Egg envelopes of Armadillidium vulgare (Latreille, 1804) (Crustacea,Isopoda Oniscidea): Ultrastructure and lectins binding

The ultrastructural study carried out on (a) oocytes of Armadillidium vulgare during vitellogenesis, (b) mature eggs taken from the ovaries during the parturial moult of the posterior half of the body, and (c) fertilized eggs collected within a few hours of their release into the brood pouch, has clearly demonstrated that before the fertilization the chorion is the only envelope present in the egg of oniscidean isopods. In the mature eggs, the chorion appears as a uniformly electron‐dense lamina, about 0.4–0.5 µm thick, which does not show any specialized area. A second envelope, described by other authors as vitelline envelope, is formed above the oolemma only right after fertilization and appears separated from the chorion by a space full of liquid. The ways in which the genesis of this envelope is realized are not yet clear; it could be interpreted rather as a fertilization membrane. The investigations carried out with the aid of a battery of FITC‐lectins have highlighted the presence at the chorion surface of unfertilized eggs of various saccharide residues distributed in uniform way. No significant change was observed in the pattern of lectins binding to the chorion of eggs taken from the brood pouch, thus demonstrating how, after the fertilization, no significant rearrangement in the distribution of saccharide residues present on the egg surface occurs in A. vulgare. The ways in which, therefore, the recognition, the binding and the entry of the peculiar sperm of oniscidean isopods into the egg occur, still remain all to be deciphered. Microsc. Res. Tech. 79:792–798, 2016. © 2016 Wiley Periodicals, Inc.     

Detection of lead in Zea mays by dual‐energy X‐ray microtomography at the SYRMEP beamline of the ELETTRA synchrotron and by atomic absorption spectroscopy

This study is related to the application of the X‐ray dual‐energy microradiography technique together with the atomic absorption spectroscopy (AAS) for the detection of lead on Zea mays stem, ear, root, and leaf samples. To highlight the places with lead intake, the planar radiographs taken with monochromatic X‐ray radiation in absorption regime with photon energy below and above the absorption edge of a given chemical element, respectively, are analyzed and processed. To recognize the biological structures involved in the intake, the dual‐energy images with the lead signal have been compared with the optical images of the same Z. mays stem. The ear, stem, root, and leaf samples have also been analyzed with the AAS technique to measure the exact amount of the hyperaccumulated lead. The AAS measurement revealed that the highest intake occurred in the roots while the lowest in the maize ears and in the leaf. It seems there is a particular mechanism that protects the seeds and the leaves in the intake process. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc. This article was published online on 1 December 2009. An error was subsequently identified. This notice is included in the online and print version to indicate that both have been corrected 19 February 2010.     

The structure and formation of the egg membranes in Nasonia vitripennis (Walker) (Hymenoptera, Pteromalidae)

A histochemical and fine structural study of the vitelline membrane and chorion was carried out. Following the uptake of protein at the oöcyte plasma membrane during yolk synthesis, the egg membranes are laid down in sequence in the intercellular space between the oöcyte and the follicle cells. The follicular epithelial cells alone contribute towards their formation. The structure of the chorion is much simpler than that of Bombus, the only other hymenopteran previously investigated.     

Egg morphology of nine species of Pollenia Robineau-Desvoidy, 1830 (Diptera: Calliphoridae)

Egg morphology of nine species of the cluster fly genus Pollenia Robineau-Desvoidy was studied with scanning electron microscopy. Funnel-shaped micropylar area, chorion covered with centrally depressed hexagons, well developed hatching pleats on the dorsal surface encompassing the median area were found in eggs of all species: P. amentaria (Scopoli), P. angustigena Wainwright, P. atramentaria (Meigen), P. labialis Robineau-Desvoidy, P. mayeri Jacentkovsky, P. pediculata Macquart, P. rudis (Fabricius), P. similis (Jacentkovsky), and P. vagabunda (Meigen). Untypical for Calliphoridae, a dual morphological structure of plastron-bearing area is described here for the first time. On the basis of our observations, all species studied share the presence of a plastron respiratory function on the entire surface between the hatching lines. Differences between species were found in the shape of the longitudinal hatching pleats and the proportion between 'island pattern' and 'hexagonal pattern' of their chorionic surface, and in the intensity of perforation of hexagons of the median area. Comparisons of Pollenia egg morphology with that of other representatives of Calliphoridae revealed its unique structure, allowing easy differentiation from other representatives of the family.     

