过度煎炸油对大鼠氧化应激及相关基因表达的影响

采用定量灌胃的方式研究过度煎炸油(DFEO)的摄入对大鼠氧化应激的影响,同时探讨其引起氧化损伤可能的机理。研究发现:干预6周后,DFEO组血清GSH–Px、T–AOC水平显著降低(ρ0.05),MDA水平显著升高(ρ0.05);肝组织切片显示出现脂肪空泡,肝窦扩张,肝细胞界限不清;同时RT–PCR结果表明,过度煎炸油会导致大鼠肝脏组织HMOX1、VEGF基因显著下调(ρ0.01)。这说明DFEO富含大量毒性物质,会影响正常的基因表达,使机体清除氧自由基能力下降,甚至破坏正常的组织细胞形态。     

抗性淀粉对过度煎炸油引起的大鼠肝损伤保护作用的研究

采用定量灌胃的方式研究抗性淀粉的补充对过度煎炸油引起的大鼠肝损伤的保护作用。选取24只成年Wistar大鼠随机均分成3组,分别灌胃生理盐水（control）、过度煎炸油（DFEO）、另外一组同时灌胃过度煎炸油和抗性淀粉（DFEO+RS）。6周后腹部主动脉取血,用于各项生化指标的检测;取肝脏组织做组织切片H&E染色观察并计算肝脏指数。结果表明,与对照组相比,DFEO组血清中ALT、ALP水平显著升高（P<0.01）,GSH-Px、T AOC水平显著降低（P<0.05）;肝组织切片显示出现脂肪空泡,肝窦扩张,肝细胞界限不清;肝指数显著下降（P<0.05）;而DFEO+RS组各项指标与对照组相比均无显著差异。这说明过度煎炸油中的含有大量毒性物质,会对肝组织细胞造成严重的损伤,而抗性淀粉对其有明显的改善效果。     

基于斑马鱼幼鱼的塑化剂肝毒性危害识别研究

目的 采用塑化剂邻苯二甲酸二乙基己酯（DEHP）和邻苯二甲酸二丁酯（DBP）作为阳性对照,建立斑马鱼幼鱼脂肪肝毒性风险评估模型进行塑化剂邻苯二甲酸丁酯苯甲酯（BBP）肝毒性危害识别。方法 分析DEHP和DBP暴露后斑马鱼肝脏脂肪信号强度及过氧化物酶体增殖物激活受体（PPAR）靶基因CD36的表达,初步构建斑马鱼幼鱼肝毒性模型;基于此模型对BBP进行肝毒性评价,此外采用肝脏表达绿色荧光转基因斑马鱼幼鱼进行肝密度分析、甘油三酯（TG）定量检测以及成鱼肝脏病理检查进一步验证模型的可靠性,应用基准剂量法（BMD）推导关键效应点。结果 DEHP、DBP表现出极显著诱发脂肪肝风险（均P<0.01）,显著上调CD36相对表达量（P<0.001,P<0.05）,提示造模成功。BBP（0.000 012 5%、0.000 025%、0.000 05%）诱发斑马鱼幼鱼显著的脂质沉积（P<0.05,P<0.001,P<0.001）、显著影响密度（P<0.05,P<0.01,P<0.01）、显著增加TG蓄积（P<0.05,P<0.05,P<0.01）、上调CD36基因表达。此外,BBP（≥0.000 012 5%）诱发斑马鱼成鱼肝脏部位脂质空泡形成、空泡间隙和数量减少,提示BBP具有明显的肝毒性。关键效应为肝脏脂肪信号强度（S）,基准剂量下限（BMDL）值为0.013 mg/L。结论 本研究成功构建了一种通过肝脏脂肪信号强度来识别塑化剂肝毒性的斑马鱼幼鱼模型,结合基准剂量法进一步为塑化剂肝毒性危害识别提供科学依据。     

