Open Circulatory System Research Articles

Changes of bacterial communities and bile acid metabolism reveal the potential “intestine-hepatopancreas axis” in shrimp

The interaction between the gut and the liver plays a significant role in individual health and diseases. Mounting evidence supports that bile acids are important metabolites in the bidirectional communication between the gut and the liver. Most of the current studies on the “gut-liver axis” have focused on higher vertebrates, however, few was reported on lower invertebrates such as shrimp with an open circulatory system. Here, microbiomic and metabolomic analyses were conducted to investigate the bacterial composition and bile acid metabolism in intestine, hemolymph and hepatopancreas of Penaeus vannamei fed diets supplemented with octanoic acid and oleic acid. After six days of feeding, the bacterial composition in intestine, hemolymph and hepatopancreas changed at different stages, with significant increases in the relative abundance of several genera such as Pseudomonas and Rheinheimera in intestine and hepatopancreas. Notably, there was a more similar bacterial composition in intestine and hepatopancreas at the genus level, which indicated the close communication between shrimp intestine and hepatopancreas. Meanwhile, higher content of some bile acids such as lithocholic acid (LCA) and α-muricholic acid (α-MCA) in intestine and lower content of some bile acids such as taurohyocholic acids (THCA) and isolithocholic acid (IsoLCA) in hepatopancreas were detected. Furthermore, Spearman correlation analysis revealed a significant correlation between bacterial composition and bile acid metabolism in intestine and hepatopancreas. The microbial source tracking analysis showed that there was a high proportion of intestine and hepatopancreas bacterial community as the source of each other. Collectively, these results showed a strong crosstalk between shrimp intestine and hepatopancreas, which suggests a unique potential “intestine-hepatopancreas axis” in lower invertebrate shrimp with an open circulatory system. Our finding contributed to the understanding of the interplay between shrimp intestine and hepatopancreas in the view of microecology and provided new ideas for shrimp farming and disease control.

Revealing Changes in Ovarian and Hemolymphatic Metabolites Using Widely Targeted Metabolomics between Newly Emerged and Laying Queens of Honeybee (Apis mellifera).

The queen bee is a central and pivotal figure within the colony, serving as the sole fertile female responsible for its reproduction. The queen possesses an open circulatory system, with her ovaries immersed in hemolymph. A continuous and intricate transportation and interchange of substances exist between the ovaries and hemolymph of queen bees. To determine the characteristic metabolites in the hemolymph and ovary, as well as understand how their rapid metabolism contributes to the process of egg-laying by queens, we reared Apis mellifera queens from three different age groups: newly emerged queen (NEQ), newly laying queen (NLQ), and old laying queen (OLQ). Using widely targeted metabolomics, our study revealed that the laying queen (NLQ and OLQ) exhibited faster fatty acid metabolism, up-regulated expression of antioxidants, and significant depletion of amino acids compared to the NEQ. This study revealed that the levels of carnitine and antioxidants (GSH, 2-O-α-D-glucopyranosyl-L-ascorbic acid, L-ascorbic acid 2-phosphate, etc.) in the NLQ and OLQ were significantly higher compared to NEQ. However, most of the differentially expressed amino acids, such as L-tryptophan, L-tyrosine, L-aspartic acid, etc., detected in NLQ and OLQ were down-regulated compared to the NEQ. Following egg-laying, pathways in the queens change significantly, e.g., Tryptophan metabolism, Tyrosine metabolism, cAMP signaling pathway, etc. Our results suggest that carnitine and antioxidants work together to maintain the redox balance of the queen. Additionally, various amino acids are responsible for maintaining the queen's egg production.

