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Abstract Background Most methods for constructing aneuploid yeast strains that have gained a specific chromosome rely on spontaneous failures of cell division fidelity. In Saccharomyces cerevisiae, extra chromosomes can be obtained when errors in meiosis or mitosis lead to nondisjunction, or when nuclear breakdown occurs in heterokaryons. We describe a strategy for constructing N+1 disomes that does not require such spontaneous failures. The method combines two well-characterized genetic tools: a conditional centromere that transiently blocks disjunction of one specific chromosome, and a duplication marker assay that identifies disomes among daughter cells. To test the strategy, we targeted chromosomes III, IV, and VI for duplication. Results The centromere of each chromosome was replaced by a centromere that can be blocked by growth in galactose, and ura3::HIS3, a duplication marker. Transient exposure to galactose induced the appearance of colonies carrying duplicated markers for chromosomes III or IV, but not VI. Microarray-based comparative genomic hybridization (CGH) confirmed that disomic strains carrying extra chromosome III or IV were generated. Chromosome VI contains several genes that are known to be deleterious when overexpressed, including the beta-tubulin gene TUB2. To test whether a tubulin stoichiometry imbalance is necessary for the apparent lethality caused by an extra chromosome VI, we supplied the parent strain with extra copies of the alpha-tubulin gene TUB1, then induced nondisjunction. Galactose-dependent chromosome VI disomes were produced, as revealed by CGH. Some chromosome VI disomes also carried extra, unselected copies of additional chromosomes. Conclusion This method causes efficient nondisjunction of a targeted chromosome and allows resulting disomic cells to be identified and maintained. We used the method to test the role of tubulin imbalance in the apparent lethality of disomic chromosome VI. Our results indicate that a tubulin imbalance is necessary for disomic VI lethality, but it may not be the only dosage-dependent effect.

The presence of pharmaceuticals in the environment is a matter of great concern. They are consistently found in the environment, raising concerns regarding human exposure through dietary intake. In this study, we observed the effect of the application of carbamazepine at 0.1, 1, 10, and 1000 μg per kg of soil contamination levels to assess stress metabolism in Zea mays L. cv. Ronaldinio at the 4th leaf, tasselling, and dent phenological stages. The transfer of carbamazepine to the aboveground and root biomass was assessed, and uptake increased dose-dependently. No direct effect on biomass production was observed, but multiple physiological and chemical changes were observed. Major effects were consistently observed at the 4th leaf phenological stage for all contamination levels, including reduced photosynthetic rate, reduced maximal and potential activity of photosystem II, decreased water potential, decreased carbohydrates (glucose and fructose) and γ-aminobutyric acid in roots, and increased maleic acid and phenylpropanoids (chlorogenic acid and its isomer, 5-O-caffeoylquinic acid) in aboveground biomass. A reduction in net photosynthesis was observed for the older phenological stages, whereas no other relevant and consistent physiological and metabolic changes related to contamination exposure were detected. Our results indicate that Z. mays can overcome the environmental stress caused by the accumulation of carbamazepine with notable metabolic changes at the early phenological stage; however, older plants adapted and only exhibited minor effects in the presence of the contaminant. The potential implications for agricultural practice could be associated with the plant's response to simultaneous stresses due to metabolite changes associated with oxidative stress., 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., (Copyright © 2023. Published by Elsevier Ltd.)

Experimental spinach plants grown in soil with (5, 10 and 20 ppm) arsenic (As) contamination were sampled in 21 days after As(V) contamination. Levels of As in spinach samples (from 0.31 ± 0.06 µg g -1 to 302.69 ± 11.83 µg g -1 ) were higher in roots and lower in leaves, which indicates a low ability of spinach to translocate As into leaves. Species of arsenic, As(III) and As(V), were represented in favor of the As (III) specie in contaminated variants, suggesting enzymatic arsenate reduction. In relation to predominant As accumulation in roots, changes in malondialdehyde levels were observed mainly in roots, where they decreased significantly with growing As contamination (from 11.97 ± 0.54 µg g -1 in control to 2.35 ± 0.43 µg g -1 in 20 ppm As). Higher values in roots than in leaves were observed in the case of 5-methylcytosine (5-mC). Despite that, a change in 5-mC by As contamination was further deepened in leaves (from 0.20 to 14.10%). In roots of spinach, expression of the CDC25 gene increased by the highest As contamination compared to the control. In the case of total phenolic content, total flavonoid content, total phenolic acids content and total antioxidant capacity were higher levels in leaves in all values, unlike the roots.

