Cytopathology of the trachea of Bombyx mori (Lepidoptera: Bombycidae) to Bombyx mori nucleopolyhedrovirus

Bombyx mori is a holometabolous insect found only in germplasm banks and morphological data related of resistance and susceptibility to diseases is important when selecting hybrids for commercial and scientific interest. This study analyzed the cytopathology of B. mori trachea to BmNPV, isolated geographically in Paraná state, Brazil. Fifth instar larvae were divided into two groups, control and inoculated; the viral suspension used was 2.4 × 10⁷ polyhedral occlusion bodies/mL. From the second to the ninth day post-inoculation, segments of silkworm organs, containing the trachea, were processed for transmission electron microscopy. Analyses of fresh hemolymph were also performed to verify the susceptibility of the hemocytes. Signs of infection were initially detected in the hemocytes and in the tracheal cells on the second and fourth post-inoculation days, respectively. The cytopathology of the trachea showed all stages of the viral cycle, which was the same as in other tissues. Virions were detected in the basal lamina, which showed structural disorganization. So, the infection time of the hemocytes prior to trachea and the presence of virus in basal lamina, suggests that the trachea was a secondary target, infected by budded virus coming from the hemolymph.     

低温诱导桑蚕体内腺体相行为的高分辨^13C固体核磁共振研究

为什么蚕在常温常压水溶液条件下即能纺出力学性能优异的蚕丝纤维, 一直是科学家们感兴趣的问题. 在过去的几十年, 人们曾用多种表征手段, 如双折射、扫描电镜、原子力显微镜和电子散射等, 在界观尺度下对蚕在吐丝过程中腺体的相行为进行研究. 发现腺体在靠近吐丝口时呈液晶态, 并认为这是导致蚕丝力学性能的重要因素. 本文则在分子水平尺度下利用核磁共振方法, 对五龄蚕活体在常温和6 ℃下存储数天后解剖的腺体进行研究. 经对化学位移及其线型的各向异性分析发现, 当将体内腺体沿吐丝方向分为3部, 即后部、中部及靠近吐丝口的前部时, 常温下, 腺体后部和中后部分子呈无规线团, 而腺体中中部、中前部和前部分子呈液晶态. 6 ℃时, 中后部分子亦呈液晶态, 前部分子排列则各向异性更大, 说明更为有序. 这种液晶态呈分形结构, 在小于纳米尺度下为无规线团, 大于纳米尺度呈有序排列. 这表明, 降温过程可使呈无规线团的丝素蛋白分子转变为液晶态, 其效果如同蚕在吐丝过程中对其腺体施加的剪切应力. 该结果对于人们探索人工合成高性能类丝素纤维的纺丝工艺和条件将有启发和指导作用.     

低温诱导桑蚕体内腺体相行为的高分辨13C固体核磁共振研究

为什么蚕在常温常压水溶液条件下即能纺出力学性能优异的蚕丝纤维,一直是科学家们感兴趣的问题. 在过去的几十年,人们曾用多种表征手段,如双折射、扫描电镜、原子力显微镜和电子散射等,在界观尺度下对蚕在吐丝过程中腺体的相行为进行研究. 发现腺体在靠近吐丝口时呈液晶态,并认为这是导致蚕丝力学性能的重要因素. 本文则在分子水平尺度下利用核磁共振方法, 对五龄蚕活体在常温和6 ℃下存储数天后解剖的腺体进行研究. 经对化学位移及其线型的各向异性分析发现,当将体内腺体沿吐丝方向分为3部,即后部、中部及靠近吐丝口的前部时,常温下,腺体后部和中后部分子呈无规线团,而腺体中中部、中前部和前部分子呈液晶态. 6 ℃时, 中后部分子亦呈液晶态,前部分子排列则各向异性更大,说明更为有序. 这种液晶态呈分形结构,在小于纳米尺度下为无规线团,大于纳米尺度呈有序排列. 这表明,降温过程可使呈无规线团的丝素蛋白分子转变为液晶态,其效果如同蚕在吐丝过程中对其腺体施加的剪切应力. 该结果对于人们探索人工合成高性能类丝素纤维的纺丝工艺和条件将有启发和指导作用.      

Susceptibility, field inhomogeneity, and chemical shift-corrected NMR microscopy: Application to the human finger in vivo

Spectroscopic proton image data recorded with the aid of a gradient-echo spectroscopic imaging pulse sequence are reported. A postdetection processing method is suggested which permits correction of artifacts due to inhomogeneity, susceptibility, and chemical-shift resonance offsets. That is, apart from the spectral information available in this way, better spatial resolutions can be achieved. The method is demonstrated by resonance-offset corrected images of the human finger in vivo. Moreover, resonance-line selective and spectroscopically resolved diffusion-weighted images and diffusivity maps rendered with the aid of the same postdetection procedure are shown.     

