Low Molecular Weight Toxins Research Articles

Evaluation of cellular immune response in rabbits after exposure to cobra venom and purified toxin fraction

Snakebite by a cobra is considered neurotoxic as the cause of neuromuscular paralysis mediated by low molecular weight toxins, which are major toxin components of cobra. However, these toxins represent a problem in generating antibodies owing to their low immunogenicity. Developing complementary strategies to improve the antibody response could be a useful approach to creating better therapeutic antivenoms with higher neutralizing potencies. To develop simple immunization strategies for more potent antivenoms by studying the effects of combining crude cobra venom and toxin fraction in a complementary way. The evaluation of specific cell immunology and cytokine mediators for relevant immune responses will be measured in a rabbit model using four simple immunization strategies. Flow cytometry will be used to quantify the number of B and T cells, and qRT-PCR will be used to ascertain the cytokine genes expressed. B cells with anti-CD20 were seen on D14, and a booster dose was insufficient to maximize the antibodies. Conversely, anti-CD5 for T cells decreased periodically but remained stable. Using a mixture of crude cobra venom and its <10 kDa fraction, peak expression of pro-inflammatory cytokine genes was seen in D42 or D58, with a rise of 4 and 6 folds. Similarly, gene expression of pro-inflammatory cytokines was greater than that of anti-inflammatory cytokines (IL-4 and IL-10), which were up-regulated after D42. Thus, immunization with both the crude and its <10 kDa fraction of cobra venom seems to have synergistic effects that boost cytokines, activate the immune system, and cause lymphocyte differentiation.

Сорбционно-структурные свойства аэрогельных материалов на основе биополимеров

Nowadays aerogel materials (AM) are successfully used as entero- and applicative sorbents to eliminate excessive amounts of heavy metals and toxins from living organisms. Natural biopolymers alginate and chitosan, as well as various lignin derivatives are an inexhaustible raw material base for the creation of AM. A significant number of sorption materials and wound coatings of various types have been developed on their basis, which is associated not only with a wide range of physicochemical properties of these polymers and their already proven biomedical activity, but also with the prevalence and renewability of raw material sources for the production of these polymers, ease of extraction, the possibility of achieving a high degree of purification and relatively low price. The key stage in the AM synthesis is the formation of a strong hydrogel which is the AM framework. One of the technological methods is to obtain interpolyelectrolyte solid hydrogel. The paper proposes two different packaging models for the formation of the structure of interpolyelectrolyte complexes (IPEC) based on biopolymers pairs: sodium alginate (ALNa)–chitosan (CT) and sodium lignosulfonate (LSNa)–CT. The first model is a block model, in which the structure is formed due to ionic bonds between the carboxyl groups of ALNa and amino groups of CT, as well as a cooperative system of hydrogen bonds and dispersion interactions. The second model is an aggregation-tubular model, the structure of which is formed through ionic bonds between sulfogroups (within the rod-shaped supramolecular structures of LSNa) and amino groups of CT, as well as hydrogen bonds and dispersion interactions. Upon the process of IPEC drying under supercritical (SC-) conditions, strong phase contacts are formed, and the changes in the gel structure become irreversible. As a result, hydrophobic micro- and mesoporous two-component AMs differing in internal structure were obtained. AM ALNa–CT are characterized by fibrillar structure, and LSNa–CT – by structural elements of spherical shape. The obtained AM ALNa–CT and LSNa–CT have high sorption activity towards water and a wide range of heavy metals and low molecular weight toxins. The purpose of the work is to study the structural and sorption properties of AM based on biopolymers of various structural organization. A significant increase in the sorption activity of AM ALNa–CT in comparison with LSNa–CT is apparently due to their different supramolecular structure. There is a combination of several sorption mechanisms such as wetting, absorption, diffusion, osmotic phenomena and chemical interaction due to the highly porous structure of AM and the presence of sorption-active centers. For citation: Brovko O.S., Palamarchuk I.A., Gorshkova N.A., Bogdanovich N.I., Ivakhnov A.D. Sorption and Structural Properties of Aerogel Materials Based on Biopolymers. Lesnoy Zhurnal = Russian Forestry Journal, 2023, no. 6, pp. 190–203. (In Russ.). https://doi.org/10.37482/0536-1036-2023-6-190-203

