Botulinum neurotoxin type C protease induces apoptosis in differentiated human neuroblastoma cells

Neuroblastomas constitute a major cause of cancer-related deaths in young children. In recent years, a number of translation-inhibiting enzymes have been evaluated for killing neuroblastoma cells. Here we investigated the potential vulnerability of human neuroblastoma cells to protease activity derived from botulinum neurotoxin type C. We show that following retinoic acid treatment, human neuroblastoma cells, SiMa and SH-SY5Y, acquire a neuronal phenotype evidenced by axonal growth and expression of neuronal markers. Botulinum neurotoxin type C which cleaves neuron-specific SNAP25 and syntaxin1 caused apoptotic death only in differentiated neuroblastoma cells. Direct comparison of translation-inhibiting enzymes and the type C botulinum protease revealed one order higher cytotoxic potency of the latter suggesting a novel neuroblastoma-targeting pathway. Our mechanistic insights revealed that loss of ubiquitous SNAP23 due to differentiation coupled to SNAP25 cleavage due to botulinum activity may underlie the apoptotic death of human neuroblastoma cells

Stopping of relativistic ions in multicomponent plasmas

Investigation of the processes of stopping of charged particles moving in different media is of significant interest for many realms of Physics, such that Nuclear Physics, Condensed Matter Physics, Plasma Physics, etc. The problem of evaluation of energy losses of relativistic protons has acquired special importance recently [1] and, due to the experimental conditions, it is necessary to estimate relativistic corrections to the asymptotic form of energy losses in non-ideal multicomponent plasmas..

Stopping of relativistic ions in multicomponent plasmas

Investigation of the processes of stopping of charged particles moving in different media is of significant interest for many realms of Physics, such that Nuclear Physics, Condensed Matter Physics, Plasma Physics, etc. The problem of evaluation of energy losses of relativistic protons has acquired special importance recently [1] and, due to the experimental conditions, it is necessary to estimate relativistic corrections to the asymptotic form of energy losses in non-ideal multicomponent plasmas..

The stopping power and straggling of strongly coupled electron fluids revisited

Measuring energy losses of beams of charged particles is an important diagnostic tool in both modern condensed matter and plasma physics..

Genetic diversity assessment in pea cultivars and lines using the SSR analysis

Background. Pea is the main leguminous crop in the Republic of Bashkortostan and widespread all over the world. The key role in the breeding of new pea cultivars is played by source material representing the phenotypic and genotypic diversity of Pisum sativum L., searched for in plant genetic resources collections. SSR markers are successfully used to study the DNA polymorphism of various genetic objects, including pea. However, the distribution of a number of microsatellite alleles in the genotypes of specific lines and cultivars of this valuable pulse crop remains practically unexplored.Materials and methods. Molecular genetic polymorphism was studied in 40 pea cultivar accessions of different ecological and geographical origin from the Vavilov Institute’s genebank of plant genetic resources or developed at regional breeding centers. Microsatellite analysis was performed using 5 SSR markers from the genomic library of microsatellites (Agrogene®, France).Results. All markers delivered good electrophoretic profiles and helped to amplify a number of alleles per locus varying from 2 (AB53) to 9 (AA355). The total number of alleles was 26, while the average number of alleles per locus was 5.2. The polymorphism information content (PIC) varied from 0.39 for locus AB53 to 0.82 for locus AA355, with the mean value of 0.60. The set of SSR markers used in the work made it possible to individualize each of the studied pea genotypes. The measured genetic distances were used to draw a dendrogram showing the distribution of genotypes according to their genetic relationship.Conclusion. Through studying the source material for pea breeding by the SSR analysis the data were obtained that provide additional information about the genetic structure of the collection and the polymorphism of the studied cultivar accessions. The results of genotyping pea cultivars and lines can be used for their genetic identification or to select parental pairs for hybridization

Development of source material for pea breeding through chemical mutagenesis and evaluation of its genetic diversity using SSR markers