Diversity of symbiotic bacteria in the digestive and circulatory systems of lobster and crayfish: important connections for efficient aquaculture. A systematic review

Abstract Lobsters and crayfishes have social, economic and ecological significance in several regions around the world. These crustaceans have complex digestive systems and open circulatory systems colonized by symbiotic microorganisms that play essential roles in the health, nutrition and disease of crustaceans, attracting considerable interest. The importance of the microbiota for the successful cultivation of animals in captivity is known, however, there is a lack of information about the symbiotic microbiota in different species of lobsters. The aim of this study was to summarize global information on microbiota studies of anatomical sites of lobsters and crayfishes looking for patterns of bacteria that colonize their intestine, hepatopancreas and haemolymph. A systematic review was conducted using “Methodi Ordinatio”. Twenty-five significant articles, from 12 countries, were identified for analysis from the Scopus database. The crustacean genera found were Cherax, Pacifastacus and Procambarus (crayfish); Homarus, Nephrops and Panulirus (lobsters). The intestinal, hepatopancreas and haemolymph microbiota show a wide variety of bacteria influenced by infections affecting the individual, stress caused by changes in the environment, availability and quality of food, period of the year, stage of development and the animal’s sex, among other aspects. The common core bacterial microbiota among crustaceans consist of the genera Acinetobacter, Aeromonas, Candidatus Bacilloplasma, Candidatus Hepatoplasma, Citrobacter, Hafnia, Photobacterium, Pseudomonas, Shewanella and Vibrio. The results broadened our understanding of the microbiota in the intestinal and circulatory systems of crustaceans, the flaws for and needs for standardization of analysis techniques, and the use of a multi-specialized approach for the protection and management of lobsters as a fisheries resource.

Drosophila as a Model to Understand Second Heart Field Development.

The genetic model system Drosophila has contributed fundamentally to our understanding of mammalian heart specification, development, and congenital heart disease. The relatively simple Drosophila heart is a linear muscular tube that is specified and develops in the embryo and persists throughout the life of the animal. It functions at all stages to circulate hemolymph within the open circulatory system of the body. During Drosophila metamorphosis, the cardiac tube is remodeled, and a new layer of muscle fibers spreads over the ventral surface of the heart to form the ventral longitudinal muscles. The formation of these fibers depends critically upon genes known to be necessary for mammalian second heart field (SHF) formation. Here, we review the prior contributions of the Drosophila system to the understanding of heart development and disease, discuss the importance of the SHF to mammalian heart development and disease, and then discuss how the ventral longitudinal adult cardiac muscles can serve as a novel model for understanding SHF development and disease.

Mechanisms and roles of the first stage of nodule formation in lepidopteran insects.

Nodule formation is a process of cellular immunity in insects and other arthropods with open circulatory systems. Based on histological observations, nodule formation occurs in 2 stages. The first stage occurs immediately after microbial inoculation and includes aggregate formation by granulocytes. The second stage occurs approximately 2-6 h later and involves the attachment of plasmatocytes to melanized aggregates produced during the first stage. The first stage response is thought to play a major role in the rapid capture of invading microorganisms. However, little is known regarding how granulocytes in the hemolymph form aggregates, or how the first stage of the immunological response protects against invading microorganisms. Since the late 1990s, our understanding of the molecules and immune pathways that contribute to nodule formation has improved. The first stage of nodule formation involves a hemocyte-induced response that is triggered by pathogen-associated molecular pattern (PAMP) recognition proteins in the hemolymph regulated by a serine proteinase cascade and cytokine (Spätzle) and Toll signaling pathways. Hemocyte agglutination proceeds through stepwise release of biogenic amine, 5-HT, and eicosanoids that act downstream of the Toll pathway. The first stage of nodule formation is closely linked to melanization and antimicrobial peptide (AMP) production, which is critical for insect humoral immunity. Nodule formation in response to artificial inoculation with millions of microorganisms has long been studied. It has recently been suggested that this system is the original natural immune system, and enables insects to respond to a single invading microorganism in the hemocoel.

Systemic coagulopathy promotes host lethality in a new Drosophila tumor model.

