Erection induced by Tx2-6 toxin of Phoneutria nigriventer spider: expression profile of genes in the nitric oxide pathway of penile tissue of mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Artificial Skin - Culturing of Different Skin Cell Lines for Generating an Artificial Skin Substitute on Cross-Weaved Spider Silk Fibres

Background: In the field of Plastic Reconstructive Surgery the development of new innovative matrices for skin repair is in urgent need. The ideal biomaterial should promote attachment, proliferation and growth of cells. Additionally, it should degrade in an appropriate time period without releasing harmful substances, but not exert a pathological immune response. Spider dragline silk from Nephila spp meets these demands to a large extent. Methodology/Principal Findings: Native spider dragline silk, harvested directly out of Nephila spp spiders, was woven on steel frames. Constructs were sterilized and seeded with fibroblasts. After two weeks of cultivating single fibroblasts, keratinocytes were added to generate a bilayered skin model, consisting of dermis and epidermis equivalents. For the next three weeks, constructs in co-culture were lifted on an originally designed setup for air/liquid interface cultivation. After the culturing period, constructs were embedded in paraffin with an especially developed program for spidersilk to avoid supercontraction. Paraffin cross-sections were stained in Haematoxylin & Eosin (H&E) for microscopic analyses. Conclusion/Significance: Native spider dragline silk woven on steel frames provides a suitable matrix for 3 dimensional skin cell culturing. Both fibroblasts and keratinocytes cell lines adhere to the spider silk fibres and proliferate. Guided by the spider silk fibres, they sprout into the meshes and reach confluence in at most one week. A well-balanced, bilayered cocultivation in two continuously separated strata can be achieved by serum reduction, changing the medium conditions and the cultivation period at the air/liquid interphase. Therefore spider silk appears to be a promising biomaterial for the enhancement of skin regeneration.

Interactions between Spider Silk and Cells - NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

Background: Several materials have been used for tissue engineering purposes, since the ideal matrix depends on the desired tissue. Silk biomaterials have come to focus due to their great mechanical properties. As untreated silkworm silk has been found to be quite immunogenic, an alternative could be spider silk. Not only does it own unique mechanical properties, its biocompatibility has been shown already in vivo. In our study, we used native spider dragline silk which is known as the strongest fibre in nature. Methodology/Principal Findings: Steel frames were originally designed and manufactured and woven with spider silk, harvesting dragline silk directly out of the animal. After sterilization, scaffolds were seeded with fibroblasts to analyse cell proliferation and adhesion. Analysis of cell morphology and actin filament alignment clearly revealed adherence. Proliferation was measured by cell count as well as determination of relative fluorescence each after 1, 2, 3, and 5 days. Cell counts for native spider silk were also compared with those for trypsin-digested spider silk. Spider silk specimens displayed less proliferation than collagen-and fibronectin-coated cover slips, enzymatic treatment reduced adhesion and proliferation rates tendentially though not significantly. Nevertheless, proliferation could be proven with high significance (p<0.01). Conclusion/Significance: Native spider silk does not require any modification to its application as a biomaterial that can rival any artificial material in terms of cell growth promoting properties. We could show adhesion mechanics on intracellular level. Additionally, proliferation kinetics were higher than in enzymatically digested controls, indicating that spider silk does not require modification. Recent findings concerning reduction of cell proliferation after exposure could not be met. As biotechnological production of the hierarchical composition of native spider silk fibres is still a challenge, our study has a pioneer role in researching cellular mechanics on native spider silk fibres.

Investigation on reproduction process in two species of crab Eriphia sebana and Ocypode saratan in the intertidal zone of Chabahar area

