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Nevin Manimala Statistics

Barcoded reciprocal hemizygosity analysis via sequencing illuminates the complex genetic basis of yeast thermotolerance

G3 (Bethesda). 2021 Dec 8:jkab412. doi: 10.1093/g3journal/jkab412. Online ahead of print.

ABSTRACT

Decades of successes in statistical genetics have revealed the molecular underpinnings of traits as they vary across individuals of a given species. But standard methods in the field can’t be applied to divergences between reproductively isolated taxa. Genome-wide reciprocal hemizygosity mapping (RH-seq), a mutagenesis screen in an inter-species hybrid background, holds promise as a method to accelerate the progress of interspecies genetics research. Here we describe an improvement to RH-seq in which mutants harbor barcodes for cheap and straightforward sequencing after selection in a condition of interest. As a proof of concept for the new tool, we carried out genetic dissection of the difference in thermotolerance between two reproductively isolated budding yeast species. Experimental screening identified dozens of candidate loci at which variation between the species contributed to the thermotolerance trait. Hits were enriched for mitosis genes and other housekeeping factors, and among them were multiple loci with robust sequence signatures of positive selection. Together, these results shed new light on the mechanisms by which evolution solved the problems of cell survival and division at high temperature in the yeast clade, and they illustrate the power of the barcoded RH-seq approach.

PMID:34878132 | DOI:10.1093/g3journal/jkab412

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Nevin Manimala Statistics

Temporal trend of tuberculosis incidence and its spatial distribution in Macapá – Amapá

Rev Saude Publica. 2021 Dec 1;55:96. doi: 10.11606/s1518-8787.2021055003431. eCollection 2021.

ABSTRACT

OBJECTIVE: To evaluate the temporal trend of tuberculosis incidence after the implementation of the rapid molecular test (RMT-TB), to identify whether tuberculosis presents seasonal variation and to classify the territory according to case density and risk areas in Macapá, Amapá.

METHODS: Ecological study of tuberculosis cases registered in the Sistema de Informação de Agravos de Notificação (SINAN – Information System for Notifiable Diseases) between 2001 and 2017. We used the Prais-Winsten test to classify the temporal trend of incidence and the interrupted time series to identify changes in the temporal trend before and after the implementation of the rapid molecular test, and to verify seasonality in the municipality. The Kernel estimator was used to classify case density and scan statistics to identify areas of tuberculosis risk.

RESULTS: A total of 1,730 cases were identified, with a decreasing temporal trend of tuberculosis incidence (-0.27% per month, 95%CI -0.13 to -0.41). The time series showed no change in level after the implementation of the GeneXpert®MTB/RIF molecular test; however, the incidence increased in the post-test period (+2.09% per month, 95%CI 0.92 to 3.27). Regarding the seasonal variation, it showed growth (+13.7%/month, 95%CI 4.71 to 23.87) from December to June, the rainy season – called amazon winter season -, and decrease (-9.21% per month, CI95% -1.37 to -16.63) in the other periods. We classified areas with high density of cases in the Central and Northern districts using Kernel and identified three protection clusters, SC1 (RR = 0.07), SC2 (RR = 0.23) and SC3 (RR = 0.36), and a high-risk cluster, SC4 (RR = 1.47), with the scan statistics.

CONCLUSION: The temporal trend of tuberculosis incidence was decreasing in the time series; however, detection increased after the introduction of RMT-TB, and tuberculosis showed seasonal behavior. The case distribution was heterogeneous, with a tendency to concentrate in vulnerable and risk territories, evidencing a pattern of disease inequality in the territory.

PMID:34878090 | DOI:10.11606/s1518-8787.2021055003431

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Nevin Manimala Statistics

Toxicity of Carnoy’s solution toward human keratinocytes: an in vitro study

Braz Oral Res. 2021 Dec 6;35:e124. doi: 10.1590/1807-3107bor-2021.vol35.0124. eCollection 2021.