Multi-drug loaded microneedles for emergency treatment of snakebite envenomation

Snakebite envenomation is a WHO classified priority neglected disease, which manifests multiple local and systemic toxic effects. Intravenous antivenom therapy is the treatment of choice in clinical settings but it cannot be employed for pre-hospital management of envenomation cases due to certain limitations. On-field first-aid measures presently include use of pressure bandage immobilization (PBI), which has sub-optimal efficacy in managing snakebite casualties. There is therefore an urgent need for developing non-immunological, therapeutic interventions that could be self-administered immediately after snakebite, alone or in tandem with PBI, to reduce the morbidity and mortality. Venom toxins enter into the bloodstream via lymphatic pathway [e.g., high molecular weight (HMw) toxins] or directly ingress into the vasculature due to cytotoxic endothelium disruption [e.g., low molecular weight (LMw) toxins]. Recent literature has identified some pharmacological targets, which when applied topically impede the systemic absorption of HMw venom toxins by modulating lymphatic transit time, whereas a few others act as direct inhibitors of LMw toxins, when injected parentally. Based on available evidences our hypothesis shall lead to deliver formulation containing combination of drugs applied topically through convenient, self-administrated microneedles, either to delay absorption and lymphatic transit of HMw venom toxins into the bloodstream or to inhibit the mechanistic pathway of LMw toxins, and could therefore be more useful for pre-hospital management following snakebite as compared to PBI alone.

Non-compartmental toxicokinetic studies of the Nigerian Naja nigricollis venom.

Snakebite envenoming (SBE) is a neglected public health problem, especially in Asia, Latin America and Africa. There is inadequate knowledge of venom toxicokinetics especially from African snakes. To mimic a likely scenario of a snakebite envenoming, we used an enzyme-linked immunosorbent assay (ELISA) approach to study the toxicokinetic parameters in rabbits, following a single intramuscular (IM) administration of Northern Nigeria Naja nigricollis venom. We used a developed and validated non-compartmental approach in the R package PK to determine the toxicokinetic parameters of the venom and subsequently used pharmacometrics modelling to predict the movement of the toxin within biological systems. We found that N. nigricollis venom contained sixteen venom protein families following a mass spectrometric analysis of the whole venom. Most of these proteins belong to the three-finger toxins family (3FTx) and venom phospholipase A2 (PLA2) with molecular weight ranging from 3 to 16 kDa. Other venom protein families were in small proportions with higher molecular weights. The N. nigricollis venom was rapidly absorbed at 0.5 h, increased after 1 h and continued to decrease until the 16th hour (Tmax), where maximum concentration (Cmax) was observed. This was followed by a decrease in concentration at the 32nd hour. The venom of N. nigricollis was found to have high volume of distribution (1250 ± 245 mL) and low clearance (29.0 ± 2.5 mL/h) with an elimination half-life of 29 h. The area under the curve (AUC) showed that the venom remaining in the plasma over 32 h was 0.0392 ± 0.0025 mg h.L−1, and the mean residence time was 43.17 ± 8.04 h. The pharmacometrics simulation suggests that the venom toxins were instantly and rapidly absorbed into the extravascular compartment and slowly moved into the central compartment. Our study demonstrates that Nigerian N. nigricollis venom contains low molecular weight toxins that are well absorbed into the blood and deep tissues. The venom could be detected in rabbit blood 48 h after intramuscular envenoming.

Recent Advances in Alternaria Phytotoxins: A Review of Their Occurrence, Structure, Bioactivity, and Biosynthesis.

