Engineering high amylose and resistant starch in maize by CRISPR/Cas9-mediated editing of starch branching enzymes

To improve the amylose content(AC)and resistant starch content(RSC)of maize kernel starch,we employed the CRISPR/Cas9 system to create mutants of starch branching enzyme I(SBEI)and starch branching enzyme IIb(SBEIIb).A frameshift mutation in SBEI(E1,a nucleotide insertion in exon 6)led to plants with higher RSC(1.07%),lower hundred-kernel weight(HKW,24.71±0.14 g),and lower plant height(PH,218.50±9.42 cm)compared to the wild type(WT).Like the WT,E1 kernel starch had irregular,polygonal shapes with sharp edges.A frameshift mutation in SBEIIb(E2,a four-nucleotide deletion in exon 8)led to higher AC(53.48%)and higher RSC(26.93%)than that for the WT.E2 kernel starch was significantly different from the WT regarding granule morphology,chain length distribution pattern,X-ray diffraction pattern,and thermal characteristics;the starch granules were more irregular in shape and comprised typical B-type crystals.Mutating SBEI and SBEIIb(E12)had a synergistic effect on RSC,HKW,PH,starch properties,and starch biosynthesis-associated gene expression.SBEIIa,SS1,SSIIa,SSIIIa,and SSIIIb were upregulated in E12 endosperm compared to WT endosperm.This study lays the foundation for rapidly improving the starch properties of elite maize lines.

Role of renin-angiotensin system/angiotensin converting enzyme-2 mechanism and enhanced COVID-19 susceptibility in type 2 diabetes mellitus

Coronavirus disease 2019(COVID-19)is a disease that caused a global pandemic and is caused by infection of severe acute respiratory syndrome coronavirus 2 virus.It has affected over 768 million people worldwide,resulting in approx-imately 6900000 deaths.High-risk groups,identified by the Centers for Disease Control and Prevention,include individuals with conditions like type 2 diabetes mellitus(T2DM),obesity,chronic lung disease,serious heart conditions,and chronic kidney disease.Research indicates that those with T2DM face a hei-ghtened susceptibility to COVID-19 and increased mortality compared to non-diabetic individuals.Examining the renin-angiotensin system(RAS),a vital regulator of blood pressure and pulmonary stability,reveals the significance of the angiotensin-converting enzyme(ACE)and ACE2 enzymes.ACE converts angiotensin-I to the vasoconstrictor angiotensin-II,while ACE2 counters this by converting angiotensin-II to angiotensin 1-7,a vasodilator.Reduced ACE2 exp-ression,common in diabetes,intensifies RAS activity,contributing to conditions like inflammation and fibrosis.Although ACE inhibitors and angiotensin receptor blockers can be therapeutically beneficial by increasing ACE2 levels,concerns arise regarding the potential elevation of ACE2 receptors on cell membranes,potentially facilitating COVID-19 entry.This review explored the role of the RAS/ACE2 mechanism in amplifying severe acute respiratory syndrome cor-onavirus 2 infection and associated complications in T2DM.Potential treatment strategies,including recombinant human ACE2 therapy,broad-spectrum antiviral drugs,and epigenetic signature detection,are discussed as promising avenues in the battle against this pandemic.

Bacterial communities and enzyme activities during litter decomposition of Elymus nutans leaf on the Qinghai-Tibet Plateau

