Invertebrate Organisms Research Articles

Zebrafish: An Attractive Model to Study Staphylococcus aureus Infection and Its Use as a Drug Discovery Tool.

Non-mammalian in vivo disease models are particularly popular in early drug discovery. Zebrafish (Danio rerio) is an attractive vertebrate model, the success of which is driven by several advantages, such as the optical transparency of larvae, the small and completely sequenced genome, the small size of embryos and larvae enabling high-throughput screening, and low costs. In this review, we highlight zebrafish models of Staphyloccoccus aureus infection, which are used in drug discovery and for studying disease pathogenesis and virulence. Further, these infection models are discussed in the context of other relevant zebrafish models for pharmacological and toxicological studies as part of early drug profiling. In addition, we examine key differences to commonly applied models of S. aureus infection based on invertebrate organisms, and we compare their frequency of use in academic research covering the period of January 2011 to January 2021.

Human type 2 diabetes mellitus-associated transcriptional disturbances in a high-sugar diet long-term exposed Drosophila melanogaster

Type 2 Diabetes mellitus (T2DM) is a multifactorial and polygenic disorder with the molecular bases still idiopathic. Experimental analyses and tests are quite limited upon human samples due to the access, variability of patient's conditions, and the size and complexity of the genome. Therefore, high-sugar diet exposure is commonly used for modeling T2DM in non-human animals, which includes invertebrate organisms like the fruit fly Drosophila melanogaster. Interestingly, high-sugar diet (HSD) induces delayed time for pupation and reduced viability in fruit fly larvae hatched from a 30% sucrose-containing medium (HSD-30%). Here we carried out an mRNA-deep sequencing study to identify differentially transcribed genes in adult fruit fly hatched and reared from an HSD-30%. Seven days after hatching, flies reared on control and HSD-30% were used to glucose and triglyceride level measurements and RNA extraction for sequencing. Remarkably, glucose levels were about 2-fold higher than the control group in fruit flies exposed to HSD-30%, whereas triglycerides levels increased 1.7-fold. After RNA-sequencing, we found that 13.5% of the genes were differentially transcribed in the dyslipidemic and hyperglycaemic insects. HSD-30% up-regulated genes involved in ribosomal biogenesis (e.g. dTOR, ERK and dS6K) and down-regulated genes involved in energetic process (e.g. Pfk, Gapdh1, and Pyk from pyruvate metabolism; kdn, Idh and Mdh2 from the citric acid cycle; ATPsynC and ATPsynẞ from ATP synthesis) and insect development. We found a remarkable down-regulation for Actin (Act88F) that likely impairs muscle development. Moreover, HSD-30% up-regulated both the insulin-like peptides 7 and 8 and down-regulated the insulin receptor substrate p53, isoform A and insulin-like peptide 6 genes, whose functional products are insulin signaling markers. All these features pointed together to a tightly correlation of the T2DM-like phenotype modeled by the D. melanogaster and an intricate array of phenomena, which includes energetic processes, muscle development, and ribosomal synthesis as that observed for the human pathology.

Fat Body-Multifunctional Insect Tissue.

Simple SummaryEfficient and proper functioning of processes within living organisms play key roles in times of climate change and strong human pressure. In insects, the most abundant group of organisms, many important changes occur within their tissues, including the fat body, which plays a key role in the development of insects. Fat body cells undergo numerous metabolic changes in basic energy compounds (i.e., lipids, carbohydrates, and proteins), enabling them to move and nourish themselves. In addition to metabolism, the fat body is involved in the development of insects by determining the time an individual becomes an adult, and creates humoral immunity via the synthesis of bactericidal proteins and polypeptides. As an important tissue that integrates all signals from the body, the processes taking place in the fat body have an impact on the functioning of the entire body.The biodiversity of useful organisms, e.g., insects, decreases due to many environmental factors and increasing anthropopressure. Multifunctional tissues, such as the fat body, are key elements in the proper functioning of invertebrate organisms and resistance factors. The fat body is the center of metabolism, integrating signals, controlling molting and metamorphosis, and synthesizing hormones that control the functioning of the whole body and the synthesis of immune system proteins. In fat body cells, lipids, carbohydrates and proteins are the substrates and products of many pathways that can be used for energy production, accumulate as reserves, and mobilize at the appropriate stage of life (diapause, metamorphosis, flight), determining the survival of an individual. The fat body is the main tissue responsible for innate and acquired humoral immunity. The tissue produces bactericidal proteins and polypeptides, i.e., lysozyme. The fat body is also important in the early stages of an insect’s life due to the production of vitellogenin, the yolk protein needed for the development of oocytes. Although a lot of information is available on its structure and biochemistry, the fat body is an interesting research topic on which much is still to be discovered.

