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The presence of vertebrate-like steroids, steroidogenic enzymes and steroid receptors has been reported exclusively in cephalopods gonads. The role played by these steroids has been also recently investigated. We here give evidence of steroidogenic activity in the brain of cephalopods. The activity of two key steroidogenic enzymes: 3beta-hydroxysteroid dehydrogenase (HSD) and 17beta-HSD is present in the lobes of the nervous system of both Sepia and Octopus. Such enzymes convert pregnenolone to progesterone and androstenedione to testosterone respectively. Binding experiments seem to assign a functional role to the androgens in the brain of cephalopods. According to the present results, the absence of any progesterone binding moiety supports the hypothesis that progesterone may be a metabolite product along the steroidogenic chain leading to androgens. The presence of steroidogenic enzymes in specific lobes of the central nervous system is discussed in terms of the possible role that steroids can play in the sexual differentiation of the brain and in influencing some coded behaviours of cephalopods, such as learning processes.

Impulsive waves impinging over steep slopes of dams or embankments may produce erosion with intense sediment transport and relevant morphological changes during fast geomorphic transients. In order to be employed as valuable tools for risk assessment and hazard mitigation, the performance of current morphodynamic models in reproducing the physical evolution of such processes has to be carefully assessed. However, despite the inherent interest of the topic, the availability of suitable experimental data is relatively scarce. For this reason, additional tests, carried out under controlled conditions, may furnish the basis for an improvement in the knowledge of the bed evolution. The obtained experimental results constitute also an important resource to evaluate the numerical models performance. To this aim, a set of laboratory experiments, in which the impact of a dam break wave on an erodible slope is reproduced, has been designed and conducted. In the paper some preliminary results are presented and discussed.

The paper analyzes the influence of resistance coefficient dependence on flow variables, viscosity and wall roughness onto the spatial development of roll-waves in the initial phase. To this aim, two models, based on time-asymptotic solutions of linearized St. Venant equations, subjected to either impulsive or oscillating perturbation, have been modified by including the dependence of the resistance coefficient on flow conditions. Independently on the perturbation type, the results show that the hypothesis of constant resistance coefficient may lead to a non-negligible underestimation of the instability spatial growth rate. Theoretical achievements are compared with results of a fully non-linear model for several combinations of Froude and Reynolds numbers and relative roughness values. The comparison shows that the spatial linear analysis with a suitable oscillating perturbation allows to accurately reproduce both spatial growth rate and celerity of the roll-waves. Such findings have to be properly accounted for in defining criteria for roll-waves occurrence in natural and manmade channels.

Summary This paper analyses conditions for roll-waves formation in homogeneous granular flows with the aim of predicting their occurrence in finite length channels. With reference to a wide class of constitutive equations of the flowing medium, the response of the linearized one-dimensional flow model to a pointwise instantaneous disturbance is studied and a closed form of the Green’s function, in both stable and unstable conditions, is provided. The exponential growth of the disturbance peak is used to predict the spatial evolution of roll-waves in dry flows of dense granular materials and a criterion for the minimum channel length necessary to appreciate roll-wave development is proposed. The comparison among theoretical and experimental data available in the literature appears promising.

Urban drainage networks (UDNs) represent important infrastructures to protect and maintain community health and safety. For these reasons, technicians and researcher are focusing more and more on topics related to vulnerability, resilience and monitoring for controlling illicit intrusions, contaminant and pathogenic spread. In the last years the complex network theory (CNT) is attracting attention as a new, useful and structured approach to analyze urban systems. The aim of this work is to evaluate potentialities of CNT approaches for UDNs vulnerability assessment and monitoring system planning. Limits and potentialities of applicability of CNT tools to UDNs are first provided evaluating the performances of standard centrality metrics. Then, it is proposed the use of tailored metrics embedding prior information, as intrinsic relevance of each node and pipe flow direction, which derive from the Horton's hierarchy and geometric data (pipe slope), respectively, without performing hydraulic simulations. The analysis is applied on two schematic literature networks of different complexity and to a real case-study. The results suggest that vulnerability/resilience, monitoring design, contaminant and pathogenic spreads can be effectively analyzed using tailored metrics. Therefore, the proposed approach represents a complementary tool respect the more complex and computationally expensive methodologies and it is particular useful for large complex networks.

