External Morphology of the egg of Spodoptera Littoralis (Lep. Noctuidae)

En el present article es descriu l'estructura i ornamentació de la superficie del còrion i de l'aparell micropilar, tal com es veu amb el microscopi electrònic d'escandallatge. La regió micropilar presenta al mig un espai circular amb 5 o 6 orificis micropilars disposats perifèricament. D'aquesta zona circular surten radialment unes 10 o 11 estructures en forma de pètal, que originen el típic micròpil en roseta dels lepidòpters. El còrion està estructurat uniformement en àrees poligonals regulars —pentagonals i hexagonals— d'uns 1.030 um2 de superfície, delimitades per un relleu on es localitzen els aeròpils, que es troben habitualment en els punts d'intersecció de tres polígons.En el presente artículo se describe la estructura y ornamentación de la superficie del corion y del aparato micropilar con el microscopio electrónico de barrido. La región micropilar presenta, en su centro, un espacio circular con 5 ó 6 orificios micropilares dispuestos perifericamente. De esta zona circular parten radialmente unas 10 ó 11 estructuras en forma de pétalo, que dan origen al típico micrópilo "en roseta" de los lepidópteros. El corion está estructurado uniformement en áreas poligonales regulares —pentagonales y hexagonales— de unos 1.030 um2 de superficie, delimitadas por un relieve, donde se localizan los aerópilos, situados habitualmente en los puntos de intersección de tres polígonos

La formació del vitel.le en els crustacis. Revisió preliminar

This is a summarized review about some aspects of the yolk formation in crustacean. At the beginning of vitellogenesis, the crustacean oocyte is characterized by a large number of vesicles in the cytoplasm. The vesicles have a multiple origin, although the majority develop directly from the rough endoplasmic reticulum and the Golgi complex (endogenous phase of vitellogenesis). In a subsequent phase, a series substances, principally glucoproteins and lipoproteins, are incorporated into the ooplasma by means of micropinocytosis (exogenus phase of vitellogenesis). The result of this process is the formation of one or several types of yolk platelet

Spermiogenesis and biflagellate spermatozzoon of the teleost fish Lampanyctus crocodilus (Myctophiformes, Myctophidae): ultrastructure and characterization of its sperm basic nuclear proteins

We undertook an ultrastructural study of the spermiogenesis of the lanternfish Lampanyctus crocodilus (Myctophiformes, Myctophidae) with special emphasis on the condensation of chromatin and the biochemical characterisation of its sperm nuclear basic proteins (SNBPs). The round head of the early spermatid of L. crocodilus develops into a curved conical-shaped head in the spermatozoon. Two flagella, present even in the spermatid, are inserted laterally at the convex side of the sperm head. Both flagella possess an axoneme with a 9 + 0 instead of the typical 9 + 2 axonemal structure. Mitochondria undergo a characteristic redistribution during spermiogenesis. A reduced number of them are present lying away from the centrioles at both ends of the concave side of the sperm head. During the chromatin condensation stages in spermiogenesis, fibrogranular structures with granules of 25 ± 5 and 50 ± 5 nm can be observed in the early spermatid and develop into larger granules of about 150 ± 50 nm in the middle spermatid. The latter granules coalesce during the transition to the advanced spermatid and spermatozoon giving rise to highly condensed chromatin in the sperm cell. Protamines are the main SNBPs associated with this chromatin; however, they are unusually large and correspond to the largest protamines described in fish to date. Small stoichiometric amounts of histones and other basic proteins coexist with these protamines in the spermatozoon

L'Espermatogènesi en els crancs (Crustacea, Brachyura) : un model atípic de condensació del nucli espermàtic

Els espermatozoides dels crustacis decàpodes es caracteritzen per tenir un nucli amb cromatina poc condensada, un acrosoma complex i un citoplasma reduït sense flagel. Es disposa d'una àmplia informació sobre la ultraestructura i variabilitat morfològica dels espermatozoides dels crustacis decàpodes, i particularment dels braquiürs, ja que la seva especificitat ha permès utilitzar-los com a caràcter filogenètic, però el procés d'espermatogènesi i la naturalesa de les proteïnes associades a la cromatina dels espermatozoides són qüestions encara no resoltes. En aquest treball hem tractat aquestes dues qüestions utilitzant com a model dues espècies de braquiürs: la cabra de mar, Maja brachydactyla, i el bou de mar, Cancer pagurus. D'aquesta manera, hem realitzat una breu descripció de la morfologia de l'aparell reproductor masculí de M. brachydactyla, hem localitzat el lloc on es desenvolupa l'espermatogènesi i hem descrit la formació dels espermatòfors que són transferits a la femella. A continuació, hem comparat el procés d'espermatogènesi de M. brachydactyla, amb els estudis fets en altres espècies, considerant de manera preferent la formació de l'acrosoma. Finalment, hem descrit diferents aspectes del nucli dels espermatozoides dels braquiürs, fent un estudi específic de les proteïnes associades al nucli de l'espermatozoide de C. pagurus.Decapods crustacean spermatozoa are characterized by a nucleus containing a low condensed chromatin, a complex acrosome and reduced cytoplasm lacking of flagellum. Large information on ultrastructure and morphological variability of the spermatozoa is available in crustacean particularly, in Brachyuran, since their specificity has been used as a phylogenetic character. However, the spermatogenesis and the nature of the proteins associated to chromatin in the nucleus of the spermatozoa are still unclear. In the present study, we have dealt both topics using two brachyuran species as model: the spider crab, Maja brachydactyla and the edible crab, Cancer pagurus. Thus, we have briefly described the morphology of the male reproductive system of M. brachydactyla in order to locate the spermatogenesis process and to describe the formation of the spermatophore transferred to the female. Finally, we have also described and compared with previous studies, the spermatogenesis process of M. brachydactyla with special attention to the acrosomal vesicle. We have finished our study describing the different characters presented in the nucleus of the spermatozoa, with special reference to the proteins associated to chromatin in C. pagurus