Molecular characterization of the brain secretory peptides,prothoracicotropic hormone (PTTH) and bombyxin,of the silkmoth Bombyx mori

Molecular characterization of the brain secretory peptides, PTTH and bombyxin, of Bombyx mori is reviewed. PTTH is a 30-kDa homodimeric glycoprotein, the monomer of which consists of 109 amino acids. Two monomers are held together by a disulfide bond. cDNA and gene coding for PTTH were cloned and the precursor protein for PTTH monomer was deduced. A novel 5-kD brain secretory peptide named bombyxin has been discovered from Bombyx brain. Bombyxin is highly homologous to vertebrate insulin-family peptides and possesses the prothoracicotropic activity when injected into brain-removed pupae of a heterologous moth, Samia cynthia ricini, though inactive to Bombyx from which it was derived. cDNA and gene coding for bombyxin were cloned, preprobombyxin protein was deduced, and posttranslational processing to generate mature bombyxin was suggested. The Bombyx genome contains highly multiple copies of the gene coding for bombyxins. Immunohistochemically, PTTH- and bombyxin-producing brain neurosecretory cells were identified.     

Applications of chemical-shift-selective NMR microscopy to the non-invasive histochemistry of plant materials

Chemical-shift-selective imaging at microscopic resolution has been applied to the study of various plant materials including orange peel, grape berries and both dried and undried fruits of fennel. It is shown that selective imaging of aromatics and carbohydrates as well as water and oil can be performed with in-plane resolution down to 13 microns and slice thicknesses of 1 mm or less, at a field strength of 4.7 Tesla. The noninvasive nature of the method gives it advantages over established methods of plant histochemistry which involve sectioning and staining to reveal different chemical constituents.     

1H NMR spin-spin relaxation and imaging in porous systems: an application to the morphological study of white portland cement during hydration in the presence of organics

Proton nuclear magnetic resonance (NMR) spin-spin relaxation and imaging have been applied to investigate white Portland cement pastes during hydration in the absence and in the presence of organic solvents. The main organic solvent investigated was methanol, alone or together with the organic waste 2-chloroaniline (2-CA), an aromatic amine representative of an important class of highly toxic compounds. For all the analysed samples, prepared with a solvent-to-cement ratio of 0.4, the decay of the echo magnetization has been fitted by adopting a model that combines an exponential component with a gaussian one. The calculated independent relaxation parameters have been discussed in terms of morphological and dynamical changes that occur during the cement hardening process and pore formation. Three kinds of water molecules: "solid-like" (chemically and physically bound), "liquid-like" (porous trapped) and "free" water, endowed with anisotropic, near isotropic and isotropic motion, respectively, were identified. Spin-echo images collected on the same samples during the hydration kinetics, allowed the changes of water and solvents spatial distribution in the porous network to be monitored, showing percolation phenomena and confirming the multimodal open channels structure of the hardened cement system. Both T(2) relaxation and imaging data indicated that a pronounced delay occurs in the cement hardening when organics are present.     

A method to distinguish between chemical shift and susceptibility effects in NMR microscopy and its application to insect larvae

We propose a simple method of distinguishing Zeeman broadening arising from susceptibility inhomogeneity and chemical shift variation, applicable to NMR microscopy. The method is based on the use of a specially built probehead in which orthogonal sample alignment is possible using the same radiofrequency (RF) coil. This allows the investigation of alignment effects in image distortion and relies on the fact that the isotropic chemical shift is invariant under reorientation, whereas the susceptibility-related local field will depend strongly on relative orientation of bounding surfaces with the external polarizing field. We apply this approach to the study of a simple phantom, and an insect larva (Spodoptera litura Fabricius), demonstrating in the latter case that susceptibility variations are sufficiently small to allow chemical shift imaging on a scale greater than 1 ppm.     

同步辐射显微技术在高分子材料结构研究中的应用

高分子材料以其优异的性能广泛应用于人类生活的每个角落,但其发展受限于研究手段.基于同步辐射先进光源的研究方法(如散射、吸收和成像等)具有高的空间、时间和能量分辨的优势,是揭示高分子材料多尺度结构形成和演化动力学最有效的工具之一.文章结合作者和国内外同行的工作,以具体案例的形式介绍了同步辐射技术在高分子材料结构研究中的应用.希望能起到抛砖引玉的作用,吸引更多的从事高分子材料结构研究的同行利用同步辐射开展科学研究,同时希望更多的进行物理学研究的同行来帮助回答高分子物理的一些基本科学问题.     