Ultramorphological characteristics of immature stages of Chrysomya albiceps (Wiedemann 1819) (Diptera: Calliphoridae), a fly specie of forensic importance

Forensic entomology is an area of science that serves as a tool in crime scene investigations. Usually, flies are the first insects to reach a dead body and can oviposit just a few hours after arrival. Therefore the knowledge of immature stages is essential for correct identification of the species found on corpses. Scanning electron microscopy gives detailed information about morphological characters helping to identify the immature forms and consequently serves as a tool in crime scene investigations. C. albiceps is a very important fly for forensic entomologists because its larvae are almost always present on a dead body and it is facultative predators and therefore can alter the composition of species present at the carcass. The aim of this study is to identify eggs, larvae, and puparia of C. albiceps using SEM. Eggs were elongated with the anterior region ending in a “Y” shape and the posterior end was tapered. The micropyle was a well‐adorned orifice with some projections around it. The first instar larva was composed of 12 segments separated by spines. Only one spiracular opening could be seen at the posterior spiracle. Body tegument was smooth and tubercles were not seen. Antennae and maxillary palps were visible. Second and third larval instars were very similar to first instar, except for the presence of anterior spiracle. However, body tegument was composed of net‐like patches and tubercles were visible. Tubercles present at the third instar larvae were robust and erect. Puparia showed a retracted cephalic region and curved tubercles. Microsc. Res. Tech. 73:779–784, 2010. © 2010 Wiley‐Liss, Inc.     

Fine structure and formation of the eggshell in scorpionfly Panorpa liui Hua (Mecoptera: Panorpidae)

The morphology and formation of the eggshell in the scorpionfly Panorpa liui Hua were examined with light microscopy and scanning and transmission electron microscopy. During oogenesis of the scorpionfly, the follicle cells multiply by mitotic divisions and diversify into four morphologically distinct subpopulations, three of which are engaged in the eggshell formation and mold various parts of chorion. The eggshell consists of three layers: An inner vitelline membrane, an outer chorion, and a precursor extrachorion. The chorion constitutes a very compact endochorion, a rough fibrillar exochorion, and a polygonal meshwork of elevated ridges. At proximal end of ovarioles the chorion of matured oocyte is covered with a loose membrane, which might be the remnant of follicle cells. The jelly substance, which acts as lubricant to protect the oviducts and ovulated eggs during ovulation, might add to the top of polygonal ridges as the outermost extrachorion after oviposition. The eggshell formation process in Panorpa is tentatively proposed. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc.     

Molecular and chemical investigations and comparisons of biomaterials for ocular surface regeneration

This study investigated and compared the ultrastructural and chemical properties of representative biomaterials for ocular surface regeneration: a human amniotic membrane (AM) in a basal plate, a human AM in reflected chorion, a preserved AM, and a human corneo‐scleral tissue. Assessments of the morphological differences in the extracellular matrices were evaluated by hematoxylin–eosin, Masson's trichrome (for total collagen), and picrosirius‐red (for newly synthesized collagen) staining. Assessments of the changes in the molecular structures and chemical compositions of the biomaterials for ocular surface regeneration were evaluated by Raman spectroscopy. A placental AM (52 %) was a dense and thick collagenous structure compared to a reflected AM (23 %). The spectroscopy did not obtain any structural information for a preserved AM. The cornea group (100 %, control) and sclera group (104 %) showed the collagen lamellae and interfibrillar spacing, and a slight inflammatory reaction with more fibrous and granulomatous tissues. There was a formation of newly synthesized collagen in a placental AM, while there were few collagen components in a reflected AM. Human AM tissues showed consistent Raman spectra and the characteristic collagen bands, similar to the corneal and scleral tissues. Therefore, these findings suggest that human placental AM and reflected AM are structurally suitable for scleral and corneal surface regeneration, respectively, while human placental or preserved AM and reflected AM are molecularly and chemically suitable for corneal and scleral surface regeneration, respectively. Microsc. Res. Tech. 77:183–188, 2014. © 2013 Wiley Periodicals, Inc.     

