I have often wondered if earthworms are the creatures who first demonstrated a functional dichotomy in evolution: they evolved to be able to clean up the battlefield after having killed foreign invaders. They have cells that, much to my wondering eyes, look very much like human natural killer cells and neutrophils when examined with cytofluorimetric analysis and microscopy. I may sound a little eccentric when I tell you that my excitement over my beloved creatures is immense--I believe they hold healing treasures for us all.

Plant and animal-derived products are crucial components in complementary and alternative medicine. Although modern medicine has provided numerous innovations and advancements, these often fail to reveal new and dependable, inexpensive treatments nor real cures that are relatively free of adverse side effects. We present evidence that hirudotherapy, which utilizes leeches, improves certain diseases, including osteoarthritis. Osteoarthritis, a disease in joints, could benefit from use of medicinal peptides found in leech saliva, components of its immune system.

Despite modern medicine's incredible innovation and resulting accumulation of valuable knowledge, many of the world's most problematic diseases such as Alzheimer Disease (AD) still lack effective cures and treatments. Western medicine has revealed many genetic, cellular, and molecular processes that characterize AD such as protein aggregation and inflammation. As the need for novel and effective treatments increases, researchers have turned towards traditional medicine as a resource. Modern, evidence based research examining traditional and complementary remedies for AD has generated promising results within the last decade. Animal based products inhibiting cellular toxicity, anti-inflammatory nutraceuticals such as omega-3 fatty acids, and plant based compounds derived from herbal medicine demonstrate viability as neuroprotective treatments and possible application in developing pharmaceuticals. Analysis of antioxidant, anti-inflammatory, and neuroprotective phytochemicals used in various traditional medicines around the world reveal potential to ameliorate and prevent the devastating neurodegeneration observed in AD.

Lumbrokinases, a group of fibrinolytic enzymes extracted from earthworm, have been widely used to prevent and treat various cardiovascular diseases. They specifically target fibrin to effectively degrade thrombi without major side effects. Plant expression systems are becoming potential alternative expression platforms for producing pharmaceutical proteins. In this work, a lumbrokinase (PI239) was produced from a plant system. Both wild-type (WT) and plant codon-optimized (OP) PI239 gene sequences were synthesized and cloned into a geminivirus-based single-vector DNA replicon system. Both vectors were independently expressed in tobacco (Nicotiana tabacum) leaves transiently by agroinfiltration. Overexpressed PI239 resulted in sudden tissue necrosis 3 days after infiltration. Remaining proteins were purified through His-tag affinity chromatography and analyzed with SDS-PAGE and Western blot methods. Purified PI239 successfully degraded artificial fibrin with relative activity of 13,400 U/mg when compared with commercial lumbrokinase product. In vitro tests demonstrated that plant-derived PI239 dissolved human blood clots and that the plant expression system is capable of producing functional PI239.

As we search for answers to modern medicine's most prevalent and challenging problems, the relationship between nutrition, immunity, and biological function of various natural compounds are preimminent. Nutritional research involving genomics provides rational capabilities for preventing disease. Scientific advances in genomic sequencing reveal opportunities for exploring diet-health relationships and potential for individual, genotype based dietary recommendations. Utilizing molecular and genetic technology to analyze impact of nutrition on genomics and metabolism reveals that nutrients may influence certain innate and/or acquired immune functions. By analyzing immune mechanisms including their cells and complex molecules, animal models have offered relevant insight that clarifies interrelations between immunity and nutrition. Plant products also provide numerous resources through bioengineering for designing novel pharmaceuticals. Having long been employed successfully in traditional and folk medicines, plant compounds exhibit anti-inflammatory, antimicrobial, and angiogenic activity. As a result, we now have a promising arsenal for successful application of bioactive compounds.

Adaptive immunity has often been considered the penultimate of immune capacities. That system is now being deconstructed to encompass less stringent rules that govern its initiation, actual effector activity, and ambivalent results. Expanding the repertoire of innate immunity found in all invertebrates has greatly facilitated the relaxation of convictions concerning what actually constitutes innate and adaptive immunity. Two animal models, incidentally not on the line of chordate evolution (C. elegans and Drosophila), have contributed enormously to defining homology. The characteristics of specificity and memory and whether the antigen is pathogenic or nonpathogenic reveal considerable information on homology, thus deconstructing the more fundamentalist view. Senescence, cancer, and immunosuppression often associated with mammals that possess both innate and adaptive immunity also exist in invertebrates that only possess innate immunity. Strict definitions become blurred casting skepticism on the utility of creating rigid definitions of what innate and adaptive immunity are without considering overlaps.

As a traditional therapy applied for thousands of years, acupuncture has recently been attracting more and more investigators throughout the world. In the theory of traditional Chinese medicine, it is proposed that acupuncture can strengthen the human body to resist diseases by puncturing needles at certain points. The characteristic that acupuncture enhances resistance is closely related with the immune system, which functions in defense, homeostasis, and surveillance. More and more research has revealed that acupuncture can regulate immunity, for example, to enhance anticancer and antistress immune function and exert anti-inflammation effects. This may be the basis of acupuncture in preventing and treating later diseases. This special issue was developed to stimulate the continuing efforts in promoting the research on acupuncture and immunity.