虾青素和/或有氧运动对D-半乳糖诱导大鼠心脏衰老的干预作用

目的 探究虾青素和/或有氧运动延缓D-半乳糖诱导大鼠心脏衰老的潜在作用机制。方法 SPF级3月龄SD雄性大鼠40只随机分为安静对照组（C组）、急性衰老模型组（D组）、急性衰老+虾青素组（DA组）、急性衰老+有氧运动组（DE组）及急性衰老+虾青素+有氧运动组（DAE组）,每组各8只。C组不进行任何干预,急性衰老各组大鼠腹腔注射100 mg/kg·d的D-半乳糖,同时分别以20 mg/kg·d 虾青素和/或运动强度为60%最大摄氧量的有氧运动进行干预,实验周期为6周。末次训练12 h后取心脏,光学显微镜观察心脏组织形态并检测相关生化指标。结果 心脏组织形态,C组正常;与C组比较,D组出现心肌细胞排列紊乱、心肌纤维断裂及炎性细胞浸润等现象;DA、DE、DAE心肌细胞排列整齐、心肌纤维断裂及炎性细胞浸润情况有所缓解,DAE组最为显著,心肌组织形态更接近于正常状态。与C组比较,D组心脏组织超氧化物歧化酶（SOD）、γ-谷氨酸半胱氨酸合酶（γ-GCS）活性及沉默信息调节因子1（SIRT1）、核因子E2相关因子2（Nrf2）、血红素氧合酶（HO-1）、B淋巴细胞瘤因子-2（Bcl-2）蛋白质表达水平、Bcl-2/Bcl-2相关X蛋白（Bax）比值显著降低（P<0.05或P<0.01）;细胞凋亡水平、Bax蛋白质表达、丙二醛（MDA）含量显著升高（P<0.05或P<0.01）。与D组比较,DA组心肌组织SIRT1、Nrf2、HO-1蛋白质表达均显著升高（P<0.05）,Bax蛋白质表达及MDA含量显著降低（P<0.05或P<0.01）;DE组心肌组织SOD活性和SIRT1、HO-1蛋白质表达显著升高（P<0.05或P<0.01）,Bax蛋白质表达及MDA含量显著降低（P<0.05或P<0.01）;DAE组心肌组织SOD、γ-GCS活性及SIRT1、Nrf2、HO-1、Bcl-2蛋白质表达和Bcl-2/Bax比值显著升高（P<0.05或P<0.01）,细胞凋亡水平、Bax蛋白质表达及MDA含量极显著降低（P<0.01）。与DA、DE组比较,DAE组心肌组织SIRT1、Bcl-2蛋白质表达及Bcl-2/Bax比值均显著升高（P<0.05或P<0.01）,细胞凋亡水平显著降低（P<0.05）;心肌组织细胞凋亡水平、SIRT1和Bcl-2蛋白质表达水平及Bcl-2/Bax比值具有协同效应。结论 虾青素和/或有氧运动干预均可通过上调SIRT1/Nrf2信号通路相关蛋白质表达,缓解和改善D-半乳糖诱导的氧化应激,降低心肌细胞凋亡水平,延缓大鼠心脏衰老。     

DHA-磷脂对肥胖小鼠脂质代谢的影响

研究了二十二碳六烯酸-磷脂（DHA-磷脂）对实验性肥胖小鼠脂质代谢的调节作用及其机制。采用Folch法和硅胶柱层析法从鸢乌贼（文字Sthenoteuthis oualaniensis）卵中分离制备得到富含DHA的磷脂。C57BL/6J雄性小鼠随机分为3组：高脂模型组,1%大豆磷脂组,1%DHA-磷脂组。连续饲喂7周后,测定小鼠血清总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-C）含量,肝脏TC、TG、磷脂（PL）含量以及肝脏脂肪合成酶（FAS）、葡萄糖-6-磷酸脱氢酶（G6PDH）、苹果酸酶（ME）、磷脂酸磷酸酶（PAP）、肉毒碱棕榈酸转移酶（CPT）、过氧化物酶体β-氧化活力。结果显示,DHA-磷脂能极显著抑制小鼠体重（P＜0.01）和肾周脂肪（P＜0.01）及精周脂肪（P＜0.01）蓄积,降低血清TC（P＜0.01）和肝脏TG（P＜0.01）水平,抑制肝脏FAS（P＜0.01）、ME（P＜0.05）、PAP（P＜0.05）的活力,提高CPT（P＜0.01）和过氧化物酶体β-氧化（P＜0.05）的活力,且效果优于大豆磷脂。DHA-磷脂具有良好的减肥作用,能够通过抑制肝脏脂质合成,促进脂质分解,有效调节肥胖小鼠的脂质代谢。     