Comparative ultrastructure of the fertilized egg envelope in Corydoras adolfoi and Corydoras sterbai,Callichthyidae, Teleostei

In teleost, the structural characteristics of fertilized egg and egg envelope are very important for classification of genus or species. The structures of fertilized egg and egg envelope from Corydoras adolfoi and Corydoras sterbai, Callichthyidae, Siluriformes in teleost were examined by scanning and transmission electron microscopes to confirm whether these morphological structures have specificities of species and family or not. The fertilized eggs of C. adolfoi and C. sterbai were non‐transparent, spherical, demersal, and strong adhesive. There were no structural differences between two species through the light microscope. The size of the fertilized eggs of C. adolfoi was 1.95 ± 0.03 mm (n = 20), and that of C. sterbai was 1.92 ± 0.03 mm (n = 20). The perivitelline space was almost not developed in both species. In both species, the adhesive protuberances structures were on the outer surface of egg envelope. And fibrous structures were specially located at attachment part of spawning bed. And the egg envelope consisted of two layers, an inner lamellae layer and an outer strong adhesive layer with high electron dense protuberances structures in cross section. Consequentially, the fertilized eggs, outer surface on the egg envelope and cross section of egg envelope have identical structure. So, these structural characteristics of fertilized eggs and egg envelope show genus Corydoras specificity.     

A model for predicting pathologist's velocity profiles when navigating virtual slides

Navigation through large microscopic images is a potential benefit for histology or pathology teaching, for improving the quality of diagnosis in pathology, or for communicating pathologists in some telemedicine applications. However, the size of this kind of images is prohibitive for navigation with conventional techniques. This article presents a soft computing model, which permits to anticipate the pathologist trajectories in diagnosis tasks when exploring virtual slides. The Bayesian strategy combines an offline model of a baseline pathologist knowledge (the prior) and a prediction online module (the likelihood) that captures a particular pathologist navigation pattern. While optimal parameters for the biologically inspired offline model are calculated using an Expectation‐Maximization strategy, prediction is carried out by a particle filter. Parameters are estimated from several series of actual navigations performed by several pathologists in different virtual slides. The present approach is compared with other conventional prediction methods and decreases the calculated MSE in about a 50% for the entire group of pathologists. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

The structure and ultrastructure of the egg capsule of Brachyptera risi (Plecoptera,Nemouroidea, Taeniopterygidae) with some remarks concerning choriogenesis

The organization of the egg capsule of the euholognathan stonefly, which represents the family Taeniopterygidae (Nemuroidea) was investigated using light and electron microscopy techniques. The presence of a complex, multilayered egg capsule, composed of a vitelline envelope, multilayered chorion, and extrachorion is described. The morphology of the eggshell of Brachyptera risi was compared with that of euholognthan and systellognathan egg coverings and the ground plan of the egg capsule in Plecoptera was discussed. Microsc. Res. Tech. 78:180–186, 2015. © 2014 Wiley Periodicals, Inc.     

Berechnungs- und anwendungsmöglichkeit einer polygonzahlPossible calculations and applications of a polygon number

The conditions derived from the degree of freedom equation show that there is a polygon number (19) for the maximum number of mobile polygons (Fig. 1, Fig. 2) with f_i = g, which is stable as long as the number of members and joints and the degrees of mobility remain the same. For these systems the amount of conditions dependent upon each other for the rigidity (17) are normally also the same, i.e. if a plain, forcibly actuated linkage with normal dimensions describes a closed curve of the highest degree in both variables of the plain, then definition of the coefficients based on the rigidity conditions is necessary and sufficient.By means of a polygon number the maximum number of elements (23) per member is determined, which can be applied for all closed linkages. Furthermore the sum of the degrees of freedom in plain, forcibly actuated kinematic linkages that can be arranged without any members crossing each other, is determined, which is the same in all polygons. In four-membered linkages (Fig. 3) this is four and in eight-membered it is five. Beyond that there are no further structures with the same sum of degrees of freedom in all polygons; the possibility exists, however, should the total polygon number be realized, that two polygons contain four (Fig. 6) or four polygons contain five and all remaining polygons contain six degrees of freedom. The sum of the degrees of freedom of six in all polygons cannot be achieved.From the analysis that was carried out, the synthesis (Fig. 3 to 6) for detecting all kinematic linkages with the same properties by reversing the process became apparent. With the aid of the number of members, joint and polygon number the systematics of the structures can be elaborated. The possibility exists, amongst others, to break the linkages down into necessary and complementary sub-systems. To detect all the structures one simply has to define all the possible complementary sub-systems. Thereby also other systems with a larger number of members, e.g. planetary gear drive, can be developed by using a search strategy.It became apparent that even with a rigid framework as a basis that is extended to mobile linkages no unknown structures can be developed.     

