Low Species Specificity Research Articles

Navigating the fungal battlefield: cysteine-rich antifungal proteins and peptides from Eurotiales.

Fungi are ubiquitous in the environment and play a key role in the decomposition and recycling of nutrients. On the one hand, their special properties are a great asset for the agricultural and industrial sector, as they are used as source of nutrients, producers of enzymes, pigments, flavorings, and biocontrol agents, and in food processing, bio-remediation and plant growth promotion. On the other hand, they pose a serious challenge to our lives and the environment, as they are responsible for fungal infections in plants, animals and humans. Although host immunity opposes invading pathogens, certain factors favor the manifestation of fungal diseases. The prevalence of fungal infections is on the rise, and there is an alarming increase in the resistance of fungal pathogens to approved drugs. The limited number of antimycotics, the obstacles encountered in the development of new drugs due to the poor tolerability of antifungal agents in patients, the limited number of unique antifungal targets, and the low species specificity contribute to the gradual depletion of the antifungal pipeline and newly discovered antifungal drugs are rare. Promising candidates as next-generation therapeutics are antimicrobial proteins and peptides (AMPs) produced by numerous prokaryotic and eukaryotic organisms belonging to all kingdom classes. Importantly, filamentous fungi from the order Eurotiales have been shown to be a rich source of AMPs with specific antifungal activity. A growing number of published studies reflects the efforts made in the search for new antifungal proteins and peptides (AFPs), their efficacy, species specificity and applicability. In this review, we discuss important aspects related to fungi, their impact on our life and issues involved in treating fungal infections in plants, animals and humans. We specifically highlight the potential of AFPs from Eurotiales as promising alternative antifungal therapeutics. This article provides insight into the structural features, mode of action, and progress made toward their potential application in a clinical and agricultural setting. It also identifies the challenges that must be overcome in order to develop AFPs into therapeutics.

Evaluation of plasma atrial natriuretic peptide concentration in healthy bottlenose dolphins (Tursiops truncates).

There are currently no standard methods for diagnosing cardiac diseases in dolphins. These diseases may consequently be overlooked and go undiagnosed. The presence and severity of cardiac diseases in humans can be determined using blood tests. Atrial natriuretic peptide (ANP) used in human cardiac examinations has low species specificity. There have already been reports of homology between dolphin and human ANP; however, its potential for clinical application in dolphins has not been tested. This study was conducted to establish a reference for ANP levels in healthy bottlenose dolphins. Healthy bottlenose dolphins (seven females; estimated to be 7-30 years of age) at an aquarium in Japan were sampled. Each animal was tested for ANP at least three times, and the mean value and standard deviation were calculated to be 43.4 ± 19.2 pg/mL. In humans, patients with high plasma ANP levels have a poor prognosis. In veterinary medicine, cutoff values for the diagnosis of mitral regurgitation and heart failure in dogs have been established and used to predict prognosis. The results of the present study may contribute to the health management of bottlenose dolphins, particularly in the early detection and treatment of cardiac diseases.

Antitumor Properties of Matrikines of Different Origins: Prospects and Problems of Their Application.

Matrikines (MKs) can be a rich source of functional nutrition components and additional therapy, thereby contributing to human health care and reducing the risk of developing serious diseases, including cancer. Currently, functionally active MKs as products of enzymatic transformation by matrix metalloproteinases (MMPs) are used for various biomedical purposes. Due to the absence of toxic side effects, low species specificity, relatively small size, and presence of various targets at the cell membranes, MKs often exhibit antitumor properties and, therefore, are promising agents for antitumor combination therapy. This review summarizes and analyzes the current data on the antitumor activity of MKs of different origins, discusses the problems and prospects for their therapeutic use, and evaluates the experimental results of studying the antitumor properties of MKs from different echinoderm species generated with the help of a complex of proteolytic enzymes from red king crab Paralithodes camtschatica. Special attention is paid to the analysis of possible mechanisms of the antitumor action of various functionally active MKs, products of the enzymatic activity of various MMPs, and the existing problems for their use in antitumor therapy.