The in vivo Tetrahymena thermophila extracellular glucose drop assay for characterization of mammalian insulin activity

Insulin activity is generally determined by an in vivo rabbit blood glucose drop assay in research and industriel laboratories. The humane experimental techniques imply the use of alternative invertebrate organisms in place of animals, known as replacement rule of the 3Rs. In this study, we report an alternative in vivo extracellular glucose drop assay using unicellular invertebrate Tetrahymena thermophila to replace the use of rabbit and mouse. This assay has four major steps; growing cells, starving cells, treatment of cells and measurement of glucose drop. In this assay, 0.2 mg/ml of human, porcine and bovine insulins dropped extracellular glucose level to 16%, 14% and 12%, respectively in ten minutes. In addition, mammalian insulins respectively increased the cell area about 19%, 15%, and 16% at 6th hour with statistically significant effect on the cell growth, but not in the cell viability. The results showed that the in vivo Tetrahymena thermophila extracellular glucose drop assay could be used as an alternative assay to replace the mouse or the rabbit insulin blood glucose drop assay.

An Interesting Molecule: γ-Aminobutyric Acid. What Can We Learn from Hydra Polyps?

Neuronal excitability is controlled primarily by γ-aminobutyric acid (GABA) in the central and peripheral nervous systems of vertebrate as well as invertebrate organisms. Besides its recognized neurotransmitter functions, GABA also plays a fundamental role in neurogenesis and synaptogenesis during embryonic development. In addition, GABAergic mechanisms are also involved in disorders of various peripheral tissues, ranging from diabetes to hypothyroidism to inflammatory responses. The discovery of the molecule and the history of its biosynthetic pathways in vertebrate and invertebrate phyla are summarized here. The occurrence and distribution of GABA, GABA-synthesizing enzymes, and receptors to GABA in the freshwater polyp Hydra vulgaris (Cnidaria: Hydrozoa), endowed with an early evolved nervous system, are discussed in relation to possible interactions with the microbiota, a stable component of Hydra polyps; their contribution to the evolution of nervous systems through microbe–neuronal interactions is proposed.

Expanding the Junction: New Insights into Non-Occluding Roles for Septate Junction Proteins during Development.

The septate junction (SJ) provides an occluding function for epithelial tissues in invertebrate organisms. This ability to seal the paracellular route between cells allows internal tissues to create unique compartments for organ function and endows the epidermis with a barrier function to restrict the passage of pathogens. Over the past twenty-five years, numerous investigators have identified more than 30 proteins that are required for the formation or maintenance of the SJs in Drosophila melanogaster, and have determined many of the steps involved in the biogenesis of the junction. Along the way, it has become clear that SJ proteins are also required for a number of developmental events that occur throughout the life of the organism. Many of these developmental events occur prior to the formation of the occluding junction, suggesting that SJ proteins possess non-occluding functions. In this review, we will describe the composition of SJs, taking note of which proteins are core components of the junction versus resident or accessory proteins, and the steps involved in the biogenesis of the junction. We will then elaborate on the functions that core SJ proteins likely play outside of their role in forming the occluding junction and describe studies that provide some cell biological perspectives that are beginning to provide mechanistic understanding of how these proteins function in developmental contexts.

Planarians (Platyhelminthes)-An Emerging Model Organism for Investigating Innate Immune Mechanisms.