The dominant plant litter plays a crucial role in carbon(C)and nutrients cycling as well as ecosystem functions maintenance on the Qinghai-Tibet Plateau(QTP).The impact of litter decomposition of dominant plants on edaphic parameters and grassland productivity has been extensively studied,while its decomposition processes and relevant mechanisms in this area remain poorly understood.We conducted a three-year litter decomposition experiment in the Gansu Gannan Grassland Ecosystem National Observation and Research Station,an alpine meadow ecosystem on the QTP,to investigate changes in litter enzyme activities and bacterial and fungal communities,and clarify how these critical factors regulated the decomposition of dominant plant Elymus nutans(E.nutans)litter.The results showed that cellulose and hemicellulose,which accounted for 95%of the initial lignocellulose content,were the main components in E.nutans litter decomposition.The litter enzyme activities ofβ-1,4-glucosidase(BG),β-1,4-xylosidase(BX),andβ-D-cellobiosidase(CBH)decreased with decomposition while acid phosphatase,leucine aminopeptidase,and phenol oxidase increased with decomposition.We found that both litter bacterial and fungal communities changed significantly with decomposition.Furthermore,bacterial communities shifted from copiotrophic-dominated to oligotrophic-dominated in the late stage of litter decomposition.Partial least squares path model revealed that the decomposition of E.nutans litter was mainly driven by bacterial communities and their secreted enzymes.Bacteroidota and Proteobacteria were important producers of enzymes BG,BX,and CBH,and their relative abundances were tightly positively related to the content of cellulose and hemicellulose,indicating that Bacteroidota and Proteobacteria are the main bacterial taxa of the decomposition of E.nutans litter.In conclusion,this study demonstrates that bacterial communities are the main driving forces behind the decomposition of E.nutans litter,highlighting the vital roles of bacterial communities in affecting the ecosystem functions of the QTP by regulating dominant plant litter decomposition.

Microplastic Can Decrease Enzyme Activities and Microbes in Soil

An in vitro study was conducted to investigate the impacts of microplastics on enzyme activities and soil bacteria. The study included four different treatments of microplastics including a control. Different levels of microplastics were applied to the soil ranging from 0% to 5%, to assess the impacts of microplastics on soil enzymes and subsequent soil bacteria. After 30 days of incubation, the soil samples were collected and growth parameters of bacteria were assessed. Activities of β-glucosidase, urease and dehydrogenase enzymes were also determined. Our results showed that the presence of microplastics in the soil significantly reduced bacterial population together with bacterial strains. The activities of β-glucosidase, urease and dehydrogenase enzymes were reduced significantly to approximately 32%, 40% and 50% in microplastics treated soils respectively. Concentration of microplastic has a role to play towards this direction;the higher the concentration of microplastic the greater is the impact on enzymes and soil bacteria. The present study on the microbial soil health vis-à-vis microplastic application indicates that the material can have negative effect on the soil bacterial population of and thus ultimately may jeopardize soil health and crop production.

Cloning and Expression of the Promoter of Maize Starch-Branching Enzyme sbeⅡb Gene

[Objective] The aim was to construct promoter of maize starch-branching enzyme sbe Ⅱb gene specifically expressed in seed.[Method] The promoter sequence of maize starch-branching enzyme sbe Ⅱb gene was amplified by LA-PCR and then cloned into pMD18-T vector.Subsequently,the promoter was cloned into pBI121 vector to construct plant expression vector pBI121-sbe Ⅱb.Recombinant plasmid pSBE-GUS was constructed by connecting GUS gene into pBI121-sbe Ⅱb and transformed into tobacco mediated by Agrobacterium tumefaciens,before detection by PCR and Southern blot.The biochemical analysis and fluorescence detection of GUS activity were performed in different parts of the tobacco by Gene Gun Method and Agrobacterium tumefaciens transformation respectively.[Result] The homology between promoter sequence cloned in this experiment and the corresponding sequence announced in the GenBank reached 98.52%.Four transformed tobaccos were obtained by PCR and Southern blot after co-culture and selective culture.A small number of blue spots appeared in leaves,and the spots could barely been seen in the stems and roots,while a large number of blue spots were found in seeds.So,it could be concluded that the promoter of sbeⅡb gene was specifically expressed in seeds.[Conclusion] Promoter of maize starch branching enzyme sbe Ⅱb specifically expressed in seeds was successfully cloned.

