Identification and location of albumin-like antigens in third-stage larva of W. bancrofti, in adult forms of Litomosoides chagasfilhoi and in the free-living nematode Caenorhabditis elegans

Antigens resembling those of host proteins have been identified on the surface of several filarial parasites, such as immunoglobulins and serum albumins. The origin of albumin-like antigens on filarial parasites remains unclear. Several authors suggested that they have been adsorbed, or that they were metabolic waste products from nutritional utilization of human albumin, or perhaps a contamination with human products. This study searched for human albumin-like antigens by Western blot and ultrastructural analyses on filarial parasites, third stage of W. bancrofti and adult females of Litomosoides chagasfilhoi, and on the free-living Caenorhabditis elegans nematode. Our results showed approximately 67kDa proteins recognized by anti-human albumin antibodies on extracts and excretory-secretory (ES) products of the third-stage W. bancrofti. Similar albumin-like proteins were also detected on the filarial parasite L. chagasfilhoi and on C. elegans extracts. The immunocytochemistry analysis showed human albumin-like antigens on similar tissues of these nematodes. These results provide evidence that these proteins have antigenic similarity and similar distribution in nematodes tissues. Our observations suggest that albumin-like antigens presented on filarial parasites are not acquired from the host, but rather are shared antigenic determinants found even in the third-stage larvae recovered from the invertebrate host.     

The ultrastructure of infective larvae (L3) of Wuchereria bancrofti after treatment with diethylcarbamazine

Although the large use of diethylcarbamazine (DEC), as the major anti-filaricide drug, its mechanism of action remains a matter of controversy. Several authors defend the hypothesis that DEC has no direct effect on nematodes. This study demonstrated that infective larvae (L3) of Wuchereria bancrofti treated in vitro with DEC presented several behaviour and morphological changes. The first alteration produced by treatment for 2 h with 3, 5, 10 microg/ml of DEC was the reduction of motility. Larvae treated with 5, 10 microg/ml DEC showed severely affected organelles, formation of several vacuoles, mainly in neurocytes and in the muscle cells, and dissolution of cytoplasm. Some larvae showed extreme cellular disorganization with abundance of large and dense mitochondria and numerous large vacuoles containing residual organelles. Lamellar bodies, probably related to an assembly of hipodermal membranes, were also observed in some damaged larvae. Thus, undoubtedly in vitro treatment with concentrations of DEC similar to therapeutic conditions, which are 1-5 microg/ml (Hawking, 1979), had a direct effect on infective larvae of W. bancrofti by causing, primarily neuromuscular alterations with subsequent damage to organelles.     

Diethylcarbamazine induces loss of microfilarial sheath of Wuchereria bancrofti

Light microscopy analyses of microfilariae of Wuchereria bancrofti treated with DEC revealed a striking loss of the microfilarial sheath. However, no effect was observed on microfilariae of Litomosoides chagasfilhoi treated with DEC. For quantitative analyses microfilariae of W. bancrofti were processed for SEM. Controls, which have not received DEC, had 29.8% of exsheathed microfilariae. Conversely, the number of exsheathed microfilariae increased as increased DEC concentrations: 5 μg/ml (75.9%), 10 μg/ml (80.1%), and 50 μg/ml (87.7%). After DEC treatment some of sheathed microfilariae showed a wrinkled surface, and in some microfilariae, sheaths were observed being liberated almost intact from the larvae surface. But, frequently residues of the lost sheath over the surface were also observed. No damage was observed in the microfilariae cuticle. The present work shows quantitative data on the loss of the microfilarial sheath of W. bancrofti after treatment with DEC. Since no loss of microfilarial sheath was observed in microfilariae of L. chagasfilhoi submitted to the same conditions, DEC may present different mechanisms of action for distinct filarial species.     