An organism responds to the invading pathogens such as bacteria, viruses, protozoans, and fungi by engaging innate and adaptive immune system, which functions by activating various signal transduction pathways. As invertebrate organisms (such as sponges, worms, cnidarians, molluscs, crustaceans, insects, and echinoderms) are devoid of an adaptive immune system, and their defense mechanisms solely rely on innate immune system components. Investigating the immune response in such organisms helps to elucidate the immune mechanisms that vertebrates have inherited or evolved from invertebrates. Planarians are non-parasitic invertebrates from the phylum Platyhelminthes and are being investigated for several decades for understanding the whole-body regeneration process. However, recent findings have emerged planarians as a useful model for studying innate immunity as they are resistant to a broad spectrum of bacteria. This review intends to highlight the research findings on various antimicrobial resistance genes, signaling pathways involved in innate immune recognition, immune-related memory and immune cells in planarian flatworms.

Interaction of Arsenophonus with Wolbachia in Nilaparvata lugens

BackgroundCo-infection of endosymbionts in the same host is ubiquitous, and the interactions of the most common symbiont Wolbachia with other symbionts, including Spiroplasma, in invertebrate organisms have received increasing attention. However, the interactions between Wolbachia and Arsenophonus, another widely distributed symbiont in nature, are poorly understood. We tested the co-infection of Wolbachia and Arsenophonus in different populations of Nilaparvata lugens and investigated whether co-infection affected the population size of the symbionts in their host.ResultsA significant difference was observed in the co-infection incidence of Wolbachia and Arsenophonus among 5 populations of N. lugens from China, with nearly half of the individuals in the Zhenjiang population harbouring the two symbionts simultaneously, and the rate of occurrence was significantly higher than that of the other 4 populations. The Arsenophonus density in the superinfection line was significantly higher only in the Maanshan population compared with that of the single-infection line. Differences in the density of Wolbachia and Arsenophonus were found in all the tested double-infection lines, and the dominant symbiont species varied with the population only in the Nanjing population, with Arsenophonus the overall dominant symbiont.ConclusionsWolbachia and Arsenophonus could coexist in N. lugens, and the co-infection incidence varied with the geographic populations. Antagonistic interactions were not observed between Arsenophonus and Wolbachia, and the latter was the dominant symbiont in most populations.

Neurotoxicity in Marine Invertebrates: An Update.

Simple SummaryThe pollution of air, soil, and sea has grown in recent decades at the same pace as human development. Climate changes add further damage to the ecosystems. Nowadays, pollutants that derive from the anthropization of the environment are indicated as “emerging pollutants”. Marine organisms, especially invertebrates, are used as model systems for ecotoxicological studies also regarding the nervous system, even if studies on pollutants’ neurotoxic effects are still few. A great leap forward in knowledge can come from integrated omics studies that bring together genomics, transcriptomics, proteomics, and metabolomics data. These studies have revealed that pollutants are dangerous for the life of marine organisms, and not only because they can be modified in the environment, but also because they can combine giving rise to new mixtures. These new combinations can be even more harmful than individual pollutants. We must not forget that many marine organisms, both invertebrates and vertebrates, become part of the human food chain. Therefore, ultimately, the pollutants that contaminate the air, soil, and sea are potentially harmful to human health. The purpose of this review is to focus on some of the pollutants that are most frequently present in marine environments and analyze their neurotoxic potential effects on some marine model organisms.Invertebrates represent about 95% of existing species, and most of them belong to aquatic ecosystems. Marine invertebrates are found at intermediate levels of the food chain and, therefore, they play a central role in the biodiversity of ecosystems. Furthermore, these organisms have a short life cycle, easy laboratory manipulation, and high sensitivity to marine pollution and, therefore, they are considered to be optimal bioindicators for assessing detrimental chemical agents that are related to the marine environment and with potential toxicity to human health, including neurotoxicity. In general, albeit simple, the nervous system of marine invertebrates is composed of neuronal and glial cells, and it exhibits biochemical and functional similarities with the vertebrate nervous system, including humans. In recent decades, new genetic and transcriptomic technologies have made the identification of many neural genes and transcription factors homologous to those in humans possible. Neuroinflammation, oxidative stress, and altered levels of neurotransmitters are some of the aspects of neurotoxic effects that can also occur in marine invertebrate organisms. The purpose of this review is to provide an overview of major marine pollutants, such as heavy metals, pesticides, and micro and nano-plastics, with a focus on their neurotoxic effects in marine invertebrate organisms. This review could be a stimulus to bio-research towards the use of invertebrate model systems other than traditional, ethically questionable, time-consuming, and highly expensive mammalian models.

