Imperative role of electron microscopy in toxicity assessment: A review

Electron microscope (EM) was developed in 1931 and since then microscopical examination of both the biological and non-biological samples has been revolutionized. Modifications in electron microscopy techniques, such as scanning EM and transmission EM, have widened their applicability in the various sectors such as understanding of drug toxicity, development of mechanism, criminal site investigation, and characterization of the nano-molecule. The present review summarizes its role in important aspects such as toxicity assessment and disease diagnosis in special reference to SARS-COV2. In the biological system, EM studies have elucidated the impact of toxicants at the ultra-structural level in various tissue in conformity to physiological alterations. Thus, EM can be concluded as an important tool in toxicity assessment and disease prognosis.     

短波紫外线照射和~(60)Co-γ辐照处理对大樱桃贮藏品质的影响

为探究不同射线辐照处理对大樱桃果实保鲜效果的影响,采用不同剂量短波紫外线(short-wave ultraviolet light,UV-C)照射(1.37、2.05、2.74 kJ/m~2)和~(60)Co-γ辐照(0.75、1.5、2.25 kGy)分别处理大樱桃果实,定期检测其冷藏期间生理品质指标和果皮超微结构变化。结果表明:贮藏15 d内,1.37 kJ/m~2照射和0.75 kGy辐照较对照组可显著延缓大樱桃果实好果率下降、抑制果实质量损失,较好保持果实质地特性,促进花青素合成,有效抑制过氧化物酶与超氧化物歧化酶活性下降,但对可溶性固形物含量影响不大,且会引起丙二醛含量增加。由于两种处理对好果率、硬度、质量损失率、花青素等感官品质核心指标具有良好的积极作用,因此对丙二醛的不利变化并不足以影响到果实的商品品质评价。电子显微镜结果显示,大樱桃经1.37 kJ/m~2照射和0.75 kGy辐照处理后,与对照组相比,能在一定程度上减轻果皮细胞膜、线粒体、叶绿体等细胞结构的损伤程度,显著延缓果皮细胞衰老。此外,在贮藏前期与末期,1.37 kJ/m~2照射和0.75 kGy辐照所发挥的作用并不相同。针对于5 d以内的短期贮藏,可采用1.37 kJ/m~2 UV-C照射,而长期贮藏则以0.75 kGy~(60)Co-γ辐照效果更好。     

Immunolocalization of corneous proteins including a serine‐tyrosine‐rich beta‐protein in the adhesive pads in the tokay gecko

Adhesive pads of geckos contain many thousands of nanoscale spatulae for the adhesion and movement along vertical or inverted surfaces. Setae are composed of interlaced corneous bundles made of small cysteine‐glycine‐rich corneous beta proteins (CBPs, formerly indicated as beta‐keratins), embedded in a matrix material composed of cytoskeletal proteins and lipids. Negatively charged intermediate filament keratins (IFKs) and positively charged CBPs likely interact within setae, aside disulphide bonds, giving rise to a flexible and resistant corneous material. Using differernt antibodies against CBPs and IFKs an updated model of the composition of setae and spatulae is presented. Immunofluorescence and ultrastructural immunogold labeling reveal that one type of neutral serine‐tyrosine‐rich CBP is weakly localized in the setae while it is absent from the spatula. This uncharged protein is mainly present in the thin Oberhautchen layer sustaining the setae, although with a much lower intensity with respect to the cysteine‐rich CBPs. These proteins in the spatula likely originate a positively charged or neutral contact surface with the substrate but the influence of lipids and cytoskeletal proteins present in setae on the mechanism of adhesion is not known. In the spatula, protein‐lipid complexes may impart the pliability for the attachment and adapt to irregular surfaces. The presence of cysteine‐glycine medium rich CBPs and softer IFKs in alpha‐layers sustaining the setae forms a flexible base for compliance of the setae to substrate and improved adhesion.     

Effect of cooking temperature on texture and flavour binding of braised sauce porcine skin

To investigate the effect of cooking temperature (55, 65, 75, 85 and 95 °C) on texture and flavour binding of braised sauce porcine skin (BSPS), sensory acceptance, microstructure and flavour-binding capacity were investigated during the processing of BSPS. Samples cooked at 85 and 95 °C showed better texture and aroma scores. Hardness and chewiness of BSPS were obviously improved at 85 and 95 °C than control group. Collagen structure was significantly destroyed over 85 °C. The porcine skin collagen heated at 85 and 95 °C showed relatively higher flavour-binding capacity than other samples. The improvement of texture of BSPS was mainly attributed to the degradation of collagen. Higher aroma scores of BSPS were related to intense binding abilities with aroma compounds at 85 and 95 °C. Cooking at 85 or 95 °C could be an optimal cooking temperature for BSPS.     

Ontogeny of the testicular excurrent duct system of male Japanese quail (Coturnix japonica): A histological,ultrastructural, and histometric study

The testicular excurrent duct system undergoes several physiological and morphological changes during the reproductive stage or breeding season in mammals, birds, and reptiles. Studies on normal age-related histomorphological changes in the excurrent duct system of Japanese quails (Coturnix japonica) remain unreported, despite the extensive use of this bird as an avian model in research studies. The current study investigated the histological, ultrastructural, and histometric changes in the testicular excurrent duct system of the Japanese quail during three reproductive stages, namely prepubertal, pubertal, and adult. Simple squamous to low cuboidal cells formed the epithelia of the rete testis in prepubertal and pubertal birds, while in adult birds the lining was low cuboidal to cuboidal. In pubertal and adult birds, the nonciliated Type I epithelial cells of the proximal efferent duct displayed a subapical endocytotic apparatus comprising coated pits, coated apical tubules, and endosomes. There was a significant increase (p ≤ .001) in epithelial heights of all ducts of the excurrent duct system in the mature, sexually active, adult birds when compared to the other age groups. The luminal and tubular diameters, and the cross-sectional areas of efferent ducts and the epididymal duct unit increased significantly (p ≤ .001) with age. It is concluded that the morphology and morphometry of the excurrent ducts of the testis of the Japanese quail change as birds mature.     