ABSTRACT

The present study aimed to characterize the chemical elements and cytotoxicity of Carnoy’s solution (CS) by comparing two different trademarked products (one Brazilian [NCS] and another imported [ICS]) using inductively coupled plasma mass spectrometry (ICP-MS) and human keratinocyte (HaCaT) cultures. For performing ICP-MS, the solutions were diluted according to calibration curves, and the chemical elements were analyzed with a spectrometer. HaCaT cells were exposed to CS concentrations ranging from 0.10% to 20% for 3 or 5 min. Cell viability was evaluated immediately (T0), 24 h (T1), and 7 days (T2) after exposure to CS using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) reduction assay. Data were analyzed using a t-test for ICP-MS and analysis of variance followed by Tukey’s post-hoc test for MTT assay, both considering statistical significance at p<0.05. ICP-MS results revealed that ICS presented significantly lower concentrations of 12 chemical elements than NCS. The results of MTT assay revealed that at T0, ICS was more cytotoxic than NCS regardless of the time of exposure (p < 0.05). At T1, the only difference between the groups was at a concentration of 0.10% after 5 min of exposure. At T2, at a concentration of 0.5%, ICS resulted in a significant reduction in cell viability compared to NCS (p < 0.05). Thus, the results showed that ICS was more cytotoxic than NCS. Collectively, our findings suggest that the individual compositions of different CS formulations should be investigated.

PMID:34878079 | DOI:10.1590/1807-3107bor-2021.vol35.0124

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Impact of irrigation protocols with some chelators and mechanical agitation on intratubular decontamination

Braz Oral Res. 2021 Dec 6;35:e127. doi: 10.1590/1807-3107bor-2021.vol35.0127. eCollection 2021.

ABSTRACT

This study evaluated main canal and intratubular decontamination using different irrigation solutions followed by adjunctive agitation steps for infected root canals. Sixty-eight lower incisors were contaminated with Enterococcus faecalis and allocated to groups according to canal treatment (n = 10): G1, NaOCl followed by ethylenediaminetetraacetic acid (EDTA); G2, a mixture of NaOCl with hydroxyethylidene bisphosphonate (HEBP); and G3, NaOCl followed by EDTA-T (EDTA with sodium lauryl ether sulfate). All three groups of teeth were agitated with passive ultrasonic irrigation (PUI) using saline solution, whereby G4, G5, and G6 were prepared as above, and agitation was performed using an XP-Endo Finisher instrument. Microbiological samples were collected from the root canals with paper points at three times: before and after chemomechanical preparation and after agitation. The colony-forming units (CFU)/mL count was determined, and bacterial intratubular viability was analyzed via confocal laser scanning microscopy using Live/Dead staining. Statistical analysis was performed using a Kruskal-Wallis test followed by Dunn tests. A Friedman test was applied for colony-counting data (α = 0.05). CFU/mL counting indicated equally effective decontamination in the experimental groups (p > 0.05). According to microscopy images, the use of irrigation solutions followed by agitation with the XP-Endo Finisher yielded better results. Moreover, NaOCl+EDTA-T followed by XP-Endo Finisher resulted in significantly lower viability than in the PUI-activated groups (p < 0.05). The cervical and medium thirds of the specimens presented similar results. Overall, NaOCl+EDTA-T exhibited the best intratubular antibacterial activity, mainly for canals that were subsequently agitated using XP-Endo Finisher.

PMID:34878082 | DOI:10.1590/1807-3107bor-2021.vol35.0127

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Pulp capping with mineral trioxide aggregate or Biodentine: a comparison of mineralized barrier formation and inflammatory and degenerative events

Braz Oral Res. 2021 Dec 6;35:e118. doi: 10.1590/1807-3107bor-2021.vol35.0118. eCollection 2021.

ABSTRACT

Biodentine in endodontic practice has been widely investigated, but comprehensive histological descriptions of degenerative and inflammatory responses are not covered in most of the studies that compare pulp capping materials. This study aimed to evaluate pulpal responses to mineral trioxide aggregate (MTA Angelus) and Biodentine, focusing on mineralized barrier formation and on inflammatory and degenerative events. 80 male Wistar rats were assigned to 5 groups, according to the materials used for pulp capping and coronal sealing (n=8 per group/period). The lower first molars were mechanically exposed, capped with either MTA or Biodentine, and restored with silver amalgam. In an additional test group, the teeth were capped and sealed with Biodentine. The teeth capped with gutta-percha and restored with silver amalgam served as positive control, whereas untreated teeth served as negative control. Pulpal responses and coronal sealing were evaluated after 14 or 21 days. Data was statistically analyzed by the Kruskal-Wallis and Dunn’s post hoc tests (p<0.05). Biodentine and MTA presented satisfactory results, showing a milder inflammatory response (p<0.0001) and more pronounced formation of mineralized barrier (p<0.0001) compared to the teeth capped with gutta-percha. As a restorative material, Biodentine kept coronal sealing in only 37.5% of the samples. Biodentine showed favorable properties in vital pulp therapy, being similar to MTA. However, it was not effective in protecting dental pulp from microleakage during the experimental period.