Malignant tumors trigger a complex network of inflammatory and wound repair responses, prompting Dvorak's characterization of tumors as "wounds that never heal."1 Some of these responses lead to profound defects in blood clotting, such as disseminated intravascular coagulopathy (DIC), which correlate with poor prognoses.2,3,4 Here, we demonstrate that a new tumor model in Drosophila provokes phenotypes that resemble coagulopathies observed in patients. Fly ovarian tumors overproduce multiple secreted components of the clotting cascade and trigger hypercoagulation of fly blood (hemolymph). Hypercoagulation occurs shortly after tumor induction and is transient; it is followed by a hypocoagulative state that is defective in wound healing. Cellular clotting regulators accumulate on the tumor over time and are depleted from the body, suggesting that hypocoagulation is caused by exhaustion of host clotting components. We show that rescuing coagulopathy by depleting a tumor-produced clotting factor improves survival of tumor-bearing flies, despite the fact that flies have an open (non-vascular) circulatory system. As clinical studies suggest that lethality in patients with high serum levels of clotting components can be independent of thrombotic events,5,6 our work establishes a platform for identifying alternative mechanisms by which tumor-driven coagulopathy triggers early mortality. Moreover, it opens up exploration of other conserved mechanisms of host responses to chronic wounds.

Differentiation and function of cardiac valves in the adult Drosophila heart

ABSTRACT Drosophila, like all insects, has an open circulatory system for the distribution of haemolymph and its components. The circulation of the haemolymph is essentially driven by the pumping activity of the linear heart. The heart is constructed as a tube into which the haemolymph is sucked and pumped forward by rhythmic contractions running from the posterior to the anterior, where it leaves the heart tube. The heart harbours cardiac valves to regulate flow directionality, with a single heart valve differentiating during larval development to separate the heart tube into two chambers. During metamorphosis, the heart is partially restructured, with the linear heart tube with one terminal wide-lumen heart chamber being converted into a linear four-chambered heart tube with three valves. As in all metazoan circulatory systems, the cardiac valves play an essential role in regulating the direction of blood flow. We provide evidence that the valves in adult flies arise via transdifferentiation, converting lumen-forming contractile cardiomyocytes into differently structured valve cells. Interestingly, adult cardiac valves exhibit a similar morphology to their larval counterparts, but act differently upon heart beating. Applying calcium imaging in living specimens to analyse activity in valve cells, we show that adult cardiac valves operate owing to muscle contraction. However, valve cell shape dynamics are altered compared with larval valves, which led us to propose our current model of the opening and closing mechanisms in the fly heart.

Identification of a novel crustacean vascular endothelial growth factor b-like in the mud crab Scylla paramamosain, and examination of its role in lipid accumulation

Vascular endothelial growth factor B (VEGFB), unlike other members of the VEGF family such as VEGFA, shows only limited potential in angiogenesis in mammals, but is involved in lipid metabolism, especially in transporting dietary lipids present in circulation through the vascular endothelium to peripheral tissues. However, the role of Vegfb in invertebrates such as decapod crustaceans with an open circulatory system is still poorly understood. Here, cDNA of a vegfb-like gene was cloned and characterized for the first time from a decapod crustacean, mud crab Scylla paramamosain. The mud crab vegfb-like gene encoded a polypeptide of 386 amino acids that possessed all the typical features of the Vegf family, and shared high homology with other crustacean Vegf orthologs. Relative expression levels of the vegfb-like gene were higher in metabolically active tissues such as gill, heart and muscle, and increased in ovary of female crabs as ovary matured. Knockdown of the vegfb-like gene in vivo reduced the contents of triglyceride in hemolymph and hepatopancreas, as well as total lipid in hepatopancreas. In addition, knockdown of the vegfb-like gene increased tissue expression levels of genes related to fatty acid transportation, allocation and hydrolysis, such as long-chain acyl-CoA synthetase 3/4 (acsl3/4), fatty acid transport protein 1 (fatp1), and hormone-sensitive lipase (hsl), and decreased the expression levels of the genes related to fatty acid synthesis like acetyl-CoA carboxylase (acc). These results in mud crab suggest that Vegfb might play important roles in regulating fatty acid absorption, utilization and allocation in crustaceans.