Investigation of the Interaction Mechanism between Lignin Structural Units and Enzyme

The effect of lignin structural units on enzymatic hydrolysis of lignocellulosic biomass was investigated,especially the inhibitory role of lignin in non-productive adsorption with enzymes.Milled wood lignin(MWL)was isolated from different hardwoods of poplar,eucalyptus and acacia.The isolated lignin samples were characterized by elemental analysis,gel permeation chromatography,nitrobenzene oxidation and fourier infrared spectroscopy.The mechanism of lignin structural units on enzymatic hydrolysis of cellulose was studied by quartz crystal microbalance(QCM).The results showed that different structural units of lignin had different adsorption capacity for enzymes.The results of nitrobenzene oxidation indicated that the S/G ratio(S:syringyl-like lignin structures;G:guaiacyl-like lignin structures)of lignin of poplar was 0.99,that of eucalyptus was 1.92 and that of acacia was 1.34.According to the results of QCM,the adsorption capacity of the three lignin films was as follows:Poplar MWL(S/G ratio 0.99)<Acacia MWL(S/G ratio 1.34)<Eucalyptus MWL(S/G ratio 1.92).Eucalyptus MWL with higher degree of condensation and S/G ratio showed stronger affinity to enzymes and more non-productive adsorption with enzymes,resulting in less adsorption between enzymes and cellulose,and lower enzymatic hydrolysis efficiency.

Metabolic enzyme inhibitory abilities, in vivo hypoglycemic ability of palmleaf raspberry fruits extracts and identification of hypoglycemic compounds

The unripe palmleaf raspberry,namely Fupenzi(FPZ),is an important medicinal and edible food.This study aims to evaluate the potential of FPZ extracts prepared with different approaches in attenuating hyperglycemia,gout,Alzheimer’s disease,and pigmentation,to obtain the enriching fraction and to identify the major active compounds.Results indicated that FPZ extracts showed weak activity against acetylcholinesterase,considerable ability against tyrosinase and xanthine oxidase,but excellent inhibition onα-glucosidase.Ultrasound-assisted 40%ethanol extract(40EUS)gave the highest phenolics content,and the bestα-glucosidase inhibition(IC_(50)=0.08μg/mL),which is 877-fold higher than that of positive control acarbose.The 40%ethanol eluting fraction of 40EUS showed the strongestα-glucosidase inhibition with the IC_(50) value of 37.79 ng/mL,it could also effectively attenuate the fasting blood glucose level and oral glucose tolerance of C57BL/6 mice.Twenty-six compounds were identified from 40%ethanol fraction by using HPLC-QTOF-MS/MS,hydrolysable tannins(including 11 ellagitannins and 4 gallotannins)were the major compounds,phenolic acids came to the second.Above results could provide important technical supporting for the further application and research of FPZ in health foods and drugs against diabetes.

基于Benders分解和分枝定界的随机交期批量流流水车间调度

针对交期随机的批量流车间调度问题,以最小化工件延期期望之和为目标,推导出工件交期符合3类经典随机分布条件下问题目标的闭式计算表达式.建立考虑换模时间与随机交期的问题数学模型,针对模型高度非线性特征对其线性化.设计一种基于逻辑的Benders分解(LBBD)与分枝定界相结合的优化算法,提出两种有效加速策略提升算法求解效率.数值实验结果验证了算法的有效性,通过随机交期与确定交期结果的比较,验证了考虑随机的必要性.

Angiotensin-converting enzyme 2 alleviates liver fibrosis through the renin-angiotensin system

The present letter to the editor is related to the study titled‘Angiotensin-converting enzyme 2 improves liver fibrosis in mice by regulating autophagy of hepatic stellate cells’.Angiotensin-converting enzyme 2 can alleviate liver fibrosis by regulating autophagy of hepatic stellate cells and affecting the renin-angiotensin system.