Mud flows represent one of the major causes of natural hazards in mountain regions. Similarly to debris flows, they consist of a hyper-concentrated mixture of water and sediments flowing down a slope and may cause serious damages to people and structures. The present paper investigates the force produced by a dam-break wave of mud impacting against a rigid wall. A power-law shear-thinning model is used to describe the rheology of the hyper-concentrated mixture. A one-dimensional shallow water model is adopted and a second-order Finite Volume scheme is employed to numerically solve the governing equations. The results indicate that depending on the fluid rheological parameters and on the bottom slope, there exists a minimum value of the wall distance above which the peak force does not exceed the asymptotic value of the hydrostatic final condition. For two different values of the channel slope, the dimensionless value of this lower bound is individuated for several values of the power-law exponent and of a dimensionless Basal Drag coefficient. An estimation of the maximum peak force for wall distance smaller than the minimum value is also provided.

The present study reports the presence of progesterone, testosterone and 17 beta-estradiol and their corresponding binding proteins in the reproductive system of Octopus vulgaris Lam (phylum Mollusca, subphy lum Cephalopoda). These sex hormones occur in testis, vas deferens, seminal vesicle, prostate and Needham's sac. The hemolymph also contains a small, but significant, amount of these hormones and their carrier proteins. Among various tissues of the reproductive system, the seminal vesicle possesses the highest concentration of progesterone (4.8 ng/g tissue). The testis is the organ which contains the highest amount of testosterone (5.2 ng/g) whereas the prostate is the organ which contains the highest amount of 17 beta-estradiol (0.92 ng/g). The presence of these hormones has been ascertained by a radioimmunoassay method, an immunoenzymatic method and by a chemical (HPLC) method. Scatchard studies indicated that vas deferens and seminal vesicle contain specific sex steroid binding molecules at affinity levels comparable to those of vertebrate steroid receptors (0.5-5.0 pmol/g protein). In addition to the presence of the hormones, the delta 5,3 beta hydroxysteroid dehydrogenase, the key enzyme of steroidogenesis, also is found in testis. From a phylogenetic point of view, these findings are very interesting because they indicate a common origin of a sex hormonal system between Mollusca Cephalopoda and Vertebrates. (C) 1996 Academic Press, Inc.

A methodological framework for the estimation of the expected value of hourly peak water demand factor and its dependence on the spatial aggregation level is presented. The proposed methodology is based on the analysis of volumetric water meter measurements with a 1-h time aggregation, preferred by water companies for monitoring purposes. Using a peculiar sampling design, both a theoretical and an empirical estimation of the expected value of the peak factor and of the related standard error (confidence bands) are obtained as a function of the number of aggregated households (or equivalently of the number of users). The proposed methodology accounts for the cross-correlation among consumption time series describing local water demand behaviours. The effects of considering a finite population is also discussed. The framework is tested on a pilot District Metering Area with more than 1000 households equipped with a telemetry system with 1-h time aggregation. Results show that the peak factor can be expressed as a power function tending to an asymptotic value greater than one for the increasing number of aggregated households. The obtained peak values, compared with several literature studies, provide useful indications for the design and management of secondary branched pipes of water distribution systems.

Climate change is exposing more and more frequently flood prone areas to potential casualties and damages. The capability of the flow to carry relevant quantities of sediments interacts with the presence of obstacles in flood-inundated areas and contributes to the increase the related hazard, constituting a relevant concern in the framework of risk analysis. Unfortunately, existing literature on this topic is rather scarce, especially for the features of sediment transport and the forces acting on rigid obstacles. In the paper, a recent two-phase shallow-water morphodynamical model, particularly suited to the analysis of fast geomorphic transients, is applied to the numerical simulation of the propagation of a dam-break wave over an erodible floodplain in presence of a rigid obstacle. The geometry of the test-case is inspired to a recent fixed-bed study reported in the literature, for which extensive experimental and numerical data concerning the flow field and the dynamic loading against the obstacle are available. Results of the numerical simulations contribute to highlight the effect of the obstacle on the changes in the bottom topography, along with the subsequent change on the loading condition on it. * Corresponding author Engineering EPiC SeriesinEngineering Volume3,2018,Pages564571 HIC 2018.13thInternational Conference onHydroinformatics G.