Alternaria is a ubiquitous fungal genus in many ecosystems, consisting of species and strains that can be saprophytic, endophytic, or pathogenic to plants or animals, including humans. Alternaria species can produce a variety of secondary metabolites (SMs), especially low molecular weight toxins. Based on the characteristics of host plant susceptibility or resistance to the toxin, Alternaria phytotoxins are classified into host-selective toxins (HSTs) and non-host-selective toxins (NHSTs). These Alternaria toxins exhibit a variety of biological activities such as phytotoxic, cytotoxic, and antimicrobial properties. Generally, HSTs are toxic to host plants and can cause severe economic losses. Some NHSTs such as alternariol, altenariol methyl-ether, and altertoxins also show high cytotoxic and mutagenic activities in the exposed human or other vertebrate species. Thus, Alternaria toxins are meaningful for drug and pesticide development. For example, AAL-toxin, maculosin, tentoxin, and tenuazonic acid have potential to be developed as bioherbicides due to their excellent herbicidal activity. Like altersolanol A, bostrycin, and brefeldin A, they exhibit anticancer activity, and ATX V shows high activity to inhibit the HIV-1 virus. This review focuses on the classification, chemical structure, occurrence, bioactivity, and biosynthesis of the major Alternaria phytotoxins, including 30 HSTs and 50 NHSTs discovered to date.

Исследование адсорбционной активности энтеросорбента на основе гидролизного лигнина

Enterosorbents based on lignin hydrolised are able to adsorb in the intestine different kinds of toxins, drugs, salts of heavy metals etc. Nowadays, such drugs are widely applied during the treatment of the adults and children from the first year of life due to its safety and proven efficacy. Objective. To study the morphology and the adsorption centers on the surface of the enterosorbent "Filtrum-STI" based on lignin hydrolised. To develop a methodology for determination the adsorption activity of the drug "Filtrum-STI" and to measure qualitatively and quantitatively its adsorption activity to low molecular weight toxins and heavy metals. Materials and methods. The morphology of the sample’s surface was investigated on an Olympus BX-51 optical microscope. IR spectra were obtained using a Thermo Nicolet Nexus 470 FT-IR infrared Fourier spectrometer. Visual identification of the adsorption activity of the enterosorbent was carried out using methylene blue and methyl orange, recommended by the RF SPh as models of low molecular weight toxins. The methodology of determination the adsorption activity to heavy metal cations was developed and the measurements of this activity were carried out using reverse complexometric titration towards Pb2+ cation. Results. Micrographs of the sample "Filtrum-STI" demonstrate the heterogeneity of its surface morphology, the tablet consists of small particles. IR spectroscopy confirmed the presence of active adsorption centers on the surface of lignin hydrolysed. Adsorption of low molecular weight toxins from aqueous solutions was obviously revealed after 15 minutes for all toxins. It is estimated that the adsorption activity of the drug is preserved at physiological pH. The mass content of adsorbed Pb2+ cations was determined by the method of reverse complexometric titration, was demonstrated that w = 19.44 ± 0.92% of cations were adsorbed. Conclusion. A methodology of the determination the adsorption activity of the enterosorbent "Filtrum-STI" was developed and successfully tested, it allowed to confirm the high adsorption properties of the studied drug to a wide range of markers in various media. Key words: enterosorbent, lignin hydrolysed, "Filtrum-STI", adsorption activity, toxins, adsorption centers

Cytokines and chemokines systemic levels are related to dialysis adequacy and creatinine clearance in patients with end-stage renal disease undergoing hemodialysis

Although the clearance of low-molecular weight toxins is modulated by dialysis dose, the relationship between dialysis adequacy and middle systemic inflammatory mediators is often overlooked. Thus, the relationship between dialysis adequacy, pro- and anti-inflammatory cytokines and chemokines in hemodialysis (HD) patients was investigated. Forty-eight HD patients (19 women and 25 men) were investigated. Age, body mass index, time in HD, nutritional status, Kt/V and blood biochemical parameters was similar in patients of both sexes (P > 0.05). Thus, patients were stratified by dialysis adequacy measured by Kt/V method (adequate Kt/V ≥ 1.2). Post-HD urea, creatinine, cytokines (IFN-γ, IL-4 and IL-10) and chemokines (CCL-2, CCL-5, CXCL-8 and CXCL-10) were higher in patients with Kt/V < 1.2 (P < 0.05). Kt/V exhibited significant correlation with CXCL-10/IP-10 serum levels. Positive correlation between creatinine with IFN-γ, CCL-2/MCP-1, and CXCL-10/IP-10, and negative correlation with IL-10 was identified in patients with Kt/V < 1.2 (P < 0.05). In patients with Kt/V ≥ 1.2, only IL-10 was positively and CXCL-10/IP-10 negatively correlated with creatinine levels (P < 0.05). Kt/V and creatinine levels exhibited variable predictive value (Kt/V = 27% to 37%, creatinine = 29% to 47%) to explain cytokines and chemokines circulating levels in patients with adequate and inadequate dialysis dose. Taken together, our findings provide evidence that in addition to modulating uremic toxins levels, such as urea and creatinine, dialysis dose is associated with circulating levels of inflammatory mediators. Thus, low Kt/V results and creatinine accumulation are potential indicators of the systemic inflammatory stress determined by up-regulation of proinflammatory cytokines and chemokines, and downregulation of anti-inflammatory cytokines.

