Venom gland size and venom complexity-essential trophic adaptations of venomous predators: A case study using spiders.

Specialized predators possess a variety of adaptations. In venomous predators, this may include the size of the venom gland and venom composition. It is expected that due to different foraging strategies, predators with a wide trophic niche (generalists) should possess larger venom glands that contain more diversified components than predators with a narrow niche (specialists). We focused on spiders, as the most diversified group of venomous predators, in which a wide variety of trophic strategies have evolved. We conducted a comparative analysis using 40 spider species, in which we measured the size of their venom gland and venom complexity using proteome profiling methods. The species were classified into three trophic groups: generalists, facultative specialists and obligatory specialists. We found that the venom glands of generalists are larger than those of obligatory specialists, which is presumably due to more frequent prey capture by the former. The complexity of venom of peptides (2-15 kDa) and proteins (15-250 kDa) was more diverse in generalists than in specialists. Multivariate analysis of venom revealed significant differences among the three trophic categories only in the complexity of peptides. Our study thus shows that venom gland size and its content have taken different pathways during the evolution of different trophic strategies in spiders. Generalists evolved larger venom glands with more complex composition, whereas obligatory specialists possess smaller glands with less diverse chemical structures.

Venom of prey-specialized spiders is more toxic to their preferred prey: A result of prey-specific toxins.

In specialized predators, a variety of adaptations have evolved to such a level of specificity that they allow very effective exploitation of focal prey. Venom is an essential adaptive trait of predatory venomous species, such as spiders, yet our knowledge of spider venom is incomplete. In agreement with the prey preference hypothesis, we expected that the venom of spider specialists should be more toxic to focal than to alternative prey, because it is composed of prey-specific toxins. Here we used spiders with three types of trophic specializations: specialists that were ant-eating, termite-eating and spider-eating. We compared the efficacy of prey capture of preferred and alternative prey (measured as paralysis latency) with that of related generalists and profiled the venom of the studied species using proteomic methods. We used 22 spider species: six myrmecophagous, two termitophagous, three araneophagous and 11 euryphagous generalist species belonging to different families. We found that ten of the eleven specialist species induced significantly shorter paralysis latency in preferred prey than in alternative prey. Generalists exhibited either similar efficiency on both prey types or slightly higher efficiency on preferred prey. Multivariate analysis of proteomic profiles (peptides and proteins) revealed significant differences between trophic specializations, particularly in peptides. Specialists appear to have venom composed of unique specific compounds as revealed by the multivariate ordination and indicator analysis. These components are likely prey-specific toxins.

Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS.

Spider venoms are highly complex mixtures containing biologically active substances with potential for use in biotechnology or pharmacology. Fingerprinting of venoms by Matrix-Assisted Laser Desorption-Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) is a thriving technology, enabling the rapid detection of peptide/protein components that can provide comparative information. In this study, we evaluated the effects of sample preparation procedures on MALDI-TOF mass spectral quality to establish a protocol providing the most reliable analytical outputs. We adopted initial sample preparation conditions from studies already published in this field. Three different MALDI matrixes, three matrix solvents, two sample deposition methods, and different acid concentrations were tested. As a model sample, venom from Brachypelma albopilosa was used. The mass spectra were evaluated on the basis of absolute and relative signal intensities, and signal resolution. By conducting three series of analyses at three weekly intervals, the reproducibility of the mass spectra were assessed as a crucial factor in the selection for optimum conditions. A sample preparation protocol based on the use of an HCCA matrix dissolved in 50% acetonitrile with 2.5% TFA deposited onto the target by the dried-droplet method was found to provide the best results in terms of information yield and repeatability. We propose that this protocol should be followed as a standard procedure, enabling the comparative assessment of MALDI-TOF MS spider venom fingerprints.

Circulating miR-17-3p, miR-29a, miR-92a and miR-135b in serum: Evidence against their usage as biomarkers in colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is one of the leading causes of cancer-related death in the world. Therefore, there is a high demand for cost-effective and non-invasive biomarkers that would enable an early detection of asymptomatic and curable disease with high sensitivity and specificity. OBJECTIVE: The main objective of this study was to investigate the potential of circulating miRNAs as biomarkers of CRC. METHODS: Total RNA enriched for small RNAs was isolated from 100~sera of patients with CRC and 30 sera of healthy donors. The expression levels of miR-17-3p, miR-29a, miR-92a and miR-135b were determined using quantitative real-time PCR. The average expression levels of particular miRNAs were normalized to miR-16 levels and statistically evaluated. RESULTS: Using Mann-Whitney U test, no significant differences were observed in miR-17-3p (P=0.18), miR-29a (P=0.14) and miR-92a (P=0.60) levels between sera of CRC patients and controls. The levels of miR-135b in serum were too low to be quantified accurately. Subsequently, we tried to correlate expression levels of analyzed miRNAs to clinical-pathological features of CRC patients. Only levels of mir-29a were correlated with the clinical stage (P=0.04). Expression levels of the other miRNAs were correlated neither with the clinical stage, nor with the grade. CONCLUSIONS: Interestingly, our results are contradictory to previous studies performed on the CRC patients from Chinese population, providing an evidence against usage of serum miR-17-3p, miR-29a, miR-92a and miR-135b as new biomarkers for early detection of CRC.