Electric Field-Driven Water Dipoles: Nanoscale Architecture of Electroporation

<div><p>Electroporation is the formation of permeabilizing structures in the cell membrane under the influence of an externally imposed electric field. The resulting increased permeability of the membrane enables a wide range of biological applications, including the delivery of normally excluded substances into cells. While electroporation is used extensively in biology, biotechnology, and medicine, its molecular mechanism is not well understood. This lack of knowledge limits the ability to control and fine-tune the process. In this article we propose a novel molecular mechanism for the electroporation of a lipid bilayer based on energetics analysis. Using molecular dynamics simulations we demonstrate that pore formation is driven by the reorganization of the interfacial water molecules. Our energetics analysis and comparisons of simulations with and without the lipid bilayer show that the process of poration is driven by field-induced reorganization of water dipoles at the water-lipid or water-vacuum interfaces into more energetically favorable configurations, with their molecular dipoles oriented in the external field. Although the contributing role of water in electroporation has been noted previously, here we propose that interfacial water molecules are the main players in the process, its initiators and drivers. The role of the lipid layer, to a first-order approximation, is then reduced to a relatively passive barrier. This new view of electroporation simplifies the study of the problem, and opens up new opportunities in both theoretical modeling of the process and experimental research to better control or to use it in new, innovative ways.</p></div

Occupancy Modeling for Improved Accuracy and Understanding of Pathogen Prevalence and Dynamics

Most pathogen detection tests are imperfect, with a sensitivity < 100%, thereby resulting in the potential for a false negative, where a pathogen is present but not detected. False negatives in a sample inflate the number of non-detections, negatively biasing estimates of pathogen prevalence. Histological examination of tissues as a diagnostic test can be advantageous as multiple pathogens can be examined and providing important information on associated pathological changes to the host. However, it is usually less sensitive than molecular or microbiological tests for specific pathogens. Our study objectives were to 1) develop a hierarchical occupancy model to examine pathogen prevalence in spring Chinook salmon Oncorhynchus tshawytscha and their distribution among host tissues 2) use the model to estimate pathogen-specific test sensitivities and infection rates, and 3) illustrate the effect of using replicate within host sampling on sample sizes required to detect a pathogen. We examined histological sections of replicate tissue samples from spring Chinook salmon O. tshawytscha collected after spawning for common pathogens seen in this population: Apophallus/echinostome metacercariae, Parvicapsula minibicornis, Nanophyetus salmincola/metacercariae, and Renibacterium salmoninarum. A hierarchical occupancy model was developed to estimate pathogen and tissue-specific test sensitivities and unbiased estimation of host- and organ-level infection rates. Model estimated sensitivities and host- and organ-level infections rates varied among pathogens and model estimated infection rate was higher than prevalence unadjusted for test sensitivity, confirming that prevalence unadjusted for test sensitivity was negatively biased. The modeling approach provided an analytical approach for using hierarchically structured pathogen detection data from lower sensitivity diagnostic tests, such as histology, to obtain unbiased pathogen prevalence estimates with associated uncertainties. Accounting for test sensitivity using within host replicate samples also required fewer individual fish to be sampled. This approach is useful for evaluating pathogen or microbe community dynamics when test sensitivity is <100%

Innate and adaptive immune responses in migrating spring-run adult chinook salmon, Oncorhynchus tshawytscha

Adult Chinook salmon (Oncorhynchus tshawytscha) migrate from salt water to freshwater streams to spawn. Immune responses in migrating adult salmon are thought to diminish in the run up to spawning, though the exact mechanisms for diminished immune responses remain unknown. Here we examine both adaptive and innate immune responses as well as pathogen burdens in migrating adult Chinook salmon in the Upper Willamette River basin. Messenger RNA transcripts encoding antibody heavy chain molecules slightly diminish as a function of time, but are still present even after fish have successfully spawned. In contrast, the innate anti-bacterial effector proteins present in fish plasma rapidly decrease as spawning approaches. Fish also were examined for the presence and severity of eight different pathogens in different organs. While pathogen burden tended to increase during the migration, no specific pathogen signature was associated with diminished immune responses. Transcript levels of the immunosuppressive cytokines IL-10 and TGF beta were measured and did not change during the migration. These results suggest that loss of immune functions in adult migrating salmon are not due to pathogen infection or cytokine-mediated immune suppression, but is rather part of the life history of Chinook salmon likely induced by diminished energy reserves or hormonal changes which accompany spawning.Keywords: Parasite burden, Salmon, Immune respons

Tolerance and Efficacy of Emamectin Benzoate and Ivermectin for the Treatment of Pseudocapillaria tomentosa in Laboratory Zebrafish (Danio rerio)

Tolerance of adult zebrafish and efficacy of emamectin benzoate and ivermectin in eliminating Pseudocapillaria tomentosa infection were evaluated. In the tolerance study, behavioral changes, fecundity, histopathology, and mortality were evaluated for in-feed administration of emamectin (0.05, 0.10, and 0.25 mg/kg) and ivermectin (0.05 and 0.10 mg/kg). All doses of emamectin were well tolerated. Ivermectin 0.05 mg/kg administration resulted in mild behavioral changes and a transient decrease in fecundity. Ivermectin 0.10 mg/kg administration resulted in severe behavioral changes and some mortality. In the efficacy study, emamectin (0.05 and 0.25 mg/kg) and ivermectin (0.05 mg/kg) were evaluated for their efficacy in eliminating P. tomentosa infection. Emamectin reduced parasite burden in infected zebrafish, and ivermectin eliminated intestinal nematode infections. Despite a small margin of safety, ivermectin 0.05 mg/kg was effective at eliminating P. tomentosa infection in adult zebrafish. Higher doses or a longer course of treatment may be needed for complete elimination of P. tomentosa infection using emamectin. In this study, we propose two possible treatments for intestinal nematode infections in zebrafish