The adoption of the Diffusive Wave (DW) instead of the Full Dynamic (FD) model in the analysis of mud flood routing within the shallow-water framework may provide a significant reduction of the computational effort, and the knowledge of the conditions in which this approximation may be employed is therefore important. In this paper, the applicability of the DW approximation of a depth-integrated Herschel–Bulkley model is investigated through linear analysis. Assuming as the initial condition a steady hypocritical decelerated flow, induced by downstream backwater, the propagation characteristics of a small perturbation predicted by the DW and FD models are compared. The results show that the spatial variation on the initial profile may preclude the application of DW model with a prescribed accuracy. Whenever the method is applicable, the rising time of the mud flood must satisfy additional constraints, whose dependence on the flow depth, along with the Froude number and the rheological parameters, is deeply analyzed and discussed.

Illicit intrusions in Sewer Systems (SSs), modifying the wastewater characteristics, may create problems to the treatment plant and/or to the final recipient water body. For this reason, the source identification (SI) problem is becoming an important issue also in SSs. For large real systems, the computational burden might make the SI methodologies impractical. In this paper a pre-screening procedure, based on the pollution matrix concept, is introduced and applied before the SI methodology. Selecting a group of possible candidate nodes and cutting consequently the scheme, a significant improvement both in terms of time and the accuracy is obtained.

A core problem associated with the water quality monitoring in the sewer system is the optimal placement of a limited number of monitoring sites. A methodology is provided for optimally design water quality monitoring stations in sewer networks. The methodology is based on information theory, formulated as a multi-objective optimization problem and solved using NSGA-II. Computer code is written to estimate two entropy quantities, namely Joint Entropy, a measure of information content, and Total Correlation, a measure of redundancy, which are maximized and minimized, respectively. The test on a real sewer network suggests the effectiveness of the proposed methodology.

Disinfection of drinking water through chlorine is commonly used in many countries for reducing pathogenic infection problems, but it may pose a risk to human health due to the formation of some byproducts, such as trihalomethanes (THMs). For an adequate management of the disinfection process, it is useful to have models capable of simulating THM concentrations in water supply systems. However, a crucial aspect is the reliability of their performances when used on real systems and the knowledge of the uncertainty of their predictions. The objective of the present paper is to investigate the simulation of THM formation through calibrated kinetic models, along with the assessment of the measurement uncertainty on the parameters' estimate and on the model predictions. In particular, a first order, a second order and a two-phase kinetic model are considered, while the effect of measurement uncertainty is mainly investigated through the first order second moment (FOSM) method. The analysis performed on a real case study shows that the three considered models have similar performances in terms of predicted concentrations, which suggests using the simplest first order one. Moreover, the FOSM results reveal that the first order model is also less affected by measurement uncertainty.

Simplified wave models - such as kinematic, diffusion and quasi-steady - are widely employed as a convenient replacement of the full dynamic one in the analysis of unsteady open-channel flows, and especially for flood routing. While their use may guarantee a significant reduction of the computational effort, it is mandatory to define the conditions in which they may be confidently applied. The present paper investigates the applicability conditions of the kinematic, diffusion and quasisteady dynamic shallow wave models for mud flows of power-law fluids. The power-law model describes in an adequate and convenient way fluids that at low shear rates fluids do not posses yield stress, such as clay or kaolin suspensions, which are frequently encountered in Chinese rivers. In the framework of a linear analysis, the propagation characteristics of a periodic perturbation of an initial steady uniform flow predicted by the simplified models are compared with those of the full dynamic one. Based on this comparison, applicability criteria for the different wave approximations for mud flood of power-law fluids are derived. The presented results provide guidelines for selecting the appropriate approximation for a given flow problem, and therefore they may represent a useful tool for engineering predictions.