安五脂素对大鼠肠缺血再灌注致肝损伤的保护作用

目的:研究安五脂素（Anwulignan）对大鼠肠缺血再灌注（intestinal ischemia/reperfusion,II/R）致远隔肝损伤的改善作用及其作用机制。方法:将SD大鼠随机均分为假手术（Sham）组、缺血再灌注（II/R）组、假手术+ Anwulignan（Sham+Anwu）组和II/R+Anwulignan（II/R+Anwu）组。假手术及II/R结束后,腹主动脉取血,取肝脏。光镜观察肝组织病理学改变,检测血清谷丙转氨酶（ALT）及谷草转氨酶（AST）活性、肝活性氧（ROS）、蛋白激酶C（PKC）含量、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽-S-转移酶（GST）活性,以及丙二醛（MDA）和8-羟基脱氧鸟苷（8-OHdG）含量。结果:光镜下Sham组大鼠肝组织结构正常,II/R组出现组织病理学改变,与II/R组比较,II/R+Anwu组损伤明显减轻。与Sham组相比,II/R组大鼠血清ALT及AST的活性,肝ROS、MDA、8-OHdG及PKC含量极显著升高（P<0.01）,肝SOD、GST活性极显著降低（P<0.01）,CAT活性显著降低（P<0.05）;而Sham+Anwu组大鼠的这些指标均无显著性变化（P>0.05）。与II/R组比较,II/R+Anwu组大鼠血清AST活性,肝MDA、8-OHdG含量极显著降低（P<0.01）,ALT活性、肝ROS以及PKC水平显著降低（P<0.05）,肝SOD、CAT及GST活性极显著升高（P<0.01）。结论:安五脂素能够提高肝组织抗氧化能力,改善II/R大鼠致肝损伤。     

多酚复合配方缓解小鼠运动疲劳

目的：探究蓝靛果多酚、山楂多酚微胶囊、乳铁蛋白和维生素C复配剂对缓解疲劳的作用。方法：以肌酸、β-羟基-β-甲基丁酸钙、1,6-二磷酸果糖、牛磺酸和维生素B₆、西瓜粉复配比为阳性对照进行小鼠力竭游泳试验，测定疲劳相关指标，并采用H&E染色观察脏器微观病理组织情况。结果：各组小鼠体重无显著差异（P>0.05），较正常组，膳食补充组脏器指数无显著差异（P>0.05）。多酚复合配方在常温和低温下均显著延长了小鼠游泳时长（分别延长了106.01%和140.94%）（P<0.01），改善了氧化应激、代谢产物积累和能量代谢，下调了炎症因子分泌（P<0.01或P<0.05），改善了小鼠肌肉中细胞核增多、肝脏中炎症细胞浸润和肾脏中肾小管上皮细胞红肿和脱落等状况。结论：多酚复合配方在低温和常温下均具有缓解运动疲劳的效果。     

簕菜提取物对酒精性肝损伤大鼠的保护作用

目的:探讨簕菜不同提取物的解酒护肝作用,为簕菜解酒护肝功能食品的研发提供依据。方法:建立白酒灌胃大鼠致酒精肝损伤模型,考察簕菜醇提物和水提物对大鼠酒精耐受、睡眠和醒酒时间的影响,检测大鼠血液中乙醇质量浓度、血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）活性,以及大鼠肝脏组织中乙醇脱氢酶（ADH）、乙醛脱氢酶（ALDH）、超氧化物歧化酶（SOD）活性及谷胱甘肽（GSH）、丙二醛（MDA）含量,切片镜检肝细胞形态变化。结果:与酒精模型组比较,除水提物低剂量组对醉酒时间无显著变化（P>0.05）外,簕菜提取物各剂量组均可显著延长酒精耐受时间,并缩短睡眠、醒酒时间（P<0.05,P<0.01）;簕菜提取物均可极显著降低血清中乙醇含量（P<0.01）;除水提低剂量组AST活力无显著变化外（P>0.05）,簕菜提取物各剂量组均能显著抑制酒精引起的大鼠血清ALT和AST活性升高（P<0.05）;极显著增加肝脏组织中ADH、ALDH、SOD活力和GSH含量（P<0.01）,降低MDA含量（P<0.01）;肝细胞镜检结果显示,簕菜提取物均能在一定程度上减轻酒精导致的肝细胞变性程度。结论:簕菜醇提物、水提物均具有较好的防醉酒和解酒护肝作用,比较而言,醇提物效果优于水提物。     