Hedgehog Signaling: Implications in Cancers and Viral Infections.

The hedgehog (SHH) signaling pathway is primarily involved in embryonic gut development, smooth muscle differentiation, cell proliferation, adult tissue homeostasis, tissue repair following injury, and tissue polarity during the development of vertebrate and invertebrate organisms. GLIoma-associated oncogene homolog (GLI) family of zinc-finger transcription factors and smoothened (SMO) are the signal transducers of the SHH pathway. Both SHH ligand-dependent and independent mechanisms activate GLI proteins. Various transcriptional mechanisms, posttranslational modifications (phosphorylation, ubiquitination, proteolytic processing, SUMOylation, and acetylation), and nuclear-cytoplasmic shuttling control the activity of SHH signaling pathway proteins. The dysregulated SHH pathway is associated with bone and soft tissue sarcomas, GLIomas, medulloblastomas, leukemias, and tumors of breast, lung, skin, prostate, brain, gastric, and pancreas. While extensively studied in development and sarcomas, GLI family proteins play an essential role in many host-pathogen interactions, including bacterial and viral infections and their associated cancers. Viruses hijack host GLI family transcription factors and their downstream signaling cascades to enhance the viral gene transcription required for replication and pathogenesis. In this review, we discuss a distinct role(s) of GLI proteins in the process of tumorigenesis and host-pathogen interactions in the context of viral infection-associated malignancies and cancers due to other causes. Here, we emphasize the potential of the Hedgehog (HH) pathway targeting as a potential anti-cancer therapeutic approach, which in the future could also be tested in infection-associated fatalities.

Disentangling the links between habitat complexity and biodiversity in a kelp-dominated subantarctic community.

Habitat complexity is one of the most important factors modulating species diversity. This feature comprises several interrelated attributes, such as number, size, and spatial arrangement of complexity‐forming elements. However, the separate and joint effects of these attributes on diversity and community structure are still not well understood. Here, we assess the relationships between several structural‐complexity attributes of the subantarctic kelp Lessonia flavicans and species richness, total abundance, and structure of kelp‐associated macrobenthic communities. We predicted that longer thalli and larger holdfasts favor greater species richness and total abundance of invertebrate organisms. To test the prediction, an observational sampling program was established in two sites of the Strait of Magellan. Uni‐ and multivariate analyses revealed both positive and negative effects of kelp structural‐complexity attributes on diversity. Holdfast diameter and maximum frond length, followed by thallus wet weight, had the strongest positive fits to species richness and total abundance; the number of stipes, on the other hand, was negatively associated with both response variables. Longer fronds were associated with greater abundances of spirorbid polychaetes. Larger holdfasts supported larger abundances of Nereididae and Terebelidae polychaetes and the limpet Nacella mytilina. Contrarily, kelps with longer fronds and more stipes supported fewer amphipods. In this way, we demonstrate that different dimensions of habitat complexity can have contrasting effects on diversity and community structure, highlighting the fundamental role of multiple dimensions of kelp habitat complexity for local biodiversity.

Differences in metallic content between marine vertebrates and invertebrates living in Oceanic Islands

The metallic content in each class of organism varies in different ways, depending on metabolism, habitat behavior, and where it is found in the trophic network. In this study, 845 specimens of different types of marine invertebrate and vertebrate organisms of the Canary Islands have been analyzed, of them the content of 20 metals and trace elements has been analyzed (Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, Pb, Sr, V and Zn) in mg / kg. In the PCoA analyzes it is clearly seen how the invertebrate and vertebrate organisms are separated according to their metallic content, there being significant differences between these two groups in each of the trace elements and metals. Invertebrate species having the highest concentration in all metals and trace elements, may have a higher concentration of metals than vertebrates because they have a very fast growth, and with it a high metabolic rate that causes higher concentrations of the elements to bioaccumulate.