PMID:34878073 | DOI:10.1590/1807-3107bor-2021.vol35.0118

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Effect of irrigation protocols on root canal wall after post preparation: a micro-CT and microhardness study

Braz Oral Res. 2021 Dec 6;35:e122. doi: 10.1590/1807-3107bor-2021.vol35.0122. eCollection 2021.

ABSTRACT

The aim of this study was to investigate the effects of different post space irrigation protocols for removing residual filling material from dentin walls, by using microcomputed tomography (micro-CT), and the influence of these protocols on dentin microhardness. Bovine incisors (n = 35) were filled with the single-cone technique and MTA Fillapex (Angelus, Londrina, PR, Brazil). Post space preparation (PSP) was performed 7 days after filling, using the Odous Touch electrical system (Odous De Deus Ind. e Com., Belo Horizonte, MG, Brazil), followed by post space irrigation using manual irrigation, passive ultrasonic irrigation, or Easy Clean, together with 2.5% sodium hypochlorite (NaOCl), or with 2.5% NaOCl and 17% EDTA (NaOCl/EDTA). Micro-CT scans were performed at three time points. The residual filling material was evaluated at three levels: cervical, middle and apical. The Knoop test was measured with four indentations around the canal lumen at three dentin depths: X (100 μm), Y (200 μm) and Z (400 μm). Statistical analysis was performed using ANOVA (p < 0.05). The effects of the activation method (p < 0.001), and the root level (p = 0.013), as well as the interaction between the irrigant and the activation method (p = 0.041), led to different percentages of residual filling material. Lower amounts of residual filling material were observed at the cervical versus the middle and apical levels (p < 0.05). No significant differences were observed in dentin microhardness (p > 0.05). The best removal of the residual filling material was performed using the Easy Clean tip and NaOCl/EDTA, regardless of the activation methods.

PMID:34878077 | DOI:10.1590/1807-3107bor-2021.vol35.0122

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Nevin Manimala Statistics

Preparation, optimization and evaluation of transdermal therapeutic system of celecoxib to treat inflammation for treatment of rheumatoid arthritis

An Acad Bras Cienc. 2021 Dec 1;93(suppl 4):e20201561. doi: 10.1590/0001-3765202120201561. eCollection 2021.

ABSTRACT

The purpose of present study was to prepare transdermal therapeutic system that could enhance dissolution of poorly aqueous soluble drug Celecoxib and thus increase its skin permeation. Solubility studies screened triacetin as oil, cremophor RH 40 as surfactant and Polyethylene Glycol 400 as co-surfactant. Pseudoternary phase diagrams were constructed to find out microemulsion region. Independent variables (oil, Smix and water) concentration was used at high (+1) and low levels (-1) that would generate 17 different combinations of microemulsions. Microemulsions were characterized, optimized and evaluated. pH, viscosity, conductivities, refractive index, droplet size and poly-dispersity-index was investigated. Prepared microemulsions were oil in water, thermodynamically stable, isotropic, transparent, deflocculated and within narrow range of size. Mathematical equations and response surface plots related the independent and dependent variables. Optimum microemulsion ME6 was further incorporated with carbomer 940 gel base to produce microemulsion based gel. ME6 and its gel showed significant difference (p<0.05) from control gel. Stability studies showed prepared MEBG of celecoxib was stable during storage period. Skin irritation studies found the gel was safe and non-irritating to skin. Anti-inflammatory studies showed significant difference (p<0.05) compared to control gel. Thus, the therapeutic system was successfully developed and optimized using Box Behnken statistical design.