西洋参枳椇子配伍对急性酒精性肝损伤的保护作用

目的:研究西洋参枳椇子组合物对酒精所致小鼠急性肝损伤的预防保护作用。方法:ICR小鼠适应饲养1周后,随机分为正常组、模型组、西洋参枳椇子组合物（Ⅰ,Ⅱ,Ⅲ）组。对小鼠连续灌胃西洋参枳椇子组合物（100 mg/kg?bw）14 d后,除正常组外,其余各组均在24 h内灌胃3次酒精（5 g/kg）诱导肝损伤。末次给予酒精6 h后,处死小鼠,检测各组小鼠血清中AST、ALT活力及TG含量,检测肝脏中GSH-Px、SOD活力及GSH和MDA含量,通过H & E染色观察肝脏病理组织变化,利用蛋白印记方法检测肝组织中p-ERK、p-JNK和p-P38蛋白的表达。结果:与模型组小鼠比较,西洋参枳椇子组合物能够明显降低由急性酒精摄入引起的血清中AST、ALT活力升高（P<0.05）,降低血清中TG水平（P<0.05）和肝组织中MDA含量（P<0.01）,提高肝组织中抗氧化酶SOD和GSH-Px活力（P<0.01）,且升高肝组织中GSH水平（P<0.01）。病理切片观察表明,模型组以出现脂肪滴为变性为主,部分出现炎性细胞浸润,各给药组对小鼠肝脏病理变化有不同程度改善作用,西洋参枳椇子组合物极显著抑制p-ERK、p-JNK和p-P38表达（P<0.01）,抑制肝细胞凋亡。结论:西洋参枳椇子组合物对急性酒精诱导的小鼠肝损伤具有较好的预防保护作用。     

羟基红花黄色素A对脂多糖诱导抑郁模型大鼠的影响

目的：通过脂多糖（LPS）诱导建立大鼠抑郁模型,研究羟基红花黄色素A的抗抑郁作用。方法：腹腔注射LPS建立大鼠抑郁模型,设置正常对照组、模型组、阳性对照组和羟基红花黄色素A不同剂量给药组,评价大鼠抑郁样行为,检测大鼠血清中TNF-α、IL-1β、IL-6、BDNF和5-HT含量,观察海马组织病理学变化,检测TLR/NLR信号通路蛋白表达情况。结果：与抑郁模型组相比,羟基红花黄色素A各剂量组大鼠水平运动、垂直运动及糖水偏好率增加,悬尾和游泳不动时间减少（P＜0.01）;血清BDNF、5-HT水平升高,TNF-α、IL-1β及IL-6水平下降（P＜0.01）;海马组织TLR2、NLRP1、NLRP3、ASC、TLR4和Caspase-1蛋白表达水平均降低（P＜0.01）。结论：羟基红花黄色素A可改善LPS诱导的大鼠抑郁样行为,其机制可能与抑制TLR/NLR信号通路有关。     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.     

Contribution of European research to risk analysis

The European Commission's, Quality of Life Research Programme, Key Action 1—Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches; harmonization of risk assessment principles, methodologies, and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries; to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women; evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here.     

低温带皮菜籽粕微粉的不同粒级部分的功能特性

为研究低温带皮菜籽粕微粉的不同粒级部分的功能特性,以经低温脱脂的带皮菜籽粕为原料,经微粉碎后筛分成212~425μm、150~212μm和106~150μm的3个不同粒级的微粉样品,检测这些样品的吸水性、吸油性、乳化性和乳化稳定性、蛋白质体外消化率。结果表明:1 3个不同粒级的微粉样品之间的粗纤维含量存在显著差异,表明三者的结构组成成分有一定差异。23个微粉样品的乳化活性和乳化稳定性随粒度级别的减小而显著增加(P0.01)。33个微粉样品的蛋白质体外消化率随粒度级别的减小而显著增加(P0.01)。4不同粒级带皮菜籽粕微粉样品的吸水性与吸油性受其结构组成物质不同和粒度的双重影响,与粒度的相关性不明显。     

Microbiology of food taints

Fresh and processed foods are often spoilt by the presence of undesirable flavours and odours caused by microbial action. The aim of this paper is to review the current knowledge of microbiologically induced taints that occur in a wide range of foodstuffs, including meats, poultry, fish, crustaceans, milk, dairy products, fruits, vegetables, cereals and cereal products. Examples have been chosen where the compounds responsible for the taint have been identified and sufficient data obtained to demonstrate the involvement of microorganisms. However, in some cases the full identity of the causative organism may not have been elucidated. The types of microorganisms covered by this review include bacteria, fungi, yeasts, actinomycetes and cyanobacteria. Although cyanobacteria do not in general infect foods, their presence in aqueous systems and water supplies can lead to off-flavours in aquatic organisms and processed foodstuffs. Several examples of each of these processes are discussed. Wherever possible, the likely biosynthetic pathway used by the microorganism to produce the offending compound in a foodstuff is indicated.     

