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Previous studies have shown oviposition deterring properties of 8 coconut free fatty acid (CFFA) compounds on fruit flies with different key deterrent components for different species. Here we evaluated oviposition deterrence of CFFA using laboratory 2-choice bioassays against Zeugodacus cucurbitae, determined key-bioactive deterrent compounds, and evaluated their behavioral mode. Unlike other reported fruit fly species, CFFA mixture increased Z. cucurbitae oviposition when directly applied on an oviposition substrate. When tested individually in subsequent tests, 4 compounds (caprylic, capric, oleic, and linoleic acids) significantly reduced the oviposition ("negative-compounds"), 1 compound (stearic acid) had no effect ("neutral-compound"), and 3 compounds (lauric, myristic, and palmitic acids) stimulated the oviposition ("positive-compounds"). The 4-component negative-compound blend was effective at reducing oviposition. However, adding stearic acid to the 4-component blend (5-component blend, 5c) further reduced oviposition. Adding any of the positive-compounds to the 5c resulted in loss of oviposition deterrence, suggesting the 5c as the key deterrent component blend. The blend was also effective in no-choice assays and when applied on cucumbers, a preferred host of Z. cucurbitae. When given a choice, Z. cucurbitae made 48.5% fewer visits, spent 39% less time, and oviposited 88.2% fewer eggs per min on 5c treated pumpkin agar than on control agar, suggesting that the 5c blend has both spatial repellency and contact deterrence. Given that all compounds are registered food additives and generally regarded as safe, this blend has potential application in behavioral control strategies, such as push-pull, to protect host fruit against Z. cucurbitae., (© 2024 Institute of Zoology, Chinese Academy of Sciences.)

Cannabis, often criticized, has active compounds that indicate multiple medical benefits. However, before the potential decriminalization and legalization of medical cannabis, pharmacists play a crucial role in the decision-making for personalized therapy and evidence-based medicine. This study investigates pharmacists' perspectives on medical cannabis, covering medical use, societal impact, research, legality, and regulation. This study was conducted through semi-structured interviews with pharmacists from different sectors. Interviews were conducted with 16 pharmacists, with 4 from each sector: industry, academia, community, and hospital. There was a consensus on the need for streamlined research before potential decriminalization and legalization of cannabis, highlighting concerns about abuse and misuse despite its benefits. With that in mind, all participants agree that there is a need to update the current legislation and regulations to better control the use and distribution of medical cannabis. Several participants also called to attention that education plays an important role in the potential decriminalization of medical cannabis, with cannabis-related stigma being a potential deterrent for not only the public but also healthcare professionals. As medical cannabis is a new subject in Malaysia, this study could ignite further multidisciplinary human-related research with strict regulations which will ultimately result in evidence-based medicine.

Here, we demonstrate that the spatial distribution of lipophilic cations governs the complexation pathways, serum stability, and biological performance of polymer-pDNA complexes (polyplexes). Previous research focused on block/statistical copolymers, whereas gradient copolymers, where the density of lipophilic cations diminishes (gradually or steeply) along polymer backbones, remain underexplored. We engineered gradient copolymers that combine the polyplex colloidal stability of block copolymers with the transfection efficiency of statistical copolymers. We synthesized length- and compositionally equivalent gradient copolymers ( G1 - G3 ) along with statistical ( S ) and block ( B ) copolymers of 2-(diisopropylamino)ethyl methacrylate and 2-hydroxyethyl methacrylate. We mapped how polymer microstructure governs pDNA loading per polyplex, pDNA conformational changes, and polymer-pDNA binding thermodynamics via static light scattering, circular dichroism spectroscopy, and isothermal titration calorimetry, respectively. While gradient steepness is a powerful design handle to improve polyplex physical properties, augment pDNA delivery capacity, and attenuate polycation-triggered hemolysis, microstructural contrasts did not elicit differences in complement activation.

Polycations are scalable and affordable nanocarriers for delivering therapeutic nucleic acids. Yet, cationicity-dependent tradeoffs between nucleic acid delivery efficiency, cytotoxicity, and serum stability hinder clinical translation. Typically, the most efficient polycationic vehicles also tend to be the most toxic. For lipophilic polycations-which recruit hydrophobic interactions in addition to electrostatic interactions to bind and deliver nucleic acids-extensive chemical or architectural modifications sometimes fail to resolve intractable toxicity-efficiency tradeoffs. Here, we employ a facile post-synthetic polyplex surface modification strategy wherein poly(l-glutamic acid) (PGA) rescues toxicity, inhibits hemolysis, and prevents serum inhibition of lipophilic polycation-mediated plasmid (pDNA) delivery. Importantly, the sequence in which polycations, pDNA, and PGA are combined dictates pDNA conformations and spatial distribution. Circular dichroism spectroscopy reveals that PGA must be added last to polyplexes assembled from lipophilic polycations and pDNA; else, PGA will disrupt polycation-mediated pDNA condensation. Although PGA did not mitigate toxicity caused by hydrophilic PEI-based polycations, PGA tripled the population of transfected viable cells for lipophilic polycations. Non-specific adsorption of serum proteins abrogated pDNA delivery mediated by lipophilic polycations; however, PGA-coated polyplexes proved more serum-tolerant than uncoated polyplexes. Despite lower cellular uptake than uncoated polyplexes, PGA-coated polyplexes were imported into nuclei at higher rates. PGA also silenced the hemolytic activity of lipophilic polycations. Our work provides fundamental insights into how polyanionic coatings such as PGA transform intermolecular interactions between lipophilic polycations, nucleic acids, and serum proteins, and facilitate gentle yet efficient transgene delivery., Competing Interests: The authors do not have any conflicts of interest to disclose., (This journal is © The Royal Society of Chemistry.)

Tape-lifting is a non-destructive method employed in the laboratory to recover and collect trace DNA evidence from crime scene exhibits with porous surfaces. The success of tape-lifting is a balance between capturing the biological material and compatibility with downstream DNA extraction processes to ensure efficient release of the tape-lifted material during DNA extraction. In this study, six commercially available low-, regular- and high-tack adhesive tapes were evaluated. The low-tack S183 tape and the highly adhesive S-Hold tape were compared for DNA recovery efficiency from different materials commonly encountered in casework. All tape-lifts were processed using PrepFiler Express™ BTA and AutoMate Express™ Forensic DNA extraction systems, DNA samples quantitated by Quantifiler TRIO, amplified using Powerplex® 21 and VeriFiler™ PLUS (VFP), and analysed on a 3500xl genetic analyser to evaluate the quality of the resultant STR profiles obtained. The more adhesive S-Hold tape recovered comparable or more DNA than the low-tack S183 tape from the majority of materials tested. However, STR profiles obtained from S183 tape-lifts were of markedly higher quality compared to S-Hold tape-lifts. This was most evident for towel, denim and printed chiffon, where S-Hold samples exhibited severe PCR inhibition, with VFP internal quality markers confirming the presence of inhibitors. The findings suggest that strong adhesion is not necessarily beneficial for tape-lifting, as the low tack S183 tape was able to efficiently recover cellular material from the surface of porous substrates commonly encountered in casework, while avoiding the co-transfer of PCR-inhibitory substances from the sampled material., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Crown Copyright © 2023. Published by Elsevier B.V. All rights reserved.)

The WRAMP structure is a protein network associated with tail-end actomyosin contractility, membrane retraction, and directional persistence during cell migration. A marker of WRAMP structures is melanoma cell adhesion molecule (MCAM) which dynamically polarizes to the cell rear. However, factors that mediate MCAM polarization are still unknown. In this study, BioID using MCAM as bait identifies the ERM family proteins, moesin, ezrin, and radixin, as WRAMP structure components. We also present a novel image analysis pipeline, Protein Polarity by Percentile ("3P"), which classifies protein polarization using machine learning and facilitates quantitative analysis. Using 3P, we find that depletion of moesin, and to a lesser extent ezrin, decreases the proportion of cells with polarized MCAM. Furthermore, although copolarized MCAM and ERM proteins show high spatial overlap, 3P identifies subpopulations with ERM proteins closer to the cell periphery. Live-cell imaging confirms that MCAM and ERM protein polarization is tightly coordinated, but ERM proteins enrich at the cell edge first. Finally, deletion of a juxtamembrane segment in MCAM previously shown to promote ERM protein interactions impedes MCAM polarization. Our findings highlight the requirement for ERM proteins in recruitment of MCAM to WRAMP structures and an advanced computational tool to characterize protein polarization.

The β-barrel assembly machinery (BAM) complex is responsible for inserting outer membrane proteins (OMPs) into the Escherichia coli outer membrane. The SecYEG translocon inserts inner membrane proteins into the inner membrane and translocates both soluble proteins and nascent OMPs into the periplasm. Recent reports describe Sec possibly playing a direct role in OMP biogenesis through interactions with the soluble polypeptide transport-associated (POTRA) domains of BamA (the central OMP component of BAM). Here we probe the diffusion behavior of these protein complexes using photoactivatable super-resolution localization microscopy and single-particle tracking in live E. coli cells of BAM and SecYEG components BamA and SecE and compare them to other outer and inner membrane proteins. To accurately measure trajectories on the highly curved cell surface, three-dimensional tracking was performed using double-helix point-spread function microscopy. All proteins tested exhibit two diffusive modes characterized by "slow" and "fast" diffusion coefficients. We implement a diffusion coefficient analysis as a function of the measurement lag time to separate positional uncertainty from true mobility. The resulting true diffusion coefficients of the slow and fast modes showed a complete immobility of full-length BamA constructs in the time frame of the experiment, whereas the OMP OmpLA displayed a slow diffusion consistent with the high viscosity of the outer membrane. The periplasmic POTRA domains of BamA were found to anchor BAM to other cellular structures and render it immobile. However, deletion of individual distal POTRA domains resulted in increased mobility, suggesting that these domains are required for the full set of cellular interactions. SecE diffusion was much slower than that of the inner membrane protein PgpB and was more like OMPs and BamA. Strikingly, SecE diffused faster upon POTRA domain deletion. These results are consistent with the existence of a BAM-SecYEG trans-periplasmic assembly in live E. coli cells., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Coconut free fatty acid (CFFA), a mixture of 8 fatty acids derived from coconut oil, is an effective repellent and deterrent against a broad array of hematophagous insects. In this study, we evaluated the oviposition deterrent activity of CFFA on spotted-wing drosophila (SWD; Drosophila suzukii), a destructive invasive pest of berries and cherries, and identified bioactive key-deterrent compounds. In laboratory 2-choice tests, CFFA deterred SWD oviposition in a dose-dependent manner with the greatest reduction (99%) observed at a 20-mg dose compared with solvent control. In a field test, raspberries treated with 20-mg CFFA received 64% fewer SWD eggs than raspberries treated with the solvent control. In subsequent laboratory bioassays, 2 of CFFA components, caprylic and capric acids, significantly reduced SWD oviposition by themselves, while 6 other components had no effect. In choice and no-choice assays, we found that a blend of caprylic acid and capric acid, at equivalent concentrations and ratio as in CFFA, was as effective as CFFA, while caprylic acid or capric acid individually were not as effective as the 2-component blend or CFFA at equivalent concentrations, indicating the 2 compounds as the key oviposition deterrent components for SWD. The blend was also as effective as CFFA for other nontarget drosophilid species in the field. Given that CFFA compounds are generally regarded as safe for humans, CFFA and its bioactive components have potential application in sustainably reducing SWD damage in commercial fruit operations, thereby reducing the sole reliance on insecticides., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Background: Bactrocera dorsalis, oriental fruit fly (OFF), is one of the most destructive agricultural pests. Although bait sprays can effectively control OFF, resistance development has been a concern. We evaluated the oviposition deterrent activity of coconut free fatty acids (CFFA), a mixture of eight coconut oil-derived fatty acids known to repel hematophagous insects and deter their feeding and oviposition, against OFF females., Results: In laboratory 72-h two-choice assays using guava-juice infused-agar as an oviposition substrate, CFFA deterred OFF oviposition in a dose-dependent manner with the greatest reduction of 87% at 20 mg dose compared to the control. When the eight CFFA components were tested individually, four compounds (caprylic, capric, oleic, and linoleic acids) significantly reduced OFF oviposition ('negative-compounds'), two (lauric and myristic acids) had no effect ('neutral-compounds'), and two (palmitic and stearic acids) stimulated OFF oviposition ('positive-compounds'). In two-choice tests, the 'negative-compounds' blend failed to elicit the same level of oviposition reduction as CFFA at equivalent concentrations found in CFFA. Adding the two 'neutral-compounds' recovered the oviposition deterrence similar to CFFA. Subsequent subtraction tests showed that four 'negative-compounds' plus lauric acid was as effective as CFFA in reducing OFF oviposition in guava-juice agar. This five-component key-deterrent blend also reduced OFF oviposition by 95 and 72% on papaya and tomato fruit, respectively., Conclusion: CFFA acts as an oviposition deterrent for OFF. Given that CFFA compounds are generally regarded as safe for humans and the environment, CFFA and its bioactive components have potential use in behavioral control strategies against OFF. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA., (© 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive motor neuron dysfunction and loss. A portion of ALS cases are caused by mutation of the proteasome shuttle factor Ubiquilin 2 ( UBQLN2 ), but the molecular pathway leading from UBQLN2 dysfunction to disease remains unclear. Here, we demonstrate that UBQLN2 regulates the domesticated gag-pol retrotransposon 'paternally expressed gene 10 (PEG10)' in human cells and tissues. In cells, the PEG10 gag-pol protein cleaves itself in a mechanism reminiscent of retrotransposon self-processing to generate a liberated 'nucleocapsid' fragment, which uniquely localizes to the nucleus and changes the expression of genes involved in axon remodeling. In spinal cord tissue from ALS patients, PEG10 gag-pol is elevated compared to healthy controls. These findings implicate the retrotransposon-like activity of PEG10 as a contributing mechanism in ALS through the regulation of gene expression, and restraint of PEG10 as a primary function of UBQLN2., Competing Interests: HB, JH, JR, SL, WC, CE, GH, JT, AM, EU, CL No competing interests declared, AW The University of Colorado, Boulder, has a patent pending for the use of PEG10 inhibitors on which the author is an inventor, (© 2023, Black, Hanson et al.)

Oxygenic photosynthesis is driven by the coupled action of the light-dependent pigment-protein complexes, photosystem I and II, located within the internal thylakoid membrane system. However, photosystem II is known to be prone to photooxidative damage. Thus, photosynthetic organisms have evolved a repair cycle to continuously replace the damaged proteins in photosystem II. However, it has remained difficult to deconvolute the damage and repair processes using traditional ensemble approaches. Here, we demonstrate an automated approach using time-lapse fluorescence microscopy and computational image analysis to study the dynamics and effects of photodamage in single cells at subcellular resolution in cyanobacteria. By growing cells in a two-dimensional layer, we avoid shading effects, thereby generating uniform and reproducible growth conditions. Using this platform, we analyzed the growth and physiology of multiple strains simultaneously under defined photoinhibitory conditions stimulated by UV-A light. Our results reveal an asymmetric cellular response to photodamage between sibling cells and the generation of an elusive subcellular structure, here named a 'photoendosome,' derived from the thylakoid which could indicate the presence of a previously unknown photoprotective mechanism. We anticipate these results to be a starting point for further studies to better understand photodamage and repair at the single-cell level., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

The 2021 annual meeting of the Entomological Society of America with the theme of "Adapt. Advance. Transform" guided the highlight compilation of urban entomology papers. Integrated pest management of urban pests relies on techniques and tools that adapt, advance, and transform over time to develop into new usable tactics and technologies; the review covers the following three themes: first, how science, researchers, and pest management professionals adapt to the changing environments; second, how urban pest management approaches and technologies advance using conventional and alternative strategies; and last, how transformation happens, leading to technological advances and sustainable pest management success. I selected papers on four groups of urban pests-ants, termites, bed bugs, and cockroaches-that were in the context of pest management in this highlights publication. Emphasis was placed on research regarding the efficacy of chemical (e.g., conventional pesticides, insect growth regulators), nonchemical, and alternative approaches (e.g., the use of heat, naturally derived compounds) for urban pests that help improve and implement sustainable pest management plans. These papers demonstrate the achievements of the urban entomology community while emphasizing the current challenges that we face from the far-reaching impact of climate change and insect pest adaptation and what should be prioritized., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cyanobacteria are photosynthetic microorganisms that play important ecological roles as major contributors to global nutrient cycles. Cyanobacteria are highly efficient in carrying out oxygenic photosynthesis because they possess carboxysomes, a class of bacterial microcompartments (BMC) in which a polyhedral protein shell encapsulates the enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and carbonic anhydrase and functions as the key component of the cyanobacterial CO 2 -concentrating mechanism (CCM). Elevated CO 2 levels within the carboxysome shell as a result of carbonic anhydrase activity increase the efficiency of RuBisCO. Yet, there remain many questions regarding the flux or exclusion of metabolites across the shell and how the activity of BMCs varies over time. These questions have been difficult to address using traditional ensemble techniques due to the heterogeneity of BMCs extracted from their native hosts or with heterologous expression. In this chapter, we describe a method to film and extract quantitative information about carboxysome activity using molecular biology and live cell, timelapse microscopy. In our method, the production of carboxysomes is first controlled by deleting the native genes required for carboxysome assembly and then re-introducing them under the control of an inducible promoter. This system enables carboxysomes to be tracked through multiple generations of cells and provides a way to quantify the total biomass accumulation attributed to a single carboxysome. While the method presented here was developed specifically for carboxysomes, it could be modified to track and quantify the activity of bacterial microcompartments in general., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Myosin heavy chain 7b (MYH7b) is an evolutionarily ancient member of the sarcomeric myosin family, which typically supports striated muscle function. However, in mammals, alternative splicing prevents MYH7b protein production in cardiac and most skeletal muscles and limits expression to a subset of specialized muscles and certain nonmuscle environments. In contrast, MYH7b protein is abundant in python cardiac and skeletal muscles. Although the MYH7b expression pattern diverges in mammals versus reptiles, MYH7b shares high sequence identity across species. So, it remains unclear how mammalian MYH7b function may differ from that of other sarcomeric myosins and whether human and python MYH7b motor functions diverge as their expression patterns suggest. Thus, we generated recombinant human and python MYH7b protein and measured their motor properties to investigate any species-specific differences in activity. Our results reveal that despite having similar working strokes, the MYH7b isoforms have slower actin-activated ATPase cycles and actin sliding velocities than human cardiac β-MyHC. Furthermore, python MYH7b is tuned to have slower motor activity than human MYH7b because of slower kinetics of the chemomechanical cycle. We found that the MYH7b isoforms adopt a higher proportion of myosin heads in the ultraslow, super-relaxed state compared with human cardiac β-MyHC. These findings are supported by molecular dynamics simulations that predict MYH7b preferentially occupies myosin active site conformations similar to those observed in the structurally inactive state. Together, these results suggest that MYH7b is specialized for slow and energy-conserving motor activity and that differential tuning of MYH7b orthologs contributes to species-specific biological roles., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Global commerce and transportation facilitate the spread of invasive species. The African big-headed ant, Pheidole megacephala (Fabricius), has achieved worldwide distribution through globalization. Since the late 19th century, Taiwan has served as a major seaport because of its strategic location. The population genetic structure of P. megacephala in Taiwan is likely to be shaped by international trade and migration between neighboring islands. In this study, we investigated the population genetics of P. megacephala colonies sampled from four geographical regions in Taiwan and elucidated the population genetic structures of P. megacephala sampled from Taiwan, Okinawa, and Hawaii. We observed a low genetic diversity of P. megacephala across regions in Taiwan. Moreover, we noted low regional genetic differentiation and did not observe isolation by distance, implying that long-distance jump dispersal might have played a crucial role in the spread of P. megacephala . We sequenced the partial cytochrome oxidase I gene and observed three mitochondrial haplotypes (TW1-TW3). TW1 and TW3 most likely originated from populations within the species' known invasive range, suggesting that secondary introduction is the predominant mode of introduction for this invasive ant. TW2 represents a novel haplotype that was previously unreported in other regions. P. megacephala populations from Taiwan, Okinawa, and Hawaii exhibited remarkable genetic similarity, which may reflect their relative geographic proximity and the historical connectedness of the Asia-Pacific region., Competing Interests: The authors declare no competing financial interests., (© 2022 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Entomopathogenic fungi allow chemical-free and environmentally safe vector management. Beauveria bassiana (Balsamo-Crivelli) Vuillemin is a promising biological control agent and an important component of integrated vector management. We investigated the mortality of Aedes albopictus (Skuse) larvae exposed to five concentrations of B. bassiana using Mycotrol ESO and adult oviposition behavior to analyze the egg-laying preferences of wild Ae. albopictus in response to different fungal concentrations. We examined the mortality of mid-instars exposed to B. bassiana concentrations of 1 × 104, 1 × 105, 1 × 106, 1 × 107, and 1 × 108 conidia/ml every 24 h for 12 d. In the oviposition behavior study, the fungus was applied to wooden paddles at 1 × 105, 1 × 107, and 1 × 109 conidia/ml, and the paddles were individually placed into quad-ovitraps. Both experiments contained control groups without B. bassiana. Kaplan-Meier survival analysis revealed that larval mortality was concentration dependent. The median lethal concentration was 2.43 × 105 conidia/ml on d 12. The median lethal time was 3.68 d at 1 × 106 conidia/ml. Oviposition monitoring revealed no significant difference in egg count between the control and treatment paddles. We observed an inverse relationship between the concentration of B. bassiana and the percentage of paddles with eggs. We concluded that concentrations above 1 × 106 conidia/ml are larvicidal, and Ae. albopictus laid similar numbers of eggs on fungus-impregnated and control wooden substrates; however, they were more likely to oviposit on substrates without B. bassiana. With these findings, we suggest that B. bassiana-infused ovitraps can be used for mosquito population monitoring while also delivering mycopesticides to adult mosquitoes., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America.)

Pseudomonas putida KT2440 is a well-studied bacterium for the conversion of lignin-derived aromatic compounds to bioproducts. The development of advanced genetic tools in P. putida has reduced the turnaround time for hypothesis testing and enabled the construction of strains capable of producing various products of interest. Here, we evaluate an inducible CRISPR-interference (CRISPRi) toolset on fluorescent, essential, and metabolic targets. Nuclease-deficient Cas9 (dCas9) expressed with the arabinose (8K)-inducible promoter was shown to be tightly regulated across various media conditions and when targeting essential genes. In addition to bulk growth data, single cell time lapse microscopy was conducted, which revealed intrinsic heterogeneity in knockdown rate within an isoclonal population. The dynamics of knockdown were studied across genomic targets in exponentially-growing cells, revealing a universal 1.75 ± 0.38 h quiescent phase after induction where 1.5 ± 0.35 doublings occur before a phenotypic response is observed. To demonstrate application of this CRISPRi toolset, β-ketoadipate, a monomer for performance-advantaged nylon, was produced at a 4.39 ± 0.5 g/L and yield of 0.76 ± 0.10 mol/mol from p -coumarate, a hydroxycinnamic acid that can be derived from grasses. These cultivation metrics were achieved by using the higher strength IPTG (1K)-inducible promoter to knockdown the pcaIJ operon in the βKA pathway during early exponential phase. This allowed the majority of the carbon to be shunted into the desired product while eliminating the need for a supplemental carbon and energy source to support growth and maintenance., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: J.C.C. is a co-founder and equity holder in Prometheus Materials Inc. All other authors declare no competing interests., (© 2022 The Authors. Published by Elsevier B.V. on behalf of International Metabolic Engineering Society.)

Mammalian cells respond to dsRNA in multiple manners. One key response to dsRNA is the activation of PKR, an eIF2α kinase, which triggers translational arrest and the formation of stress granules. However, the process of PKR activation in cells is not fully understood. In response to increased endogenous or exogenous dsRNA, we observed that PKR forms novel cytosolic condensates, referred to as dsRNA-induced foci (dRIFs). dRIFs contain dsRNA, form in proportion to dsRNA, and are enhanced by longer dsRNAs. dRIFs enrich several other dsRNA-binding proteins, including ADAR1, Stau1, NLRP1, and PACT. Strikingly, dRIFs correlate with and form before translation repression by PKR and localize to regions of cells where PKR activation is initiated. We hypothesize that dRIF formation is a mechanism that cells use to enhance the sensitivity of PKR activation in response to low levels of dsRNA or to overcome viral inhibitors of PKR activation.

The differential separation method is key to recovering a DNA profile of the sperm donor from sexual assault samples. However, low numbers of spermatozoa from the perpetrator are often swamped by the victim's epithelial cells or lost during the separation process, with the separation process labor-intensive, time-consuming, and operator-dependent. The self-sealing filter of the i-sep ® DL spin column allows direct lysis of the substrate throughout the differential separation process while preventing intact sperm cells from passing through, maximizing DNA recovery, and separation of non-sperm and sperm cells present. This study investigated the efficacy of a modified differential separation method, which incorporated the i-sep ® DL spin column in comparison with the conventional pellet-based differential separation method. Using semen dilution series and mock post-coital samples, the sensitivity, reproducibility, repeatability, and efficiency of sperm DNA recovery of the pellet-based differential to the i-sep ® method were evaluated side by side. The i-sep ® differential method was more sensitive in capturing sperm fraction DNA, with informative semen donor alleles detected from high dilutions of semen inputs where the pellet method has been unsuccessful. The i-sep ® differential method reduces manual handling, generating repeatable, and reproducible results between operators. Re-extraction of samples previously processed by the pellet or i-sep ® differential method showed that the pellet method failed to recover 15-88% of sperm fraction DNA, while the i-sep ® differential method was able to recover >99% in the initial extraction. The i-sep ® method is robust for processing sexual assault samples, overcoming the challenges of sperm DNA losses encountered by pellet-based methods., (© 2022 Commonwealth of Australia. Journal of Forensic Sciences © 2022 American Academy of Forensic Sciences.)

Frankliniella occidentalis is among the most economically significant pests of greenhouse crops, whose resistance to conventional insecticides has created demand for biopesticides such as essential oils. We assessed the fumigant toxicity of linalool against F. occidentalis , F. insularis , and Solanum lycopersicum . Thrips were fumigated with polyacrylamide hydrogels containing either ( R )-linalool, ( S )-linalool, racemic linalool, or a binary mixture of ( R )-linalool with one of twelve adjuvants (i.e., peppermint, cedarwood, neem, clove, coconut, jojoba, soybean, olive, α-terpineol, 1,8-cineole, trans-anethole, or ( R )-pulegone). Solanum lycopersicum seedlings were exposed to ( R )-linalool or a mixture of ( R )-linalool and peppermint oil via conditioned hydrogels or foliar spray. For F. insularis , ( R )-linalool was more toxic than ( S )-linalool, with LC 50 values of 11.7 mg/L air and 16.7 mg/L air, respectively. Similarly for F. occidentalis , ( R )-linalool was more toxic than ( S )-linalool, with LC 50 values of 29.0 mg/L air and 34.9 mg/L air, respectively. Peppermint oil and α-terpineol were the only synergists, while the other adjuvants exhibited varying degrees of antagonism. All seedling treatments demonstrated phytotoxicity, but symptoms were most severe for foliar sprays and mixtures containing peppermint oil. While hydrogels conditioned in linalool may be a favorable substitute to conventional insecticides, the cross-resistance demonstrated herein indicates that expectations should be metered.

Tools for refined cell-specific targeting have significantly contributed to understanding the characteristics and dynamics of distinct cellular populations by brain region. While advanced cell-labeling methods have accelerated the field of neuroscience, specifically in brain mapping, there remains a need to quantify and analyze the data. Here, by modifying a toolkit that localizes electrodes to brain regions (SHARP-Track; Slice Histology Alignment, Registration, and Probe-Track analysis), we introduce a post-imaging analysis tool to map histological images to established mouse brain atlases called SHARCQ (Slice Histology Alignment, Registration, and Cell Quantification). The program requires MATLAB, histological images, and either a manual or automatic cell count of the unprocessed images. SHARCQ simplifies the post-imaging analysis pipeline with a step-by-step GUI. We demonstrate that SHARCQ can be applied for a variety of mouse brain images, regardless of histology technique. In addition, SHARCQ rectifies discrepancies in mouse brain region borders between atlases by allowing the user to select between the Allen Brain Atlas or the digitized and modified Franklin-Paxinos Atlas for quantifying cell counts by region. SHARCQ produces quantitative and qualitative data, including counts of brain-wide region populations and a 3D model of registered cells within the atlas space. In summary, SHARCQ was designed as a neuroscience post-imaging analysis tool for cell-to-brain registration and quantification with a simple, accessible interface. All code is open-source and available for download (https://github.com/wildrootlab/SHARCQ)., (Copyright © 2022 Lauridsen et al.)

With heat treatments to control drywood termites (Blattodea: Kalotermitidae), the presence of heat sinks causes heat to be distributed unevenly throughout the treatment areas. Drywood termites may move to galleries in heat sink areas to avoid exposure to lethal temperatures. Our studies were conducted in Crytotermes brevis -infested condominiums in Honolulu, Hawaii to reflect real-world condominium scenarios; either a standard heat treatment performed by a heat remediation company, or an improved heat treatment was used. For improved treatments, heated air was directed into the toe-kick voids of C. brevis infested cabinets to reduce heat sink effects and increase heat penetration into these difficult-to-heat areas. Eight thermistor sensors placed inside the toe-kick voids, treatment zone, embedded inside cabinets' sidewalls, and in a wooden cube recorded target temperatures of above 46 °C or 50 °C for 120 min. Pre-treatment and follow-up inspections were performed at 6 months posttreatment to monitor termite inactivity using visual observations and by recording the numbers of spiked peaks on a microwave technology termite detection device (Termatrac). In improved treatment condominiums, significantly higher numbers of spiked peaks were recorded at pre-treatment as compared to 6 months posttreatment. Efficacious heat treatment protocols using the improved methods are proposed.

Argentine ants are one of the most common nuisance pest ants treated by pest management professionals (PMPs) in southern and western urban residential areas of the United States. Two new technologies (spraying with a pheromone adjuvant and using a biodegradable hydrogel bait delivery method) were used to develop a unique low-impact integrated pest management (IPM) protocol for Argentine ants in urban residential settings. The IPM protocol included a one-time perimeter spray treatment with 0.03% fipronil (mixed with a pheromone adjuvant) at the beginning of the ant season to achieve a quick knockdown. The initial spray application was followed by a biodegradable hydrogel baiting with 1% boric acid as a maintenance treatment. This low-impact IPM protocol was compared with two other conventional methods: (1) one initial fipronil application and one pyrethroid spray application for maintenance, or (2) one initial fipronil application and one essential oil insecticide spray application for maintenance. Based on Argentine ant foraging activity, the protocols were compared for their control efficacy. Insecticide use information and treatment time were also recorded and compared among different treatment protocols. Our results provided empirical data to support the effectiveness and economic feasibility of the low-impact IPM protocol for managing Argentine ants in urban residential settings., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Here, we review the literature on the development and application of hydrogel compounds for insect pest management. Researchers have used hydrogel compounds for the past few decades to achieve the controlled release of various contact insecticides, but in recent years, hydrogel compounds have also been used to absorb and deliver targeted concentrations of toxicants within a liquid bait to manage insect pests. The highly absorbent hydrogel acts as a controlled-release formulation that keeps the liquid bait available and palatable to the target pests. This review discusses the use of various types of hydrogel compounds in pest management based on different environmental settings (e.g., agricultural, urban, and natural areas), pest systems (e.g., different taxa), and modes of insecticide delivery (e.g., spray vs bait). Due to their unique physicochemical properties, hydrogel compounds have great potential to be developed into new and efficacious pest management strategies with minimal environmental impact. We will also discuss the future research and development of hydrogels in this review., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

Carboxysomes, prototypical bacterial microcompartments (BMCs) found in cyanobacteria, are large (~1 GDa) and essential protein complexes that enhance CO 2 fixation. While carboxysome biogenesis has been elucidated, the activity dynamics, lifetime, and degradation of these structures have not been investigated, owing to the inability of tracking individual BMCs over time in vivo. We have developed a fluorescence-imaging platform to simultaneously measure carboxysome number, position, and activity over time in a growing cyanobacterial population, allowing individual carboxysomes to be clustered on the basis of activity and spatial dynamics. We have demonstrated both BMC degradation, characterized by abrupt activity loss followed by polar recruitment of the deactivated complex, and a subclass of ultraproductive carboxysomes. Together, our results reveal the BMC life cycle after biogenesis and describe the first method for measuring activity of single BMCs in vivo., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

In unison, fingerprinting and DNA analysis have played a pivotal role in forensic investigations. Fingerprint powders that are available on the market can come in a range of colors and with specific properties. This study evaluated the efficiency of DNA extraction from samples coated with 3 brands of fingerprint powders: Lightning, Sirchie, and SupraNano, covering a range of colors and properties. A total of 23 fingerprint powders were tested using the Chelex, Promega DNA IQ™, and Applied Biosystems™ PrepFiler™ DNA extraction protocols. The DNA IQ™ and PrepFiler™ methods extracted higher yields of DNA in comparison to Chelex, which also accounted for better quality of PowerPlex x00AE; 21 DNA profiles recovered. There were no signs of degradation or inhibition in the quantification data, indicating that samples returning low DNA yield was due to interference during DNA extraction and not PCR inhibition. DNA profiles were recovered from the majority of fingerprint powders with only a single powder, Sirchie Magnetic Silver, failing to produce a profile using any of the methods tested. A link was observed between the DNA extraction chemistry, fingerprint powder property, that is, nonmagnetic, magnetic and aqueous, and the brand of fingerprint powder. Overall, the DNA IQ™ method was favorable for nonmagnetic fingerprint powders, while magnetic fingerprint powders produced more DNA profiles when extracted with the PrepFiler™ chemistry. This study highlights the importance of screening DNA extraction chemistries for the type of fingerprint powder used, as there is not a single DNA extraction method that suits all fingerprint powder brands and properties., (© 2019 American Academy of Forensic Sciences.)

Photosynthetic organisms regulate their responses to many diverse stimuli in an effort to balance light harvesting with utilizable light energy for carbon fixation and growth (source-sink regulation). This balance is critical to prevent the formation of reactive oxygen species that can lead to cell death. However, investigating the molecular mechanisms that underlie the regulation of photosynthesis in cyanobacteria using ensemble-based measurements remains a challenge due to population heterogeneity. Here, to address this problem, we used long-term quantitative time-lapse fluorescence microscopy, transmission electron microscopy, mathematical modelling and genetic manipulation to visualize and analyse the growth and subcellular dynamics of individual wild-type and mutant cyanobacterial cells over multiple generations. We reveal that mechanical confinement of actively growing Synechococcus sp. PCC 7002 cells leads to the physical disassociation of phycobilisomes and energetic decoupling from the photosynthetic reaction centres. We suggest that the mechanical regulation of photosynthesis is a critical failsafe that prevents cell expansion when light and nutrients are plentiful, but when space is limiting. These results imply that cyanobacteria must convert a fraction of the available light energy into mechanical energy to overcome frictional forces in the environment, providing insight into the regulation of photosynthesis and how microorganisms navigate their physical environment.

The repair of DNA damage requires the ordered recruitment of many different proteins that are responsible for signaling and subsequent repair. A powerful and widely used tool for studying the orchestrated accumulation of these proteins at damage sites is laser microirradiation in live cells, followed by monitoring the accumulation of the fluorescently labeled protein in question. Despite the widespread use of this approach, there exists no rigorous method for characterizing the recruitment process quantitatively. Here, we introduce a diffusion model that explicitly accounts for the unique sizes and shapes of individual nuclei and uses two variables: D eff , the effective coefficient of diffusion, and F, the fraction of mobile protein that accumulates at sites of DNA damage. Our model quantitatively describes the accumulation of three test proteins, poly-ADP-ribose polymerases 1 and 2 (PARP1/2) and histone PARylation factor 1. D eff for PARP1, as derived by our approach, is 6× greater than for PARP2 and in agreement with previous literature reports using fluorescence correlation spectroscopy and fluorescence recovery after photobleaching. Our data indicate that histone PARylation factor 1 arrives at sites of DNA damage independently of either PARP. Importantly, our model, which can be applied to existing data, allows for the direct comparison of the coefficient of diffusion for any DNA repair protein between different cell types, obtained in different laboratories and by different methods, and also allows for the interrogation of cell-to-cell variability., (Copyright © 2019. Published by Elsevier Inc.)

Background: Insecticide sprays used for ant control cause environmental contamination. Liquid bait is a safe and effective alternative, but it requires bait stations to dispense the toxicant. We developed a biodegradable hydrogel to deliver liquid bait obviating the need for bait stations., Results: Alginate hydrogel beads with preferred rigidity and maximum hydration in 25% sucrose solution were engineered by optimizing a crosslinking process. The moisture content of the substrate on which the beads were placed and the relative atmospheric humidity significantly influenced water loss dynamics of the hydrated hydrogel beads. Laboratory choice studies indicated that hydrated hydrogel beads had reduced palatability to foraging ants when they lost ≥50% water. An enzyme-linked immunosorbent assay (ELISA) indicated that the insecticide thiamethoxam added to sucrose solution was absorbed into the hydrogel beads. Hydrogel beads conditioned in sucrose solution with 1 mg L -1 thiamethoxam provided complete control of all castes of Argentine ant Linepithema humile (Mayr) colony by 14 days post treatment in the laboratory trial and provided a 79% reduction in ant activity after 8 weeks in the field trial., Conclusion: Alginate hydrogel beads provided an effective delivery system for liquid baits laced with low concentrations of insecticide to control Argentine ants. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Background: Accumulating evidence indicate that circulating microRNAs (miRNAs) are useful independent non-invasive biomarkers, with unique miRNA signatures defined for various pathophysiological conditions. However, there are no established universal housekeeping miRNAs for the normalisation of miRNAs in body fluids. We have previously identified an oestrogen-responsive miRNA, miR-494, in regulating the anticoagulant, Protein S, in HuH-7 liver cells. Moreover, increased thrombotic risk associated with elevated circulating oestrogen levels is frequently observed in pregnant women and oral contraceptive users. In order to identify other oestrogen-responsive miRNAs, including miR-494, that may be indicative of increased thrombotic risk in plasma, we used nanoString analysis to identify robust and stable endogenous reference miRNAs for the study of oestrogen-responsive miRNAs in plasma., Results: We compared the plasma miRNA expression profile of individuals with: (1) Low circulating oestrogens (healthy men and non-pregnant women not taking oral contraceptives), (2) High circulating synthetic oestrogens, (women taking oral contraceptives) and (3) High circulating natural oestrogens (pregnant females >14 weeks gestation). From the nanoString analyses, 11 candidate reference miRNAs which exhibited high counts and not significantly differentially expressed between groups were selected for validation using realtime quantitative polymerase chain reaction (RT-qPCR) and digital droplet PCR (DDPCR) in pooled plasma samples, and the stability of their expression evaluated using NormFinder and BestKeeper algorithms. Four miRNAs (miR-25-5p, miR-188-5p, miR-222-3p and miR-520f) demonstrated detectable stable expression between groups and were further analysed by RT-qPCR in individual plasma samples, where miR-188-5p and miR-222-3p expression were identified as a stable pair of reference genes. The miRNA reference panel consisting of synthetic spike-ins cel-miR-39 and ath-miR159a, and reference miRNAs, miR-188-5p and miR-222-3p was useful in evaluating fold-change of the pregnancy-associated miRNA, miR-141-3p, between groups., Conclusion: The miRNA reference panel will be useful for normalising qPCR data comparing miRNA expression between men and women, non-pregnant and pregnant females, and the potential effects of endogenous and synthetic oestrogens on plasma miRNA expression.

In contrast to events at the cell leading edge, rear-polarized mechanisms that control directional cell migration are poorly defined. Previous work described a new intracellular complex, the Wnt5a-receptor-actomyosin polarity (WRAMP) structure, which coordinates the polarized localization of MCAM, actin, and myosin IIB in a Wnt5a-induced manner. However, the polarity and function for the WRAMP structure during cell movement were not determined. Here we characterize WRAMP structures during extended cell migration using live-cell imaging. The results demonstrate that cells undergoing prolonged migration show WRAMP structures stably polarized at the rear, where they are strongly associated with enhanced speed and persistence of directional movement. Strikingly, WRAMP structures form transiently, with cells displaying directional persistence during periods when they are present and cells changing directions randomly when they are absent. Cells appear to pause locomotion when WRAMP structures disassemble and then migrate in new directions after reassembly at a different location, which forms the new rear. We conclude that WRAMP structures represent a rear-directed cellular mechanism to control directional migration and that their ability to form dynamically within cells may control changes in direction during extended migration., (© 2017 Connacher et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

Background: Reproductive division of labor is one of the key features of social insects. Queens are adapted for reproduction while workers are adapted for foraging and colony maintenance. In many species, however, workers retain functional ovaries and can lay unfertilized male eggs or trophic eggs. Here we report for the first time on the occurrence of physogastric workers and apparent worker reproduction in the invasive yellow crazy ant Anoplolepis gracilipes (Fr. Smith). We further examined the reproductive potential and nutritional role of physogastric workers through multidisciplinary approaches including morphological characterization, laboratory manipulation, genetic analysis and behavioral observation., Results: Egg production with two types of eggs, namely reproductive and trophic eggs, by physogastric workers was found. The reproductive egg was confirmed to be haploid and male-destined, suggesting that the workers produced males via arrhenotokous parthenogenesis as no spermatheca was discovered. Detailed observations suggested that larvae were mainly fed with trophic eggs. Along with consumption of trophic eggs by queens and other castes as part of their diet, the vital role of physogastric workers as "trophic specialist" is confirmed., Conclusion: We propose that adaptive advantages derived from worker reproduction for A. gracilipes may include 1) trophic eggs provisioned by physogastric workers likely assist colonies of A. gracilipes in overcoming unfavorable conditions such as paucity of food during critical founding stage; 2) worker-produced males are fertile and thus might offer an inclusive fitness advantage for the doomed orphaned colony.

One of the prime limiting factors of optical imaging in biological applications is the diffusion of light by tissue, which prevents focusing at depths greater than the optical diffusion limit (typically ?1??mm). To overcome this challenge, wavefront shaping techniques that use a spatial light modulator (SLM) to correct the phase of the incident wavefront have recently been developed. These techniques are able to focus light through scattering media beyond the optical diffusion limit. However, the low speeds of typically used liquid crystal SLMs limit the focusing speed. Here, we present a method using a digital micromirror device (DMD) and an electro-optic modulator (EOM) to measure the scattering-induced aberrations, and using a liquid crystal SLM to apply the correction to the illuminating wavefront. By combining phase modulation from an EOM with the DMD’s ability to provide selective illumination, we exploit the DMD’s higher refresh rate for phase measurement. We achieved focusing through scattering media in less than 8 ms, which is sufficiently short for certain in vivo applications, as it is comparable to the speckle correlation time of living tissue.

To evaluate the effects of the juvenile hormone analogue pyriproxyfen on colonies of the Pharaoh ant Monomorium pharaonis (L.), peanut oil containing different concentrations (0.3, 0.6, or 0.9%) of pyriproxyfen was fed to monogynous (1 queen, 500 workers, and 0.1 g of brood) and polygynous (8 queens, 50 workers, and 0.1 g of brood) laboratory colonies of M. pharaonis. Due to its delayed activity, pyriproxyfen at all concentrations resulted in colony elimination. Significant reductions in brood volume were recorded at weeks 3 - 6, and complete brood mortality was observed at week 8 in all treated colonies. Brood mortality was attributed to the disruption of brood development and cessation of egg production by queens. All polygynous colonies exhibited significant reduction in the number of queens present at week 10 compared to week 1. Number of workers was significantly lower in all treated colonies compared to control colonies at week 8 due to old-age attrition of the workers without replacement. At least 98.67 ± 1.33% of workers were dead at week 10 in all treated colonies. Thus, treatment with slow acting pyriproxyfen at concentrations of 0.3 - 0.9% is an effective strategy for eliminating Pharaoh ant colonies.

Budding and relocation of nests are important characteristics of the Pharaoh ant, Monomorium pharaonis (L.), an important pest of artificial structures. Pharaoh ant colony movements induced by several types of disturbances were evaluated in the laboratory. The percentages of workers and brood in the source and new nest sites were determined at Days 0, 1, 3, and 5 following physical disturbance (temporal removal of nestmates), chemical disturbance (application of pyrethroid insecticide), invasion by heterospecific ants, food depletion, and moisture depletion in the laboratory. All disturbances were performed in the source nest, which was connected to an empty new nest site. Almost all workers moved and carried the entire brood to the new nest site when subjected to physical disturbance, chemical disturbance, and ant invasion on Day 1, whereas only <5% of workers were present in the new nest site in the undisturbed control. After these disturbances, the brood was never relocated back to the original nest site in this 5-d study. When subjected to food depletion, ∼60% of the brood were found in the new nest site and ∼40% of the brood remained in the original nest on Day 5, resulting in a polydomous population. In contrast, moisture depletion did not show any significant effect on colony movement. These results provide useful information about the causes of Pharaoh ant colony budding and guidance about how to develop effective control and prevention strategies., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Non-invasively focusing light into strongly scattering media, such as biological tissue, is highly desirable but challenging. Recently, ultrasonically guided wavefront shaping technologies have been developed to address this limitation. So far, the focusing resolution of most implementations has been limited by acoustic diffraction. Here, we introduce nonlinear photoacoustically guided wavefront shaping (PAWS), which achieves optical diffraction-limited focusing in scattering media. We develop an efficient dual-pulse excitation approach to generate strong nonlinear photoacoustic (PA) signals based on the Grueneisen relaxation effect. These nonlinear PA signals are used as feedback to guide iterative wavefront optimization. As a result, light is effectively focused to a single optical speckle grain on the scale of 5-7 µm, which is ~10 times smaller than the acoustic focus with an enhancement factor of ~6,000 in peak fluence. This technology has the potential to benefit many applications that desire highly confined strong optical focus in tissue.

Optical-resolution photoacoustic flowmetry (PAF) allows noninvasive single-cell flow measurements. However, its operational depth is limited by optical diffusion, which prevents focusing beyond shallow depths in scattering media, as well as reducing the measurement signal-to-noise ratio (SNR). To overcome this limitation, we used binary-amplitude wavefront shaping to enhance light focusing in the presence of scattering. Here, the transmission modes that contributed constructively to the intensity at the optical focus were identified and selectively illuminated, resulting in a 14-fold intensity increase and a corresponding increase in SNR. This technique can potentially extend the operational depth of optical-resolution PAF beyond 1 mm in tissue.

Time-reversed ultrasonically encoded (TRUE) optical focusing in turbid media was previously implemented using both analog and digital phase conjugation. The digital approach, in addition to its large energy gain, can improve the focal intensity and resolution by iterative focusing. However, performing iterative focusing at each focal position can be time-consuming. Here, we show that by gradually moving the focal position, the TRUE focal intensity is improved, as in iterative focusing at a fixed position, and can be continuously scanned to image fluorescent targets in a shorter time. In addition, our setup is, to the best of our knowledge, the first demonstration of TRUE focusing using a digital phase conjugate mirror in a reflection mode, which is more suitable for practical applications.