Development of an aquatic exposure assessment model for Imidacloprid in sewage treatment plant discharges arising from use of veterinary medicinal products

BackgroundImidacloprid is an active ingredient included in plant protection, biocidal and veterinary medicinal products (VMPs). VMPs containing Imidacloprid are formulated as spot-on products or collars and designed to protect pets, predominantly dogs and cats, from parasite infestation. Monitoring data collected under the Water Framework Directive between 2016 and 2018 showed detectable and varying levels of Imidacloprid in the UK surface water bodies. The aim of the work was to investigate the potential contribution of VMPs by developing a model for predicting the emissions from sewage treatment plants from the use of dog and cat spot-on and collar VMPs. Due to the absence of appropriate exposure models for VMPs, the model was built based on the principles of environmental exposure assessment for biocidal products.ResultsThree emission paths were considered to be the most likely routes for repeated emissions to waterways from the use of spot-on and collar VMPs, i.e., transfer to pet bedding followed by washing, washing/bathing of dogs, and walking dogs in the rain. The developed model was used to calculate the Imidacloprid concentrations in surface water after discharge from wastewater treatment plants. Realistic worst-case input parameters were deduced from sales and survey data and experimental studies. Modelled total concentrations in surface water for each pathway ranged from 0.84 to 4.8 ng/L. The calculated concentrations did not exceed the ecological thresholds for the most sensitive aquatic invertebrate organisms and were found to be much lower than the UK monitoring data for river water. For example, the calculated concentration from the bathing/washing of dogs was < 3% of the highest levels of Imidacloprid measured in surface waters.ConclusionIn conclusion, a model has been successfully built and applied. The modelled data indicate that these VMPs make only a very small contribution to the levels of Imidacloprid observed in the UK water monitoring programme. Further, calculated concentrations do not exceed ecotoxicological threshold values indicating acceptable chronic safety to aquatic organisms.

Transcriptome Analysis Elucidates the Key Responses of Bryozoan Fredericella sultana during the Development of Tetracapsuloides bryosalmonae (Myxozoa).

Bryozoans are sessile, filter-feeding, and colony-building invertebrate organisms. Fredericella sultana is a well known primary host of the myxozoan parasite Tetracapsuloides bryosalmonae. There have been no attempts to identify the cellular responses induced in F. sultana during the T. bryosalmonae development. We therefore performed transcriptome analysis with the aim of identifying candidate genes and biological pathways of F. sultana involved in the response to T. bryosalmonae. A total of 1166 differentially up- and downregulated genes were identified in the infected F. sultana. Gene ontology of biological processes of upregulated genes pointed to the involvement of the innate immune response, establishment of protein localization, and ribosome biogenesis, while the downregulated genes were involved in mitotic spindle assembly, viral entry into the host cell, and response to nitric oxide. Eukaryotic Initiation Factor 2 signaling was identified as a top canonical pathway and MYCN as a top upstream regulator in the differentially expressed genes. Our study provides the first transcriptional profiling data on the F. sultana zooid’s response to T. bryosalmonae. Pathways and upstream regulators help us to understand the complex interplay in the infected F. sultana. The results will facilitate the elucidation of innate immune mechanisms of bryozoan and will lay a foundation for further analyses on bryozoan-responsive candidate genes, which will be an important resource for the comparative analysis of gene expression in bryozoans.

Eobania vermiculata as a potential indicator of nitrate contamination in soil

The effects of nitrates were analysed on the land snail Eobania vermiculata, a good bioindicator to assess the effects of certain pollutants in soil. It is known that the molluscs are very sensitive to contamination substances and can be used as sentinel organism for environmental pollution assessment. The nitrates are present in fertilizers and in food additives and their excess can not only be harmful to the environment but also dangerous for the humans. Indeed, in the mammals the nitrates are converted into nitrites and can cause a series of complications as the formation of methaemoglobin and cancers. In this study, adult organisms of E. vermiculata were exposed to soil containing 2000 mg/L of nitrates for 30 days to evaluate the stool microbiome and the histological changes at the level of the foot. Eggs of these snails were similarly treated to observe their hatching, survival and development. Histological changes were observed at level of the foot of adult snails exposed to nitrate and in their stools was evident an increase of bacteria, especially those that have a high ability to exploit nitrates and nitrogen as nutrients. Instead, the treated eggs showed changes in hatching, hypopigmentation of newborn snails and a decrease of their survival in time. The overall information obtained from these endpoints can provide important information regarding the quality of the environment. In addition, they also showed that the invertebrate organism E. vermiculata despite being a simple organism is very useful and efficient for ecotoxicological studies.