Long-Term Spinal Ventral Root Reimplantation,but not Bone Marrow Mononuclear Cell Treatment,Positively Influences Ultrastructural Synapse Recovery and Motor Axonal Regrowth

We recently proposed a new surgical approach to treat ventral root avulsion, resulting in motoneuron protection. The present work combined such a surgical approach with bone marrow mononuclear cells (MC) therapy. Therefore, MC were added to the site of reimplantation. Female Lewis rats (seven weeks old) were subjected to unilateral ventral root avulsion (VRA) at L4, L5 and L6 levels and divided into the following groups (n = 5 for each group): Avulsion, sealant reimplanted roots and sealant reimplanted roots plus MC. After four weeks and 12 weeks post-surgery, the lumbar intumescences were processed by transmission electron microscopy, to analyze synaptic inputs to the repaired α motoneurons. Also, the ipsi and contralateral sciatic nerves were processed for axon counting and morphometry. The ultrastructural results indicated a significant preservation of inhibitory pre-synaptic boutons in the groups repaired with sealant alone and associated with MC therapy. Moreover, the average number of axons was higher in treated groups when compared to avulsion only. Complementary to the fiber counting, the morphometric analysis of axonal diameter and “g” ratio demonstrated that root reimplantation improved the motor component recovery. In conclusion, the data herein demonstrate that root reimplantation at the lesion site may be considered a therapeutic approach, following proximal lesions in the interface of central nervous system (CNS) and peripheral nervous system (PNS), and that MC therapy does not further improve the regenerative recovery, up to 12 weeks post lesion.     

采前氯吡苯脲处理对采后‘秦美’猕猴桃细胞超微结构的影响

研究‘秦美’猕猴桃盛花期后28?d用0、10、20?mg/L氯吡苯脲（N-(2-chloro-4-pyridyl)-N’-phenylurea，CPPU）蘸果处理对采后冷藏期猕猴桃果实细胞超微结构的影响。结果表明：CPPU处理加速了猕猴桃果实细胞壁及内部结构的降解，且CPPU质量浓度越大，受损程度越大；10?mg/L?CPPU处理加速了猕猴桃果实淀粉颗粒及胞间质的降解，促使细胞壁弯曲变形及细胞间隙出现，造成猕猴桃果实硬度迅速下降；而20?mg/L?CPPU处理使猕猴桃果实细胞壁严重变形，线粒体严重空泡化，内部结构消失，淀粉颗粒完全降解，细胞间的黏合力丧失。据此认为，CPPU处理加快了猕猴桃果实在贮藏过程中细胞壁、线粒体及淀粉颗粒的降解速度，损坏了细胞器及膜系统的完整性，从而使猕猴桃果实硬度及耐藏性下降，贮藏寿命缩短，品质下降。因此，猕猴桃生产中不建议使用CPPU处理。     

The Effect of µ/m‐Calpain on Protein Degradation of Chicken Breast Meat

This study was designed to determine the effects of µ/m‐calpain on the degradation of cytoskeletal proteins in pectoralis major. Four chickens were slaughtered and the breasts were removed and stored for 12 hr at 4 °C. Each sample was divided into three groups and respectively immersed in control reagent, calpain inhibitor, and caspase inhibitor at 4 °C. The samples were used to evaluate troponin‐T and desmin degradation, calpain activity, and myofibril ultrastructure at 12 hr, day 1, day 3, and day 7. Casein zymography revealed that µ‐calpain could not be detected in all samples after 12 hr postmortem. The calpain inhibitor inhibited µ/m‐calpain activity and reduced troponin‐T and desmin degradation during 7 day postmortem. The caspase inhibitor inhibited µ/m‐calpain activity and, troponin‐T and desmin degradation before day 3 postmortem. The findings suggest that, µ/m‐calpain had an effect on cytoskeletal protein degradation after 12 hr postmortem.     

Bioinspired design of hybrid composite materials

ABSTRACT

Mimicking the natural design motifs of structural biological materials is a promising approach to achieve a unique combination of strength and toughness for engineering materials. In this study, we proposed a 2D computational model, which is a two-hierarchy hybrid composite inspired by the ultrastructural features of bone. The model is composed of alternating parallel array of two subunits (A & B) mimicking ‘mineralized collagen fibril’ and ‘extrafibrillar matrix’ of bone at ultrastructural level. The subunit-A is formed by short stiff platelets embedded within a soft matrix. The subunit-B consists of randomly distributed stiff grains bonded by a thin layer of tough adhesive phase. To assess the performance of the bioinspired design, a conventional unidirectional long-fiber composite made with the same amount of hard and soft phases was studied. The finite element simulation results indicated that the toughness, strength and elastic modulus of the bioinspired composite was 312%, 83%, and 55% of that of the conventional composite, respectively. The toughness improvement was attributed to the prevalent energy-dissipating damage of adhesive phase in subunit-B and crack-bridging by subunit-A, the two major toughening mechanisms in the model. This study exemplifies some insights into natural design of materials to gain better material performance.