The Storm Water Management Model (SWMM) is considered one of the standard tools for modeling sewer systems, but it does not have the programmer's toolkit. In this work a toolkit library is presented, which has been built for running wastewater quality simulations from an outside environment. An example of the developed Toolkit use is furnished applying it to a pollution source identification (PSI) problem, expressed as an optimization problem. The presented example shows the capability of the Toolkit in providing a useful tool for performing network analysis from other applications and demonstrates the good performance of the proposed PSI methodology.

"The occurrence of roll-waves in mud-flows is commonly investi-gated starting from the formulation of the marginal stability threshold of a linearized one-dimensional viscoplastic (shear-thinning) flow model. Since for this non-newtonian rheological model such a threshold may oc-cur in hypocritical flow, the downstream boundary condition may have a non-negligible effect on the spatial growth\/decay of the perturbation. The paper presents the solution of the 1D linearized flow of a Herschel & Bulkley fluid in a channel of finite length, in the neighbourhood of a hypocritical base uniform flow. Both linearly stable and unstable condi-tions are considered. The analytical solution is found applying the Laplace transform method and obtaining the first-order analytical expressions of the upstream and downstream channel response functions in the time domain. The effects of both the yield stress and the rheological law exponent are discussed, recovering as particular cases both power-law and Bingham fluids. The theoretical achievements may be used to extend semi-empirical criteria commonly employed for predicting roll waves occurrence in clear-water even to mud-flows."

In this study we have investigated the role of 17beta-estradiol and progesterone in the reproduction of the crayfish Cherax albidus by using vitellogenin (VTG) as a biomarker. Early-vitellogenic (EV), full-vitellogenic (FV), and non-vitellogenic (NV) females of Cherax albidus were treated with 17beta-estradiol, progesterone, or both for 4 weeks. Levels of VTG mRNA in the hepatopancreas were detected by RT-PCR. The PCR product was sequenced and showed 97% homology with Cherax quadricarinatus VTG. 17beta-estradiol was more effective than progesterone and 17beta-estradiol plus progesterone in increasing the vitellogenin transcript in the hepatopancreas of EV and FV females. On the contrary, progesterone was more effective than 17beta-estradiol and 17beta-estradiol plus progesterone in increasing the vitellogenin concentration in the hemolymph of EV and FV females. Hepatopancreas histology and fatty acid composition of females injected with hormones showed major modifications. No effects were registered in NV females. In conclusion, 17beta-estradiol and progesterone influence VTG synthesis, although our data indicate that they act through different pathways and are not effective until the proper hormonal environment is established, as demonstrated by their inefficacy in NV females.

To examine the neurochemistry underlying the firing of the RPeD1 neuron in the respiratory central pattern generator of the pond snail, Lymnaea stagnalis, we examined electrophysiologically and pharmacologically either "active" or "silent" preparations by intracellular recording and pharmacology. GABA inhibited electrical firing by hyperpolarizing RPeD1, while picrotoxin, an antagonist of GABA(A) receptors, excited silent cells and reversed GABA-induced inhibition. Action potential activity was terminated by 1 mM glutamate (Glu) while silent cells were depolarized by the GluR agonists, AMPA, and NMDA. Kainate exerted a complex triphasic effect on membrane potential. However, only bath application of AMPA desensitized the firing. These data indicate that GABA inhibits RPeD1 via activation of GABA(A) receptors, while Glu stimulates the neuron by activating AMPA-sensitive GluRs.

Waste water management plant protection is a major concern for water cycle management entities. The rapid identification and possible localization of anomalous or even malicious waste liquids immissions may allow for undertaking pollution risk mitigation actions (e.g. using of ancillary basins) and reduce maintainance costs. Pervasive monitoring of the transport network is hence needed although economic and technical issues prevent its implementation. The SIMONA project is aimed to design, deploy and test an integrated, intelligent, pervasive monitoring infrastructure based on a network of low cost/low mainteinance quali-quantitative multisensor nodes. A scalable data processing facility permit the ingestion and the processing of the data stream while a set of models provide for quali-quantitative forecasting increasing the manager situational awareness about the smart infrastructure. All the information is made available via a GIS based Web HCI.