Arsenic represents a serious health threat in localities with a high arsenic-polluted environment and can easily get into the human food chain through agronomy production in areas affected by arsenic contamination. Onion plants that were grown in controlled conditions in arsenic-contaminated soil (5, 10, and 20 ppm) were harvested 21 days after contamination. Arsenic levels (from 0.43 ± 0.03 µg g -1 to 1761.11 ± 101.84 µg g -1 ) in the onion samples were high in the roots and low in the bulbs and leaves, which is probably caused by a reduced ability of the onions to transport arsenic from roots to bulbs and leaves. Arsenic species As(V) and As(III) in As(V)-contaminated soil samples were represented strongly in favor of the As(III) species. This indicates the presence of arsenate reductase. Levels of 5-methylcytosine (5-mC) (from 5.41 ± 0.28% to 21.17 ± 1.33%) in the onion samples were also higher in the roots than in the bulbs and leaves. Microscopic sections of the roots were examined, and the most damage was found in the 10 ppm As variant. Photosynthetic parameters pointed to a significant decrease in photosynthetic apparatus activity and the deterioration of the physiological state of plants as arsenic content increased in the soil.

Point-of-care antigen tests are an important tool for SARS-CoV-2 detection. Antigen tests are less sensitive than real-time reverse transcriptase PCR (rRT-PCR). Data on the performance of the BinaxNOW antigen test compared to rRT-PCR and viral culture by symptom and known exposure status, timing during disease, or exposure period and demographic variables are limited. During 3 to 17 November 2020, we collected paired upper respiratory swab specimens to test for SARS-CoV-2 by rRT-PCR and Abbott BinaxNOW antigen test at two community testing sites in Pima County, Arizona. We administered a questionnaire to capture symptoms, known exposure status, and previous SARS-CoV-2 test results. Specimens positive by either test were analyzed by viral culture. Previously we showed overall BinaxNOW sensitivity was 52.5%. Here, we showed BinaxNOW sensitivity increased to 65.7% among currently symptomatic individuals reporting a known exposure. BinaxNOW sensitivity was lower among participants with a known exposure and previously symptomatic (32.4%) or never symptomatic (47.1%) within 14 days of testing. Sensitivity was 71.1% in participants within a week of symptom onset. In participants with a known exposure, sensitivity was highest 8 to 10 days postexposure (75%). The positive predictive value for recovery of virus in cell culture was 56.7% for BinaxNOW-positive and 35.4% for rRT-PCR-positive specimens. Result reporting time was 2.5 h for BinaxNOW and 26 h for rRT-PCR. Point-of-care antigen tests have a shorter turnaround time than laboratory-based nucleic acid amplification tests, which allows for more rapid identification of infected individuals. Antigen test sensitivity limitations are important to consider when developing a testing program.

