Prime editing (PE) is a powerful genome engineering approach that enables the introduction of base substitutions, insertions and deletions into any given genomic locus. However, the efficiency of PE ...varies widely and depends not only on the genomic region targeted, but also on the genetic background of the edited cell. Here, to determine which cellular factors affect PE efficiency, we carry out a focused genetic screen targeting 32 DNA repair factors, spanning all reported repair pathways. We show that, depending on cell line and type of edit, ablation of mismatch repair (MMR) affords a 2-17 fold increase in PE efficiency, across several human cell lines, types of edits and genomic loci. The accumulation of the key MMR factors MLH1 and MSH2 at PE sites argues for direct involvement of MMR in PE control. Our results shed new light on the mechanism of PE and suggest how its efficiency might be optimised.
Landfill leachates are high-strength complex mixtures containing dissolved organic matter, ammonia, heavy metals, and sulfur species, among others. The problem of leachate treatment has subsisted for ...some time, but an efficient and cost-effective universal solution capable of ensuring environmental resources protection has not been found. Aerobic granular sludge (AGS) has been considered a promising technology for biological wastewater treatment in recent years. Granules’ layered structure, with an aerobic outer layer and an anaerobic/anoxic core, enables the presence of diverse microbial populations without the need for support media, allowing simultaneous removal of different pollutants in a single unit. Besides, its strong and compact arrangement provides higher tolerance to toxic pollutants and the ability to withstand large load fluctuations. Furthermore, its good that settling properties allow high biomass retention and better sludge separation. Nevertheless, AGS-related research has focused on carbon-nitrogen-phosphorus removal, mainly from sanitary sewage. This review aims to summarize and analyze the main findings and problems reported in the literature regarding AGS application to landfill leachate treatment and identify the knowledge gaps for future applications.
The present paper describes fundamentals, advantages and limitations of the Box-Behnken design (BBD) for the optimization of analytical methods. It establishes also a comparison between this design ...and composite central, three-level full factorial and Doehlert designs. A detailed study on factors and responses involved during the optimization of analytical systems is also presented. Functions developed for calculation of multiple responses are discussed, including the desirability function
, which was proposed by Derringer and Suich in 1980. Concept and evaluation of robustness of analytical methods are also discussed. Finally, descriptions of applications of this technique for optimization of analytical methods are presented.
To reduce dependence on wood and deforestation, alternative materials have been considered. This research evaluated particleboards panels of Pinus elliotti mixed with residues of polyethylene ...terephthalate (PET) in flakes with 50% of each material, glued with 10% of castor oil-based polyurethane resin (PUR). The temperature during board production was varied to evaluate its influence on the physical and mechanical properties of the boards. The study concludes that the increase in temperature did not result in a significant improvement in the particleboard properties, but their results showed application potential according to the normative standards.
Healthcare-related infections caused by resistant microorganisms are a severe public health problem and are becoming increasingly prevalent in the hospital environment, especially Pseudomonas ...aeruginosa. This work aimed to evaluate the resistance profile of Pseudomonas aeruginosa to antimicrobials before the COVID-19 pandemic and during the pandemic period. Bacteria strains were obtained from tracheal aspiration, sputum, and bronchoalveolar lavage for diagnosis and phenotypic characterization. Matrix assisted laser-desorption ionization-time of flight mass spectrometry (MALD-TOF MS) was used to identify strains. Automated Phoenix and VITEK® 2 Compact system and the disc diffusion method were performed to determine the antimicrobial susceptibility profile. A total of 41,000 medical reports from adult patients with pneumonia were analyzed. Of these, 951 patients were positive for P. aeruginosa, of which 373 were related to the pre-pandemic period and 578 to the pandemic period. Older men (≥60 years) were more prevalent in both periods. P. aeruginosa strains were resistant to imipenem in both periods: 38.8 and 42.5%, respectively, followed by meropenem (34.2 and 39.2%), ciprofloxacin (33.6 and 36.7%), and levofloxacin (34.9 and 43.5%). Intensive care units had the highest percentage of affected patients (62 and 65%) compared with other sectors, with a prevalence of 71% in the public network before COVID-19 and 59% during the pandemic. Our data showed a prevalence of P. aeruginosa in elderly patients in both the pre-pandemic and pandemic periods. In addition, an increase in P. aeruginosa resistance to beta-lactams, quinolones, carbapenems, and cephalosporins was observed during the COVID-19 pandemic compared with the period before the pandemic, especially in ICUs.
