Matrix metalloproteinase-activated anthrax lethal toxin demonstrates high potency in targeting tumor vasculature

Anthrax lethal toxin (LT), a virulence factor secreted by Bacillus anthracis, is selectively toxic to human melanomas with the BRAF V600E activating mutation because of its proteolytic activities toward the mitogen-activated protein kinase kinases (MEKs). To develop LT variants with lower in vivo toxicity and high tumor specificity, and therefore greater potential for clinical use, we generated a mutated LT that requires activation by matrix metalloproteinases (MMPs). This engineered toxin was less toxic than wild-type LT to mice because of the limited expression of MMPs by normal cells. Moreover, the systemically administered toxin produced greater anti-tumor effects than wild-type LT toward human xenografted tumors. This was shown to result from its greater bioavailability, a consequence of the limited uptake and clearance of the modified toxin by normal cells. Furthermore, the MMP-activated LT had very potent anti-tumor activity not only to human melanomas containing the BRAF mutation but also to other tumor types, including lung and colon carcinomas regardless of their BRAF status. Tumor histology and in vivo angiogenesis assays showed that this anti-tumor activity is due largely to the indirect targeting of tumor vasculature and angiogenic processes. Thus, even tumors genetically deficient in anthrax toxin receptors were still susceptible to the toxin therapy in vivo. Moreover, the modified toxin also displayed lower immunogenicity compared with the wild-type toxin. All these properties suggest that this MMP-activated anti-tumor toxin has potential for use in cancer therapy.     

Intracranial administration of the G‐protein coupled estrogen receptor 1 antagonist,G‐15, selectively affects dimorphic characteristics of the song system in zebra finches (Taeniopygia guttata)

In zebra finches (Taeniopygia guttata), estradiol contributes to sexual differentiation of the song system but the receptor(s) underlying its action are not exactly known. Whereas mRNA and/or protein for nuclear estrogen receptors ERα and ERβ are minimally expressed, G‐protein coupled estrogen receptor 1 (GPER1) has a much greater distribution within neural song regions and the syrinx. At present, however, it is unclear if this receptor contributes to dimorphic development of the song system. To test this, the specific GPER1 antagonist, G‐15, was intracranially administered to zebra finches for 25 days beginning on the day of hatching. In males, G‐15 significantly decreased nuclear volumes of HVC and Area X. It also decreased the muscle fiber sizes of ventralis and dorsalis in the syrinx. In females, G‐15 had no effect on measures within the brain, but did increase fiber sizes of both muscle groups. In sum, these data suggest that GPER1 can have selective and opposing influences on dimorphisms within the song system, but since not all features were affected additional factors are likely involved. © 2018 Wiley Periodicals, Inc. Develop Neurobiol, 2018     

Anthrax Lethal Toxin Downregulates Claudin-5 Expression in Human Endothelial Tight Junctions

Vascular leakage pathologies such as pleural effusion and hemorrhage are hallmarks of anthrax pathogenesis. We previously reported that anthrax lethal toxin (LT), the major virulence factor of anthrax, reduces barrier function in cultured primary human microvascular endothelial cells. Here, we show that LT-induced barrier dysfunction is accompanied by the reduced expression of the endothelial tight junction (TJ) protein claudin-5 but no change in the expression of other TJ components occludin, ZO-1, ZO-2, or the adherens junction (AJ) protein VE-cadherin. The downregulation of claudin-5 correlated temporally and dose-dependently with the reduction of transendothelial electrical resistance. LT-induced loss of claudin-5 was independent of cell death and preceded the appearance of actin stress fibers and altered AJ morphology. Pharmacological inhibition of MEK-1/2, two kinases that are proteolytically inactivated by LT, showed a similar reduction in claudin-5 expression. We found that LT reduced claudin-5 mRNA levels but did not accelerate the rate of claudin-5 degradation. Mice challenged with LT also showed significant reduction in claudin-5 expression. Together, these findings support a possible role for LT disruption of endothelial TJs in the vascular leakage pathologies of anthrax.     

Influence of the nature of quantum dot surface cations on interactions with DNA

Quantum dots are semiconductor nanoparticles that are approximately 1-10nm in diameter, similar to small proteins, and their photoluminescence is sensitive to the presence and nature of adsorbates. We have deployed these nanomaterials as luminescent probes of DNA structure. Sequence dependent conformational flexibility of DNA is of great interest due to its implications for drug-DNA and DNA-protein interactions. The counterion atmosphere surrounding DNA plays an important role in its structure, dynamics, and packaging. In this paper, we investigate the effect that various monovalent and divalent cations have on the binding of 4.5 nm CdS quantum dots to oligonucleotides that have sequence-directed intrinsic structure.     

Increased level of long non coding RNA H19 is correlated with the downregulation of miR-326 and BCL-2 genes in pediatric acute lymphoblastic leukemia,a possible hallmark for leukemogenesis

Molecular Biology Reports - Long non-coding RNAs (lncRNAs) and their role in competitive endogenous RNA (ceRNA) networks have emerged as fundamental debates in the biological processes of...     