Toxicity of Tagetes erecta Essential Oil Based Combinatorial Formulations on Various Metabolic Enzymes in Indian White Termite Odontotermes obesus

Plant essential oils and their constituents have proven to be very effective against insects, especially termites. They are the best alternative to synthetic pesticides that are harmless to the environment and human health. In the present study, different enzymes, namely alkaline phosphatase, acid phosphatase, glutamate-oxaloacetate transaminase, glutamate-pyruvate transaminase and acetylcholinesterase, were evaluated in Indian termite Odontotermes obesus in a combination preparation based on Tagetes erecta essential oil. For the study of anti-termite effects against worker termites were treated topically with 40% and 80% of the 24-hour LD50 values of various combination formulations. Subsequent bioassays at 40% and 80% of LD50 of combinatorial preparations S-AST-A, S-AST-B, and S-AST-C resulted in a significant (p > 0.05) decrease in glutamate-oxaloacetate transaminase. After 16 hours of treatment, they were 87.92%, 80.17%, 89.30%, 79.17%, 81.92% and 73.17% respectively. In vivo exposures of 40% and 80% of the LD50 of the combination formulation caused a highly significant (p > 0.05) reduction (p > 0.05) of all test enzymes tested compared to controls. The effects of different oil formulations exhibited time- and dose-dependent responses, resulting in physiological changes in the concentrations of various enzymes. The combined mixture of Tagetes erecta essential oils has significantly better anti-termite ability compared to inorganic insecticides. Findings from this study will help support termite control in fields, gardens and homes in a sustainable way, without the downsides of insecticide resistance and pollution. These could potentially be used to produce commercial formulations for use against pests.

Toxic Effects of Citrus maxima Based Combinatorial Formulations on Important Metabolic Enzymes in Indian White Termite Odontotermes obesus

In the present study, Citrus maxima essential crude oil extract was used to prepare combinatorial formulations and workers of Indian white termite Odontotermes obesus were treated topically with 40% and 80% of 24 hr LD50 values of these formulations. In subsequent bioassays levels of various enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and acetylcholinesterase were evaluated to determine the anti-termite efficacy of Citrus maxima essential oil based combinatorial formulations. S-RET-A, S-RET-B and S-RET-C caused significant (p > 0.05) decrease in glutamate oxaloacetate transaminase i.e. 87.47%, 86.81% and 81.77% & 82.04%, 79.39% and 74.75% respectively at 16 h treatment. In vivo exposure of 40% and 80% of LD50 of combinatorial formulations caused very significant (p > 0.05) reduction in all the test enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase expect acetylcholinesterase levels after 16 h all tested treatments in comparison to control. Both dose-response and time period were found important in physiological alteration in levels of various enzymes. Combinatorial mixtures of Citrus essential oils have shown synergistic activity against termites. The research findings of the present study would help termite control in crop fields, gardens and houses in a sustainable way.

Molecular identification of the key starch branching enzyme-encoding gene SBE2.3 and its interacting transcription factors in banana fruits

Starch branching enzyme(SBE)has rarely been studied in common starchy banana fruits.For the first time,we report here the molecular characterization of seven SBE(MaSBE)and six SBE(MbSBE)genes in the banana A-and B-genomes,respectively,which could be classified into three distinct subfamilies according to genome-wide identification.Systematic transcriptomic analysis revealed that six MaSBEs and six MbSBEs were expressed in the developing banana fruits of two different genotypes,BaXi Jiao(BX,AAA)and Fen Jiao(FJ,AAB),among which MaSBE2.3 and MbSBE2.3 were highly expressed.Transient silencing of MaSBE2.3 expression in banana fruit discs led to a significant decrease in its transcription,which coincides with significant reductions in total starch and amylopectin contents compared to those of empty vector controls.The suggested functional role of MaSBE2.3 in banana fruit development was corroborated by its transient overexpression in banana fruit discs,which led to significant enhancements in total starch and amylopectin contents.A number of transcription factors,including three auxin response factors(ARF2/12/24)and two MYBs(MYB3/308),that interact with the MaSBE2.3 promoter were identified by yeast one-hybrid library assays.Among these ARFs and MYBs,MaARF2/MaMYB308 and MaARF12/MaARF24/MaMYB3 were demonstrated via a luciferase reporter system to upregulate and downregulate the expression of MaSBE2.3,respectively.