Mechanism of elastic energy storage of honey bee abdominal muscles under stress relaxation

Energy storage of passive muscles plays an important part in frequent activities of honey bee abdomens due to the muscle distribution and open circulatory system. However, the elastic energy and mechanical properties of structure in passive muscles remain unclear. In this article, stress relaxation tests on passive muscles from the terga of the honey bee abdomens were performed under different concentrations of blebbistatin and motion parameters. In stress relaxation, the load drop with the rapid and slow stages depending on stretching velocity and stretching length reflects the features of myosin-titin series structure and cross-bridge-actin cyclic connections in muscles. Then a model with 2 parallel modules based on the 2 feature structures in muscles was thus developed. The model described the stress relaxation and stretching of passive muscles from honey bee abdomen well for a good fitting in stress relaxation and verification in loading process. In addition, the stiffness change of cross-bridge under different concentrations of blebbistatin is obtained from the model. We derived the elastic deformation of cross-bridge and the partial derivatives of energy expressions on motion parameters from this model, which accorded the experimental results. This model reveals the mechanism of passive muscles from honey bee abdomens suggesting that the temporary energy storage of cross-bridge in terga muscles under abdomen bending provides potential energy for springback during the periodic abdomen bending of honey bee or other arthropod insects. The finding also provides an experimental and theoretical basis for the novel microstructure and material design of bionic muscle.

Modelling Cancer Metastasis in Drosophila melanogaster.

Cancer metastasis, the process by which tumour cells spread throughout the body and form secondary tumours at distant sites, is the leading cause of cancer-related deaths. The metastatic cascade is a highly complex process encompassing initial dissemination from the primary tumour, travel through the blood stream or lymphatic system, and the colonisation of distant organs. However, the factors enabling cells to survive this stressful process and adapt to new microenvironments are not fully characterised. Drosophila have proven a powerful system in which to study this process, despite important caveats such as their open circulatory system and lack of adaptive immune system. Historically, larvae have been used to model cancer due to the presence of pools of proliferating cells in which tumours can be induced, and transplanting these larval tumours into adult hosts has enabled tumour growth to be monitored over longer periods. More recently, thanks largely to the discovery that there are stem cells in the adult midgut, adult models have been developed. We focus this review on the development of different Drosophila models of metastasis and how they have contributed to our understanding of important factors determining metastatic potential, including signalling pathways, the immune system and the microenvironment.

Іnfluence of some heavy metals to the pulmonary and direct diffusive respiration of the great ramshorn Planorbarius corneus allospecies (Mollusca: Gastropoda: Planorbidae) from the Ukrainian river system

The vicaristic genetic allospecies (“western” and “eastern”) of superspecies complex Planorbarius corneus s. lato from Ukrainian river network have the combined bimodal respiration mode. These molluscs use their true lungs for atmospheric oxygen breathing, and the diffusive (surface) respiration is used for soluble oxygen consumption. Oxygen transportation to different tissues and organs and carbon dioxide from them to lungs occurs due to respiratory pigment hemoglobin in their liquid internal medium (hemolymph), circulating in vessels, lacunas and sinuses of these animals’ open circulatory system. The aim of present study was to clarify the features and difference levels of lung and surface respiration for “western” and “eastern” allospecies under the impact of heavy metal ions in water environment (MPC: 0.5, 1, 2, 3). It was established that the toxins could be ranged by their toxicity level for activation of molluscs’ respiratory function: Cu2+>Zn2+>Ni2+ >Mn2+. The “eastern” allospecies appeared to be more sensitive and less durable for all aforementioned toxins by both lung and direct diffusive respiration, comparing to the “western” allospecies.

Ventricular Infusion and Nanoprobes Identify Cerebrospinal Fluid and Glymphatic Circulation in Human Nerves.