同步辐射显微技术在高分子材料结构研究中的应用

高分子材料以其优异的性能广泛应用于人类生活的每个角落,但其发展受限于研究手段.基于同步辐射先进光源的研究方法(如散射、吸收和成像等)具有高的空间、时间和能量分辨的优势,是揭示高分子材料多尺度结构形成和演化动力学最有效的工具之一.文章结合作者和国内外同行的工作,以具体案例的形式介绍了同步辐射技术在高分子材料结构研究中的应用.希望能起到抛砖引玉的作用,吸引更多的从事高分子材料结构研究的同行利用同步辐射开展科学研究,同时希望更多的进行物理学研究的同行来帮助回答高分子物理的一些基本科学问题.     

The regenerative cells during the metamorphosis in the midgut of bees

The midgut epithelium of bees is formed by the digestive cells, responsible for enzyme secretion and nutrient absorption and for small regenerative cells that are placed in nests scattered among the digestive cells. During metamorphosis, the larval midgut epithelium degenerates and a new adult midgut epithelium is built during larval differentiation of regenerative cells. The present work focuses on the midgut epithelial modifications during the post-embryonic development of the stingless bee Melipona quadrifasciata anthidioides worker and the occurrence of regenerative cell proliferation during midgut metamorphosis in order to test the hypothesis that adult midgut epithelium of worker bees results from regenerative cell proliferation during the pupal stage. Regenerative cell proliferation was detected during larval lifespan. Larval aging is followed by an increase in the number and the size of the nests of regenerative cells. Larval epithelium degeneration begins 2 days after the start of defecation process and in this period the nests of regenerative cells are in contact by means of cytoplasmic extension which have many septate desmosomes and gap junctions. The BrdU immunoreactive regenerative cells were found in the prepupae 12 h after BrdU injection, suggesting that regenerative cell population increase during this larval period. Regenerative cell proliferation results in the increase of the regenerative cell population and not in the formation of new digestive cells because the proliferation of regenerative cells would not be enough to reestablish the nests of regenerative cells and at the same time form new adult digestive cells. In this sense the hypothesis that digestive adult cells originate from regenerative cell proliferation during midgut metamorphosis in M. quadrifasciata anthidioides was rejected.     

Application of the solid state NMR to the study of the alcohol/alkane mixtures adsorption onto graphite

The mixing of molecules adsorbed from solution to different interfaces has both industrial and academic relevance and the mixing behaviour at the interface is a key to understanding for example, that the surface tension of a mixture of two surfactants is lower than either of the two pure materials and many other effects. In this paper, we report, for the first time, the application of Solid State NMR to the study of alkane/alcohol mixtures, in a range of relative size ratio between 0 and 0.35, adsorbed onto graphite at high, multilayer coverage. Moreover, this paper evaluated, for the first time, the utility of the combined used of 1H and 2H NMR for: (i) determining the surface composition and (ii) making a theoretical approach to the sorption isotherm. A variety of preferential adsorption behaviour is reported. Preferential adsorption of the longer molecule (decane vs. heptanol) from a mixture has been observed. However, if both components are of similar length, the alcohol is preferentially adsorbed (heptanol vs. octane and octanol vs. octane). Finally, a linear relation between the relative size ratio and the amount of alcohol at monolayer coverage is observed.     

Application of interference microscopy to the study of hologram build-up in LiNbO3 crystals

Interference microscopy was applied to direct microscopic observation of temporal evolution of phase holograms in LiNbO₃:Fe photorefractive crystals. First a hologram was recorded in the sample, and diffraction efficiency was monitored during hologram build-up using inactinic laser light. Thus kinetics of hologram build-up could be determined. The initial hologram was erased using white light. Then a series of write-erase cycles were performed with increasing exposure times. Holograms were observed by interference microscope after each exposure. The time elapsed between the exposure and the microscopic observation was negligible compared to the relaxation time of the hologram. The obtained temporal evolution of the grating profile gives a deeper insight into the physical mechanism of hologram formation in photorefractive materials than simple diffraction efficiency measurements. A congruently grown sample of LiNbO₃ doped with 10⁻³ mol/mol Fe in melting was studied by this method. Sample thickness was set to 300 μm to allow correct microscopic observation. Plane-wave holograms were recorded in the samples using an Ar-ion laser at λ = 488.0 nm of grating constants of 3, 6.5 and 8.8 μm.     

Superspin in NMR: Application to the ABX system

Superspin is defined to be the angular momentum associated with operators on a nuclear spin system. These operators correspond to magnetizations and the effect of a pulse on these magnetizations is most easily calculated using this formalism. The derivation of the superspin characteristics of the magnetizations in an ABX system is given in detail.     

Application of field-ion microscopy techniques to metallurgical problems

The invention of field-ion microscopy and related techniques by E.W. Müller has created a new field of high-resolution analyses. While much of its application to metallurgy in the past was oriented towards instrumentation and technique, today there is an increasing trend towards using the techniques for problem-related studies. Some examples of the application of field-ion microscopy in studies of grain boundaries, ordering and clustering, and precipitation will be reviewed.     