A Fiber Optic Sensor for the Simultaneous Measurement of Dual-parameter Based on Hydrogel-immobilized Enzyme Complex

A novel fiber optic sensor based on hydrogel-immobilized enzyme complex was developed for the simultaneous measurement of dual-parameter,the leap from a single parameter detecting fiber optic sensor to a fiber optic sensor that can continuously detect two kinds of parameters was achieved.By controlling the temperature from high to low,the function of fiber sulfide sensor and fiber DCP sensor can be realized,so as to realize the continuous detection of dual-parameter.The different variables affecting the sensor performance were evaluated and optimized.Under the optimal conditions,the response curves,linear detection ranges,detection limits and response times of the dual-parameter sensor for testing sulfide and DCP were obtained,respectively.The sensor displays high selectivity,good repeatability and stability,which have good potentials in analyzing sulfide and DCP concentration of practical water samples.

Hepatic angiotensin-converting enzyme 2 expression in metabolic dysfunction-associated steatotic liver disease and in patients with fatal COVID-19

BACKGROUND Metabolic dysfunction-associated steatotic liver disease(MASLD),characterised by hepatic lipid accumulation,causes inflammation and oxidative stress accompanied by cell damage and fibrosis.Liver injury(LI)is also frequently reported in patients hospitalised with coronavirus disease 2019(COVID-19),while preexisting MASLD increases the risk of LI and the development of COVID-19-associated cholangiopathy.Mechanisms of injury at the cellular level remain unclear,but it may be significant that severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)which causes COVID-19,uses angiotensin-converting expression enzyme 2(ACE2),a key regulator of the‘anti-inflammatory’arm of the renin-angiotensin system,for viral attachment and host cell invasion.AIM To determine if hepatic ACE2 levels are altered during progression of MASLD and in patients who died with severe COVID-19.METHODS ACE2 protein levels and localisation,and histological fibrosis and lipid droplet accumulation as markers of MASLD were determined in formalin-fixed liver tissue sections across the MASLD pathological spectrum(isolated hepatocellular steatosis,metabolic dysfunction-associated steatohepatitis(MASH)+/-fibrosis,end-stage cirrhosis)and in post-mortem tissues from patients who had died with severe COVID-19,using ACE2 immunohistochemistry and haematoxylin and eosin and picrosirius red staining of total collagen and lipid droplet areas,followed by quantification using machine learning-based image pixel classifiers.RESULTS ACE2 staining is primarily intracellular and concentrated in the cytoplasm of centrilobular hepatocytes and apical membranes of bile duct cholangiocytes.Strikingly,ACE2 protein levels are elevated in non-fibrotic MASH compared to healthy controls but not in the progression to MASH with fibrosis and in cirrhosis.ACE2 protein levels and histological fibrosis are not associated,but ACE2 and liver lipid droplet content are significantly correlated across the MASLD spectrum.Hepatic ACE2 levels are also increased in COVID-19 patients,especially those showing evidence of LI,but are not correlated with the presence of SARS-CoV-2 virus in the liver.However,there is a clear association between the hepatic lipid droplet content and the presence of the virus,suggesting a possible functional link.CONCLUSION Hepatic ACE2 levels were elevated in nonfibrotic MASH and COVID-19 patients with LI,while lipid accumulation may promote intra-hepatic SARS-CoV-2 replication,accelerating MASLD progression and COVID-19-mediated liver damage.

Timosaponin AⅢ induces drug-metabolizing enzymes by activating constitutive androstane receptor (CAR) via dephosphorylation of the EGFR signaling pathway

The current study aimed to assess the effect of timosaponin AⅢ(T-AⅢ)on drug-metabolizing enzymes during anticancer therapy.The in vivo experiments were conducted on nude and ICR mice.Following a 24-day administration of T-AⅢ,the nude mice exhibited an induction of CYP2B10,MDR1,and CYP3A11 expression in the liver tissues.In the ICR mice,the expression levels of CYP2B10 and MDR1 increased after a three-day T-AⅢ administration.The in vitro assessments with HepG2 cells revealed that T-AⅢ induced the expression of CYP2B6,MDR1,and CYP3A4,along with constitutive androstane receptor(CAR)activation.Treatment with CAR siRNA reversed the T-AⅢ-induced increases in CYP2B6 and CYP3A4 expression.Furthermore,other CAR target genes also showed a significant increase in the expression.The up-regulation of murine CAR was observed in the liver tissues of both nude and ICR mice.Subsequent findings demonstrated that T-AⅢ activated CAR by inhibiting ERK1/2 phosphorylation,with this effect being partially reversed by the ERK activator t-BHQ.Inhibition of the ERK1/2 signaling pathway was also observed in vivo.Additionally,T-AⅢ inhibited the phosphorylation of EGFR at Tyr1173 and Tyr845,and suppressed EGF-induced phosphorylation of EGFR,ERK,and CAR.In the nude mice,T-AⅢ also inhibited EGFR phosphorylation.These results collectively indicate that T-AⅢ is a novel CAR activator through inhibition of the EGFR pathway.