The present study with headline investigation on reproduction in two species of Crab Eriphia sebana and Ocypode saratan was carried out in the intertidal zone of Chabahar in thirteen month from December 2004 to December 2005. Checked samplings have been taken, 45 number Crab monthly from any four stations by manual or use trap. During this study the following subjects were measured: Temperature range and salinity, Measurable coast granule, Determination of sex ratio, Relations carapace width with carapace length, Carapace width with total body weight, Gonad weight, gonadosomatic index, condition factor, gastrosomatic index, investigation content in stomach, LM50, Growth parameters, plenty distribution length and width and gonad weight and total body weight. Studied on measurable coast granule were expressed that Ocypode saratan in Desalination station, were nest in soils equable sand and this quantity were confirmed in Pozm station. Sex ratio were assign in desalination area and Pozm M: F 0/44:0/56 and in Tiss and Chabahar M: F 0/45:0/55. Carapace length and carapace width (cm) and body weight (g) Furthest were designated in Ocypode saratan within carapace width sequential: female: 5/42-6/15-105/13 and male: 5/53-6/25-108/91 and in Eriphia sebana within Tiss area sequential: female: 5/12-5/94-110/21 and male 5/14-60/01-114/37. Have been linear relationship between carapace length and carapace width and equaled CW = a CL + b. Weight growth in two species were be modal and equaled BW= aCLb and increased Crab weight by built up carapace width. Maximum gonad weight in Ocypode saratan within desalination area in female have been outcome 3/39 and in male 0/84g and in Eriphia sebana extreme within Tiss during may in female were be 4/18 and in male 1/1g. Stomach content in Eriphia sebana were involved a black until half-purplish liquid and yellowish in Ocypode saratan. Stomach contents identifiable were being in four groups: Molluscoid, Crustacean, Plankton and Fish. Carapace width during the first year of maturation have been LM50:3/77 in Desalination area and LM50:3/92 in Pozm for Ocypode saratan and LM50:4/26 in Tiss and LM50:4/62 in Chabahar. Ability spawning in Eriphia sebana within Tiss has been CW=4/17cm and in Ocypode saratan within Desalination area CW=4/23cm. Maximun value of Loo for Eriphia sebana was equal 59/67 and growth factor K=0/68 within Tiss and Loo =61/64 , K=0/65 for Ocypode saratan within Desalination area. Maximun GSI and GI have been within Desalination area and Tiss and minimum within Pozm and Chabahar. The maturity stages of two species were classifed into six stages. Review on GSI, CF have been showed that relation with temperature and salinity and definer in two species have been spawned in two period that Maximun in spring premier than autumn.

Mixture of time-dependent growth models with an application to blue swimmer crab length-frequency data

Understanding how aquatic species grow is fundamental in fisheries because stock assessment often relies on growth dependent statistical models. Length-frequency-based methods become important when more applicable data for growth model estimation are either not available or very expensive. In this article, we develop a new framework for growth estimation from length-frequency data using a generalized von Bertalanffy growth model (VBGM) framework that allows for time-dependent covariates to be incorporated. A finite mixture of normal distributions is used to model the length-frequency cohorts of each month with the means constrained to follow a VBGM. The variances of the finite mixture components are constrained to be a function of mean length, reducing the number of parameters and allowing for an estimate of the variance at any length. To optimize the likelihood, we use a minorization–maximization (MM) algorithm with a Nelder–Mead sub-step. This work was motivated by the decline in catches of the blue swimmer crab (BSC) (Portunus armatus) off the east coast of Queensland, Australia. We test the method with a simulation study and then apply it to the BSC fishery data.

Crabbing into an uncertain future: the blue swimming crab fisher in coastal town of Eastern Philippines

This research project sought to find out the socio-economic status of the small-scale fishers of the blue swimming crab (Portunuspelagicus) in Samar, considering the diminishing volume of catch of the species in the recent years. Using a blend of quantitative and qualitative methods, the study employed an interview schedule, focus group discussion (FGD) and observation in collecting data not only from the fishers but also from other sectors directly involved in the blue swimming crab industry.

Spermathecae of the mangrove crab Ucides cordatus: a histological and histochemical view

Ucides cordatus is the most commercially, important mangrove crab in Brazil. In spite of its economic importance, there are few studies of its reproduction, in particular the female reproductive system. The present study describes the histology and histochemistry of the spermathecae of U. cordatus. Adult females were can lit monthly from July 2004 through June 2005, at Iguape, State of São Paulo. The crabs were anaesthetized, and their spermathecae removed and fixed in Davidson's fluid, following the histological routine for paraffin. The slides were stained with HE, xylidine Ponceau, PAS, alcian blue (pH 1.0 and 2.5), Sudan black B and picrosirius-haematoxylin. Histologically, the spermathecae possesses a capsule of conjunctive tissue, rich in collalgen fibres, which surrounds the secretory columnar epithelium. In the lumen, individual sperm packets are not observed; the spermatophores are intermixed with the seminal fluid and secretions of the spermathecae itself. A large proportion of the free spermatozoids and spermatophores are arranged in homogeneous masses in the proximal part of the spermathecae. The secretion produced by the columnar epithelium appears to promote the movement of the gametes to the fertilization chamber, in a ventral position, allowing fertilization of the oocytes. Histochemically, the secretion produced by the columnar epithelium was strongly positive for neutral polysaccharides, positive for acid polysaccharides, and weakly positive for proteins and lipids. Tills secretion forms a glycoprotein matrix which is associated with maintenance of the spermatophores, which can remain stored for long periods.