The study of unsteady shallow-layer flows, as those occurring during floods, may be performed through either fully dynamic models or using simplified momentum equations (e.g. kinematic, diffusion and quasi-steady approximations). Among the latter, kinematic approximation may provide a significant reduction of the computational effort, which represents a key advantage in time-critical applications. The proper application of this simplified model is however generally subjected to some limitations, which the modeler has to know and accurately verify to avoid errors. The present paper aims to investigate the applicability range of the kinematic approximation to shallow flows, with special concern to unsteady mud-flows, such as the hyper-concentrated floods. In the present analysis the rheological model proposed by O'Brien et al. (1993), broadly appreciated and widespread in the technical community, is considered. This model relies on a quadratic relationship linking the shear rate to the total shear stress, accounting for turbulent, dispersive and cohesive yield stresses. To provide relatively simple applicability criteria, linear analysis is applied to compare the celerity of a small perturbation of an initial steady uniform flow as predicted by the simplified model with those of the full dynamic model. Based on this comparison, applicability criteria for the kinematic approximation for mud-flows are derived. In particular, the proposed criteria rely on the comparison between the flood rising-time and a threshold time-scale: whenever the former is larger than the latter, the estimation of the propagation celerity of the simplified model is guaranteed with a prescribed accuracy. These criteria are presented considering the effect of all model parameters, based on the particular case of a pyroclastic mud, typically encountered in volcanic areas of southern Italy. The achieved results are discussed, aiming to guide the choice of the appropriate simplified model for mud flood analysis

Nitric oxide (NO) signaling is involved in numerous physiological processes in mollusks, e.g., learning and memory, feeding behavior, neural development, and defence response. We report the first molecular cloning of NOS mRNA from a cephalopod, the cuttlefish Sepia officinalis (SoNOS). SoNOS was cloned using a strategy that involves hybridization of degenerate PCR primers to highly conserved NOS regions, combined with RACE procedure. Two splicing variants of SoNOS, differing by 18 nucleotides, were found in the nervous system and the ink gland of Sepia. In situ hybridization shows that SoNOS is expressed in the immature and mature cells of the ink gland and in the regions of the nervous system that are related to the ink defence system.

The concerted action of many neuropeptides has been implicated in the nervous control of specific behaviors in many molluscs. In the present study, the presence of amidated tetrapeptide Ala-Pro-Gly-Trp-NH2 (APGWamide) in those lobes that are involved in the control of reproductive behavior in Octopus vulgaris has been investigated. APGWamide immunoreactivity was mainly confined to the posterior olfactory lobule and in the inferior frontal system. These areas are involved in Octopus in the processing of either chemotactile sense or olfaction. From these lobes, immunoreactive fibers reached other lobes of the central nervous system (CNS) which could be indirectly involved in the reproductive behavior. APGWamide immunoreactivity was also present in the glandular cells of the oviducal gland in the female reproductive system. These results constitute the first detailed immunolocalization of APGWamide in cephalopods and open a new insight into the possible effects that both distant and close chemical stimuli can exert on neuropeptidergic circuitries, which may affect the reproductive behavior of cephalopods.

As a part of continuous research on the neurobiology of the cephalopods in general, and the neuroendocrine control of reproduction in Octopus vulgaris in particular, the presence, the molecular analysis and the effect of FMRFamide on the screening-pigment migration in the visual system have been analysed. FMRFamide immunoreactive fibres are present in the outer plexiform layer of the retina as well as in the plexiform zone of the deep retina. These fibres presumably come from optic and olfactory lobes. We isolated an incomplete Octopus FMRFamide cDNA which encodes an amino terminal truncated precursor containing several FMRFamide-related peptides (FaRPs) showing a high degree of identity with the FaRPs encoded in the precursor of Sepia officinalis, except for the presence of an Rpamide related peptide, present only in cnidarians. Finally, stimulation of isolated retina demonstrated that the effect of this tetrapeptide, coupled with dopamine, is the induction of an extreme adaptation of the retina to the light condition. This situation de facto inhibits sexual maturation. Our results on the effect of FMRFamide on the retina confirm the suggested hypothesis that this peptide plays an inhibitory role on the activity of optic gland.