Human astrovirus MLB replication in vitro: persistence in extraintestinal cell lines

MLB astroviruses were identified 10 years ago in feces from children with gastroenteritis of unknown etiology and have been unexpectedly detected in severe cases of meningitis/encephalitis, febrile illness of unknown etiology, and respiratory syndromes. The aim of this study was to establish a cell culture system supporting MLB astrovirus replication. We used two clinical strains to infect several cell lines, an MLB1 strain from a gastroenteritis case, and an MLB2 strain associated with a neurologic infection. Efforts to propagate the viruses in the Caco-2 cell line were unsuccessful. In contrast, we identified two human nonintestinal cell lines, Huh-7 and A549, permissive for both genotypes. After serial passages in the Huh-7.5 cell line, the adapted strains were able to establish persistent infections in the Huh-7.5, Huh-7AI, and A549 cell lines, with high viral loads (up to 10 log10 genome copies/ml) detected by quantitative reverse transcription-PCR (RT-qPCR) in the culture supernatant. Immunofluorescence assays demonstrated infection in about 10% of cells in persistently infected cultures. Electron microscopy revealed particles of 32 to 33 nm in diameter after negative staining of cell supernatants and capsid arrays in ultrathin sections with a particularly high production in Huh-7.5 cells. Interferon (IFN) expression by infected cells and effect of exogenous IFN varied depending on the type of infection and the cell line. The availability of a cell culture system to propagate MLB astroviruses represents a key step to better understand their replicative cycle, as well as a source of viruses to conduct a wide variety of basic virologic studie

Pathogenicity and virulence of Hepatitis A virus.

Hepatitis A is an acute infection of the liver, which is mostly asymptomatic in children and increases the severity with age. Although in most patients the infection resolves completely, in a few of them it may follow a prolonged or relapsed course or even a fulminant form. The reason for these different outcomes is unknown, but it is generally accepted that host factors such as the immunological status, age and the occurrence of underlaying hepatic diseases are the main determinants of the severity. However, it cannot be ruled out that some virus traits may also contribute to the severe clinical outcomes. In this review, we will analyze which genetic determinants of the virus may determine virulence, in the context of a paradigmatic virus in terms of its genomic, molecular, replicative, and evolutionary feature

The Critical role of codon composition on the translation efficiency robustness of the Hepatitis A virus capsid

Hepatoviruses show an intriguing deviated codon usage, suggesting an evolutionary signature. Abundant and rare codons in the cellular genome are scarce in the human hepatitis A virus (HAV) genome, while intermediately abundant host codons are abundant in the virus. Genotype-phenotype maps, or fitness landscapes, are a means of representing a genotype position in sequence space and uncovering how genotype relates to phenotype and fitness. Using genotype-phenotype maps of the translation efficiency, we have shown the critical role of the HAV capsid codon composition in regulating translation and determining its robustness. Adaptation to an environmental perturbation such as the artificial induction of cellular shutoff not naturally occurring in HAV infection involved movements in the sequence space and dramatic changes of the translation efficiency. Capsid rare codons, including abundant and rare codons of the cellular genome, slowed down the translation efficiency in conditions of no cellular shutoff. In contrast, rare capsid codons that are abundant in the cellular genome were efficiently translated in conditions of shutoff. Capsid regions very rich in slowly translated codons adapt to shutoff through sequence space movements from positions with highly robust translation to others with diminished translation robustness. These movements paralleled decreases of the capsid physical and biological robustness, and resulted in the diversification of capsid phenotypes. The deviated codon usage of extant hepatoviruses compared with that of their hosts may suggest the occurrence of a virus ancestor with an optimized codon usage with respect to an unknown ancient host

DNA-interacting proteins in the spermiogenesis of the mollusc Murex brandaris

Sperm chromatin of Murex brandaris (a neogastropod mollusc) undergoes a series of structural transitions during spermiogenesis. The DNA-interacting proteins responsible for these changes as well as the mature protamines present in the ripe sperm nucleus have been characterized. The results reveal that spermiogenic nuclear proteins are protamine precursors that are subjected to a substantial number of small N-terminal deletions that gradually modify their overall charge. The composition of mature protamines is remarkably simple in turn, promoting an efficient and extremely tight packaging of DNA. The pattern of spermiogenic chromatin condensation in M. brandaris clearly departs from that corresponding to vertebrate chromatin

Improving virus production through quasispecies genomic selection and molecular breedings

Virus production still is a challenging issue in antigen manufacture, particularly with slow-growing viruses. Deep-sequencing of genomic regions indicative of efficient replication may be used to identify high-fitness minority individuals suppressed by the ensemble of mutants in a virus quasispecies. Molecular breeding of quasispecies containing colonizer individuals, under regimes allowing more than one replicative cycle, is a strategy to select the fittest competitors among the colonizers. A slow-growing cell culture-adapted hepatitis A virus strain was employed as a model for this strategy. Using genomic selection in two regions predictive of efficient translation, the internal ribosome entry site and the VP1-coding region, high-fitness minority colonizer individuals were identified in a population adapted to conditions of artificially-induced cellular transcription shut-off. Molecular breeding of this population with a second one, also adapted to transcription shut-off and showing an overall colonizer phenotype, allowed the selection of a fast-growing population of great biotechnological potential