Project Flappy Crab : an edugame for music learning

This chapter discusses some possibilities of gamification and remixing processes for music education. It also analyzes the concepts of gamification, mashup/remix and presents its possible usage in education—music teaching—through the development of the project/educational game FLAPPY CRAB. The chapter begins with a brief introduction to the concepts listed above, trying to consider them in the school context. After that, we will make the summary presentation of the music educational game FLAPPY CRAB, a clone of the GEARS Studios Flappy Bird, developed for mobile devices and other platforms with the UNITY 3D© game engine. In this chapter we’ll talk yet, albeit briefly, about the game engine used in the development of this educational application. This educational game aims to assess the possible impacts that its use has on learning and skill development related to auditory memory, qualitative discrimination of musical sound height (pitch—in the range of an octave with a central point in 440 Hz), visual identification of musical notation symbols and its relative organization according to the grammar rules of traditional music spelling. The game has been tested by a group of approximately 30 teenagers over a period of about 6 months, over which data was collected. In this chapter, we will present a review of the preliminary data collected to date.

Growth of the mangrove crab Ucides cordatus (Brachyura, Ocypodidae)

During monthly samplings between September 1998 and August 2000. 3,660 specimens of Ucides cordatus (Linnaeus, 1763) (2054 males and 1606 females) were obtained and examined for size (CW carapace width) to determine growth-age equations for each sex. This species showed a slower growth, with a marked seasonal oscillation, in females as compared to males, suggesting application of the seasonal and nonseasonal von Bertalanffy growth model, respectively. CW&PROP; and k constant were closely similar for the two sexes (CW&PROP; (male) = 90.3 mm: CW&PROP; (female) = 88.6 mm; k(male) = 0.28; k(female) = 0.26). The age at sexual maturity was estimated to be around 3 years, while the age at legal size (CW = 60 mm) was 3.8 and 4.7 years for males and females, respectively. In the laboratory, juvenile stages did not show differences in growth rates under the same temperature and photoperiod conditions.

Growth of the speckled swimming crab, Arenaeus cribrarius (Lamarck, 1818) (Crustacea, Brachyura, Portunidae), in Ubatuba (SP), Brazil

A total of 2629 individuals of Arenaeus cribrarius (1293 males and 1336 females) were captured in Ubatuba (SP), from August 1996 to July 1997. Individuals were distributed in 5 mm size class carapace width (CW), to verify sex-specific growth-age equations. The Von Bertalanffy model was chosen to determine the growth rate and expressed by CW=120.52[1-e(-1.80t)] for males and CW=100.81[1-e(-1.60t)] for females. The age estimated for the first juvenile stage (t(o)) was 6.1 and 8.3 days for males and females, respectively. The maximum age determined was 1.8 years for males and 2 years for females, which correspond to a maximum size of 115.8 and 96.7 mm, respectively. The maximum size (CWmax) estimated using 95% of asymptotic size was 114.5 mm for males and 95.8 mm for females. Males have a precocious sexual maturity (5 months) when compared to females (6.8 months). The growth rate and size of A. cribrarius are higher than other portunid species, with great interest for aquaculture.