Immunorecognition capacity of Indian polyvalent antivenom against venom toxins from two populations of Echis carinatus

Clinicians report low efficacy of Indian polyvalent antivenom (PAV), with >20 vials required for treatment of a snakebite envenoming. We hypothesize that the antivenom efficacy could be reduced due to insufficient antibodies against some venom toxins. To test this, we used third-generation antivenomics to reveal bound and unbound venom toxins of Echis carinatus venom from Goa (ECVGO) and Tamil Nadu (ECVTN). We used 60, 120, 180, 240, 300, and 360 μg of venom and passed through mini-columns containing ~5 mg Antivenom bound to CNBr beads. The non-retained (unbound) and retained (bound) toxins were identified using reverse-phase HPLC and tandem mass spectrometry. Low molecular weight toxins - Short disintegrins (5.3 kDa) and DIS domain of P-II SVMP from ECVGO and ECVTN showed poor binding with antivenom. The immunorecognition sites of antivenom saturated at the lower antivenom-venom ratio for ECVGO than for ECVTN. The immunoretained capacity of antivenom against ECVTN was 140.6 μg and ECVGO was 125.1 μg. The amount of immunoretained toxins quantified can further be used to estimate the efficacy of antivenom by correlating it with in-vivo studies. The unbound toxins identified from this study could be targeted to improve the effectiveness of antivenom.

Snake venom proteomics and antivenomics of two Sundaic lance-headed pit vipers: Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia)

Envenomation by two medically important Sundaic pit vipers, Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia), causes hemotoxic syndrome with a potentially fatal outcome. Research on the compositions and antigenicity of these pit viper venoms is however lacking, limiting our understanding of the pathophysiology and treatment of envenomation. This study investigated the venom proteomes of both species through a protein decomplexation strategy, applying C18 reverse-phase high-performance liquid chromatography followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein identification through nano-electrospray ionization liquid chromatography-tandem mass spectrometry (nano-ESI-LCMS/MS) of trypsin-digested peptides. The venom antigenicity was profiled against the Thai Green Pit Viper Antivenom (GPVAV, a hetero-specific antivenom), using indirect enzyme-linked immunosorbent assay (ELISA). The venom proteomes of T. wiroti and T. puniceus consisted of 10 and 12 toxin families, respectively. The major proteins were of diverse snake venom serine proteases (19–30% of total venom proteins), snake venom metalloproteinases (17–26%), disintegrins(9–16%), phospholipases A2 (8–28%) and C-type lectins (~8%). These were putative snake toxins implicated in hemorrhage and coagulopathy, consistent with clinical hemotoxicity. GPVAV showed strong immunorecognition toward high and medium molecular weight proteins (e.g., SVMP and PLA2) in both venoms, while a lower binding activity was observed toward small proteins such as disintegrins. Conserved antigenicity in the major hemotoxins supported toxicity cross-neutralization by GPVAV and indicated that the immunorecognition of low molecular weight toxins may be optimized for improved binding efficacy. Taken together, the study provides insights into the pathophysiology and antivenom treatment of envenomation caused by T. wiroti and T. puniceus in the region.