Ultrastructural characterization of the hemocytes of Culex quinquefasciatus (DIPTERA: Culicidae)

Six hemocytes cell types from Culex quinquefasciatus were identified by light and transmission electron microscopy: They are prohemocytes (9.3%), spherulocytes (1.6%), adipohemocytes (0.8%), oenocytoids (4.6%), plasmatocytes (43.4%) and granulocytes (40.3%). The prohemocytes were the smallest hemocytes encountered in the hemolymph, displaying a large and centrally located nucleus, almost filling the whole cell. The spherulocytes, which were small hemocytes, presented small and numerous spherules with a lamellar pattern and an electron-dense core. Rare adipohemocytes were observed in the C. quinquefasciatus hemolymph, presenting large nucleus with an evident nucleolus, cytoplasm containing rough endoplasmic reticulum (RER), mitochondriae and lipid inclusions. C. quinquefasciatus oenocytoids showed homogeneous cytoplasm with several granules, completely or partially filled with amorphous material. These cells showed abundant smooth endoplasmic reticulum (SER) and dense mitochondriae. By light microscopy analysis we identified two morphological types of plasmatocytes, granular and agranular. However, ultrastructural investigation revealed that the granular cells contained lipid inclusion between RER membranes, instead of membrane-bounded granules. The granulocytes presented a fusiform or circular profile and displayed a unique and very complex process of granules formation, including organization of polysomes inside vesicles that protrude from the Golgi system, synthesis of a proteinaceous material, condensation of the granule matrix and recycling of endoplasmic membranes. Intense endocytic pathways were also observed in the granulocytes.     

Cellular localisation of calcium ions during potato hypersensitive response to Potato virus Y

Ca(2+) is one of the most universal and versatile signalling molecules and is involved in almost every aspect of cellular processes. Accumulating evidence suggests that Ca(2+) serves as a messenger in many growth and developmental processes and in plant responses to biotic and abiotic stresses. Numerous signals have been shown to induce transient elevation of cytoplasmic [Ca(2+)](cyt) in plants. The calcium free ions were detected cytochemically in Solanum tuberosum cv. Rywal tissues as a hypersensitive response (HR) from 10h to 5 days after a Potato virus Y (PVY) infection. Calcium was detected in vivo by its reaction with Alizarin S Red, producing an intense red staining in contact with calcium free ions. Calcium was found in the necrotic area of the epidermal and mesophyll cells 3 days after the PVY infection (when morphological symptoms on potato leaves appeared). Calcium ions were detected cytochemically in HR also by its reaction with potassium pyroantimonate. Inoculation with PVY(NTN) and also PVY(N) Wi induced a rapid hypersensitive response during which highly localised increased accumulation of electron-dense deposits of calcium pyroantimonate were detected. Calcium deposition was observed in necrotic and non-necrotic areas, starting from 10h after PVY infection. The deposits were present along ER cisternae, chloroplasts and mitochondria envelopes connected with PVY particles. The precipitates of calcium antimonate were detected near the nuclear envelope, inside karyolymph and along tracheary elements, especially when virus particles were present inside.     

Ultrastructural analysis of microfilariae of Wuchereria bancrofti obtained from persistent carriers after repeated courses of diethylcarbamazine

Single dose of diethylcarbamazine (DEC) used in control programs is effective in breaking the transmission of filariasis. In order to investigate the effect of aggressive therapy on Wuchereria bancrofti (Wb) microfilariae, DEC was given to 29 patients who were positive for the circulating filarial antigen (CFA) assay but did not have clinical manifestations of filariasis, at 6 mg/kg/day for 12 days and again six months later using the same dosing regimen. For each patient, microfilarial density and serum CFA were followed up for two years. Ultrastructural analyses on Wb microfilariae obtained after repeated treatment with DEC were also performed. Microfilaremia and antigenemia decreased significantly after 12 months but returned to the initial levels after 24 months. This could indicate, as shown by other authors, that aggressive repeated therapy with DEC alone is ineffective in eradicating adult W. bancrofti, particularly in infected but asymptomatic individuals. The objective of the present study was to analyze the microfilaremic and antigenemic behavior and ultrastructural changes caused by different DEC concentrations in vitro in Wb microfilariae obtained from individuals who were sensitive and refractory to treatment. After in vitro treatment of the microfilariae using 5 and 10 μg/ml of DEC for 1 h, ultrastructural analysis revealed low levels of cell damage compared with embryos obtained from individuals from a different area who had never received DEC treatment before. The results obtained suggest that microfilariae from patients who receive repeated aggressive therapy are less sensitive to DEC in vitro.     