The standard metabolic rate of a land snail (Cepaea hortensis) is a repeatable trait and influences winter survival

Phenotypic selection on physiological parameters is an underrepresented topic in studies of evolutionary biology. There is especially a lack of studies involving invertebrate organisms. We studied the repeatability of the standard metabolic rate (SMR) and the effect of individual variation in SMR on the subsequent winter survival in a terrestrial shell-bearing mollusc, the white-lipped snail (Cepaea hortensis) in mid-Norway. SMR was measured twice during the autumn and – after an experimental overwintering at controlled conditions – twice during the following spring. We found a significant repeatability of SMR over all three time periods tested, with a clear effect of time, with a high repeatability of 0.56 over 4 days during spring, 0.44 over 12 days in the autumn and 0.17 over 194 days from autumn to spring. That SMR is a repeatable physiological trait across the winter period during which a possible selection might occur, suggests that SMR could be a potential target of natural selection. We indeed found that the autumn SMR significantly influenced the probability of survival during the winter period, with a combination of a positive linear (P = .011) and a quadratic stabilizing (P = .001) effect on SMR. Our results hence support the view that metabolic rate is an important physiological component influencing the fitness of an organism.

Analysis of total hydrocarbon and heavy metal accumulation in sediment, water and associated organisms of Mangrove ecosystem in the Niger Delta

Background /Objectives: This study\'s objectives are (1) to determine the total hydrocarbon (THC) and heavy metals in sediment and water samples collected from mangrove forest at different locations; (2) to compare the THC and heavy metals across different mangrove associated species; (3) to compare the relationship between THC and heavy metals in different organisms across different locations and (4) to determine the THC and heavy metal concentration between vertebrates and invertebrate organisms in mangrove forest. Methods/ Statistical analysis: It is hypothesized that chemical contamination will bioaccumulate across multiple mangrove associated organisms. Physicochemical analysis for Cadmium (Cd), Zinc (Zn), Lead (Pb), Iron (Fe) and total hydrocarbon (THC), was carried out on sediment, water, crabs, fish, insect, anadara and bird droppings, and was measured by spectrophotometric method using HACH DR 890 colorimeter and microwave accelerated reaction system (MARS Xpress, North Carolina). Findings: There was a significant difference in chemical composition between mangrove associated species (F4, 145 = 2.83, P=0.03). Anadara has the highest THC (36.44±9/4mg/kg) and Iron (6.97±1.32mg/kg) concentrations while bird droppings had the highest Lead concentration (10.83±1.27mg/kg). Fish had the highest Cadmium (4.20±1.01mg/kg) and Zinc (15.88±5.53mg/kg) concentrations. The order of organisms contamination is anadara>fish>bird droppings>crab>insect. The order of metal concentration is THC>Zn>Pb>Fe>Cd. In contrast, there was no significant difference between vertebrate and invertebrate organisms (F1, 148 = 0.08, P=0.78). Vertebrates have higher chemical composition (Cd, Pb and Zn) as compared to invertebrates (Fe and THC). Application/Improvement: Concentration of THC and heavy metals in most mangroves species were above the FAO/WHO standards. This implies that there is a horizontal THC and heavy metal contamination across trophic levels, which is detrimental to public health. Keywords: Bioaccumulation; heavy metals; invertebrates; Niger delta; pollution; vertebrates

Biological stenciling of mineralization in the skeleton: Local enzymatic removal of inhibitors in the extracellular matrix.