Background:Growing evidence suggests that cerebrospinal fluid circulates in human nerves. Several conditions encountered by the plastic surgeon may be related to dysregulation of this system, including nerve transection, stretch injuries, and peripheral neuropathy. The purpose of this study was to show how ventricular infusion and nanoprobes identify CSF and glymphatic circulation in neural sheaths of human nerves.Methods:The technique of ventricular infusion using buffered saline was developed in 2017. The technique was used in a series of eight fresh cadavers before dissection of the median nerve, and combined with fluorescent imaging and nanoprobe injections in selected specimens.Results:Eight cadaver specimens underwent ventricular infusion. There were six female and two male specimens, ages 46–97 (mean 76.6). Ventricular cannulation was performed successfully using coordinates 2 cm anterior to coronal suture and 2.5 cm lateral to sagittal suture. Depth of cannulation ranged from 44 to 56 mm (mean 49.7). Ventricular saline infusion complemented by nanoprobe injection suggests CSF flows in neural sheaths, including pia meninges, epineurial channels, perineurium, and myelin sheaths (neurolemma).Conclusions:Ventricular infusion and nanoprobes identify CSF flow in neural sheaths of human nerves. CSF flow in nerves is an open circulatory system that occurs via channels, intracellular flow, and cell-to-cell transport associated with glial cells. Neural sheaths, including neurolemma, may participate in glucose and solute transport to axons. These techniques may be used in anatomic dissection and live animal models, and have been extended to the central nervous system to identify direct ventricle-to-pia meninges CSF pathways.

Evolución filogenética del corazón

In 1859 Charles Darwin in his “The Origin of Species” formulated three facts that explain the principle of evolution: the presence of a common ancestor, the existence of advantageous genetic mutations for survival and the elimination of unfavorable ones or Natural Selection. The embryological development of the mammalian heart improves the need for external oxygen uptake, transport and release in the tissues, which exists in all multicellular living beings from the most primitive and enables the function of all organs and systems. In invertebrates, such as insects, arthropods and molluscs, the open circulatory system reaches a development similar to that of human Carnegie stages 10-13: endocardial tubes, straight tube, or early stages of the bulboventricular loop. In vertebrates, the closed system of fish develops hearts with three and four chambers equivalent to the torsion phase of the loop in mammals, or the fifth week of human embryonic development, with a model closer to that of mammals appearing in reptiles with differences such as the incomplete septation of the ventricles and the development of two aortic trunks in crocodilians. Birds and mammals have reached a similar degree of embryonic development of their hearts, with four septate chambers, the same as the Cono Truncus one, and which separate the systemic and pulmonary circulation. All the embryological development of the human heart is completed between days 20 and 60 of gestation, concluding in the fetal phase its growth together with the rest of the organism.

Hazard Identification Related to the Presence of Vibrio spp., Biogenic Amines, and Indole-Producing Bacteria in a Non-Filter Feeding Marine Gastropod (Tritiamutabilis) Commercialized on the Italian Market.

Tritia mutabilis is a carrion-feeder edible marine gastropod with an open circulatory system. Therefore, biological, and chemical contaminants associated with the feed can reach all body tissues. The aim of the present study was to investigate the possible association of these characteristics with some food safety hazards. Vibrio spp. load, and the prevalence of pathogenic V. parahaemolyticus, V. vulnificus, and V. cholerae, were investigated. Moreover, biogenic amines (BAs) and indole-producing bacteria (IPB), markers of seafood decomposition, were quantified for the first time in an edible carrion-feeder. Overall, 49 batches were analyzed (38 from retail, and 11 from primary production). The Vibrio spp. load resulted of 5.64 ± 0.69 log10 CFU g−1 at retail, and 5.27 ± 0.74 at harvest but all batches resulted negative for pathogenic Vibrio. Histamine, putrescine, cadaverine, and tyramine were detected both at harvest and at the retail level. Their sum (BAs Index) showed a mean value of 50.45 and 65.83 mg Kg−1 in batches at harvest and at retail, respectively. IPB were detected at harvest and upon refrigeration for three days (T1–T3). The mean load resulted in 2.52 ± 0.85 log10 MPN g−1 at T0, 3.31 ± 1.23 at T3 in batches immediately refrigerated, and 3.22 ± 1.18 at T3 in batches previously immersed in clean seawater. Our results contribute to identifying food-borne hazards for T. mutabilis that may be related to the retention of biogenic amines and indole-producing bacteria due to carrion feeding.