Detection of tracheal branching with computerized tomography:The relationship between the angles and age-gender

BACKGROUND The data obtained on the anatomical knowledge of the tracheobronchial system can be used for diagnosis,treatment and interventional interventions in areas such as anesthesia,thoracic surgery,pulmonary physiology.AIM To determine the tracheobronchial branching angles in pediatric and adult populations by using the multislice computed tomography(CT)and minimum intensity projection(MinIP)technique,which is a non-invasive method.METHODS Our study was carried out retrospectively.Patients who underwent contrast and non-contrast CT examination,whose anatomically and pathophysiologically good tracheobronchial system and lung parenchyma images were obtained,were included in the study.Measurements were made in the coronal plane of the lung parenchyma.In the coronal plane,right main bronchus-left main bronchus angle,right upper lobe bronchus-intermedius bronchus angle,right middle lobe bronchus-right lower lobe bronchus angle,left upper lobe bronchus-left lower lobe bronchus angle were measured.RESULTS The study population consisted of 1511 patients,753 pediatric(mean age:13.4±4.3;range:1-18 years)and 758 adults(mean age:54.3±17.3;range:19-94 years).In our study,tracheal bifurcation angle was found to be 73.3°±13.7°(59.6°-87°)in the whole population.In the pediatric group,the right-left main coronal level was found to be higher in boys compared to girls(74.6°±12.9°vs 71.2°±13.9°,P=0.001).In the adult group,the right-left main coronal level was found to be lower in males compared to females(71.9°±12.9°vs 75.8°±14.7°,P<0.001).CONCLUSIONS Our study,with the number of 1511 patients,is the first study in the literature with the largest number of patient populations including pediatric and adult demographic data,measuring the angle values of the tracheobronchial system using multislice CT and MinIP technique.Study data will not only be a guide during invasive procedures,but it can also guide studies to be done with imaging methods.

Investigation and Benefits of the Use of Enzymes for Unbleached Kraft Recycling Wet Strength Paper at Lab and Industrial Scale

The deterioration of recycled fibers especially unbleached kraft with high wet strength resin content due to the irreversible structural changes caused by drying and added chemicals makes the raw material difficult to repulp.The mechanical effect in the pulper over time with chemical treatment has a negative impact on the recycled fibers.At lab scale,different compositions of enzymatic treatment C022L were under investigation to select the most efficient laccase Lacc1,Lacc2 or Lacc3 and to observe the impact of lipases during repulping at low and high consistencies.Pulp disintegration at different times was evaluated to define the level of rejects and to analyze the morphology of fibers after treatments.These results were more significant for Lacc2,by increasing the pulp consistency to 15%in the pulper.Combining lipases with CELODASE 022L appeared to decrease the efficiency of enzymes.The results showed a high reduction of energy power with the enzymatic treatment and a significant reduction of fines level in fibers’suspension.The most efficient version of C022L was used at industrial scale to compare directly with the standard conditions used in a paper mill.