Research of the adsorption of a methylene blue enterosorbents of various nature

The results of the adsorption study of methylene blue dye as the low molecular weight toxins marker from its aqueous solutions by carbon, silicon and polymer enterosorbents are presented. It has been shown that the activated carbons modification by water-soluble cellulose polyelectrolyte makes it possible to increase their adsorption capacity. The applicability of Langmuir, Freundlich, Temkin and Redlich – Peterson adsorption models for the approximation of experimental adsorption isotherms of methylene blue on the enterosorbents has been estimated. It has been established that the three-parameter Redlich – Peterson equation describes this process better than others. This was evident from the comparison of the regression coefficients, constants and parameters values of these equations.

Multiplex Immunoassay Techniques for On-Site Detection of Security Sensitive Toxins.

Biological toxins are a heterogeneous group of high molecular as well as low molecular weight toxins produced by living organisms. Due to their physical and logistical properties, biological toxins are very attractive to terrorists for use in acts of bioterrorism. Therefore, among the group of biological toxins, several are categorized as security relevant, e.g., botulinum neurotoxins, staphylococcal enterotoxins, abrin, ricin or saxitoxin. Additionally, several security sensitive toxins also play a major role in natural food poisoning outbreaks. For a prompt response to a potential bioterrorist attack using biological toxins, first responders need reliable, easy-to-use and highly sensitive methodologies for on-site detection of the causative agent. Therefore, the aim of this review is to present on-site immunoassay platforms for multiplex detection of biological toxins. Furthermore, we introduce several commercially available detection technologies specialized for mobile or on-site identification of security sensitive toxins.

Evaluation of Mass Sensitive Micro-Array biosensors for their feasibility in multiplex detection of low molecular weight toxins using mycotoxins as model compounds

Mycotoxins produced by Fusarium species including trichothecenes, zearalenone and fumonisins, can co-contaminate food and feed throughout the supply chain, including cereal grains and animal feeds. There is an increasing demand to enhance global food security by improving the monitoring of mycotoxins throughout our food supply chain. For time and cost-efficient analysis, rapid tests capable of detecting multiple toxins from a single sample are ideal. Considering these current trends in mycotoxin testing, this project examined the feasibility of using both a portable and non-portable mass-based biosensor for multiplex mycotoxin detection. The biosensor was a mass sensitive microarray (MSMA) which consisted of 4 × 16 miniaturized mass sensitive transducer pixels based on solidly mounted resonator (SMR) technology. Functionalisation of individual pixels on the sensor surface using nano-spotting technology for the simultaneous and semi-quantitative detection of three regulated mycotoxins: T2-toxin (T2) zearalenone (ZEN), and fumonisin B1 (FumB1) was examined. With the integration of portable and non-portable microfluidic devices for antibody and standard sample injections, competitive inhibition assays were developed followed by singleplex and multiplex calibration curves. The characteristics and performance of the MSMA were evaluated including sensitivity which was determined as the concentration causing 50% inhibition. Sensitivity of singleplex assays using the portable microfluidic device (PMD) were 1.3 ng/ml, 2.0 ng/ml and 6.8 ng/ml for T2, FumB1 and ZEN, respectively. Sensitivity of the multiplex assay again using the PMD was 6.1 ng/ml, 3.6 ng/ml and 2.4 ng/ml for T2, FumB1 and ZEN, respectively. The PMD was an easy to use and highly sensitive screening tool which has been demonstrated for the multiplex detection of three regulated mycotoxins. Analysis was in real time and results were fully digital. Since detection of analytes was by mass it was both a label-free and cost-efficient method proposed method of analysis for mycotoxins.

P1055REMARKABLE REMOVALS OF BETA-2-MICROGLOBULIN AND PHOSPHATE WITH SHORT-DAILY HOME HEMODIALYSIS USING LOW DIALYSATE FLOW RATE