Estimating abundance and spatial distribution patterns of the bubble crab Dotilla fenestrata (Crustacea : Brachyura)

The bubble crab Dotilla fenestrata forms very dense populations on the sand flats of the eastern coast of Inhaca Island, Mozambique, making it an interesting biological model to examine spatial distribution patterns and test the relative efficiency of common sampling methods. Due to its apparent ecological importance within the sandy intertidal community, understanding the factors ruling the dynamics of Dotilla populations is also a key issue. In this study, different techniques of estimating crab density are described, and the trends of spatial distribution of the different population categories are shown. The studied populations are arranged in discrete patches located at the well-drained crests of nearly parallel mega sand ripples. For a given sample size, there was an obvious gain in precision by using a stratified random sampling technique, considering discrete patches as strata, compared to the simple random design. Density average and variance differed considerably among patches since juveniles and ovigerous females were found clumped, with higher densities at the lower and upper shore levels, respectively. Burrow counting was found to be an adequate method for large-scale sampling, although consistently underestimating actual crab density by nearly half. Regression analyses suggested that crabs smaller than 2.9 mm carapace width tend to be undetected in visual burrow counts. A visual survey of sampling plots over several patches of a large Dotilla population showed that crab density varied in an interesting oscillating pattern, apparently following the topography of the sand flat. Patches extending to the lower shore contained higher densities than those mostly covering the higher shore. Within-patch density variability also pointed to the same trend, but the density increment towards the lowest shore level varied greatly among the patches compared.

Population structure and growth of the hermit crab Pagurus brevidacylus (Anomura : Paguridae) from the northern coast of São Paulo, Brazil

The hermit crab Paguras brevidactylus (Crustacea: Anomura: Paguridea) from the infralittoral area of Anchieta Island, Ubatuba, was characterized by population Structure (size, sex ratio, reproduction and recruitment) and growth. Animals were collected monthly during 1999 by SCUBA diving. A total of 1525 individuals was collected (633 males and 892 females), 695 of them were ovigerous females. Overall sex ratio was 0.7:1 in favour of females. The crabs showed a unimodal distribution with males significantly larger than females. Ovigerous females were collected during all months and in high percentages from 1.0 mm of shield length, demonstrating intense and Continuous reproduction. The longevity was approximately 24 months for males and 18 for females, which showed larger growth rate and reached sexual maturity earlier (two months) than males. The low number of males in this Population may be due to the longer life span. Moreover, the sexual dimorphism favours males during the intra- and interspecific fights by shell, food, reproduction and territory. Females demonstrated a short life cycle and intense reproduction.

DEVELOPMENT OF A NUCLEIC ACID-BASED SPECIFIC GROWTH MODEL FOR JUVENILE BLUE CRAB, CALLINECTES SAPIDUS

The evaluation and identification of habitats that function as nurseries for marine species has the potential to improve conservation and management. A key assessment of nursery habitat is estimating individual growth. However, the discrete growth of crustaceans presents a challenge for many traditional in situ techniques to accurately estimate growth over a short temporal scale. To evaluate the use of nucleic acid ratios (R:D) for juvenile blue crab (Callinectes sapidus), I developed and validated an R:D-based index of growth in the laboratory. R:D based growth estimates of crabs collected in the Patuxent River, MD indicated growth ranged from 0.8-25.9 (mg·g-1·d-1). Overall, there was no effect of size on growth, whereas there was a weak, but significant effect of date. These data provide insight into patterns of habitat-specific growth. These results highlight the complexity of the biological and physical factors which regulate growth of juvenile blue crabs in the field.

Population structure and sexual maturity of the calico box crab Hepatus epheliticus Linnaeus (Brachyura, Hepatidae) from Yucatan Peninsula, Mexico

The calico box crab Hepatus epheliticus is an abundant species from shallow and continental shelf waters of the Atlantic coast of USA and Mexico. Information about population structure and sexual maturity is absent, even though this crab is caught to be used as bait for the octopus fishery in the Campeche Bank, Mexico. In order to achieve such information, a total of 768 individuals were collected from January to March 2010 through baited traps installed in the Yucatan Peninsula, Mexico. Our results showed that sex ratio is biased towards more males than females (1:0.55), contradicting to that reported in other brachyuran crabs. The absence of ovigerous females suggests that they did not enter into the traps during embryogenesis. Males reached a larger maximum size than females (64.0 +/- 6.15 and 58.4 +/- 5.60 mm carapace width, respectively). The general scheme of growth being positive allometric throughout ontogeny of both sexes. Males presented a transition phase from juveniles to adult corresponding to the puberty moult. The estimation of the onset of functional sexual maturity revealed a steady situation for the population, with 21.5 and 13.8% of males and females, respectively, morphologically immature at the time of catch. This study constitutes the first report on population structure and sexual maturity in a population of the calico box crab H. epheliticus.