Examination of the cytotoxic and embryotoxic potential and underlying mechanisms of next-generation synthetic trioxolane and tetraoxane antimalarials

Semisynthetic artemisinin-based therapies are the first-line treatment for P. falciparum malaria, but next-generation synthetic drug candidates are urgently required to improve availability and respond to the emergence of artemisinin-resistant parasites. Artemisinins are embryotoxic in animal models and induce apoptosis in sensitive mammalian cells. Understanding the cytotoxic propensities of antimalarial drug candidates is crucial to their successful development and utilization. Here, we demonstrate that, similarly to the model artemisinin artesunate (ARS), a synthetic tetraoxane drug candidate (RKA182) and a trioxolane equivalent (FBEG100) induce embryotoxicity and depletion of primitive erythroblasts in a rodent model. We also show that RKA182, FBEG100 and ARS are cytotoxic toward a panel of established and primary human cell lines, with caspase-dependent apoptosis and caspase-independent necrosis underlying the induction of cell death. Although the toxic effects of RKA182 and FBEG100 proceed more rapidly and are relatively less cell-selective than that of ARS, all three compounds are shown to be dependent upon heme, iron and oxidative stress for their ability to induce cell death. However, in contrast to previously studied artemisinins, the toxicity of RKA182 and FBEG100 is shown to be independent of general chemical decomposition. Although tetraoxanes and trioxolanes have shown promise as next-generation antimalarials, the data described here indicate that adverse effects associated with artemisinins, including embryotoxicity, cannot be ruled out with these novel compounds, and a full understanding of their toxicological actions will be central to the continuing design and development of safe and effective drug candidates which could prove important in the fight against malaria

Water Governance in Decentralising Indonesia

Under new democratic regimes in the country of the South, governance innovation is often found at the regional level. This article, using the concept of institutional capacity, shows that powerful efforts affecting regional water resource coordination emerge locally. The paper analyzes fresh water cooperation in the urban region of Cirebon, Indonesia. It is shown that city and their surrounding regions in decentralizing Indonesia show signs of increasing institutional capacity between local actors. An informal approach and discretionary local decision-making, influenced by logic of appropriateness and tolerance are influential. At the same time, these capacities are compromised by significant inequality and a unilateral control of water resources, and they are being challenged by a strong authoritarian political culture inherited from a history of centralized government. The paper points to the need to establish greater opportunities for water governance at the regional level to transcend inter-local rivalry, and thus improve decentralized institutional capacity further

Landscape characteristics and coho salmon (Oncorhynchus kisutch) distributions: explaining abundance versus occupancy

Distribution of fishes, both occupancy and abundance, is often correlated with landscape-scale characteristics (e.g., geology, climate, and human disturbance). Understanding these relationships is essential for effective conservation of depressed populations. We used landscape characteristics to explain the distribution of coho salmon (Oncorhynchus kisutch) in the Oregon Plan data set, one of the first long-term, probabilistic salmon monitoring data sets covering the full range of potential habitats. First we compared data structure and model performance between the Oregon Plan data set and two published data sets on coho salmon distribution. Most of the variation in spawner abundance occurred between reaches but much also occurred between years, limiting potential model performance. Similar suites of landscape predictors are correlated with coho salmon distribution across regions and data sets. We then modeled coho salmon spawner distribution using the Oregon Plan data set and determined that landscape characteristics could not explain presence vs. absence of spawners but that the percentage of agriculture, winter temperature range, and the intrinsic potential of the stream could explain some variation in abundance (weighted average R² = 0.30) where spawners were present. We conclude that the previous use of nonrandom monitoring data sets may have obscured understanding of species distribution, and we suggest minor modifications to large-scale monitoring programs

Construction of an In Silico Structural Profiling Tool Facilitating Mechanistically Grounded Classification of Aquatic Toxicants.

The performance of chemical safety assessment within the domain of environmental toxicology is often impeded by a shortfall of appropriate experimental data describing potential hazards across the many compounds in regular industrial use. In silico schemes for assigning aquatic-relevant modes or mechanisms of toxic action to substances, based solely on consideration of chemical structure, have seen widespread employment─including those of Verhaar, Russom, and later Bauer (MechoA). Recently, development of a further system was reported by Sapounidou, which, in common with MechoA, seeks to ground its classifications in understanding and appreciation of molecular initiating events. Until now, this Sapounidou scheme has not seen implementation as a tool for practical screening use. Accordingly, the primary purpose of this study was to create such a resource─in the form of a computational workflow. This exercise was facilitated through the formulation of 183 structural alerts/rules describing molecular features associated with narcosis, chemical reactivity, and specific mechanisms of action. Output was subsequently compared relative to that of the three aforementioned alternative systems to identify strengths and shortcomings as regards coverage of chemical space