Schlieren diagnostics of pulsed electron beam ablation of polymethyl-methacrylate

The investigation of the interaction of pulsed electron beams with PMMA (polymethylmethacrylate) targets is reported. The electron beam of some 10^–8 s in duration is produced in a pulsed low-pressure gas discharge. The beam power density of up to 10⁸ W/cm² leads to a surface plasma formation similar to that of the pulsed laser ablation process. The propagation of the ablated material and the shock wave inside the PMMA target are observed by means of Schlieren diagnostics. An electron density gradient of over 3×10¹⁹ cm^–4 has been observed in the expanding plasma up to 1.5 s after the plasma formation. During the early stage of expansion, the expansion velocity of the plasma plume as determined by the steep electron density gradient is around 10⁵ cm/s. The pressure behind the shock front inside the PMMA target as determined from the shock velocity exceeds 0.3 Gpa.     

Ultrastructural localization of Trypanosoma cruzi lysosomes by aryl sulphatase cytochemistry

Lysosomes of trypanosomatid protozoa are poorly known. In this work we have cytochemically detected the lysosomal enzyme aryl sulphatase in the trypanosomatids Trypanosoma cruzi and Crithidia fasciculata, by using p-nitrocatecholsulphate as substrate. Positive reaction was located exclusively inside membrane-bound cytoplasmic vesicles distributed throughout the cell body. Electron-dense reaction was either dispersed homogeneously through the vesicular matrix or located at the vesicle periphery, apposed to the membrane, with fine granular deposits occasionally found at the vesicular matrix. Trypomastigote and epimastigote forms of T. cruzi lacked electron-dense deposits at the plasma membrane, thus indicating that aryl sulphatase was not secreted to the environment. Furthermore, no positive reaction was detected in epimastigote reservosomes, which are organelles considered as pre-lysosomal compartments. Thus, our data show that reservosomes and lysosomes are organelles that can be distinguished by the cytochemical localization of aryl sulphatase in T. cruzi epimastigotes and trypomastigotes. Positive reaction in cytoplasmic vesicles of C. fasciculata choanomastigotes confirmed the specificity of the reaction for lysosomes in other trypanosomatid species.     

Ultrastructural analysis of Wuchereria bancrofti (Nematoda: Filarioidea) body wall

Bancroftian filariasis constitutes the principal mosquito-borne nematode infection of humans and the surface of adult of Wuchereria bancrofti seems to be especially important in the intricate interplay between host and parasite. The study of the parasite's surface structure might help to understand the localization and function of various organelles. W. bancrofti adult worms were recovered from untreated patients during hydrocele repair surgery and studied by transmission electron microscopy. The body wall of adult parasite is composed of cuticle, hypodermis and muscular layer. Cuticle is the external layer and shows transverse cuticular striation. It is composed by an epicuticle, cortical layers, median layer, fibrous layers and basal layer. The epicuticle is the most external cuticular layer and appears as a single laminar electron-dense layer. The cortical external region is more electron-dense and granular in appearance than the inner cortical layer. Electron-dense structures, called bosses are randomly distributed filling the cuticular striation. The median layer is formed by an electron-dense and continuous thick line. The fibrous layer is subdivided in inner and external layers connected by projections. The basal layer includes a large quantity of membranous projections directed toward the hypodermis. The hypodermis is a syncytium where some cellular organelles are observed. The somatic musculature is meromyarian. The muscle fibers consist of contractile and non-contractile regions and the contractile region is composed of myofilaments separated by dense body. This is the first study of W. bancrofti adult worms obtained from untreated patients and studied by transmission electron microscopy.     