Abstract Background and Aims Short-daily hemodialysis (HD) with low-dialysate volume is an appealing portable dialysis approach for home use. Although this type of HD has proved being effective for the volume control and the clearance of low molecular-weight uremic toxins, limited data are available on the impact on the removal rates of other uremic toxins like β2-microglobulin (β2M) or phosphate (P), whose clearance is limited by sequestration into compartments, poor diffusion, high time-dependency, or protein binding. We evaluated the impact of short-daily HD with slow dialysate flow rate on the removal of solutes of different molecular weights and distribution volumes. Method Single-session and weekly balances of β2M, P, urea, and creatinine were prospectively assessed with total dialysate collection and serum measurements before and after 341 dialysis sessions (mean dialysate volume: 30963 ± 862 mL; mean length of dialysis session: 153 ± 8 min) in 31 stable patients (female; 9, 29 %; mean age: 55.6 ± 13.6 y; dry weight: 74.9 ± 13.3 kg) undergoing short-daily home HD with NxStage cycler, between July 2014 and October 2019. The mean blood flow rate was 365 ± 17 mL/min, whereas the mean dialysate flow rate was 194 ± 12 mL/min. Results Single-session β2M, P, urea, and creatinine removals were 0.138 ± 0.050 g, 0.610 ± 0.161 g, 18.89 ± 6.07 g and 1.07 ± 0.31 g, respectively, whereas the reduction rates (%) were 38.0 ± 13.0, 46.8 ± 8.6, 48.2 ± 7.0 and 46.6 ± 6.6, for β2M, P, urea and creatinine, respectively. The estimated weekly β2M, P, urea and creatinine removals in HDD patients dialyzing 5-6 days per week were comparable with 4-h in-center thrice-weekly on-line hemodiafiltration according to previous studies (Table 1). Conclusion Treating patients with short-daily HD with low-dialysate volume at a 5-6 days per week prescription may achieve an efficient weekly β2M and P removal.

An Electrochemical Fiveplex Biochip Assay Based on Anti-Idiotypic Antibodies for Fast On-Site Detection of Bioterrorism Relevant Low Molecular Weight Toxins.

Modern threats of bioterrorism force the need for multiple detection of biothreat agents to determine the presence or absence of such agents in suspicious samples. Here, we present a rapid electrochemical fiveplex biochip screening assay for detection of the bioterrorism relevant low molecular weight toxins saxitoxin, microcystin-LR, T-2 toxin, roridin A and aflatoxin B1 relying on anti-idiotypic antibodies as epitope-mimicking reagents. The proposed method avoids the use of potentially harmful toxin-protein conjugates usually mandatory for competitive immunoassays. The biochip is processed and analyzed on the automated and portable detection platform pBDi within 13.4 min. The fiveplex biochip assay revealed toxin group specificity to multiple congeners. Limits of detection were 1.2 ng/mL, 1.5 ng/mL, 0.4 ng/mL, 0.5 ng/mL and 0.6 ng/mL for saxitoxin, microcystin-LR, T-2 toxin, roridin A or aflatoxin B1, respectively. The robustness of the fiveplex biochip for real samples was demonstrated by detecting saxitoxin, microcystin-LR, HT-2 toxin, roridin A and aflatoxin B1 in contaminated human blood serum without elaborate sample preparation. Recovery rates were between 52–115% covering a wide concentration range. Thus, the developed robust fiveplex biochip assay can be used on-site to quickly detect one or multiple low molecular weight toxins in a single run.

Quantitative proteomic analysis of venom from Southern India common krait (Bungarus caeruleus) and identification of poorly immunogenic toxins by immune-profiling against commercial antivenom

ABSTRACTObjectives: To study the venom proteome composition of Southern India (SI) Common Krait (Bungarus caeruleus) and immunological cross-reactivity between venom against commercial antivenom.Methods: Proteomic analysis was done by nano LC-MS/MS and toxins were quantitated by label-free analysis. The immunological cross-reactivity of venom towards polyvalent antivenom (PAV) was assessed by ELISA, Immunoblotting, and immuno-chromatographic methods.Results: A total of 57 enzymatic and non-enzymatic proteins belonging to 12 snake venom protein families were identified. The three finger toxins (3FTx) (48.3%) and phospholipase A2 (PLA2) (37.6%) represented the most abundant non-enzymatic and enzymatic proteins, respectively. β-bungarotoxin (12.9%), a presynaptic neurotoxin, was also identified. The venom proteome composition is well correlated with its enzymatic activities, reported pharmacological properties, and clinical manifestations of krait envenomation. Immuno-cross-reactivity studies demonstrated better recognition of high molecular weight proteins (>45 kDa) of this venom by PAVs compared to low molecular weight (<15 kDa) toxins such as PLA2 and 3FTxs.Conclusion: The poor recognition of <15 kDa mass SI B. caeruleus venom proteins is of grave concern for the successful treatment of krait envenomation. Therefore, emphasis should be given to improve the immunization protocols and/or supplement of antibodies raised specifically against the <15 kDa toxins of this venom.

