thus,mutant proteins prove pathogenic by compromising claudin-10 TJ strand assembly.Additional ex vivo investigations indicate their insertion into TJs to occur in a tissue-specific manner.
基金funded by the research project "Current impacts of global changes on evolution of karst (based on the integrated monitoring of model karst geosystems in Bulgaria)"Science Research Fund (Grant No. DN14/10/20.12.2017)+1 种基金funded by the South-west University of Blagoevgrad (grants RP-A 10/15, RP-A 13/17)SRF (SRF 02/70)
文摘This research reveals relationships between climate variables and inter-annual dynamics in the area of the glacieret located in the cirque Golemiya Kazan in the Pirin Mountains. The study period is 1993–2017. The correlations are identified using statistical methods. Also, a statistical model is constructed, including some climate variables as predictors. Despite the evident decrease of the glacieret's size in the period from the 1950 s onwards, the long-term trends for the last decades have been insignificant. The main climatic factors influencing the inter-annual dynamics in the area of the glacieret are air temperature, precipitation, zonal and meridional winds and relative humidity. With respect to the dynamics in the area of the glacieret, the important trends in the different climate variables are those of the warm period air temperatures and zonal(u) wind. They also determine to a great extent its future development by acting in two opposite directions – rising temperatures in the warm period will lead to a rapid decrease of its area by the end of the melting season, while the change of wind directionfrom west to east in the warm period will increase its area. The influence of the zonal wind in the warm period is explained mainly by the location of the glacieret in the cirque. Generally, the glacieret is tilted downwards from west to east. Thus, westerly winds facilitate blowing away the snow from the surface of the glacieret, assisting its melting in the warm period. Easterly winds do not have such an effect. The combination of the opposite effects of these two most important climate variables leads to the most likely scenario for the future development of the glacieret, according to which by the middle of this century it is expected to turn into a semi-permanent snow patch, which disappears after some summers, and by the end of the century to completely melt every year before the end of the melting season.
基金the European Research Council(European Union’s Horizon H2020 research and innovation program grant agreements No 724881Research conducted at MAX IV is supported by the Swedish Research Council under Contract 2018-07152the Swedish Governmental Agency for Innovation Systems under Contract 2018-04969,and Formas under Contraa 2019-02496.
文摘New 4^(th)-generation synchrotron sources,with their increased bilince,promise to greatly improve the performances of coherent X-ray microscopy.This perspective is of major interest for crystal microscopy,which aims at revealing the 3D crystalline structure of matter at the nanoscale,an approach strongly limited by the available coherent flux.Our results,based on Bragg ptychography experiments performed at the frst 4-generation synchrotron source,demonstrate the possibility of retrieving a high-quality image of the crystalline sample,with unprecedented quality.Importantly,the larger available coherent flux produces datasets with enough information to overcome experimental limitations,such as strongly deteriorated scanning conditions.We show this achievement would not be posible with 30-generation sources,a limit that has inhibited the development of this otherwise powerful microscopy method,so far.Hence,the advent of next-generation synchrotron sources not only makes Bragg ptychography suitable for high throughput studies but also strongly relaxes the associated experimental constraints,making it compatible with a wider range of experimental set-ups at the new synchrotrons.
文摘Aim： To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application. Methods： Semen samples from 87 unselected infertile patients were used to perform the following assays： （i） detection of active caspase-3 （n = 17）; （ii） integrity of the mitochondrial membrane potential （MMP） （n = 17）; （iii） externalization of phosphatidylserine （EPS） （n = 16）; and （iv） detection of intact acrosomes via CD46 （n = 37）. After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14. Results： Differences of up to ± 5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA^TM showed mean differences 〈 5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences 〉 5% at day 3. The CD46-FITC labeling displayed absolute differences 〈 5% CD46-positive spermatozoa at days 3, 7, 10 and 14. Conclusion： Although immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.
文摘Soil types, humus types and vegetation as well as their hypsometric variation were analysed in terms of sequences in the northern part of the high mountains of the Pirin National Park at altitudes between looo and 2400 m a.s.1. The study area is characterised by a large variety of natural parameters like petrology （mainly marble and granite）, morphology （different slope deposits, exposition） and the orographic climate gradient. Statistical analyses using these parameters provided a basis for the soil group classification of the sites. Based on a Digital Terrain Model （DTM） and a geological map of the Pirin National Park, the results of these statistical analyses were used to generate a ＂map of potential soil groups＂ （regionalisation using GIS）. Six potential soil groups could be determined. The resulting map exhibits a confidence level of 68 % on 74.4 % of the covered area. Rendzic Leptosols, in combination with Folic Histosols and Histi-lithic Leptosols occur in the alpine and subalpine regions on calcareous substrates. With decreasing altitude they are replaced by a mosaic of Rendzic Leptosols, Phaeozems and an increasing occurrence of Cambic Umbrisols. Umbrisols found on silicatic substrates in the alpine region are replaced by Cambic Umbrisols with decreasing altitude as well. Hence, pedogenesis is characterised by increasing browning and depth of the soil profiles with decreasing altitude. The pH-level is slightly acidic to neutral in lower zones and on calcareous rocky bases. Acidification increases in the subalpine zone. Soil pH decreases down to 4 on silicate subtrates. Typical humic values in mineral topsoils are 10 to 12 %, and in organic layers of the soils above 2000 m a.s.1, they are even more. The C：N ratio closely ranges around 20 （median）.
基金European Research Council(851201)Ministerio de Ciencia e Innovación(PID2022-142340NB-I00,PID2019-106910GB-I00)Austrian Science Fund(10.55776/ZK91)。
文摘L.G.and S.M.also thank Valentina Shumakova,Elizaveta Gangrskaia,Alessandra Bellissimo,Audrius Pugzlys,and Andrius Baltu?ka for fruitful discussions and computational support by the Vienna Scientific Cluster(VSC 4).The authors thankfully acknowledge the computer resources at MareNostrum and SCAYLE,and the technical support provided by Barcelona Supercomputing Center(RES-FI-2022-3-0041).R.M.-H.,L.S.-T.,L.P.,E.C.J.,and C.H.-G.acknowledge funding from the European Research Council(ERC)under the European Union’s Horizon 2020 research and innovation program(Grant Agreement No.851201)and support from Ministerio de Ciencia e Innovación(PID2022-142340NB-I00,PID2019-106910GB-I00).This research was funded in whole or in part by the Austrian Science Fund(FWF)(10.55776/ZK91).For open access purposes,the author has applied a CC BY public copyright license to any author accepted manuscript version arising from this submission.
文摘血管胰岛素抵抗是肥胖的一个特点,可导致血管异常与疾病的发生。然而,目前对肥胖相关的血管胰岛素抵抗及功能异常的潜在分子机制仍了解很少。该文假设TRAF3交互蛋白2(TRAF3 interacting protein 2,TRAF3IP2),一个已知在心血管病激活病理应激通路的促炎症接头分子,与肥胖相关的血管胰岛素抵抗与功能异常有因果联系。研究人员通过体外在内皮细胞、离体动脉中,体内在喂食诱发肥胖的TRAF3IP2敲除小鼠模型中进行基因调控.
文摘In on-shell Zγ production, deviations from the Standard Model for the Zγ＂ and ZγZ＊ couplings may be described in terms of eight parameters, hV （i = 1, 4; V = 7, Z） [1]. The parameters hiγ describe the Zγγ＊ couplings and the parameters hz the Ziγ,ZγZ＊couplings.