Inflammasome sensor Nlrp1b-dependent resistance to anthrax is mediated by caspase-1, IL-1 signaling and neutrophil recruitment

Bacillus anthracis infects hosts as a spore, germinates, and disseminates in its vegetative form. Production of anthrax lethal and edema toxins following bacterial outgrowth results in host death. Macrophages of inbred mouse strains are either sensitive or resistant to lethal toxin depending on whether they express the lethal toxin responsive or non-responsive alleles of the inflammasome sensor Nlrp1b (Nlrp1b(S/S) or Nlrp1b(R/R), respectively). In this study, Nlrp1b was shown to affect mouse susceptibility to infection. Inbred and congenic mice harboring macrophage-sensitizing Nlrp1b(S/S) alleles (which allow activation of caspase-1 and IL-1β release in response to anthrax lethal toxin challenge) effectively controlled bacterial growth and dissemination when compared to mice having Nlrp1b(R/R) alleles (which cannot activate caspase-1 in response to toxin). Nlrp1b(S)-mediated resistance to infection was not dependent on the route of infection and was observed when bacteria were introduced by either subcutaneous or intravenous routes. Resistance did not occur through alterations in spore germination, as vegetative bacteria were also killed in Nlrp1b(S/S) mice. Resistance to infection required the actions of both caspase-1 and IL-1β as Nlrp1b(S/S) mice deleted of caspase-1 or the IL-1 receptor, or treated with the Il-1 receptor antagonist anakinra, were sensitized to infection. Comparison of circulating neutrophil levels and IL-1β responses in Nlrp1b(S/S),Nlrp1b(R/) (R) and IL-1 receptor knockout mice implicated Nlrp1b and IL-1 signaling in control of neutrophil responses to anthrax infection. Neutrophil depletion experiments verified the importance of this cell type in resistance to B. anthracis infection. These data confirm an inverse relationship between murine macrophage sensitivity to lethal toxin and mouse susceptibility to spore infection, and establish roles for Nlrp1b(S), caspase-1, and IL-1β in countering anthrax infection.     

Persian sturgeon insulin-like growth factor I: molecular cloning and expression during various nutritional conditions

The effects of different periods of starvation (1, 2, 3, and 4 weeks) and subsequent re-feeding (over a 4 week) on the compensatory growth performance and insulin-like growth factor I (IGF-I) mRNA expression in liver and white muscle were investigated in juvenile Persian sturgeon (Acipenser persicus). First, a fragment of 617 nucleotides coding for IGF-I was cloned from liver, which included an open reading frame of 486 nucleotides, encoding a 162 amino acid preproIGF-I. This is composed of a 45 aa for signal peptide, a 117 aa for the mature peptide comprising the B, C, A, and D domains, and a 47 aa for E domain. The mature Persian sturgeon IGF-I exhibits high sequence identities with other sturgeon species and teleost, ranging between 68 and 95 %. The pattern of IGF-I mRNA expression in the liver and white muscle was measured in response to different periods of starvation and subsequent re-feeding. Nutritional status influenced IGF-I mRNA expression pattern in both liver and muscle. IGF-I mRNA expression in the liver increased during starvation, before decreasing after re-feeding. Furthermore, white muscle IGF-I mRNA expression showed better responses to nutritional status and decreased following starvation and increased by re-feeding. However, changes in the expression of IGF-I mRNA were not significantly different between any of the treatments in both tissues. These data suggest that muscle and liver IGF-I mRNA expression do not have a regulatory role for somatic growth induced by compensatory growth in Persain sturgeon.     

In vitro spectroscopic investigation of groove binding interaction of Fe3O4@CaAl-LDH@L-Dopa with calf thymus DNA

Abstract

The principal goal of this study is to evaluate the interaction of Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles with calf thymus DNA. The magnetic nanoparticles were previously prepared by a chemical co-precipitation method, and the surface of the Fe₃O₄ nanoparticles was coated with CaAl layered double hydroxides. The antiparkinsonian drug “L-Dopa” was carried by this core–shell nanostructure to achieve the drug delivery system with suitable properties for biological applications. Also, the interaction of Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles with CT-DNA was studied using, UV–Visible spectroscopy, viscosity, circular dichroism (CD), and fluorescence spectroscopy techniques. The results of investigations demonstrated that Fe₃O₄@CaAl-LDH@L-Dopa and Fe₃O₄@CaAl-LDH nanoparticles have interacted via minor groove binding and intercalated to CT-DNA, respectively.     

In vitro effects of gossypol on testicular lactic dehydrogenase-X and other dehydrogenases

Thein vitro inhibition of several rat testis dehydrogenases by gossyPol was examined. Inclusion of the coenzyme (substrate for NADP⁺-isocitrate dehydrogenase) in the Preincubation mixture containing the enzyme and gossyPol, Protected the enzymes against inhibition by gossyPol. Lactic dehydrogenase-X was amongst the least Protected enzymes. This, couPled with its lowK _i for gossyPol makes it one of the most vulnerable target enzymesin vivo for gossyPol action. The inhibition kinetics for lactic dehydrogenase-X were comPetitive when NADH was Present during Preincubation, but non-comPetitive when the coenzyme was excluded during Preincubation. In the latter condition, the enzyme seems to undergo Progressive inactivation with time causing a nonreversible tyPe of inhibition.