Morphology of the fertilizable mature egg in the Acanthopagrus latus,A. schlegeli and Sparus sarba (Teleostei: perciformes: sparidae)

Through appropriate scanning electron microscope (SEM) processing, the shape and the surface structures of just‐mature eggs are preserved integrally for observation and measuring to guarantee accurate comparison and prediction on the phylogenetic relationships among three species of teleost, i.e. Acanthopagrus latus, Acanthopagrus schlegeli and Sparus sarba of Sparidae. Eggs in all three species can be categorized as type III eggs. Specifically, a regular pattern is found in the arrangement of micropyle and its surrounding pore canals from the inspected surface structures of the mature eggs in three species. Other than these similarities in egg surface structures, several differences in micropyle structures, micropyle region and size of pore canals are also observed in eggs of respective species. Comprehension and accurate data are statistically computed and analyzed through hierarchical cluster analysis to provide solid species‐specific evidences and standardized empirical reference for accurate species identification and phylogeny determination.     

Comparison of the tracheal systems of Anopheles sinensis and Aedes togoi larvae using synchrotron X‐ray microscopic computed tomography (respiratory system of mosquito larvae using SR‐μCT)

Mosquito‐borne diseases, such as malaria, dengue fever, and Zika virus, are serious global health issues. Vector control may be an important strategy in reducing the mortality caused by these diseases. The respiratory system of mosquito larvae in the water has to inhale atmospheric oxygen as aquatic organisms. In this study, the three‐dimensional (3D) structures of the dorsal longitudinal trunks (DLTs) of the tracheal systems of Anopheles sinensis and Aedes togoi were compared using synchrotron X‐ray microscopic computed tomography. DLT respiratory frequencies were also investigated. Interestingly, the larvae of the two mosquito species exhibit tracheal systems that are both morphologically and functionally distinct. A. sinensis hangs horizontally under the water surface, and has a smaller DLT volume than A. togoi. In contrast, A. togoi hangs upside down using a siphon by fixing its tip to the water surface. The frequency of peristaltic movement in A. togoi is higher than that of A. sinensis. These differences in the structures and breathing behaviors of the respiratory systems of mosquito larvae provide new insights into the tracheal systems of mosquito larvae, which should help develop novel effective control strategies targeting mosquito larvae.     

Time‐series investigation of fused vesicles in microvessel endothelial cells with atomic force microscopy

Vesicles or caveolae within endothelial cells, fusing together to form vacuolar organelles, are implicated in macromolecular transport and cellular element transmigration across the blood–brain barrier (BBB) during inflammation and ischemia. Vacuolar organelles have been described by transmission electron microscopy and immunofluorescence, but the details of their dynamics have not been well addressed yet. Herein, by using tapping mode atomic force microscopy (AFM), we observed the time‐series changes of fused vesicles within the serum‐free cultured rat cerebral microvessel endothelial cells. The fused vesicles were certainly proved by fluorescent staining of Fm4‐64 combining simultaneous AFM imaging, as well as the field emission scanning electron microscopy technique. And energy dispersive spectrum results additionally implied that there may be specific structure and compositions around the vesicle region. These results indicate that increased vesicles in BBB may contribute to the formation of fused vesicles and a higher probability to construct the trans‐endothelial channel across endothelium layer. Furthermore, the AFM application may open up a new approach to investigate the details of transcellular process by fused vesicles. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

To b or not to b: The ongoing saga of peptide b ions

Modern soft ionization techniques readily produce protonated or multiply protonated peptides. Collision‐induced dissociation (CID) of these protonated species is often used as a method to obtain sequence information. In many cases fragmentation occurs at amide bonds. When the charge resides on the C‐terminal fragment so‐called y ions are produced which are known to be protonated amino acids or truncated peptides. When the charge resides on the N‐terminal fragment so‐called b ions are produced. Often the sequence of y and b ions are essential for peptide sequencing. The b ions have many possible structures, a knowledge of which is useful in this sequencing. The structures of b ions are reviewed in the following with particular emphasis on the variation of structure with the number of amino acid residues in the b ion and the effect of peptide side chain on b ion structure. The recent discovery of full cyclization of larger b ions results in challenges in peptide sequencing. This aspect is discussed in detail. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 28:640–654, 2009     

MRT letter: A novel tegumental gland in female imagoes of the neotropical termite Cornitermes cumulans (Isoptera,termitidae, syntermitinae)

In general, the exocrine glands of social insects are structures involved in the chemical communication associated with social life. Here, we report the discovery of an unknown tegumental gland that is present in the female imagoes of Cornitermes cumulans and occurs next to the well‐developed tergal glands that have previously been described. The tegumental glands release their secretion in the intersegmental membrane and are composed of bicellular units, a secretory cell and a canal cell, that are closely located to the epidermal cells in the inferior part of the eighth and ninth tergites. The ultrastructure of the glandular cells showed abundant smooth endoplasmic reticulum, suggesting that the secretion may be pheromonal, although its function is still unknown. These exocrine structures are facing the tergal glands, and we hypothesized that they act synergistically with the tergal glands to generate short‐range attraction during tandem behavior. Microsc. Res. Tech., 2010. © 2010 Wiley‐Liss, Inc.     