This special issue of eCAM has brought together several unique investigators interested in invertebrate animal models. Investigators would benefit by examining advances and applications of findings derived from useful terrestrial and marine invertebrate animal models. Results derived from well-defined evidence-based approaches are usually relevant to humans; the advantages are numerous, inexpensive, and noncontroversial and are not subject to rigid, ethical, or moral guidelines. In the historical context, these animals have been crucial in many cultures before the advent of industrial medicines thus providing the only source of aid. Now, there is a move to return to or at least to include these practices lest they disappear since, in certain cultures, they represent the sole source of useful health remedies. However, evidence-based medicine emphasizes the importance of subjecting animal products to rigorous analysis. Results are then standardized and serve to educate populations including those that use exclusively western practices. eCAM stands ready to accept the importance and contributions of bees as valuable sources of beneficial activities (pollination) and products (honey and propolis) that for centuries have enhanced human lives as food and medicine. Others include corals, millipedes, maggots, and earthworms. Corals are marine invertebrates that live in compact colonies. Humans have learned that they are useful in the manufacture of jewelry and chemical compounds that are useful against cancer, AIDS, and pain and serve as skeletons during bone grafting. Millipedes are arthropods (the same group as shrimps, crabs, butterflies, and bees) which in some cultures are used during pregnancy and as cures for fever, wounds, earaches, and hemorrhoids and food among the Bobo people of Burkina Faso when crushed; sometimes they are boiled and eaten in tomato sauce. Since the time of Napoleon and even centuries earlier, maggots have been used especially in patients with severe intractable wounds caused by nerve degeneration that results from diabetes. In fact, leeches and maggots are now valuable components of the emerging field of biotherapy, the therapeutic use of living creatures to clean gangrenous tissue often found in ulcers, burns, and postoperative infections. Earthworms are experiencing growing popularity due to their many applications including sources of high protein as food and the production of an anticlotting agent known as lumbrokinase that dissolves blood clots in patients. Aside from observations and usefulness by the layman, certain techniques are valuable as molecular biologists strive to understand the minute structure of molecules in order to improve therapies. Let us not forget the contributions of earthworms as tillers of the soil which enhance agricultural output. Earthworms keep the soil aerated by their constant churning which makes for improved yield of plants regardless of the proposed use. Chih-Yang Huang Edwin L. Cooper Catherine Fang-Yeu Poh Wei-Wen Kuo Tung-Sheng Chen Ronald Sherman

Over the last few years, we have pursued the use and exploitation of invertebrate immune systems, most notably their humoral products, to determine what effects their complex molecules might exert on humans, specifically their potential for therapeutic applications. This endeavor, called "bioprospecting," is an emerging necessity for biomedical research. In order to treat the currently "untreatable," or to discover more efficient treatment modalities, all options and potential sources must be exhausted so that we can provide the best care to patients, that is, proceed from forest and ocean ecosystems through the laboratory to the bedside. Here, we review current research findings that have yielded therapeutic benefits, particularly as derived from soft and hard corals. Several applications have already been demonstrated, including anti-inflammatory properties, anticancer properties, bone repair, and neurological benefits.

Recent evidence supports that prokaryotes exhibit adaptive immunity in the form of CRISPR (Clustered Regularly Interspersed Short Palindromic Repeats) and Cas (CRISPR associated proteins). The CRISPR–Cas system confers resistance to exogenous genetic elements such as phages and plasmids by allowing for the recognition and silencing of these genetic elements. Moreover, CRISPR–Cas serves as a memory of past exposures. This suggests that the evolution of the immune system has counterparts among the prokaryotes, not exclusively among eukaryotes. Mathematical models have been proposed which simulate the evolutionary patterns of CRISPR, however large gaps in our understanding of CRISPR–Cas function and evolution still exist. The CRISPR–Cas system is analogous to small RNAs involved in resistance mechanisms throughout the tree of life, and a deeper understanding of the evolution of small RNA pathways is necessary before the relationship between these convergent systems is to be determined. Presented in this review are novel RNAi therapies based on CRISPR–Cas analogs and the potential for future therapies based on CRISPR–Cas system components.

Previous work exploring the interrelationships between sex steroids (e.g. androgens, testosterones and 11-ketotestosterones) and social behavior in teleosts suggest that mirror-elicited aggression in cichlid fish may not trigger a hormonal response. Using the Mozambique tilapia (Oreochromis mossambicus) to analyze immune responses as a result of social stress, we measured levels of cortisol and melatonin using Enzyme-Linked Immunosorbent Assay (ELISA) assays. In this work, we demonstrated that cortisol concentrations are significantly lower yet the levels of melatonin remain unchanged in tilapia that are fighting their mirror image. Our results suggested that in tied fights, certain hormone levels remain unchanged (e.g. androgens) due to the lack of melatonin induction.