Study of pyrolytic carbon films for non-emitting grid applications

Pyrolytic carbon films were prepared on a hot substrate inside UHV system back-filled with 6 × 10⁻⁵ Torr of CH₄. The resulting films showed two different chemical states of carbon; carbides formed on W and Ta substrates and graphitic carbon layers were yound on ?. Ir substrate. Alter BaO deposition, the carbides showed a work function too low to be useful as grids. On the other hand, the work function of pyrolytic carbon films were high both before and after Ba or BaO coverage. Study of the degradation mechanisms of carbon films, formed by e-bean techniques, however, showed that they also reacted rapidly with W and T₂ resulting in a carbide formation. Thus it suggested that in the application of pyrolytic graphite grids, contact with carbide-forming metals should be avoided. Interaction of the pyrolytic carbon with BaO also resulted in the removal of carbon. However, this “leeching” phenomenon projects a life for pyrolytic graphite grids much longer than actually observed in the tube tests. The diffusion of Ba and BaO through pyrolytic carbon layer was evident in this workm but its precise relation to life failure rate is not yet understood.     

Structure and ultrastructure of spermatozoa of Chrysomya megacephala (Diptera: Calliphoridae)

The spermatozoa of Chrysomya megacephala are similar to those described for other Brachycera. In this species, the spermatozoa are long and thin, measuring about 590 μm in length, of which the head region measures approximately 60 μm. The head includes a monolayered acrosome with electron-lucid material, and the shape of the nucleus, in cross-sections, varies from circular to oval with completely condensed chromatin. The centriole was observed in the zone of flagellar implantation, below the “peg” region. In the region of overlap, the followings structures are observed: nucleus, centriolar adjunct, mitochondrial derivatives and axoneme. The two mitochondrial derivatives are of different lengths but similar diameter. The axoneme is of a conventional insectan type with a 9 + 9 + 2 microtubular arrangement, with accessory tubules flanked by the electron-dense intertubular material. The male internal reproductive tract consists of testis, vas deferens, seminal vesicle, accessory glands and ejaculatory duct.     

Identification of mechanisms responsible for fetal weight reduction in mice following ultrasound exposure

This study challenges existing interpretations of the effects of ultrasound on fetal development. The results demonstrated that a lower mean fetal weight in mice observed after intrauterine exposure to 2 MHz ultrasound resulted from maternal distress, and was not simply a direct effect of ultrasound interaction with the fetuses. At day 8 of gestation continuous wave ultrasound was applied to different regions of the maternal abdomen at spatial average intensities between 0.5 and 5.5 W cm-2 for 3 min. Insonating the uterus directly resulted in resorptions, abortions and gross developmental abnormalities, with rare instances of fetal weight reduction which were apparently associated with maternal distended bladder syndrome (DBS). When the ultrasound beam deliberately avoided the uterus, by a dorsal approach centred on the vertebral column, mean fetal weight at day 18 was significantly lower in all mothers showing DBS. Fetuses of insonated mothers not affected by DBS were not significantly different from the sham-insonated group whether the uterus was directly exposed or not. The fetal weight reduction endpoint was therefore found to be more closely correlated with maternal damage (DBS) than with ultrasound intensity.     

Laser-induced microexplosion confined in the bulk of a sapphire crystal: evidence of multimegabar pressures

Extremely high pressures (approximately 10 TPa) and temperatures (5 x 10(5) K) have been produced using a single laser pulse (100 nJ, 800 nm, 200 fs) focused inside a sapphire crystal. The laser pulse creates an intensity over 10(14) W/cm2 converting material within the absorbing volume of approximately 0.2 microm3 into plasma in a few fs. A pressure of approximately 10 TPa, far exceeding the strength of any material, is created generating strong shock and rarefaction waves. This results in the formation of a nanovoid surrounded by a shell of shock-affected material inside undamaged crystal. Analysis of the size of the void and the shock-affected zone versus the deposited energy shows that the experimental results can be understood on the basis of conservation laws and be modeled by plasma hydrodynamics. Matter subjected to record heating and cooling rates of 10(18) K/s can, thus, be studied in a well-controlled laboratory environment.     