Application of transmission electron microscopy to the clinical study of viral and bacterial infections: present and future

Transmission electron microscopy has had a profound impact on our knowledge and understanding of viruses and bacteria. The 1000-fold improvement in resolution provided by electron microscopy (EM) has allowed visualization of viruses, the existence of which had previously only been suspected as the causative agents of transmissible infectious disease. Viruses are grouped into families based on their morphology. Viruses from different families look different and these morphological variances are the basis for identification of viruses by EM. Electron microscopy initially came to prominence in diagnostic microbiology in the late 1960s when it was used in the rapid diagnosis of smallpox, by differentiating, on a morphological basis, poxviruses from the less problematic herpesviruses in skin lesions. Subsequently, the technique was employed in the diagnosis of other viral infections, such as hepatitis B and parvovirus B19. Electron microscopy has led to the discovery of many new viruses, most notably the various viruses associated with gastroenteritis, for which it remained the principal diagnostic method until fairly recent times. Development of molecular techniques, which offer greater sensitivity and often the capacity to easily process large numbers of samples, has replaced EM in many areas of diagnostic virology. Hence the role of EM in clinical virology is evolving with less emphasis on diagnosis and more on research, although this is likely only to be undertaken in specialist centres. However, EM still offers tremendous advantages to the microbiologist, both in the speed of diagnosis and the potential for detecting, by a single test, any viral pathogen or even multiple pathogens present within a sample. There is continuing use of EM for the investigation of new and emerging agents, such as SARS and human monkeypox virus. Furthermore, EM forms a vital part of the national emergency response programme of many countries and will provide a frontline diagnostic service in the event of a bioterrorism incident, particularly in the scenario of a deliberate release of smallpox virus. In the field of bacteriology, EM is of little use diagnostically, although some bacterial pathogens can be identified in biopsy material processed for EM examination. Electron microscopy has been used, however, to elucidate the structure and function of many bacterial features, such as flagellae, fimbriae and spores and in the study of bacteriophages. The combined use of EM and gold-labelled antibodies provides a powerful tool for the ultrastructural localisation of bacterial and viral antigens.     

Application of129Xe NMR to polymer blends

¹²⁹Xe NMR experiments on two different polymer blends are described. The first system, a blend of polypropylene (PP) and the copolymer of polypropylene and polyethylene (EP), shows separate domains of polypropylene and of the copolymer. The latter phase forms rubbery domains in the polypropylene matrix. The second system is the compatible blend of polyvinylidenefluoride and polymethylmethacrylate. For the incompatible blend two Xe resonances are found, one for Xe absorbed in the matrix and one for Xe in the rubbery EP phase. The chemical shift of the Xe absorbed in the rubbery phase can be related to the polyethylene content of the copolymer. The line widths and chemical shifts are affected by the polymer motions as a function of the temperature. Although the system has two very different glass transition temperatures, it is striking to see that upon approaching the polypropylene glass transition temperature the Xe resonance of Xe in the rubbery phase is also affected. Due to the dipolar interaction between Xe spins and polymer proton spins, cross-polarization experiments can be performed. This allows the measurement of correlated NMR spectra. The compatible blend shows only one line with a chemical shift proportional to the composition.     

FMRFamide-like midgut endocrine cells during the metamorphosis in Melipona quadrifasciata anthidioides (Hymenoptera, Apidae)

The FMRFamide, gastrin and cholecystokinin (CCK) occurrence in endocrine cells of insects has been described by several authors, although their functions are still not well defined for this group of animals. In the present study, the occurrence of endocrine cells producing FMRFamide, gastrin 1 and CCK-8 in the midgut (ventriculus) of Melipona quadrifasciata anthidioides (Hymenoptera, Apidae), before, during and after the metamorphosis, were investigated by means of pre-embedding immunofluorescence techniques. FMRFamide reactivity was found in the endocrine cells as well as in the nervous fibers and neurons of the intestine of these bees. 'Open' and 'closed' types of FMRFamide-like cells were observed in last instar larvae. In the black eyed pupae the producing cells of FMRFamide seemed to be immature, and, in the workers, where the FMRFamide producing cells were more abundant, the production of this substance seemed to occur only in the open cells. Reactivity of the nervous fibers and neurons were observed, during the prepupae, white eyed pupae, and pink eyed pupae. The same did not occur with the midgut endocrine cells. There were no immunoreactivity observations for gastrin 1 and for CCK-8. The FMRFamide-like cells were present in the midgut of these insects during or close to the period that they were eating, which indicates that the FMRFamide may be involved in the control of the digestive process.