Chemical, biochemical, and immunohistochemical evidence is reported demonstrating the presence in the brain of the cuttlefish Sepia officinalis of a Ca2+-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate, occurring in neurons and fibers functionally related to the inking system. Nitric oxide synthase activity was concentrated for the most part in the cytosolic fraction and was masked by other citrulline-forming enzyme(s). The labile nitric oxide synthase could be partially purified by ammonium sulfate precipitation of tissue extracts, followed by affinity chromatography on 2′, 5′-ADP-agarose and calmodulin-agarose. The resulting activity, immunolabeled at 150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by antibodies to rat neuronal nitric oxide synthase, depended on NADPH and tetrahydro-L-biopterin, and was inhibited by NG-nitro-L-arginine. NMDAR2/3 subunit-immunoreactive proteins migrating at 170 kDa could also be detected in brain extracts, along with glutamate (whole brain : 0.32 ± 0.03 μmol of glutamate/mg of protein ; optic lobes : 0.22 ± 0.04 ; vertical complex : 0.06 ± 0.06 basal lobes : 0.58 ± 0.04 ; brachial lobe : 0.77 ± 0.06 ; pedal lobe : 1.04 ± 0.08 ; palliovisceral lobe : 0.86 ± 0.05). Incubation of intact brains with 1.5 mM glutamate or NMDA or the nitric oxide donor 2-(N,N-diethylamino)diazenolate-2-oxide caused a fivefold rise in the levels of cyclic GMP, indicating operation of the glutamate-nitric oxide-cyclic GMP signaling pathway. Immunohistochemical mapping of Sepia CNS showed specific localization of nitric oxide synthase-like and NMDAR2/3-like immunoreactivities in the lateroventral palliovisceral lobe, the visceral lobe, and the pallial and visceral nerves, as well as in the sphincters and wall of the ink sac.

Cephalopods are endowed with the most sophisticated nervous system among invertebrates and exhibit a repertoire of complex behaviors, such as spatial and observational learning. Cephalopod eyes supply a wide range of information which are utilized for these learning behaviors. Although our understanding of vertebrate physiology greatly benefited from the sub-cellular analysis of cephalopod nervous system, as shown by the discovery of the ionic bases of action potentials and of the Ca2+ requirement for neurotransmitter release Surprisingly, the cellular basis by which the visual system drives the sophisticated repertoire of cephalopod behaviors are still poorly understood. In this review, we will describe the current knowledge about cephalopod phototransduction. Light excites cephalopod photoreceptors by either inducing Ca2+ release from intracellular stores or activating cation-permeable channels by an as yet unknown mechanism. A 92 kDa protein, which is homologous to the Drosophila transient receptor potential (TRP) gene, is the most likely mediator of light-induced currents in cephalopods. A number of models which explain the mechanism whereby cephalopod TRP channel is gated by light will be discussed.

In several species of cephalopod molluscs there is good evidence for the presence of L-glutamate in the central and peripheral nervous system and evidence for both classes of ionotropic receptor, AMPA/kainate and NMDA. The best evidence for glutamate being a transmitter in cephalopods comes from pharmacological, immunohistochemical and molecular investigations on the giant fibre system in the squid stellate ganglion. These studies confirm there are AMPA/kainate-like receptors on the third-order giant axon. In the (glial) Schwann cells associated with the giant axons both classes of glutamate receptor occur. Glutamate is an excitatory transmitter in the chromatophores and in certain somatic muscles and its action is mediated primarily via AMPA/kainate-like receptors, but at some chromatophores there are NMDA-like receptors. In the statocysts the afferent crista fibres are also glutamatergic, acting at non-NMDA receptors. In the brain (of Sepia) a neuronal NOS is activated by glutamate with subsequent production of nitric oxide and elevation of cGMP levels. This signal transduction pathway is blocked by D-AP-5, a specific antagonist of the NMDA receptor. Recently immunohistochemical analysis has demonstrated (in Sepia and Octopus) the presence of NMDAR2A /B – like receptors in motor centres, in the visual and olfactory systems and in the learning system. Physiological experiments have shown that glutamatergic transmission is involved in long term potentation (LTP) in the vertical lobe of Octopus, a brain area involved in learning. This effect seems to be mediated by non-NMDA receptors. Finally in the CNS of Sepia NMDA-mediated nitration of tyrosine residues of cytoskeletal protein such as α-tubulin, has been demonstrated.