Loss of the nuclear receptor corepressor SLIRP compromises male fertility

Nuclear receptors (NRs) and their coregulators play fundamental roles in initiating and directing gene expression influencing mammalian reproduction, development and metabolism. SRA stem Loop Interacting RNA-binding Protein (SLIRP) is a Steroid receptor RNA Activator (SRA) RNA-binding protein that is a potent repressor of NR activity. SLIRP is present in complexes associated with NR target genes in the nucleus; however, it is also abundant in mitochondria where it affects mitochondrial mRNA transcription and energy turnover. In further characterisation studies, we observed SLIRP protein in the testis where its localization pattern changes from mitochondrial in diploid cells to peri-acrosomal and the tail in mature sperm. To investigate the in vivo effects of SLIRP, we generated a SLIRP knockout (KO) mouse. This animal is viable, but sub-fertile. Specifically, when homozygous KO males are crossed with wild type (WT) females the resultant average litter size is reduced by approximately one third compared with those produced by WT males and females. Further, SLIRP KO mice produced significantly fewer progressively motile sperm than WT animals. Electron microscopy identified disruption of the mid-piece/annulus junction in homozygous KO sperm and altered mitochondrial morphology. In sum, our data implicates SLIRP in regulating male fertility, wherein its loss results in asthenozoospermia associated with compromised sperm structure and mitochondrial morphology.Shane M. Colley, Larissa Wintle, Richelle Searles, Victoria Russell, Renee C. Firman, Stephanie Smith, Kathleen DeBoer, D. Jo Merriner, Ben Genevieve, Jacqueline M. Bentel, Bronwyn G. A. Stuckey, Michael R. Phillips, Leigh W. Simmons, David M. de Kretser, Moira K. O, Bryan, Peter J. Leedma

A nomenclature for restriction enzymes, DNA methyltransferases, homing endonucleases and their genes

A nomenclature is described for restriction endonucleases, DNA methyltransferases, homing endonucleases and related genes and gene products. It provides explicit categories for the many different Type II enzymes now identified and provides a system for naming the putative genes found by sequence analysis of microbial genomes

Jury System in France Cour de Assies: Could it be adopted in the Indonesian Criminal Court?

This qualitative article provides findings on the possibility of implementing the France Cour de Assies jury system in Indonesian criminal court. The researcher explains the reasons for adopting the jury system in the French assize court, the position of the jury, the procedure for appointing jurors, explaining the advantages of the jury. The comparative perspective allows the researchers to reveal what is less known and gain better insight into two more things: the relationship between criminal justice and public involvement in modern society; and the meaning of criminal justice. Researchers use secondary data comprising books, journals, or references about the French Cour de Assies, including legislation, judicial decisions, customs, and doctrines. The adoption of the jury system in Indonesia is essential, considering that Indonesia is a democratic country; therefore, it must include the involvement of the public in the judiciary. It is possible to involve the public in case of examination in court as a jury, which is the obligation of a democratic country. This article suggests that the jury system in the Cour de Assies France can be accepted because, in Indonesian criminal justice, it is necessary to involve the public in determining if someone is guilty

Motor step size and ATP coupling efficiency of the dsDNA translocase EcoR124I

The Type I restriction-modification enzyme EcoR124I is an archetypical helicase-based dsDNA translocase that moves unidirectionally along the 3′–5′ strand of intact duplex DNA. Using a combination of ensemble and single-molecule measurements, we provide estimates of two physicochemical constants that are fundamental to a full description of motor protein activity—the ATP coupling efficiency (the number of ATP consumed per base pair) and the step size (the number of base pairs transported per motor step). Our data indicate that EcoR124I makes small steps along the DNA of 1 bp in length with 1 ATP consumed per step, but with some uncoupling of the ATPase and translocase cycles occurring so that the average number of ATP consumed per base pair slightly exceeds unity. Our observations form a framework for understanding energy coupling in a great many other motors that translocate along dsDNA rather than ssDNA

Identification of ejaculated proteins in the house mouse (Mus domesticus) via isotopic labeling

<p>Abstract</p> <p>Background</p> <p>Seminal fluid plays an important role in successful fertilization, but knowledge of the full suite of proteins transferred from males to females during copulation is incomplete. The list of ejaculated proteins remains particularly scant in one of the best-studied mammalian systems, the house mouse (<it>Mus domesticus</it>), where artificial ejaculation techniques have proven inadequate. Here we investigate an alternative method for identifying ejaculated proteins, by isotopically labeling females with ¹⁵N and then mating them to unlabeled, vasectomized males. Proteins were then isolated from mated females and identified using mass spectrometry. In addition to gaining insights into possible functions and fates of ejaculated proteins, our study serves as proof of concept that isotopic labeling is a powerful means to study reproductive proteins.</p> <p>Results</p> <p>We identified 69 male-derived proteins from the female reproductive tract following copulation. More than a third of all spectra detected mapped to just seven genes known to be structurally important in the formation of the copulatory plug, a hard coagulum that forms shortly after mating. Seminal fluid is significantly enriched for proteins that function in protection from oxidative stress and endopeptidase inhibition. Females, on the other hand, produce endopeptidases in response to mating. The 69 ejaculated proteins evolve significantly more rapidly than other proteins that we previously identified directly from dissection of the male reproductive tract.</p> <p>Conclusion</p> <p>Our study attempts to comprehensively identify the proteins transferred from males to females during mating, expanding the application of isotopic labeling to mammalian reproductive genomics. This technique opens the way to the targeted monitoring of the fate of ejaculated proteins as they incubate in the female reproductive tract.</p