Constitutive expression and discovery of antimicrobial peptides in Zygogramma bicolorata (Coleoptera: Chrysomelidae).

The expression, identification, and discovery of less toxic antimicrobial peptides (AMPs) are significant in managing infectious pathogens. AMPs triggered in response to the immune system have evolved to defend against pathogens and wounding. The protein composition of Zygogramma bicolorata hemolymph is of diagnostic importance as the open circulatory systems of the insects involve signaling through hemolymph. They have conserved many ancestral vertebrate genes that may help better understand the evolution of innate immunity. The present work describes the isolation, purification, identification, and bioinformatics analysis of AMPs from the immunized hemolymph of Z. bicolorata. Thirty-nine peptides were isolated from reverse-phase high-performance liquid chromatography and sequenced via mass spectrometry analysis. The immunization process recorded a threefold higher protein concentration in immunized hemolymph when compared with nonimmunized one. For the first time, the proteomic study on Z. bicolorata hemolymph unveils the three novel proteins in the family Chrysomelidae with no homology in the database, indicating its novelty and the expression of the rest of 36 well-known proteins, including heat-shock, immune, structural, signaling proteins, and others speak for its method validity. Combining the expression of novel AMPs, detoxifying enzymes, hemolytic, and cytotoxic assays, and this work can elucidate new pathways to immune response mechanisms. Its molecular basis also holds the potential applicability in the future drug development process against pathogenic fungi such as Aspergillus niger and Candida albicans.

Depuration of a Freshwater Clam (Batissa Violacea) from Rewa River in Fiji Using a Bio-Filter Set-Up in Closed and Open Water Circulatory System

The effectiveness of the freshwater bivalve Batissa violacea depuration was tested in closed and open water circulatory system over a 48 h period. The closed circulatory system included a sand biofilter. Microbial levels were assessed every 4 h using Total Aerobic Plate Count (TPC) for heterotrophs and Most Probable Number (MPN) for coliforms. TPC and coliform loads in bivalve tissue reduced rapidly to low and undetectable levels in a closed circulatory system while open system showed a slower reduction. Both TPC and coliform loads remained above detectable levels throughout the depuration period. Closed system showed similar patterns of logarithmic reduction of TPC and coliforms in all cases with R2>0.95 and p<0.001. Similar results were observed for tank water however, reduction of TPC and coliforms were slower. Biofilm formation was observed in the interior walls of the aquarium tanks over 48 h in all cases. Physicochemical parameters did not show any significant change. The reduction in TPC and coliform load in B. violacea suggests that biofilter in a closed water circulatory system is a simple, cost-effective, water conserving and effective way to significantly reduce the spoilage and coliform bacterial load that is accumulated in the clams.

The Serine Protease Homolog, Scarface, Is Sensitive to Nutrient Availability and Modulates the Development of the Drosophila Blood-Brain Barrier.

The adaptable transcriptional response to changes in food availability not only ensures animal survival but also lets embryonic development progress. Interestingly, the CNS is preferentially protected from periods of malnutrition, a phenomenon known as “brain sparing.” However, the mechanisms that mediate this response remain poorly understood. To get a better understanding of this, we used Drosophila melanogaster as a model, analyzing the transcriptional response of neural stem cells (neuroblasts) and glia of the blood–brain barrier (BBB) from larvae of both sexes during nutrient restriction using targeted DamID. We found differentially expressed genes in both neuroblasts and glia of the BBB, although the effect of nutrient deficiency was primarily observed in the BBB. We characterized the function of a nutritional sensitive gene expressed in the BBB, the serine protease homolog, scarface (scaf). Scaf is expressed in subperineurial glia in the BBB in response to nutrition. Tissue-specific knockdown of scaf increases subperineurial glia endoreplication and proliferation of perineurial glia in the blood–brain barrier. Furthermore, neuroblast proliferation is diminished on scaf knockdown in subperineurial glia. Interestingly, reexpression of Scaf in subperineurial glia is able to enhance neuroblast proliferation and brain growth of animals in starvation. Finally, we show that loss of scaf in the blood–brain barrier increases sensitivity to drugs in adulthood, suggesting a physiological impairment. We propose that Scaf integrates the nutrient status to modulate the balance between neurogenesis and growth of the BBB, preserving the proper equilibrium between the size of the barrier and the brain.SIGNIFICANCE STATEMENT The Drosophila BBB separates the CNS from the open circulatory system. The BBB glia are not only acting as a physical segregation of tissues but participate in the regulation of the metabolism and neurogenesis during development. Here we analyze the transcriptional response of the BBB glia to nutrient deprivation during larval development, a condition in which protective mechanisms are switched on in the brain. Our findings show that the gene scarface reduces growth in the BBB while promoting the proliferation of neural stem, assuring the balanced growth of the larval brain. Thus, Scarface would link animal nutrition with brain development, coordinating neurogenesis with the growth of the BBB.