Radically enhanced molecular recognition

The tendency for viologen radical cations to dimerize has been harnessed to establish a recognition motif based on their ability to form extremely strong inclusion complexes with cyclobis(paraquat-p-phenylene) in its diradical dicationic redox state. This previously unreported complex involving three bipyridinium cation radicals increases the versatility of host–guest chemistry, extending its practice beyond the traditional reliance on neutral and charged guests and hosts. In particular, transporting the concept of radical dimerization into the field of mechanically interlocked molecules introduces a higher level of control within molecular switches and machines. Herein, we report that bistable and tristable [2]rotaxanes can be switched by altering electrochemical potentials. In a tristable [2]rotaxane composed of a cyclobis(paraquat-p-phenylene) ring and a dumbbell with tetrathiafulvalene, dioxynaphthalene and bipyridinium recognition sites, the position of the ring can be switched. On oxidation, it moves from the tetrathiafulvalene to the dioxynaphthalene, and on reduction, to the bipyridinium radical cation, provided the ring is also reduced simultaneously to the diradical dication

Life Histories, Demography, and Distribution of a Fluvial Bull Trout Population

To describe the life histories and demography of a fluvial population of Bull Trout Salvelinus confluentus, we PIT-tagged and radio-tagged Bull Trout captured in Mill Creek, a tributary of the Walla Walla River (Washington–Oregon), during 1998–2009. Adult abundance declined 63% during 2006–2010, driven primarily by a 10-fold reduction in subadult-to-adult returns. Larger subadults and fall–winter emigrants survived at higher rates, but they were a small proportion of the subadult migrants. The survival rates of larger, generally older adults were also more than 40% greater than those of smaller adults. Changes in abundance influenced other characteristics of the population. For example, adult upstream movement into spawning areas during 1999–2005 peaked in late July, whereas the smaller runs observed during 2006–2010 peaked in early September, and the relationship between fish size and migration timing shifted. Unlike many adfluvial populations, more than 90% of the adults in Mill Creek spawned annually. Bull Trout that spawned in main-stem Mill Creek were primarily larger migratory adults; however, about 20% of the large adults were strictly or intermittently resident, remaining in the spawning area year-round. The downstream extent of individuals' migratory distributions varied greatly—from just downstream of the spawning area to the mouth of the Walla Walla River and potentially hundreds of kilometers into the Columbia River. Despite a large sample size of radio-tagged fish, radiotelemetry substantially underestimated the distribution and range that were evident from PIT tag detections. Life history terms such as “migratory,” “resident,” and “fluvial” and their associations with body size, movement, and distribution are useful for describing general patterns, but they fail to reflect the diversity and complexity within and among populations. For Bull Trout in Mill Creek, that life history diversity, including small, resident adult forms in the tributaries and a continuum of distribution for large adults, maximizes the use of available habitat and likely contributes to the population's persistence

Predicting Crappie Recruitment in Ohio Reservoirs with Spawning Stock Size, Larval Density, and Chlorophyll Concentrations

Stock-recruit models typically use only spawning stock size as a predictor of recruitment to a fishery. In this paper, however, we used spawning stock size as well as larval density and key environmental variables to predict recruitment of white crappies Pomoxis annularis and black crappies P. nigromaculatus, a genus notorious for variable recruitment. We sampled adults and recruits from 11 Ohio reservoirs and larvae from 9 reservoirs during 1998-2001. We sampled chlorophyll as an index of reservoir productivity and obtained daily estimates of water elevation to determine the impact of hydrology on recruitment. Akaike's information criterion (AIC) revealed that Ricker and Beverton-Holt stock-recruit models that included chlorophyll best explained the variation in larval density and age-2 recruits. Specifically, spawning stock catch per effort (CPE) and chlorophyll explained 63-64% of the variation in larval density. In turn, larval density and chlorophyll explained 43-49% of the variation in age-2 recruit CPE. Finally, spawning stock CPE and chlorophyll were the best predictors of recruit CPE (i.e., 74-86%). Although larval density and recruitment increased with chlorophyll, neither was related to seasonal water elevation. Also, the AIC generally did not distinguish between Ricker and Beverton-Holt models. From these relationships, we concluded that crappie recruitment can be limited by spawning stock CPE and larval production when spawning stock sizes are low (i.e., CPE , 5 crappies/net-night). At higher levels of spawning stock sizes, spawning stock CPE and recruitment were less clearly related. To predict recruitment in Ohio reservoirs, managers should assess spawning stock CPE with trap nets and estimate chlorophyll concentrations. To increase crappie recruitment in reservoirs where recruitment is consistently poor, managers should use regulations to increase spawning stock size, which, in turn, should increase larval production and recruits to the fishery.This research was funded by Federal Aid in Sport Fish Restoration Project F-69-P, administered jointly by the U.S. Fish and Wildlife Service and Ohio Department of Natural Resources, Division of Wildlife, and the Department of Evolution, Ecology, and Organismal Biology at Ohio State University