Analysis for organic residues from aids to polymerization used to make plastics intended for food contact

Polymers intended for food contact use have been analysed for organic residues which could be attributed to a range of substances employed as polymerization aids (e.g. initiators and catalysts). A wide range of polymers was extracted with solvents and the extracts analysed by gas chromatography-mass spectrometry (GC-MS). The overwhelming majority of substances identified were not derived from aids to polymerization but were oligomers, additives and adventitious contaminants. However, a small number of substances were identified as initiator residues. These included tetramethylsuccinonitrile (TMSN) which was observed in two polymers and it derived from recombination of two azobisisobutyronitrile (AIBN) initiator radicals. Methyl benzoate, benzoic acid, biphenyl and phenyl benzoate were detected in one poly(methyl methacrylate) sample and in two polyvinylchlorides and they are thought to be derived from benzoyl peroxide initiator. TMSN was subsequently targeted for analysis of poly-(methyl methacrylate) plastics using proton nuclear magnetic resonance spectrometry (¹     

Tests of potential functional barriers for laminated multilayer food packages. Part II: Medium molecular weight permeants

Experiments were performed to characterize the kinetics of the permeation of different medium molecular weight model permeants: bisphenol A, warfarin and anthracene, from liquid paraffin, through a surrogate potential functional barrier (25 microns-thick orientated polypropylene--OPP) into the food simulants olive oil and 3% (w/v) acetic acid. The characterization of permeation kinetics generally observed the permeation models previously reported to explain the experimental permeation results obtained for a low molecular weight group of model permeants. In general, the model permeants exhibited behaviour consistent with their relative molecular weights with respect to (a) the time taken to attain steady-state permeation into the food simulant in which they were more soluble, (b) their subsequent steady-state permeation rates, and (c) their partition between liquid paraffin and the OPP membrane.     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. I: Interlaboratory studies of methods of analysis

This paper describes the first part of a project undertaken to develop mussel reference materials for Paralytic Shellfish Poisoning (PSP) toxins. Two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX) in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the second part of the project: the certification exercise. In the first study, 18 laboratories were asked to measure STX and dc-STX in rehydrated lyophilized mussel material and to identify as many other PSP toxins as possible with a method of their choice. In the second interlaboratory study, 15 laboratories were additionally asked to determine quantitatively STX and dc-STX in rehydrated lyophilized mussel and in a saxitoxin-enriched mussel material. The first study revealed that three out of four postcolumn derivatization methods and one pre-column derivatization method sufficed in principle to determine STX and dc-STX. Most participants (13 of 18) obtained acceptable calibration curves and recoveries. Saxitoxin was hardly detected in the rehydrated lyophilized mussels and results obtained for dc-STX yielded a CV of 58% at a mass fraction of 1.86 mg/kg. Most participants (14 out of 18) identified gonyautoxin-5 (GTX-5) in a hydrolysed extract provided. The first study led to provisional criteria for linearity, recovery and separation. The second study revealed that 6 out of 15 laboratories were able to meet these criteria. Results obtained for dc-STX yielded a CV of 19% at a mass fraction of 3.49mg/kg. Results obtained for STX in the saxitoxin-enriched material yielded a CV of 19% at a mass fraction of 0.34mg/kg. Saxitoxin could not be detected in the PSP-positive material. Hydrolysis was useful to confirm the identity of GTX5 and provided indicative information about C1 and C2 toxins in the PSP-positive material. The methods used in the second interlaboratory study showed sufficiently consistent analysis results to undertake a certification exercise to assign certified values for STX and dc-STX in lyophilized mussel.     

Focus on China's Paper Industry

In the English section of this issue, 〈China Paper Newsletters〉 will introduce ＂National Development and Reform Commission Issued Announcement for Selection of Major Preliminary Research Projects for the ＇13th Five-Year Plan＇＂, ＂2013 Annual Report of China＇s Paper Industry＂, and news of projects and other policies.     

Listen to downstream & search for innovation

正Nowadays,textile enterprises are all taking efforts in transformation and upgrading,like improving producing capacity and optimizing production structure to face market downturn.It claimed a higher request to the standard of textile equipments.In the upcoming of ITMA ASIA+CITME 2014exhibition,this magazine have interviewed several branch associations and a series of relative enterprises,to summarize industrial developing status