Survey for snake-tick (Ixodida) association in some districts of West Bengal, India

During a study, two tick species viz. Amblyomma gervaisi and Am. helvolum were collected from snakes covering 15 species belonging to 12 genera. The period of extended survey of snake- tick association was undertaken from 2009 to 2015 covering both natural and captive habitats from six districts of West Bengal, India. It was observed that Am. gervaisi mostly prefers Naja naja, Daboia russelii and Ptyas mucosa as host, whereas Am. helvolum prefers N. kaouthia and Bungarus fasciatus. Naja kaouthia is preferred as host by both Am. gervaisi and Am. helvolum. Eryx johnii and Gongilophis conicus are found to be infested by both Am. gervaisi and Am. helvolum, whereas Python molurous is only infested by Am. helvolum. No ticks are found to associate with six snake species viz., Xenochrophis piscator, Lycodon aulicus, L. jara, Ahaetulla nasuta, Chrysopelea ornata, and Boiga trigonata. It is noted that tick loads on host snakes in natural habitats is greater than captivity, with few negligible exceptions. Statistical analysis of tick prevalence and abundance are done stating that Am. gervaisi is dominant snake infesting tick compared to Am. helvolum in West Bengal. Species population of both Am. gervaisi and Am. helvolum denote female's domination with low species specificity.

Coevolutionary analyses of the relationships between piroplasmids and their hard tick hosts.

Host–parasite coevolution is a key driver of biological diversity. To examine the evolutionary relationships between piroplasmids and their hard tick hosts, we calculated the molecular clock and conducted phylogenetic analyses of both groups. Based on our results, we conclude that the divergence time of piroplasmids (∼56 Mya) is later than divergence time of their hard tick hosts (∼86 Mya). From analyses of the evolution of both piroplasmid and vector lineages and their association, we know that hard ticks transmit piroplasmids with high genus specificity and low species specificity.

Heterologous Radioimmunoassay for Llama and Alpaca Luteinizing Hormone with a Monoclonal Antibody, an Equine Standard and a Human Tracer

A radioimmunoassay for llama and alpaca LH was developed using a human I125LH tracer from a commercial kit, equine LH diluted in human LH free serum as standard, and a monoclonal antibody (518B7) specific for LH but with low species specificity. A 60-min delay in the addition of the tracer and overnight incubation gave a sensitivity of 0.8 microgram L-1. The intra-assay coefficient of variation was 37% at 1 microgram L-1, declined to 15% at 4 micrograms L-1 and was below 6% for concentrations up to 32 micrograms L-1. The inter-assay coefficients of variation for 3 control samples were 20% (2.8 micrograms L-1), 16% (7.1 micrograms L-1) and 9.8% (19 micrograms L-1). In an attempt to increase sensitivity, all tubes were preincubated for 4 h at room temperature before adding the tracer, and the sample volume was increased from 50 microL to 100 microL (in the standard curve the increased volume was compensated for by human LH free serum). With this protocol, the assay sensitivity was 0.5 microgram L-1. The assay was validated clinically and demonstrated increased concentrations of LH after mating in llamas and alpacas. Furthermore, the assay was used to monitor LH responses to a single dose of GnRH in llamas (adult males and females at different ages).

Radioimmunoassay of Bovine, Ovine and Porcine Luteinizing Hormone with a Monoclonal Antibody and a Human Tracer

A radioimmunoassay for bovine (bLH), ovine (oLH) and porcine (pLH) luteinizing hormone was developed using a human 125ILH tracer from a commercial kit and a monoclonal antibody (518B7) specific for LH but with low species specificity. Standard curves demonstrated similar binding kinetics when bLH, oLH and pLH were incubated with tracer and antibody for 2 h at room temperature. A 30-min delay in the addition of the tracer gave sufficient sensitivity when analysing pLH. Separation of antibody-bound LH from free hormone was achieved by using second antibody-coated micro Sepharose beads. The assay was validated and the performance compared with that of an RIA currently in use for determination of bLH and oLH (coefficient of correlation: 0.99 and 0.98). Regardless of the standards used, intra-assay coefficients of variation were < 10% for LH concentrations exceeding 1 microgram/L. The inter-assay coefficients of variation were < 15%. The assay was used for clinical evaluation demonstrating the pre-ovulatory LH surge in two cyclic cows, LH pulsatility in an oophorectomized ewe and LH response to GnRH injection in a boar.