Development and neuronal dependence of cutaneous sensory nerve formations: Lessons from neurotrophins

Null mutations of genes from the NGF family of NTs and their receptors (NTRs) lead to loss/reduction of specific neurons in sensory ganglia; conversely, cutaneous overexpression of NTs results in skin hyperinnervation and increase or no changes in the number of sensory neurons innervating the skin. These neuronal changes are paralleled with loss of specific types of sensory nerve formations in the skin. Therefore, mice carrying mutations in NT or NTR genes represent an ideal model to identify the neuronal dependence of each type of cutaneous sensory nerve ending from a concrete subtype of sensory neuron, since the development, maintenance, and structural integrity of sensory nerve formations depend upon sensory neurons. Results obtained from these mouse strains suggest that TrkA positive neurons are connected to intraepithelial nerve fibers and other sensory nerve formations depending from C and Aδ nerve fibers; the neurons expressing TrkB and responding to BDNF and NT‐4 innervate Meissner corpuscles, a subpopulation of Merkell cells, some mechanoreceptors of the piloneural complex, and the Ruffini's corpuscles; finally, a subpopulation of neurons, which are responsive to NT‐3, support postnatal survival of some intraepithelial nerve fibers and Merkel cells in addition to the muscle mechanoreceptors. On the other hand, changes in NTs and NTRs affect the structure of non‐nervous structures of the skin and are at the basis of several cutaneous pathologies. This review is an update about the role of NTs and NTRs in the maintenance of normal cutaneous innervation and maintenance of skin integrity. Microsc. Res. Tech. 2010. © 2009 Wiley‐Liss, Inc.     

Infrared consequence spectroscopy of gaseous protonated and metal ion cationized complexes

In this article, the new and exciting techniques of infrared consequence spectroscopy (sometimes called action spectroscopy) of gaseous ions are reviewed. These techniques include vibrational predissociation spectroscopy and infrared multiple photon dissociation spectroscopy and they typically complement one another in the systems studied and the information gained. In recent years infrared consequence spectroscopy has provided long‐awaited direct evidence into the structures of gaseous ions from organometallic species to strong ionic hydrogen bonded structures to large biomolecules. Much is being learned with respect to the structures of ions without their stabilizing solvent which can be used to better understand the effect of solvent on their structures. This review mainly covers the topics with which the author has been directly involved in research: structures of proton‐bound dimers, protonated amino acids and DNA bases, amino acid and DNA bases bound to metal ions and, more recently, solvated ionic complexes. It is hoped that this review reveals the impact that infrared consequence spectroscopy has had on the field of gaseous ion chemistry. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 28:586–607, 2009     

Gloss phenomena and image analysis of atomic force microscopy in molecular and cell biology

Proper sample preparation, scan setup, data collection and image analysis are key factors in successful atomic force microscopy (AFM), which can avoid gloss phenomena effectively from unreasonable manipulations or instrumental defaults. Fresh cleaved mica and newly treated glass cover were checked first as the substrates for all of the sample preparation for AFM. Then, crystals contamination from buffer was studied separately or combined with several biologic samples, and the influence of scanner, scan mode and cantilever to data collection was also discussed intensively using molecular and cellular samples. At last, images treatment and analysis with off‐line software had been focused on standard and biologic samples, and artificial glosses were highly considered for their high probability. SCANNING 31: 49–58, 2009. © 2009 Wiley Periodicals, Inc.     

Differences between nanoscale structural and electrical properties of AZO:N and AZO used in polymer light‐emitting diodes

Conducting atomic force microscopy and scanning surface potential microscopy were adopted to investigate the nanoscale surface electrical properties of N‐doped aluminum zinc oxide (AZO:N) films that were prepared by pulsed laser deposition (PLD) at various substrate temperatures. Experimental results demonstrated that when the substrate temperature is 150°C and the N₂O background pressure is 150 mTorr, the N‐dopant concentration on the surface is optimal. In addition, the root‐mean‐square roughness value of the film surface, the low contact current (<400 nA) conducting region as a percentage of the total area, and the mean work function value are 1.43 nm, 96.9%, and 4.88 eV, respectively, all of which are better than those of the optimal AZO film made by PLD. This result indicates that N‐doped AZO films are better for use as window materials in polymer light‐emitting diodes. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.