Peperomia serpens (Piperaceae), popularly known as “carrapatinho”, is an epiphyte herbaceous liana grown wild on different host trees in the Amazon rainforest. Its leaves are largely used in ...Brazilian folk medicine to treat inflammation, pain and asthma.
This study investigated the effects of essential oil of Peperomia serpens (EOPs) in standard rodent models of pain and inflammation.
The antinociceptive activity was evaluated using chemical (acetic acid and formalin) and thermal (hot plate) models of nociception in mice whereas the anti-inflammatory activity was evaluated by carrageenan- and dextran-induced paw edema tests in rats croton oil-induced ear edema, as well as cell migration, rolling and adhesion induced by carrageenan in mice. Additionally, phytochemical analysis of the EOPs has been also performed.
Chemical composition of the EOPs was analyzed by gas chromatography and mass spectrometry (GC/MS). Twenty-four compounds, representing 89.6% of total oil, were identified. (E)-Nerolidol (38.0%), ledol (27.1%), α-humulene (11.5%), (E)-caryophyllene (4.0%) and α-eudesmol (2.7%) were found to be the major constituents of the oil. Oral pretreatment with EOPs (62.5–500mg/kg) significantly reduced the writhing number evoked by acetic acid injection, with an ED50 value of 188.8mg/kg that was used thereafter in all tests. EOPs had no significant effect on hot plate test but reduced the licking time in both phases of the formalin test, an effect that was not significantly altered by naloxone (0.4mg/kg, s.c.). EOPs inhibited the edema formation induced by carrageenan and dextran in rats. In mice, EOPs inhibited the edema formation by croton oil as well as the leukocyte and neutrophil migration, the rolling and the adhesion of leukocytes.
These data show for the first time that EOPs has a significant and peripheral antinociceptive effect that seems unrelated to interaction with the opioid system. EOPs also displays a significant anti-inflammatory effect in acute inflammation models. This effect seems to be related to components which inhibit the production of several inflammatory mediators. These results support the widespread use of Peperomia serpens in popular medicine to treat inflammation and pain.
Growing clinical evidence has implicated complement as a pivotal driver of COVID-19 immunopathology. Deregulated complement activation may fuel cytokine-driven hyper-inflammation, thrombotic ...microangiopathy and NET-driven immunothrombosis, thereby leading to multi-organ failure. Complement therapeutics have gained traction as candidate drugs for countering the detrimental consequences of SARS-CoV-2 infection. Whether blockade of terminal complement effectors (C5, C5a, or C5aR1) may elicit similar outcomes to upstream intervention at the level of C3 remains debated. Here we compare the efficacy of the C5-targeting monoclonal antibody eculizumab with that of the compstatin-based C3-targeted drug candidate AMY-101 in small independent cohorts of severe COVID-19 patients. Our exploratory study indicates that therapeutic complement inhibition abrogates COVID-19 hyper-inflammation. Both C3 and C5 inhibitors elicit a robust anti-inflammatory response, reflected by a steep decline in C-reactive protein and IL-6 levels, marked lung function improvement, and resolution of SARS-CoV-2-associated acute respiratory distress syndrome (ARDS). C3 inhibition afforded broader therapeutic control in COVID-19 patients by attenuating both C3a and sC5b-9 generation and preventing FB consumption. This broader inhibitory profile was associated with a more robust decline of neutrophil counts, attenuated neutrophil extracellular trap (NET) release, faster serum LDH decline, and more prominent lymphocyte recovery. These early clinical results offer important insights into the differential mechanistic basis and underlying biology of C3 and C5 inhibition in COVID-19 and point to a broader pathogenic involvement of C3-mediated pathways in thromboinflammation. They also support the evaluation of these complement-targeting agents as COVID-19 therapeutics in large prospective trials.