Processing Effects on Selected Antioxidant Activities and Metabolizing Enzyme Inhibition of M. Koneigii (Curry Leaves) Extracts

Curry leaves, scientifically termed Murraya koenigii, are renowned in South Asian cuisine for their flavor enhancement and potential health benefits, including antioxidative, anti-inflammatory, and antidiabetic properties. This study aimed to evaluate the impact of thermal processing methods on curry leaves by analysing Total Phenolic Content (TPC), Total Flavonoid Content (TFC), antioxidant activity, and metabolizing enzyme inhibition. Fresh curry leaves were subjected to thermal treatments: Oven-dried at 60˚C and Air-dried at 25˚C for 2 weeks. Extracts were prepared using Ethanol and water solvents. Results indicated that Air-dried leaves exhibited significantly higher TPC (5132.65 mg GAE/100 g) and TFC (243.13 mg CE/100 g) compared to Fresh and Oven-dried leaves. Antioxidant assays show that oven-dried curry leaves at 60˚C displayed higher results in NORS, FRAP, and TEAC assays compared to Fresh and Air-dried leaves. Ethanol extracts showed better extraction of bioactive compounds than aqueous extracts. Moreover, Lipase inhibition activity was notably high, indicating potential health benefits. This study provides valuable insights into the effects of processing methods on curry leaf extracts, emphasizing the importance of solvent selection for optimal extraction of bioactive compounds.

ON THE EMPTY BALLS OF A CRITICAL OR SUBCRITICAL BRANCHING RANDOM WALK

Let{Z_(n)}_(n)≥0 be a critical or subcritical d-dimensional branching random walk started from a Poisson random measure whose intensity measure is the Lebesugue measure on R^(d).Denote by R_(n):=sup{u>0:Z_(n)({x∈R^(d):∣x∣<u})=0}the radius of the largest empty ball centered at the origin of Z_(n).In this work,we prove that after suitable renormalization,Rn converges in law to some non-degenerate distribution as n→∞.Furthermore,our work shows that the renormalization scales depend on the offspring law and the dimension of the branching random walk.This completes the results of Révész[13]for the critical binary branching Wiener process.

Dietary manganese supplementation inhibits abdominal fat deposition possibly by regulating gene expression and enzyme activity involved in lipid metabolism in the abdominal fat of broilers

Excessive abdominal fat deposition seriously restricts the production efficiency of broilers.Several studies found that dietary supplemental manganese(Mn)could effectively reduce the abdominal fat deposition of broilers,but the underlying mechanisms remain unclear.The present study aimed to investigate the effect of dietary supplementation with the inorganic or organic Mn on abdominal fat deposition,and enzyme activity and gene expression involved in lipid metabolism in the abdominal fat of male or female broilers.A total of 4201-d-old AA broilers(half males and half females)were randomly allotted by body weight and gender to 1 of 6 treatments with 10 replicates cages of 7 chicks per cage in a completely randomized design involving a 3(dietary Mn addition)×2(gender)factorial arrangement.Male or female broilers were fed with the Mn-unsupplemented basal diets containing 17.52 mg Mn kg^(-1)(d 1-21)and 15.62 mg Mn kg^(-1)(d 22-42)by analysis or the basal diets supplemented with 110 mg Mn kg^(-1)(d 1-21)and 80 mg Mn kg^(-1)(d 22-42)as either the Mn sulfate or the Mn proteinate with moderate chelation strength(Mn-Prot M)for 42 d.The results showed that the interaction between dietary Mn addition and gender had no impact(P>0.05)on any of the measured parameters;abdominal fat percentage of broilers was decreased(P<0.003)by Mn addition;Mn addition increased(P<0.004)adipose triglyceride lipase(ATGL)activity,while Mn-Prot M decreased(P<0.002)the fatty acid synthase(FAS)activity in the abdominal fat of broilers compared to the control;Mn addition decreased(P<0.009)diacylglycerol acyltransferase 2(DGAT2)mRNA expression level and peroxisome proliferator-activated receptor γ(PPARγ)mRNA and protein expression levels,but up-regulated(P<0.05)the ATGL mRNA and protein expression levels in the abdominal fat of broilers.It was concluded that dietary supplementation with Mn inhibited the abdominal fat deposition of broilers possibly via decreasing the expression of PPARγand DGAT2 as well as increasing the expression and activity of ATGL in the abdominal fat of broilers,and Mn-Prot M was more effective in inhibiting the FAS acitivity.