Electrochemical Biochip Assays Based on Anti-idiotypic Antibodies for Rapid and Automated On-Site Detection of Low Molecular Weight Toxins.

Phycotoxins and mycotoxins, such as paralytic shellfish poisoning toxins, type A trichothecenes, and aflatoxins are among the most toxic low molecular weight toxins associated with human poisoning incidents through the consumption of naturally contaminated food. Therefore, there is an utmost need for rapid and sensitive on-site detection systems. Herein, an electrochemical biochip for fast detection of saxitoxin, T-2 toxin as well as aflatoxin M1 and their corresponding congeners, respectively, using a portable and fully automated detection platform (pBDi, portable BioDetector integrated) was developed. Toxin analysis is facilitated upon the biochip via an indirect competitive immunoassay using toxin-specific antibodies combined with anti-idiotypic antibodies. The developed biochips enable detection in the low ng/mL-range within 17 min. Moreover, the assays cover a wide linear working range of 2–3 orders of magnitude above the limit of detection with an inter-chip coefficient of variation lower than 15%. The broad specificity of the employed antibodies which react with a large number of congeners within the respective toxin group allows efficient screening of contaminated samples for the presence of these low molecular weight toxins. With respect to the analysis of human urine samples, we focused here on the detection of saxitoxin, HT-2 toxin, and aflatoxin M1, all known as biomarkers of acute toxin exposure. Overall, it was proved that the developed biochip assays can be used to rapidly and reliably identify severe intoxications caused by these low molecular weight toxins.

Preparation and Properties of Biosorbents on the Base of Fruit Seed Shell

The process of obtaining of new biosorbents from apricot seed shells by oxidative-organosolvent processing in acetic acid - hydrogen medium has been studied. The structural and sorption properties of the materials obtained using physical and chemical methods were investigated. The effect of the reactants ratio in solution, as well as the duration of the modification process on the yield, content of cellulose, lignin, and mineral components and on sorption properties of the products was studied. The dependence between the mineral content and the adsorption pore volume was shown. The positive effect of hydrogen peroxide application in the process of modifying of plant waste on cellulose content was estimated. Increasing the duration of modification increases the contribution of oxidative transformation of lignin into soluble products and decreases the yield of the lignocelluloses’ materials. The regularities of sorption of marker of low molecular weight toxins and non-steroidal anti-inflammatory drugs on obtained products were investigated. It has been found that the highest sorption capacity of obtained lignocelluloses’ sorbents towards methylene blue and Sodium Diclofenac corresponds to the samples with polysaccharide content 60%. It has been also found that the sorption equilibrium occurs within 120 min of contact. The obtained results demonstrate the feasibility of application of such lignocelluloses’ carriers in the production of prolonged action drugs.

An overview of Phoneutria nigriventer spider venom using combined transcriptomic and proteomic approaches.

Phoneutria nigriventer is one of the largest existing true spiders and one of the few considered medically relevant. Its venom contains several neurotoxic peptides that act on different ion channels and chemical receptors of vertebrates and invertebrates. Some of these venom toxins have been shown as promising models for pharmaceutical or biotechnological use. However, the large diversity and the predominance of low molecular weight toxins in this venom have hampered the identification and deep investigation of the less abundant toxins and the proteins with high molecular weight. Here, we combined conventional and next-generation cDNA sequencing with Multidimensional Protein Identification Technology (MudPIT), to obtain an in-depth panorama of the composition of P. nigriventer spider venom. The results from these three approaches showed that cysteine-rich peptide toxins are the most abundant components in this venom and most of them contain the Inhibitor Cysteine Knot (ICK) structural motif. Ninety-eight sequences corresponding to cysteine-rich peptide toxins were identified by the three methodologies and many of them were considered as putative novel toxins, due to the low similarity to previously described toxins. Furthermore, using next-generation sequencing we identified families of several other classes of toxins, including CAPs (Cysteine Rich Secretory Protein—CRiSP, antigen 5 and Pathogenesis-Related 1—PR-1), serine proteinases, TCTPs (translationally controlled tumor proteins), proteinase inhibitors, metalloproteinases and hyaluronidases, which have been poorly described for this venom. This study provides an overview of the molecular diversity of P. nigriventer venom, revealing several novel components and providing a better basis to understand its toxicity and pharmacological activities.