Rapid antigen tests, such as the Abbott BinaxNOW COVID-19 Ag Card (BinaxNOW), offer results more rapidly (approximately 15-30 minutes) and at a lower cost than do highly sensitive nucleic acid amplification tests (NAATs) (1). Rapid antigen tests have received Food and Drug Administration (FDA) Emergency Use Authorization (EUA) for use in symptomatic persons (2), but data are lacking on test performance in asymptomatic persons to inform expanded screening testing to rapidly identify and isolate infected persons (3). To evaluate the performance of the BinaxNOW rapid antigen test, it was used along with real-time reverse transcription-polymerase chain reaction (RT-PCR) testing to analyze 3,419 paired specimens collected from persons aged ≥10 years at two community testing sites in Pima County, Arizona, during November 3-17, 2020. Viral culture was performed on 274 of 303 residual real-time RT-PCR specimens with positive results by either test (29 were not available for culture). Compared with real-time RT-PCR testing, the BinaxNOW antigen test had a sensitivity of 64.2% for specimens from symptomatic persons and 35.8% for specimens from asymptomatic persons, with near 100% specificity in specimens from both groups. Virus was cultured from 96 of 274 (35.0%) specimens, including 85 (57.8%) of 147 with concordant antigen and real-time RT-PCR positive results, 11 (8.9%) of 124 with false-negative antigen test results, and none of three with false-positive antigen test results. Among specimens positive for viral culture, sensitivity was 92.6% for symptomatic and 78.6% for asymptomatic individuals. When the pretest probability for receiving positive test results for SARS-CoV-2 is elevated (e.g., in symptomatic persons or in persons with a known COVID-19 exposure), a negative antigen test result should be confirmed by NAAT (1). Despite a lower sensitivity to detect infection, rapid antigen tests can be an important tool for screening because of their quick turnaround time, lower costs and resource needs, high specificity, and high positive predictive value (PPV) in settings of high pretest probability. The faster turnaround time of the antigen test can help limit transmission by more rapidly identifying infectious persons for isolation, particularly when used as a component of serial testing strategies., Competing Interests: All authors have completed and submitted the International Committee of Medical Journal Editors form for disclosure of potential conflicts of interest. No potential conflicts of interest were disclosed.

This study analyzes the effects of acetaminophen (APAP) as a contaminant on physiological characteristics of lettuce plants ( Lactuca sativa L.). Experiments were provided in an experimental greenhouse with semi-controlled conditions. The effect of different amounts of contaminant was evaluated by using regression analysis. Plants were grown in five concentrations of APAP: 0 µM, 5 µM, 50 µM, 500 µM, and 5 mM for 14 days in two variants, acute and chronic. The obtained results show that the monitored parameters were demonstrably influenced by the experimental variant. Plants are more sensitive to chronic contamination compared to acute. Significant ( p < 0.05) deviation in photosynthesis and fluorescence was observed compared to the control in different variants. The highest doses of APAP reduced the intensity of photosynthesis by a maximum of more than 31% compared to the control. A reduction of 18% was observed for the fluorescence parameters. Pronounced correlation was described between chlorophyll fluorescence parameters and yield mainly under APAP conditions. The amount of chlorophyll was influenced by exposure to APAP.

Purpose: The purpose of this study was to assess the rate of postoperative deep vein thrombosis (DVT) as a function of oral anticoagulation therapy after total hip replacement surgery., Methods: A total of 125 patients completed the study. All the patients received sequential gradient pneumatic compression over elastic stockings until hospital discharge. In addition, all the patients underwent postoperative heparin therapy followed by oral warfarin therapy, adjusted in dose to maintain a goal international normalized ratio (INR) level of 2.0 to 3.0. Warfarin therapy and compression stockings were continued for 1 month after surgery. Bilateral duplex scanning was performed 1 and 4 weeks after surgery to assess the rate of DVT., Results: Nineteen of the 125 patients had DVT develop (15.2%). Of those thromboses, six (31.6%) and 13 (68%) were detected 1 week and 1 month after surgery, respectively. The rate of proximal DVT was 2.4% (3 of 125) 1 week after surgery and rose to 8.2% (10 of 122) 1 month after surgery. Most DVT cases (64%; 12 of 19) were asymptomatic. The patients in whom DVT developed had significantly lower INR values during the second to fourth postoperative weeks than did those patients without thrombosis, and no differences in INR values were found during the first postoperative week., Conclusion: The risk of the development of DVT extends beyond hospital discharge in patients who undergo total hip replacement, despite a regimen of prolonged oral anticoagulation therapy. This is particularly true in patients whose INR values did not reach therapeutic range during the first postoperative month. Therefore, thrombosis prophylaxis regimens on the basis of the administration of warfarin should try to maintain INR values within therapeutic range during the entire first postoperative month to minimize the incidence of DVT.