Characterization of a monoclonal antibody which detects luteinizing hormone from diverse mammalian species

The present study describes the development and characterization of a monoclonal antibody (518B7) generated against bovine LH (bLH). Although 518B7 was extremely specific for LH, very low species specificity was observed. A RIA using this antibody and radioiodinated equine LH (eLH) showed good sensitivity for all mammalian LH preparations tested, with the exception of human LH (15%, relative to the eLH reference standard). Activities of most mammalian LH's ranged between approximately 50–200%. Much less activity was detected with reptilian LH (<1.5%). Amphibian and avian LH fractions were essentially inactive. The reactivities of LH alpha and beta subunits from a variety of mammals clearly showed that the antibody reacts with the beta subunit. Sensitive RIAs were also developed utilizing 125I-bovine and 125I-rat LH. Interestingly, all hormone preparations which showed sufficient reactivity for statistical analysis within the dose ranges used in the present study (0.01–1000 ng/tube) produced a displacement curve parallel to the reference standard. We have also validated the use of 518B7 in detecting LH in serum. Parallel dilution curves relative to purified LH reference standards were observed with equine and bovine serum samples and equine pituitary extract. High (average 94%) recoveries were also seen with bovine serum with known amounts of exogenously added bLH. Similar patterns of LH secretion were detected with a RIA based upon 125I-bLH and 518B7 and a previously described polyclonal antibody-based RIA in bovine serum samples during estrus. Thus, a monoclonal antibody for LH has been produced which can be used to develop sensitive and specific RIAs in many different mammalian species. This antibody can be readily produced in amounts sufficient to provide a stable source of a high quality LH antibody, which may find wide applications in endocrinological research dealing with both domestic and laboratory animals.

The immunological specificity of myosins from cross-striated muscles as revealed by quantitative microcomplement fixation and enzyme inhibition by antisera

The immunological properties of myosins, especially their muscle-type, class and species specificity, are still controversial. It is the opinion of the authors that the lack of agreement might at least in part be due to the use of contaminated myosins as immunogens and inappropriate methods. We, therefore, purified myosins to a very high degree (≈99%) and induced antisera in guinea-pigs. Studies of quantitative microcomplement fixation and enzyme-inhibition by antisera yielded the following results: myosins of cross-striated muscle have an absolute class, a very pronounced muscle-type and a low species specificity. It can be shown that even a very small contamination of myosins with other proteins could seriously hamper the experiments and that the results obtained depend significantly on the immunological methods employed.

Immunochemical Studies on Proteins Important in Blood Coagulation

SummaryThis report draws attention to the fact that only minute quantities of purified serum proteins (1 —15 μg/kg body weight) are required to stimulate antibody production. Injection of amounts greater than 1 mg/kg either is a waste or favors the production of non-specific antisera. Potent antisera can be produced in rabbits with a single intramuscular injection using either Al(OH)3 gel or Amphojel as an adjuvant.A number of purified plasma protein preparations which appear homogenous by ultra centrifugation or biological activity studies actually contain several antigenic components and are immunologically heterogeneous, when checked with highly specific antisera.Fibrinogen prepared by alcohol fractionation and tannic acid purification was found to be free of prothrombin, antihemophilic factor and Profibrinolysin. Both Immunoelectrophoresis and double diffusion in agar indicated a homogeneous preparation.Fibrinogens prepared from human, canine and bovine plasmas exhibited a low species specificity.

The specificity of anti-kidney antibody determined by its effect upon tissue culture explants.

While investigating the pathogenesis of experimental nephritis produced by rabbit anti-rat-kidney antibody, it occurred to us that the effects of antikidney antibody on kidney tissue might readily be visualized in tissue cultures. The specificity of tissue antigens has previously been investigated by the usual immunologic procedures [1] and the effects of antibodies [2,3] upon tissue explants has long been known. The growth and function of tissue explants have previously been used to study the specificity of tissue antibodies [4,5], but the antisera used were of low, uncertain potency, and only a few different tissues were examined in testing the toxic effects. The following study shows that rabbit anti-rat-kidney gamma globulin is toxic for tissue explants of brain and heart muscle, as well as kidney. These effects are of low species specificity and may be observed in cultures prepared from chick embryo tissues, as well as those prepared from rat tissues. In addition, the serum of patients with glomerular nephritis was found to contain a substance with similar effects and specificity.