PMID:34878046 | DOI:10.1590/0001-3765202120201561

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Evaluation of quantitative polymerase chain reaction for the detection of Toxoplasma gondii oocysts shed by cats

Rev Bras Parasitol Vet. 2021 Dec 1;30(4):e016621. doi: 10.1590/S1984-29612021091. eCollection 2021.

ABSTRACT

Felines are definitive hosts of Toxoplasma gondii and can shed oocysts in their feces, contaminating the environment. Sporulated oocysts are highly resistant to the environment and have higher infectivity, which are attributed to many toxoplasmosis outbreaks. The aim of the present study was to evaluate a quantitative polymerase chain reaction (qPCR) technique for the detection of T. gondii oocysts shed by cats. Twelve cats from a previous vaccine experiment were challenged orally with 600 cysts of the TgDoveBr8 strain on day 72. Fecal samples were collected daily using the centrifugal flotation technique, with microscopic examination (Sheather technique) and qPCR for 20 days after the challenge. Cats from all groups shed oocysts in their feces. Five negative cats in the Sheather were positive according to qPCR on the 3rd day post-inoculation (dpi). Oocysts were detected on the 4th dpi using the Sheather; however, there was no statistical difference between the two methods (p=0.1116). In addition, there was no statistically significant difference in oocyst shedding between the groups according to the Sheather technique (p=0.6534) and qPCR (p=0.9670). In conclusion, these results demonstrate that qPCR can be used as an alternative to the Sheather to detect and quantify T. gondii oocysts.

PMID:34878069 | DOI:10.1590/S1984-29612021091

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Nevin Manimala Statistics

Knowledge of primary care nurses before and after educational intervention on diabetic foot

Rev Gaucha Enferm. 2021 Dec 6;42:e20200452. doi: 10.1590/1983-1447.2021.20200452. eCollection 2021.

ABSTRACT

OBJECTIVE: To compare the knowledge of the nurses about the diabetic foot before and after the educational intervention.

METHOD: Quasi-experimental study, before-and-after type, carried out with 53 nurses, from March to June 2016, in the municipality of Campina Grande, Paraíba. The Questioning Methodology was used to develop the educational intervention. Knowledge was investigated using an instrument designed by the researchers. For data analysis before and after the intervention, the McNemar and Wilcoxon tests were performed.

RESULTS: The knowledge of the nurses about the diabetic foot turned out to be deficient, with a mean of correct answers in the pre-test of 23.8 (SD±12.8) and after 41.9 (SD±9,2), with a statistically significant difference (p<0.01).

CONCLUSION: It was verified a significant increase in the knowledge after the intervention, highlighting the items related to evaluate the loss of plantar protective sensitivity, essential for the prevention of diabetic foot.

PMID:34878024 | DOI:10.1590/1983-1447.2021.20200452

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Nevin Manimala Statistics

Central Sterile Services Department: psychosocial risks related to the prescribed organization of nursing work

Rev Gaucha Enferm. 2021 Dec 6;42:e20200433. doi: 10.1590/1983-1447.2021.20200433. eCollection 2021.

ABSTRACT

OBJECTIVE: The aim of this study is to investigate the characteristics of work-flow organization of the Central Sterile Services Department and analyze whether nursing workers are subject to psychosocial risks.

METHOD: Mixed study, carried out between November 2017 and July 2019 with nursing workers of the Department of Central Sterile Services of a university hospital. Sampling for the quantitative stage of this study consisted of 36 professionals who have filled the Prescribed Work Organization Scale form, followed by statistical analysis. Qualitative sampling was obtained through collective interview with 19 members of the nursing team.

RESULTS: Of the 36 nursing workers, 68.3% were female, 69.4% were over 45 years of age and 75% of the professionals assessed reported as being exposed to a medium psychosocial risk with the emergence of two categories. The aspects related to pleasure at work were highlighted as being the importance of the unit, indirect patient care, professional recognition, and teamwork. The main reported suffering triggers were the lack of human resources, supplies, maintenance, communication, and the devaluation of the unit.

CONCLUSION: The prevalence of medium psychosocial risks indicated the need for actions aiming at improving work processes.

PMID:34878021 | DOI:10.1590/1983-1447.2021.20200433