Many aspects of molluscan reproduction rely on peptides. Egg laying, secretory activity, sexual behavioral repertoires, and chemical attraction are some of the key actions of sexual behavior controlled by peptides in mollusks. In this chapter a detailed analysis of peptides involved in molluscan reproduction is reported. In particular, the discovery, the structure of the gene, and the distribution and biological actions of egg laying hormones, GnRH, APGWamide, vasopressin/oxytocin, and pheromones are discussed. Only those peptides for which the primary structure is known are reported in this chapter.

In this study, we have identified a 28-kDa protein resembling the linker H1 in the testis and prostate of the reproductive system of Octopus vulgaris. This protein, OvH1, was partially purified by reverse phase high-pressure liquid chromatography (HPLC) of the perchloric acid extract from testis nuclei. It showed electrophoretic mobility, CD spectrum and amino acid composition highly comparable with those of the mammalian histone. Moreover, it was microheterogeneous, as resulted from prostate and testis HPLC and mass spectrometry analyses. Such analysis showed that in testis there are two H1 subfractions, which do not appear in the prostate. Amino acid composition of the major testis specific variant (OvH1t) showed high similarity with rat testis specific H1t. The histone-like nature of OvH1 was confirmed by its ability to bind DNA as tested both by circular dichroism and protection of the nucleic acid toward deoxyribonuclease I activity. The circular dichroism spectra of Octopus DNA in the absence and presence of increasing amounts of the protein showed a dose-dependent effect, leading to a progressive compactness of the polynucleotide. OvH1/DNA complexes were also resistant to nuclease digestion. The presence of H1 in the testis and prostate of the reproductive system of Octopus is discussed in light of the fact that there is a similarity between its behavior and that of vertebrates. Mol. Reprod. Dev. 63: 355–365, 2002. © 2002 Wiley-Liss, Inc.

In this study for the first time we have characterized a progesterone receptor in the reproductive system of the female of Octopus vulgaris. Scatchard analysis revealed that one binding component with high affinity and low capacity for the ligand was present only in the nuclear extract. Competition experiments showed that the progesterone receptor was strictly specific for progesterone. DNA-cellulose binding and DEAE-Sephacel both confirmed the presence of one 3H-progesterone binding component which eluted at a salt concentration of 0.14 +/- 0.05 M NaCl and 0.15 +/- 0.05 M NaCl respectively. By using monoclonal antibodies against chicken progesterone receptor (subunits A and B), we have localized on Western Blot one band of about 70 kDa. Immunoreactivity for progesterone binding molecules has been localized in the nuclei of the follicle cells of the ovary, of the proximal portion of the oviduct and of the outer region of the nidimental gland. These data, taken together, provide evidence that in Octopus vulgaris the progesterone receptor has biochemical and immunohistochemical characteristics resembling those of progesterone receptor in vertebrates.

Free d -aspartate ( d -Asp) has been previously found in the nervous system of Octopus vulgaris (Mollusca: Cepalopoda) and has recently also been found in many endocrine tissues of the rat. The present study examined whether this enantiomer also occurs in the reproductive system and the brain of the octopus. In this mollusk, d -aspartate was present in both the male and the female reproductive systems. In males, it was found at high concentrations in the prostate, vas deferens, Needham's sac, and testis. In females, a high concentration was found in the oviduct, accessory nidamental gland, and ovary. The concentration varied between 0.4 and 2.9 μmol/g wet tissue, which corresponds to a percentage of d -Asp/total d + l -Asp between 7 and 33%. No appreciable quantities of d -Asp were found in the digestive, excretory, circulatory, and muscular systems, indicating that in the octopus this enantiomer may play a role in both the nervous and the reproductive systems.