The Role of Bacterial Natural Products in Predator Defense

Bacterially produced natural products, i.e., low molecular weight metabolites, or derivatives thereof, constitute most of the commercially available antibiotics as well as a large proportion of anticancer drugs. While indispensable as therapeutically active compounds, the ecological roles of many of these bacterial natural products remain poorly understood. Here, we discuss these metabolites in light of ­microbial predator defense: soil bacteria are constantly threatened by a variety of predators and the secretion of low molecular weight toxins enables the producing bacteria to kill or deter the predator. Conversely, a deeper understanding of these microbial predator–prey interactions can lead to the discovery of novel compounds, which in turn can be of therapeutic use.

The impact of endogenous intoxication on biochemical indicators of blood of cows with calves

The notion “endotoxin” is conditional, for any normal metabolit has a toxic impact of its excessive accumulation in organism. Endotoxins include products of breakdown of tissue proteins, peroxides and other products of free radical oxidation, low molecular weight toxins, toxins of microorganisms. Current knowledge on the mechanism of endotoxin action on the organism of pregnant cows is based on the key role of the immune system: it provides animals with protection against the infections, eliminates extraneous elements of endogenous and exogenous origin. It is defined that under endogenous intoxication, toxic metabolites suppress the activity of immune system of organism, decreases the resistability of an organism to negative effects of environmental factors. Animals under endogenous intoxication had the following clinical symptoms: swelling of mammal gland, edemas of external genitals, anemia of mucous membranes, functional disorders of proventriculuses and intestines, animals were stressed. It was defined that under endogenous intoxication in 8–9 months of pregnancy, blood of highly-productive cows had a tendency of decrease in the number of erythrocytes by 21.0%, in the level of hemoglobin – by 24.3%, increase in the number of leucocytes by 7.6% compared to the value for blood of cows with physiological pregnancy process. The development of endogenous intoxication among pregnant cows causes disorders in protein-synthesizing function of liver, which is indicated by decrease in the level of total protein and its fractions. It was defined that endogenous intoxication decreased the content of total protein in blood by 18.4%. Over the period of study, among pregnant cows under the development of endogenous intoxication, heightened activity of amino transferaces in the blood was observed. In the ninth month of pregnancy, the activity of amino transferaces in blood serum of cows from experimental group increased by 28.6%, and the activity of aspartate aminotransferace increased by 20.1% relatively. The activity of amino transferaces and decrease in the level of total protein and its fractions in blood serum is one of the first biochemical studies on the diagnosing endotoxin manifestations and indicates destructive processes in liver. It was defined that pregnant cows under the development of endogenous intoxication suffer disorders in detoxicating function of liver and disorders in filtrating function of kidneys. The developing endogenous intoxication of pregnant cows suppresses the activity of enzymes in their glutathione system of antioxidant protection, which is indicated by decrease in the activity of glutathione peroxidase by 27.5 %, glutathione reductase by 42.9%, glucose-6-phosphate dehydrogenase by 11.2% in blood serum. The lowest activity of enzymes of glutathione system in blood of highly-productive cows under endogenous intoxication was observed in their ninth month of pregnancy, which is related to increase in activation of processes of lipid peroxidation and misbalance between the activity of antioxidant system and intensity of lipid peroxidation. Reliable increase in the content of lipid hydroperoxides by 70.9% and in concentration of malondialdehide by 54.8% was observed. Misbalance towards generation of active forms of oxygen and their metabolites, exhaustion of antioxidant system and disorders in balance cause oxidative stress.