Background. Pea (Pisum sativum L.) is a valuable leguminous crop of worldwide importance. The main problem of modern plant breeding is a decrease in the genetic diversity of crops, including pea. One of the ways to increase genetic polymorphism is the use of chemically induced mutagenesis. Sodium azide (NaN3) is a highly effective chemical mutagen successfully used in mutation breeding to increase the productivity of cultivated plants and enrich them with new useful traits. We used it to obtain new pea breeding material.Materials and methods. Experiments were carried out to obtain pea mutants using sodium azide at the concentrations of 1, 5 and 10 mM and the exposure time of 3 and 9 h. Molecular genetic polymorphism of the М2 plants and the original cultivar was assessed using 10 SSR markers from the microsatellite genomic library (Agrogene®, France).Results. Optimal concentrations of sodium azide and the duration of seed treatment with it were identified: 1–5 mM for 3 h. Sixteen mutant populations were obtained; in ten of them a change in the leaf type was found. An analysis of the yield structure components revealed a significant superiority (p < 0.05) over the initial cultivar ‘Pamyati Khangildina’ in the mutant populations No. 1, No. 5, No. 9, No. 10, No. 15 and No. 16 in the number of seeds per pod, No. 9 and No. 16 in the weight of 1000 seeds, and No. 16 in the weight of seeds per plant. A dendrogram constructed on the basis of the SSR analysis data showed the degree of differences between the M2 populations of pea plants and the initial cultivar ‘Pamyati Khangildina’.Conclusion. The obtained mutant populations are planned to be used in pea breeding as sources of high seed numbers in pods, seed yield, seed weight per plant, and large seed size. A microsatellite analysis with 10 SSR markers revealed differences among the M2 mutant populations at the genetic level and made it possible to identify them

Two complementary approaches for intracellular delivery of exogenous enzymes.

Intracellular delivery of biologically active proteins remains a formidable challenge in biomedical research. Here we show that biomedically relevant enzymes can be delivered into cells using a new DNA transfection reagent, lipofectamine 3000, allowing assessment of their intracellular functions. We also show that the J774.2 macrophage cell line exhibits unusual intracellular uptake of structurally and functionally distinct enzymes providing a convenient, reagent-free approach for evaluation of intracellular activities of enzymes

Recent developments in engineering non-paralytic botulinum molecules for therapeutic applications

This review discusses the expanding application of botulinum neurotoxin in treating neurological conditions. The article specifically explores novel approaches to using non-paralytic botulinum molecules. These new molecules, such as BiTox or el-iBoNT, offer an alternative for patients who face limitations in using paralytic forms of botulinum neurotoxin due to concerns about muscle function loss. We highlight the research findings that confirm not only the effectiveness of these molecules but also their reduced paralytic effect. We also discuss a potential cause for the diminished paralytic action of these molecules, specifically changes in the spatial parameters of the new botulinum molecules. In summary, this article reviews the current research that enhances our understanding of the application of new botulinum neurotoxins in the context of common conditions and suggests new avenues for developing more efficient molecules

ADDRESSING THE GROWING BURDEN OF NCDS: RETURN TO ALMA-ATA AND PRIMARY HEALTHCARE APPROACH

Last year was the 40th anniversary of the Declaration of Alma-Ata. The conference organized by WHO and the United Nations Children’s Fund proclaimed the ambitious goal - Health for All by the Year of 2000 and introduced the Primary Health Care approach that was considered as the means to achieve the goal. At the same time, some authors think that the main Alma-Ata deficiency was the fault to clearly define the difference between primary medical care and PHC approach, which involves universal coverage, inter-sectoral collaboration, community-based curative and preventive services. This short report discusses the excessive alcohol consumption and its dynamics throughout years in Kazakhstan and other post USSR countries, linking it to high rates of non-communicable diseases (NCDs) within these countries. It also emphasizes the importance of intersectoral approach in tackling excessive alcohol consumption that may well lead to an improved management of NCDs

Control of Autophagosome Axonal Retrograde Flux by Presynaptic Activity Unveiled Using Botulinum Neurotoxin Type A

Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity