Development of an Immunocapture-Based Polymeric Optical Fiber Sensor for Bacterial Detection in Water.

Conventional methods for pathogen detection in water rely on time-consuming enrichment steps followed by biochemical identification strategies, which require assay times ranging from 24 hours to a week. However, in recent years, significant efforts have been made to develop biosensing technologies enabling rapid and close-to-real-time detection of waterborne pathogens. In previous studies, we developed a plastic optical fiber (POF) immunosensor using an optoelectronic configuration consisting of a U-Shape probe connected to an LED and a photodetector. Bacterial detection was evaluated with the immunosensor immersed in a bacterial suspension in water with a known concentration. Here, we report on the sensitivity of a new optoelectronic configuration consisting of two POF U-shaped probes, one as the reference and the other as the immunosensor, for the detection of Escherichia coli. In addition, another methos of detection was tested where the sensors were calibrated in the air, before being immersed in a bacterial suspension and then read in the air. This modification improved sensor sensitivity and resulted in a faster detection time. After the immunocapture, the sensors were DAPI-stained and submitted to confocal microscopy. The histograms obtained confirmed that the responses of the immunosensors were due to the bacteria. This new sensor detected the presence of E. coli at 104 CFU/mL in less than 20 min. Currently, sub-20 min is faster than previous studies using fiber-optic based biosensors. We report on an inexpensive and faster detection technology when compared with conventional methods.

The role of arbuscular mycorrhizal fungi in the alleviation of cadmium stress in cereals: A multilevel meta-analysis.

The symbiotic relationships between crop species and arbuscular mycorrhizal fungi (AMF) are crucial for plant health, productivity, and environmental sustainability. The roles of AMF in reducing crop stress caused by cadmium (Cd) toxicity and in the remediation of Cd-contaminated soil are not fully understood. Here we report on a meta-analysis that sought to identify the functions of AMF in cereals under Cd stress. A total of 54 articles published between January 1992 and September 2022 were used to create the dataset, which provided 7216 data sets on mycorrhizal cereals under Cd stress examined. AMF effects on colonization rate, biomass, physiological level, nutritional level, and plant Cd level were measured using the logarithmic response ratio (Ln R). The results showed that AMF overall greatly reduced 5.14 - 33.6 % Cd stress on cereals in greenhouse experiments under controlled conditions. AMF colonization significantly stimulated crop biomass by 65.7 %, boosted the formation of photosynthetic pigments (23.2 %), and greatly increased plant nitrogen (24.8 %) and phosphorus (58.4 %) uptake. The dilution effect of mycorrhizal plants made the Cd concentration decline by 25.2 % in AMF plants compared to non-mycorrhizal ones. AMF also alleviated Cd stress by improving osmotic regulators (soluble protein, sugar, and total proline, from 14.8 to 36.0 %) and lowering the membrane lipid peroxidation product (MDA, 12.9 %). Importantly, the results from the random forest and model selection analysis demonstrated that crop type, soil characteristics, chemical form, and Cd levels were the main factors determining the function of AMF in alleviating Cd stress. Additionally, there was a significant interaction between AMF colonization rate and Cd addition, but their interactive effect was less than the colonization rate alone. This meta-analysis demonstrated that AMF inoculation could be considered as a promising strategy for mitigation of Cd stress in cereals.

The Potential of Allelochemicals from Microalgae for Biopesticides.

Improvements in agricultural productivity are required to meet the demand of a growing world population. Phytopathogens, weeds, and insects are challenges to agricultural production. The toxicity and widespread application of persistent synthetic pesticides poses a major threat to human and ecosystem health. Therefore, sustainable strategies to control pests are essential for agricultural systems to enhance productivity within a green paradigm. Allelochemicals are a less persistent, safer, and friendly alternative to efficient pest management, as they tend to be less toxic to non-target organisms and more easily degradable. Microalgae produce a great variety of allelopathic substances whose biocontrol potential against weeds, insects, and phytopathogenic fungi and bacteria has received much attention. This review provides up-to-date information and a critical perspective on allelochemicals from microalgae and their potential as biopesticides.

Arbuscular Mycorrhizal Fungi Alleviate Low Phosphorus Stress in Maize Genotypes with Contrasting Root Systems.

Soil available phosphorus (P) is one of the main factors limiting plant growth and yield. This study aimed to determine the role of arbuscular mycorrhizal fungi (AMF) in P-use efficiency in two maize genotypes with contrasting root systems in response to low P stress. Maize genotypes small-rooted Shengrui 999 and large-rooted Zhongke 11 were grown in rhizoboxes that were inoculated with or without AMF (Funneliformis mosseae) under low P (no added P) or optimal P (200 mg kg-1) for 53 days. Low P stress significantly inhibited shoot and root growth, photosynthesis, tissue P content, and root P concentration in both genotypes. Shengrui 999 was more tolerant to P stress with less reduction of these traits compared to Zhongke 11. Shengrui 999 had a higher AMF infection rate than Zhongke 11 at both P levels. Under P deficit, inoculation with AMF significantly promoted plant growth and P uptake in both genotypes with more profound effects seen in Zhongke 11, whilst Shengrui 999 was more dependent on AMF under optimal P. Low P stress inhibited the growth and physiological attributes of both genotypes. The small-rooted Shengrui 999 was more tolerant to low P than Zhongke 11. Inoculation with AMF alleviates low P stress in both genotypes with a more profound effect on Zhongke 11 at low P and on Shengrui 999 at high P conditions.

Aeromonas allosaccharophila Strain AE59-TE2 Is Highly Antagonistic towards Multidrug-Resistant Human Pathogens, What Does Its Genome Tell Us?

Multidrug-resistant bacteria are of critical importance and a problem for human health and food preservation; the discovery of new antimicrobial substances to control their proliferation is part of the solution. This work reports on 57 antagonistic Aeromonas strains, of which 38 strains were antagonistic towards problematic human pathogens. The genome of the most antagonistic strain was sequenced and identified as Aeromonas allosaccharophila. Its genome was fully annotated and mined for genes that might explain that activity. Strain AE59-TE was antagonistic toward clinically relevant gram-negative and gram-positive multidrug-resistant bacteria, including Klebsiella pneumoniae KPC, Escherichia coli ESBL, Salmonella typhimurium, and Staphylococcus aureus MRSA. Strain AE59-TE2 was identified by multilocus sequence analysis. Genome mining identified four genes homologous to the bacteriocin, zoocin A from Streptococcus equi and a gene 98% similar to cvpA linked to colicin V production. A. allosaccharophila strain AE59-TE2 produced antimicrobial activity against a broad range of bacteria, including important gram-negative bacteria, not typically targeted by bacteriocins. Herewere described novel zoocin genes that are promising for industrial applications in the food and health sectors. Interesting and important antagonistic activity is described combined with the first detailed genomic analysis of the species Aeromonas allosaccharophila.

The Genome Sequence of Brucella intermedia DF13, a 2,4-Dichlorophenoxyacetic Acid-Degrading Soil Bacterium Isolated in Brazil.

Brucella intermedia/Ochrobactrum intermedium strain DF13 was isolated from Brazilian soil and is able to degrade 2,4-dichlorophenoxyacetic acid (2,4-D). Here, we report on its genome sequence, with 4,570,268 bp and a 57.8% G+C content.

The Genome of Enterobacter hormaechei Strain MG02, a 2,4-Dichlorophenoxyacetic Acid-Degrading Bacterium Isolated from Brazilian Soil.

Enterobacter hormaechei strain MG02 was isolated from a mixed culture collected from soil with a history of pesticide application. This strain degrades 2,4-dichlorophenoxyacetic acid (2,4-D). Here, we report on its genome, which has 4,923,875 bp and 55.4% G+C content.

The Effects of Coronavirus Disease 2019 on Pediatric Anesthesiologists: A Survey of the Members of the Society for Pediatric Anesthesia.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has affected the personal and professional lives of all health care workers. Anesthesiologists frequently perform virus-aerosolizing procedures (eg, intubation and extubation) that place them at increased risk of infection. We sought to determine how the initial COVID-19 outbreak affected members of the Society for Pediatric Anesthesia (SPA) on both personal and professional levels. Specifically, we examined the potential effects of gender and age on personal stress, burnout, sleep deprivation, anxiety, depression, assessed job satisfaction, and explored financial impact. METHODS: After receiving approval from the SPA Committees for Research and Quality and Safety and the Colorado Multiple Institutional Review Board, we e-mailed a questionnaire to all 3245 SPA members. The survey included 22 questions related to well-being and 13 questions related to effects of COVID-19 on current and future practice, finances, retirement planning, academic time and productivity, and clinical and home responsibilities. To address low initial response rates and quantify nonresponse bias, we sent a shortened follow-up survey to a randomly selected subsample (n = 100) of SPA members who did not respond to the initial survey. Response differences between the 2 cohorts were determined. RESULTS: A total of 561 (17%) members responded to the initial questionnaire. Because of COVID-19, 21.7% of respondents said they would change their clinical responsibilities, and 10.6% would decrease their professional working time. Women were more likely than men to anticipate a future COVID-19-related job change (odds ratio [OR] = 1.92, 95% confidence interval [CI], 1.12-2.63; P = .011), perhaps because of increased home responsibilities (OR = 2.63, 95% CI, 1.74-4.00; P < .001). Additionally, 14.2% of respondents planned to retire early, and 11.9% planned to retire later. Women and non-White respondents had higher likelihoods of burnout on univariate analysis (OR = 1.75, 95% CI, 1.06-2.94, P = .026 and OR = 1.82, 95% CI, 1.08-3.04, P = .017, respectively), and 25.1% of all respondents felt socially isolated. In addition, both changes in retirement planning and future occupational planning were strongly associated with total job satisfaction scores (both P < .001). CONCLUSIONS: The COVID-19 pandemic has affected the personal and professional lives of pediatric anesthesiologists, albeit not equally, as women and non-Whites have been disproportionately impacted. The pandemic has significantly affected personal finances, home responsibilities, and retirement planning; reduced clinical and academic practice time and responsibilities; and increased feelings of social isolation, stress, burnout, and depression/anxiety.

Development of Microalgae Biodiesel: Current Status and Perspectives.

Microalgae are regarded as a promising source of biodiesel. In contrast with conventional crops currently used to produce commercial biodiesel, microalgae can be cultivated on non-arable land, besides having a higher growth rate and productivity. However, microalgal biodiesel is not yet regarded as economically competitive, compared to fossil fuels and crop-based biodiesel; therefore, it is not commercially produced. This review provides an overall perspective on technologies with the potential to increase efficiency and reduce the general costs of biodiesel production from microalgae. Opportunities and challenges for large-scale production are discussed. We present the current scenario of Brazilian research in the field and show a successful case in the research and development of microalgal biodiesel in open ponds by Petrobras. This publicly held Brazilian corporation has been investing in research in this sector for over a decade.

Genome Sequence of Pseudomonas sp. Strain LAP_36, A Rhizosphere Bacterium Isolated from King George Island, Antarctica.

Pseudomonas sp. strain LAP_36 was isolated from rhizosphere soil from Deschampsia antarctica on King George Island, South Shetland Islands, Antarctica. Here, we report on its draft genome sequence, which consists of 8,794,771 bp with 60.0% GC content and 8,011 protein-coding genes.

Sulphate-reducing bacterial community structure from produced water of the Periquito and Galo de Campina onshore oilfields in Brazil.

Sulphate-reducing bacteria (SRB) cause fouling, souring, corrosion and produce H2S during oil and gas production. Produced water obtained from Periquito (PQO) and Galo de Campina (GC) onshore oilfields in Brazil was investigated for SRB. Produced water with Postgate B, Postgate C and Baars media was incubated anaerobically for 20 days. DNA was extracted, 16S rDNA PCR amplified and fragments were sequenced using Illumina TruSeq. 4.2 million sequence reads were analysed and deposited at NCBI SAR accession number SRP149784. No significant differences in microbial community composition could be attributed to the different media but significant differences in the SRB were observed between the two oil fields. The dominant bacterial orders detected from both oilfields were Desulfovibrionales, Pseudomonadales and Enterobacteriales. The genus Pseudomonas was found predominantly in the GC oilfield and Pleomorphominas and Shewanella were features of the PQO oilfield. 11% and 7.6% of the sequences at GC and PQO were not classified at the genus level but could be partially identified at the order level. Relative abundances changed for Desulfovibrio from 29.8% at PQO to 16.1% at GC. Clostridium varied from 2.8% at PQO and 2.4% at GC. These data provide the first description of SRB from onshore produced water in Brazil and reinforce the importance of Desulfovibrionales, Pseudomonadales, and Enterobacteriales in produced water globally. Identifying potentially harmful microbes is an important first step in developing microbial solutions that prevent their proliferation.

Coral microbiome manipulation elicits metabolic and genetic restructuring to mitigate heat stress and evade mortality.

Beneficial microorganisms for corals (BMCs) ameliorate environmental stress, but whether they can prevent mortality and the underlying host response mechanisms remains elusive. Here, we conducted omics analyses on the coral Mussismilia hispida exposed to bleaching conditions in a long-term mesocosm experiment and inoculated with a selected BMC consortium or a saline solution placebo. All corals were affected by heat stress, but the observed "post-heat stress disorder" was mitigated by BMCs, signified by patterns of dimethylsulfoniopropionate degradation, lipid maintenance, and coral host transcriptional reprogramming of cellular restructuration, repair, stress protection, and immune genes, concomitant with a 40% survival rate increase and stable photosynthetic performance by the endosymbiotic algae. This study provides insights into the responses that underlie probiotic host manipulation. We demonstrate that BMCs trigger a dynamic microbiome restructuring process that instigates genetic and metabolic alterations in the coral host that eventually mitigate coral bleaching and mortality.

Decolorization and detoxification of different azo dyes by Phanerochaete chrysosporium ME-446 under submerged fermentation.

Azo dyes are widely used in the textile industry due to their resistance to light, moisture, and oxidants. They are also an important class of environmental contaminant because of the amount of dye that reaches natural water resources and because they can be toxic, mutagenic, and carcinogenic. Different technologies are used for the decolorization of wastewater containing dyes; among them, the biological processes are the most promising environmentally. The aim of this study was to evaluate the potential of Phanerochaete chrysosporium strain ME-446 to safely decolorize three azo dyes: Direct Yellow 27 (DY27), Reactive Black 5 (RB5), and Reactive Red 120 (RR120). Decolorization efficiency was determined by ultraviolet-visible spectrophotometry and the phytotoxicity of the solutions before and after the fungal treatment was analyzed using Lactuca sativa seeds. P. chrysosporium ME-446 was highly efficient in decolorizing DY27, RB5, and RR120 at 50 mg L-1, decreasing their colors by 82%, 89%, and 94% within 10 days. Removal of dyes was achieved through adsorption on the fungal mycelium as well as biodegradation, inferred by the changes in the dyes' spectral peaks. The intensive decolorization of DY27 and RB5 corresponded to a decrease in phytotoxicity. However, phytotoxicity increased during the removal of color for the dye RR120. The ecotoxicity tests showed that the absence of color does not necessarily translate to an absence of toxicity.

A synthetic small molecule stalls pre-mRNA splicing by promoting an early-stage U2AF2-RNA complex.

Dysregulated pre-mRNA splicing is an emerging Achilles heel of cancers and myelodysplasias. To expand the currently limited portfolio of small-molecule drug leads, we screened for chemical modulators of the U2AF complex, which nucleates spliceosome assembly and is mutated in myelodysplasias. A hit compound specifically enhances RNA binding by a U2AF2 subunit. Remarkably, the compound inhibits splicing of representative substrates and stalls spliceosome assembly at the stage of U2AF function. Computational docking, together with structure-guided mutagenesis, indicates that the compound bridges the tandem U2AF2 RNA recognition motifs via hydrophobic and electrostatic moieties. Cells expressing a cancer-associated U2AF1 mutant are preferentially killed by treatment with the compound. Altogether, our results highlight the potential of trapping early spliceosome assembly as an effective pharmacological means to manipulate pre-mRNA splicing. By extension, we suggest that stabilizing assembly intermediates may offer a useful approach for small-molecule inhibition of macromolecular machines.

A Survey of the Society for Pediatric Anesthesia on the Use, Monitoring, and Antagonism of Neuromuscular Blockade.

BACKGROUND: Although the package insert clearly states that "the safety and efficacy of sugammadex in pediatric patients have not been established," we hypothesized that sugammadex is used widely in pediatric anesthetic practice supplanting neostigmine as the primary drug for antagonizing neuromuscular blockade (NMB). Additionally, we sought to identify the determinants by which pediatric anesthesiologists choose reversal agents and if and how they assess NMB in their practice. Finally, because of sugammadex's effects on hormonal contraception, we sought to determine whether pediatric anesthesiologists counseled postmenarchal patients on the need for additional or alternative forms of contraception and the risk of unintended pregnancy in the perioperative period. METHODS: We e-mailed a questionnaire to all 3245 members of the Society of Pediatric Anesthesia (SPA) requesting demographic data and attitudes regarding use of NMB agents, monitoring, and antagonism practices. To address low initial response rates and quantify nonresponse bias, we sent a shortened follow-up survey to a randomly selected subsample (n = 75) of SPA members who did not initially respond. Response differences between the 2 cohorts were determined. RESULTS: Initial questionnaire response rate was 13% (419 of 3245). Overall, 163 respondents (38.9%; 95% confidence interval [CI], 34.2-43.8) used sugammadex as their primary reversal agent, and 106 (25.2%; 95% CI, 21.2-30.0) used it exclusively. Respondents with ≤5 years of practice used sugammadex as their primary reversal agent more often than those with ≥6 years of practice (odds ratio [OR]: 2.08; 95% CI, 1.31-3.31; P = .001). This increased utilization remained after controlling for institutional restriction and practice type (adjusted OR [aOR]: 2.20; 95% CI, 1.38-3.54; P = .001). Only 40% of practitioners always assess NMB (train-of-four), and use was inversely correlated with years of practice (Spearman ρ = -0.11, P = .04). Anesthesiologists who primarily used sugammadex assess NMB less routinely (OR: 0.56; 95% CI, 0.34-0.90; P = .01). A slim majority (52.8%) used sugammadex for pediatric postmenarchal girls; those with less experience used it more commonly (P < .001). Thirty-eight percent did not discuss its effects on hormonal contraception with the patient and/or family, independent of anesthesiologist experience (P = .33) and practice location (P = .38). No significant differences were seen in demographics or practice responses between initial and follow-up survey respondents. CONCLUSIONS: Sugammadex is commonly used in pediatric anesthesia, particularly among anesthesiologists with fewer years of practice. Failure to warn postmenarchal adolescents of its consequences may result in unintended pregnancies. Finally, pediatric anesthesia training programs should emphasize objective monitoring of NMB, particularly with sugammadex use.

Physiological and molecular responses of Setaria viridis to osmotic stress.

Drought-tolerant species, such as Setaria viridis, a C4 model plant, make physiological and biochemical adjustments water limitation and recover from the stress upon its release. We investigated S. viridis (A10.1 accession) responses to continuing osmotic stress. The osmotic stress was imposed using polyethylene glycol (PEG) 8000 (7.5%) for 10 days. Morphological traits and stomatal conductance were measured daily for the 10 days. On days 6 and 10, the following traits were measured separately for root and shoot: relative water content (RWC), osmotic potential (OP), electrolytic leakage (EL), and proline content. qPCR analysis was used to evaluate the expression of five selected genes in roots (SvLEA, SvDREB1C, SvPIP2-1, SvHSP20, and SvP5CS2), and chlorophyll a fluorescence was measured on three key days. The morphological data demonstrated a drastic reduction in shoot biomass as an effect of water deficit caused by the osmotic stress. Shoot biomass reduction could be associated with putative ABA-dependent signaling involved in SvDREB1C expression. Stomatal conductance and photosynthesis were severely affected up until day 6, however, stomatal conductance and some photosynthetic parameters such as FV/FM, ABS/RC, and DI0/RC showed total or slight recovery on day 10. Root EL decreased in treated plants suggesting an investment in membrane protection by osmoregulator expression such as dehydrin (SvLEA) and proline (SvP5CS2) genes. Our data suggest that S. viridis exhibited a partial recovery from an imposed and constant osmotic stress within 10 days.

A two-step probing method to compare lysine accessibility across macromolecular complex conformations.

Structural models of large and dynamic molecular complexes are appearing in increasing numbers, in large part because of recent technical advances in cryo-electron microscopy. However, the inherent complexity of such biological assemblies comprising dozens of moving parts often limits the resolution of structural models and leaves the puzzle as to how each functional configuration transitions to the next. Orthogonal biochemical information is crucial to understanding the molecular interactions that drive those rearrangements. We present a two-step method for chemical probing detected by tandem mass-spectrometry to globally assess the reactivity of lysine residues within purified macromolecular complexes. Because lysine side chains often balance the negative charge of RNA in ribonucleoprotein complexes, the method is especially useful for detecting changes in protein-RNA interactions. By probing the E. coli 30S ribosome subunit, we established that the reactivity pattern of lysine residues quantitatively reflects structure models derived from X-ray crystallography. We also used the strategy to assess differences in three conformations of purified human spliceosomes in the context of recent cryo-electron microscopy models. Our results demonstrate that the probing method yields powerful biochemical information that helps contextualize architectural rearrangements of intermediate resolution structures of macromolecular complexes, often solved in multiple conformations.

Fungal communities in oil contaminated mangrove sediments - Who is in the mud?

Mangroves are ecosystems located in tropical and subtropical regions of the world and are vital for coastal protection. Their unique characteristics make them hotspots for carbon cycling and biological diversity. Studies on isolated filamentous fungi and environmental and anthropogenic factors that influence sediments offer new understandings on how to preserve mangroves. Here we report on the filamentous fungi isolated from four mangroves. We correlated fungal community composition with sediment texture, polycyclic aromatic hydrocarbons concentration (oil pollution), pH, salinity, organic matter, total and thermotolerant coliforms (sewage pollution). In total we identified 34 genera and 97 species. The most polluted sites had highest species richness whereas the best preserved site showed the lowest species richness. Oil spill and sewage pollution were identified as the drivers of fungal community composition in the most polluted sites. We found very distinct fungal communities with no >5 species shared between any two mangrove sites.

Enantioselective Synthesis of Thailanstatin A Methyl Ester and Evaluation of in Vitro Splicing Inhibition.

Thailanstatin A has been isolated recently from the fermentation broth of B. thailandensis MSMB43. We describe here an enantioselective convergent synthesis of thailanstatin A methyl ester and evaluation of its splicing activity. Synthesis of both highly functionalized tetrahydropyran rings were carried out from commercially available tri- O-acetyl-d-glucal as the key starting material. Our convergent synthesis involved the synthesis of both tetrahydropyran fragments in a highly stereoselective manner. The fragments were then coupled using cross-metathesis as the key step. The synthesis of the diene subunit included a highly stereoselective Claisen rearrangement, a Cu(I)-mediated conjugate addition of MeLi to set the C-14 methyl stereochemistry, a reductive amination reaction to install the C16-amine functionality, and a Wittig olefination reaction to incorporate the diene unit. The epoxy alcohol subunit was synthesized by a highly selective anomeric allylation, a Peterson olefination, and a vanadium catalyzed epoxidation that installed the epoxide stereoselectively. Cross-metathesis of the olefins provided the methyl ester derivative of thailanstatin A. We have carried out in vitro splicing studies of the methyl ester derivative, which proved to be a potent inhibitor of the spliceosome.

Prp8 positioning of U5 snRNA is linked to 5' splice site recognition.

Prp8 is an essential protein that regulates spliceosome assembly and conformation during pre-mRNA splicing. Recent cryo-EM structures of the spliceosome model Prp8 as a scaffold for the spliceosome's catalytic U snRNA components. Using a new amino acid probing strategy, we identified a dynamic region in human Prp8 that is positioned to stabilize the pre-mRNA in the spliceosome active site through interactions with U5 snRNA. Mutagenesis of the identified Prp8 residues in yeast indicates a role in 5' splice site recognition. Genetic interactions with spliceosome proteins Isy1, which buttresses the intron branch point, and Snu114, a regulatory GTPase that directly contacts Prp8, further corroborate a role for the same Prp8 residues in substrate positioning and activation. Together the data suggest that adjustments in interactions between Prp8 and U5 snRNA help establish proper positioning of the pre-mRNA into the active site to enhance 5' splice site fidelity.