Characteristics of Co-filled carbon nanotubes

The Co-filled carbon nanotubes (CNTs) film was produced on silicon substrate by electron cyclotron resonance microwave plasma chemical vapor deposition (ECR-CVD). The effects of different plasma powers of 200, 300, 400 and 500 W, on the morphology, structure and electrical properties of the CNTs film, were studied. The results showed that the surface density of the vertical nanotubes decreased when the plasma power was higher than 200 W. When plasma power of 300 W was used, the ends of the metal-filled carbon nanotubes (MF-CNTs) became straighter and more uniform. The Co-filled CNTs grown at 300 and 400 W had a current discharge at the applied voltages of 30 and 40 V, respectively. In addition, the surface morphology and the structure of the CNTs film were examined using scanning electron microscopy (SEM) and high-resolution field emission gun transmission electron microscopy (TEM). Energy dispersive X-ray spectroscopy (EDXS) analyses were performed to identify the composition of the material inside the CNTs.     

聚变堆第一壁连续W/Cu 梯度材料的热工性能优化

针对不同体积分布指数p的W/Cu连续功能梯度材料的偏滤器第一壁结构,采用有限元软件计算了 8MW•m⁻²稳态运行热加载以及等离子体破裂条件下1GW•m⁻²热流冲击下的力学响应。相同稳态加载条件下,W/Cu 连续功能梯度材料的最优分布指数与分层梯度材料存在较大差异,其最优等效应力比分层梯度材料要小26%,表现出更优异的性能。在热冲击响应过程中,连续梯度W/Cu材料塑性损伤随p值不同也存在较大变化,其最优p值与其稳态运行时热应力最优p值存在一定差异,从第一壁应用条件考虑,应综合选取,最佳p值在1.2附近。综合来看,连续梯度W/Cu材料具有更连续变化的热物理属性及力学性能,在聚变堆第一壁结构设计中具有更大的应用潜力。     

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针对不同体积分布指数p的W/Cu连续功能梯度材料的偏滤器第一壁结构,采用有限元软件计算了8MW.m?2稳态运行热加载以及等离子体破裂条件下1GW.m?2热流冲击下的力学响应。相同稳态加载条件下,W/Cu连续功能梯度材料的最优分布指数与分层梯度材料存在较大差异,其最优等效应力比分层梯度材料要小26%,表现出更优异的性能。在热冲击响应过程中,连续梯度W/Cu材料塑性损伤随p值不同也存在较大变化,其最优p值与其稳态运行时热应力最优p值存在一定差异,从第一壁应用条件考虑,应综合选取,最佳p值在1.2附近。综合来看,连续梯度W/Cu材料具有更连续变化的热物理属性及力学性能,在聚变堆第一壁结构设计中具有更大的应用潜力。     

超短超强激光脉冲辐照超薄碳膜电离状态研究

为了进一步理解极端条件下物质的电离特性, 特别是超短超强激光脉冲辐照超薄靶时等离子体的形成与分布, 本文以超薄碳膜为例, 细致研究了超短超强激光脉冲辐照下原子的离化过程. 分析和比较了强激光场直接作用电离和靶内静电场电离等两种场致电离形式, 在碰撞电离可以忽略的情况下, 发现更多的电离份额是来自靶内静电场的电离方式. 研究了激光脉冲强度对电离的影响, 发现激光脉冲强度越强, 电离速度越快, 产生的高价态离子所占比例也越高.当激光强度为1×10²⁰ W/cm²时, 尽管该强度高于电离生成C⁺⁶所需要的激光强度阈值, 但该激光脉冲并不能将整个靶电离成C⁺⁶离子, 对此本文进行了详细的分析. 在研究激光脉冲宽度的影响时, 发现激光脉宽越小, 电离速度越快, 但越小的激光脉冲电离获得的高价态离子越少.     

钨珠在穿靶过程中破损问题的数值研究

 使用二维有限元法,研究了直径为6 mm和8.5 mm的预制破片（钨珠、钢珠）以不同速度在稠密气体中飞行时的破损问题,初步确定了珠子破损的阈值速度。研究表明,破损的程度与珠子的速度、材料、大小有密切关系。同时还研究了破损了的破片对6 mm厚钢靶的穿透能力。