•Deregulated complement activation underpins the pathophysiology of severe COVID-19•Complement inhibition abrogates COVID-19 hyper-inflammation leading to resolution of SARS-CoV-2-associated ARDS•Divergent profiles of C3 vs C5 inhibition point to a broader impact of C3 inhibition on NET-driven thromboinflammation
Aim
To evaluate the incidence of root dentinal microcracks after two extraction techniques through micro‐computed tomographic (micro‐CT) analysis.
Methodology
Thirty pairs of premolars, extracted for ...orthodontic reasons, were assigned to two experimental groups, following a split‐mouth design (n = 30): traumatic technique group (TTG) – teeth extracted following a conventional protocol using forceps, and an atraumatic technique group (ATG) – teeth extracted following an atraumatic technique protocol in which the root was gently detached from the periodontal ligament using a periotome. All teeth were immediately immersed in distilled water and scanned in a micro‐CT device within 72 h following the extraction. The assessment of the images was completed on cross‐section images by two masked evaluators. The Mann–Whitney test was used to assess the significant differences between the groups regarding the number of cross‐sections displaying microcracks whilst the Fischer’s exact test was used to assess differences in the frequency of specimens with defects (P < 0.05).
Results
A total of 52 750 micro‐CT cross‐sectional images were evaluated. A total of 352 cross‐sections had at least one dentinal defect. Microcracks were observed in five teeth of the TTG group (n = 352 slices) and in two teeth of the ATG group (n = 103 slices). The statistical analysis used was unable to detect a significant difference between the groups in terms of the number of dentinal microcracks (P = 0.233) and in the frequency of teeth displaying microcracks (P = 0.424).
Conclusion
A small number of sound teeth without endodontic treatment had dentinal microcracks regardless of the extraction technique applied. No difference was noticed between the traumatic and atraumatic techniques in the incidence of microcracks.
Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and ...microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil.
Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak.
The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non-vectorial ZIKV transmission routes, needs further investigation.
We report the detection and characterization of the transiting sub-Neptune TOI-1759 b, using photometric time series from the Transiting Exoplanet Survey Satellite (TESS) and near-infrared ...spectropolarimetric data from the Spectro-Polarimètre Infra Rouge (SPIRou) on the Canada-France-Hawaii Telescope. TOI-1759 b orbits a moderately active M0V star with an orbital period of 18.849975 ± 0.000006 days, and we measured a planetary radius and mass of 3.06 ± 0.22
R
⊕
and 6.8 ± 2.0
M
⊕
. Radial velocities were extracted from the SPIRou spectra using both the cross-correlation function and the line-by-line methods, optimizing the velocity measurements in the near-infrared domain. We analyzed the broadband spectral energy distribution of the star and the high-resolution SPIRou spectra to constrain the stellar parameters and thus improve the accuracy of the derived planet parameters. A least squares deconvolution analysis of the SPIRou Stokes V polarized spectra detects Zeeman signatures in TOI-1759. We modeled the rotational modulation of the magnetic stellar activity using a Gaussian process regression with a quasi-periodic covariance function and find a rotation period of 35.65
−0.15
+0.17
days. We reconstructed the large-scale surface magnetic field of the star using Zeeman-Doppler imaging, which gives a predominantly poloidal field with a mean strength of 18 ± 4 G. Finally, we performed a joint Bayesian Markov chain Monte Carlo analysis of the TESS photometry and SPIRou radial velocities to optimally constrain the system parameters. At 0.1176 ± 0.0013 au from the star, the planet receives 6.4 times the bolometric flux incident on Earth, and its equilibrium temperature is estimated at 433 ± 14 K. TOI-1759 b is a likely gas-dominated sub-Neptune with an expected high rate of photoevaporation. Therefore, it is an interesting target to search for neutral hydrogen escape, which may provide important constraints on the planetary formation mechanisms responsible for the observed sub-Neptune radius desert.
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