The effects of the juvenile hormone analog pyriproxyfen (at concentrations of 0.1, 0.5, and 1%) on egg production, number of ovarioles, and length of oocytes were examined in queens of the Pharaoh ant Monomorium pharaonis (L.). Pyriproxyfen significantly reduced egg production in queens from week 3 onwards. Queens that were exposed to 1% pyriproxyfen stopped producing eggs at week 8. After 8 wk, ovaries were dissected from all queens, and the number of ovarioles and the length of the largest oocytes were recorded. The ovaries of queens in treated colonies were smaller than those in untreated queens, and the number of ovarioles in the ovaries was significantly lower in all pyriproxyfen-treated queens. Queens treated with the highest concentrations of pyriproxyfen tended to have significantly shorter oocytes than untreated queens. Histological studies of the ovaries revealed that pyriproxyfen caused vacuolation in the ovarioles, thickening of the tunica propria, development of small eggs, and underdevelopment of nurse cells and the follicular epithelium. Exposure to pyriproxyfen reduced egg production and induced severe morphological changes in the ovaries of queens, and the effects increased with increased concentration of pyriproxyfen.

Phase distortions due to scattering in random media restrict optical focusing beyond one transport mean free path. However, scattering can be compensated for by applying a correction to the illumination wavefront using spatial light modulators. One method of obtaining the wavefront correction is by iterative determination using an optimization algorithm. In the past, obtaining a feedback signal required either direct optical access to the target region, or invasive embedding of molecular probes within the random media. Here, we propose using ultrasonically encoded light as feedback to guide the optimization dynamically and non-invasively. In our proof-of-principle demonstration, diffuse light was refocused to the ultrasound focal zone, with a focus-to-background ratio of more than one order of magnitude after 600 iterations. With further improvements, especially in optimization speed, the proposed method should find broad applications in deep tissue optical imaging and therapy.

This study focuses on the larvicidal, oviposition, and ovicidal effects of a crude extract of Artemisia annua against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus. Dried cells of Artemisia annua from cell suspension cultures were extracted using hexane. The extract showed moderate larvicidal effects against mosquitoes. At 24-h post treatment, the LC50 values for Anopheles sinensis, Aedes aegypti, and Culex quinquefasciatus were recorded as 244.55, 276.14, and 374.99 ppm, respectively. The percentage mortality of larvae was directly proportional to the tested concentration. Anopheles sinensis was found to be the most susceptible species, whereas Culex quinquefasciatus was the most tolerant to the Artemisia annua extract. The results indicated that the Artemisia annua extract showed concentration-dependent oviposition deterrent activity and had a strong deterrent effect. At 500 ppm, the percentage effective repellency was more than 85% compared with the control group for all the species, with oviposition activity index values of -0.94, -0.95, and -0.78 for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. In the ovicidal assay, the percentage hatchability of eggs after treatment with 500 ppm of Artemisia annua extract was significantly lower than the control, with values of 48.84 ± 4.08, 38.42 ± 3.67, and 79.35 ± 2.09% for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. Artemisia annua was found to be more effective against Aedes aegypti and Anopheles sinensis compared with Culex quinquefasciatus. This study indicated that crude extract of A. annua could be a potential alternative for use in vector management programs.

Background: Acquired protein S (PS) deficiency is highly associated with elevated circulating estrogen levels resulting from pregnancy, oral contraceptives, and estrogen replacement therapy; however, the mechanism of estrogen-mediated acquired PS deficiency remains poorly understood. Increasing evidence indicates that estrogen receptor signaling can indirectly modulate the expression of target genes at the post-transcriptional level by modulating the expression of microRNAs (miRNAs), and miRNAs have also been demonstrated to be involved in the regulation of hemostasis., Objectives: To investigate the mechanism of estrogen-mediated downregulation of PROS1 expression by the microRNA miR-494., Methods: Computational analyses of the PROS1 3'-untranslated region (UTR) were performed to identify putative miRNA-binding sites, and direct targeting of the PROS1 3'-UTR by miR-494 was determined with dual luciferase reporter assays in HuH-7 cells. Reporter vectors containing the PROS1 3'-UTR sequence with deleted miR-494-binding sites were also analyzed with luciferase reporter assays. The effects of estrogen on miR-494 and PROS1 mRNA levels in HuH-7 cells were determined by quantitative real-time PCR, and estrogen-mediated changes to secreted PS levels in culture supernatant of HuH-7 cells were measured with an ELISA., Results: The PROS1 3'-UTR sequence contains three putative miR-494-binding sites. miR-494 directly targets PROS1, and miR-494 levels are upregulated following estrogen treatment in HuH-7 liver cells in association with downregulated PROS1 mRNA and PS levels., Conclusions: The results from this study provide the first evidence for miRNA downregulation of PROS1 by miR-494, and suggest that miR-494 is involved in the mechanism of estrogen-mediated downregulation of PS expression., (© 2013 International Society on Thrombosis and Haemostasis.)

A 68 year old man with significant cardiorespiratory risks factors presented with a ruptured thoracic aortic aneurysm (TAA). This was treated by emergency thoracic endovascular aneurysm repair (TEVAR) under general anaesthesia (GA). An incidental abdominal aortic aneurysm (AAA) was not treated. Eight months later, he presented with ruptured AAA. Due to the patient's compromised respiratory system, he underwent endovascular aneurysm repair (EVAR) under local anaesthesia (LA). He had a smoother post-operation recovery compared to the first repair under GA.

We describe theoretical and experimental demonstration for optical detection of ultrasound using a spectral hole engraved in cryogenically cooled rare-earth ion-doped solids. Our method utilizes the dispersion effects due to the spectral hole to perform phase-to-amplitude modulation conversion. Like previous approaches using spectral holes, it has the advantage of detection with large étendue. The method also has the benefit that high sensitivity can be obtained with moderate absorption contrast for the spectral holes.

A method for characterizing the phase response of spatial light modulators (SLMs) by using a Sagnac interferometer is proposed and demonstrated. The method represents an improvement over conventional diffraction-based or interferometric techniques by providing a simple and accurate phase measurement while taking advantage of the inherent phase stability of a Sagnac interferometer. As a demonstration, the phase response of a commercial liquid crystal on a silicon SLM is characterized and then linearized by using a programmable lookup table. The transverse phase profile over the SLM surface is also measured.

The androgen receptor (AR) is the most widely expressed steroid hormone receptor in human breast cancers and androgens including 5alpha-dihydrotestosterone are potent inhibitors of breast cancer cell proliferation. The extracellular signal-regulated mitogen activated protein kinase (ERK/MAPK) pathway is hyperactivated in a proportion of breast tumors and can interact with steroid hormone receptor signaling by altering receptor phosphorylation, turnover, ligand, and cofactor interactions. To examine the effects of ERK/ MAPK hyperactivity on AR levels, MCF-7 cells were stably transfected with a plasmid encoding a constitutively active MEK1 protein to create MCF-7-DeltaMEK1 cells. Treatment of MCF-7-DeltaMEK1 with androgens caused a transient increase in AR protein levels, similar to that observed in untransfected MCF-7 cells treated with androgens. Androgens also inhibited the proliferation of MCF-7-DeltaMEK1 cells by 50-60% following 8 days of treatment in association with increased accumulation of cells in the G1 phase of the cell cycle. These results indicate that although ERK/MAPK hyperactivation in breast cancer cells is associated with reduced estrogen receptor (ERalpha) levels and antiestrogen resistance, AR levels are maintained and breast cancer cells remain susceptible to the growth inhibitory effects of androgens.