Preliminary Studies on the Humoral Immune Response of Mulberry Silkworm, Bombyx mori Larvae Fed on Different Mulberry Varieties

Diseases infecting silkworms cause 30-40% cocoon crop loss. The silkworm, Bombyx mori L. defense against pathogens is provided by open circulatory system through humoral and cellular response. Three mulberry varieties, Morus alba var. Rosa, M. alba var. Morittiana and M. alba var. Laevigata, were used to study the effect of host plant and bacterial contagious infection on the humoral immune of B. mori larvae. The specific activity of related enzymes in haemolymph of fifth instar larvae was estimated. Results indicated that mulberry varieties and bacterial contagious infection had significant effects on enzymes activity values in larval haemolymph. Moreover, M. alba var. laevigata was superior in the humoral immune response.

Nonenzymatic Spontaneous Oxidative Transformation of 5,6-Dihydroxyindole.

Melanin is an important phenolic skin pigment found throughout the animal kingdom. Tyrosine and its hydroxylated product dopa provide the starting material for melanin biosynthesis in all animals. Through a set of well-established reactions, they are converted to 5,6-dihydroxyindole (DHI) and DHI-2-carboxylic acid (DHICA). Oxidative polymerization of these two indoles produces the brown to black eumelanin pigment. The steps associated with these transformations are complicated by the extreme instability of the starting materials and the transient and highly reactive nature of the intermediates. We have used mass spectral studies to explore the nonenzymatic mechanism of oxidative transformation of DHI in water. Our results indicate the facile production of not only dimeric and trimeric products but also higher oligomeric forms of DHI upon exposure to air in solution, even under nonenzymatic conditions. Such instantaneous polymerization of DHI avoids toxicity to self-matter and ensures the much-needed deposition of melanin at (a) the wound site and (b) the infection site in arthropods. The rapid deposition of DHI melanin is advantageous for arthropods given their open circulatory system; the process limits blood loss during wounding and prevents the spread of parasites by encapsulating them in melanin, limiting the damage.

Fine structure of the intercalated disc and cardiac junctions in the black widow spider Latrodectus mactans

Arthropods have an open circulatory system with a simple tubular heart, so it has been estimated that the contractile pumping structure of the cardiac muscle will be less efficient than that of vertebrates. Nevertheless, certain arthropods are known to have far superior properties and characteristics than vertebrates, so we investigated the fine structural features of intercalated discs and cardiac junctions of cardiac muscle cells in the black widow spider Latrodectus mactans. Characteristically, the spider cardiac muscle has typical striated features and represents a functional syncytium that supports multiple connections to adjacent cells by intercalated discs. Histologically, the boundary lamina of each sarcolemma connects to the basement membrane to form an elastic sheath, and the extracellular matrix allows the cells to be anchored to other tissues. Since the intercalated disc is also part of sarcolemma, it contains gap junctions for depolarization and desmosomes that keep the fibers together during cardiac muscle contraction. Furthermore, fascia adherens and macula adherens (desmosomes) were also identified as cell junctions in both sarcolemma and intercalated discs. To enable the coordinated heartbeat of the cardiac muscle, the muscle fibers have neuronal innervations by multiple axons from the motor ganglion.