Biomineralization is remarkably diverse and provides myriad functions across many organismal systems. Biomineralization processes typically produce hardened, hierarchically organized structures usually having nanostructured mineral assemblies that are formed through inorganic-organic (usually protein) interactions. Calcium‑carbonate biomineral predominates in structures of small invertebrate organisms abundant in marine environments, particularly in shells (remarkably it is also found in the inner ear otoconia of vertebrates), whereas calcium-phosphate biomineral predominates in the skeletons and dentitions of both marine and terrestrial vertebrates, including humans. Reconciliation of the interplay between organic moieties and inorganic crystals in bones and teeth is a cornerstone of biomineralization research. Key molecular determinants of skeletal and dental mineralization have been identified in health and disease, and in pathologic ectopic calcification, ranging from small molecules such as pyrophosphate, to small membrane-bounded matrix vesicles shed from cells, and to noncollagenous extracellular matrix proteins such as osteopontin and their derived bioactive peptides. Beyond partly knowing the regulatory role of the direct actions of inhibitors on vertebrate mineralization, more recently the importance of their enzymatic removal from the extracellular matrix has become increasingly understood. Great progress has been made in deciphering the relationship between mineralization inhibitors and the enzymes that degrade them, and how adverse changes in this physiologic pathway (such as gene mutations causing disease) result in mineralization defects. Two examples of this are rare skeletal diseases having osteomalacia/odontomalacia (soft bones and teeth) - namely hypophosphatasia (HPP) and X-linked hypophosphatemia (XLH) - where inactivating mutations occur in the gene for the enzymes tissue-nonspecific alkaline phosphatase (TNAP, TNSALP, ALPL) and phosphate-regulating endopeptidase homolog X-linked (PHEX), respectively. Here, we review and provide a concept for how existing and new information now comes together to describe the dual nature of regulation of mineralization - through systemic mineral ion homeostasis involving circulating factors, coupled with molecular determinants operating at the local level in the extracellular matrix. For the local mineralization events in the extracellular matrix, we present a focused concept in skeletal mineralization biology called the Stenciling Principle - a principle (building upon seminal work by Neuman and Fleisch) describing how the action of enzymes to remove tissue-resident inhibitors defines with precision the location and progression of mineralization.

A new terrestrial trace fossil Feoichnus martini n. isp. from the Upper Cretaceous Two Medicine Formation (USA)

AbstractA new trace fossil, Feoichnus martini new ichnospecies, from the Two Medicine Formation is here described. This ichnotaxon is reported from the upper Campanian deposits of the Egg Mountain locality (Montana) and consists of a hemispherical to hemiellipsoidal structure with a truncated upper edge, and a regular, rounded lower edge marked by a lined border composed of stained layers. The trace maker likely impregnated the border using organic fluids. The simple lined wall observed in F. martini n. isp. suggests that the structure was produced by an invertebrate soil-dwelling organism, likely an insect. Specimens are preserved as casts in calcitic Inseptisols alongside an abundant vertebrate fossil record composed by dinosaurian and nondinosaurian fossil remains, dinosaur eggs and nest structures, and pervasive insect bioturbation. Feoichnus martini n. isp. represents an additional, minor component of the impoverished Celliforma ichnofacies reported at Egg Mountain and expands the paleogeographical distribution of the ichnogenus Feoichnus Krause et al., 2008 to the Upper Cretaceous deposits of Montana, USA.UUID: http://zoobank.org/7c1a5026-7f27-4f12-a9fb-1eb8daa93baf

Fruit Fly as a Model Organism for Blood-Brain Barrier Penetration and Infectious Disease in the Nanomedical Niche

Drosophila melanogaster, a.k.a. the common fruit fly, is a simple organism that may give a rapid, high-throughput response in regard to the therapeutic efficacy of nanoparticles and drugs, while circumventing the high environmental and monetary cost of today’s typical in vivo assays involving more complex animals, along with the immeasurable suffering imposed onto them. Here we give the progress report on our effort to turn D. melanogaster into a model organism for the in vivo testing of Blood-Brain Barrier (BBB) penetration of nano-particles and the treatment of infectious disease. We show that orally ingested superparamagnetic nanoparticles successfully cross the BBB in D. melanogaster and localize to the optic lobes of the third instar larval brain, while causing no adverse effects to the invertebrate organisms. We also show that both orally ingested calcium phosphate nanoparticles and biofilm-forming P. aeruginosa localize to the Drosophila crop, the food storage organ of the fly, which shrinks in response to infection. The model does not induce mortality consequential to infection and the effects of the internalization and proliferation of the microbes are evaluable by measuring the crop parameters, including fluorescence intensity and size. Continued development of these two models could simplify the preclinical testing of medical treatments and of pharmaceutical agents for neurological and infectious disease, while ensuring robust and reliable levels of statistical significance at low cost.