DNA methylation patterns suggest the involvement of DNMT3B and TET1 in osteosarcoma development.

DNA methylation may be involved in the development of osteosarcomas. Osteosarcomas commonly arise during the bone growth and remodeling in puberty, making it plausible to infer the involvement of epigenetic alterations in their development. As a highly studied epigenetic mechanism, we investigated DNA methylation and related genetic variants in 28 primary osteosarcomas aiming to identify deregulated driver alterations. Methylation and genomic data were obtained using the Illumina HM450K beadchips and the TruSight One sequencing panel, respectively. Aberrant DNA methylation was spread throughout the osteosarcomas genomes. We identified 3146 differentially methylated CpGs comparing osteosarcomas and bone tissue samples, with high methylation heterogeneity, global hypomethylation and focal hypermethylation at CpG islands. Differentially methylated regions (DMR) were detected in 585 loci (319 hypomethylated and 266 hypermethylated), mapped to the promoter regions of 350 genes. These DMR genes were enriched for biological processes related to skeletal system morphogenesis, proliferation, inflammatory response, and signal transduction. Both methylation and expression data were validated in independent groups of cases. Six tumor suppressor genes harbored deletions or promoter hypermethylation (DLEC1, GJB2, HIC1, MIR149, PAX6, and WNT5A), and four oncogenes presented gains or hypomethylation (ASPSCR1, NOTCH4, PRDM16, and RUNX3). Our analysis also revealed hypomethylation at 6p22, a region that contains several histone genes. Copy-number changes in DNMT3B (gain) and TET1 (loss), as well as overexpression of DNMT3B in osteosarcomas provide a possible explanation for the observed phenotype of CpG island hypermethylation. While the detected open-sea hypomethylation likely contributes to the well-known osteosarcoma genomic instability, enriched CpG island hypermethylation suggests an underlying mechanism possibly driven by overexpression of DNMT3B likely resulting in silencing of tumor suppressors and DNA repair genes.

Analysis of the Mutational Landscape of Osteosarcomas Identifies Genes Related to Metastasis and Prognosis and Disrupted Biological Pathways of Immune Response and Bone Development.

Osteosarcoma (OS) is the most prevalent type of bone tumor, but slow progress has been achieved in disentangling the full set of genomic events involved in its initiation and progression. We assessed by NGS the mutational spectrum of 28 primary OSs from Brazilian patients, and identified 445 potentially deleterious SNVs/indels and 1176 copy number alterations (CNAs). TP53 was the most recurrently mutated gene, with an overall rate of ~60%, considering SNVs/indels and CNAs. The most frequent CNAs (~60%) were gains at 1q21.2q21.3, 6p21.1, and 8q13.3q24.22, and losses at 10q26 and 13q14.3q21.1. Seven cases presented CNA patterns reminiscent of complex events (chromothripsis and chromoanasynthesis). Putative RB1 and TP53 germline variants were found in five samples associated with metastasis at diagnosis along with complex genomic patterns of CNAs. PTPRQ, KNL1, ZFHX4, and DMD alterations were prevalent in metastatic or deceased patients, being potentially indicative of poor prognosis. TNFRSF11B, involved in skeletal system development and maintenance, emerged as a candidate for osteosarcomagenesis due to its biological function and a high frequency of copy number gains. A protein-protein network enrichment highlighted biological pathways involved in immunity and bone development. Our findings reinforced the high genomic OS instability and heterogeneity, and led to the identification of novel disrupted genes deserving further evaluation as biomarkers due to their association with poor outcomes.

Detection of mosaicism for segmental and whole chromosome imbalances by targeted sequencing.

Mosaic segmental and whole chromosome copy number alterations are postzygotic variations known to be associated with several disorders. We have previously presented an efficient targeted sequencing approach to simultaneously detect point mutations and copy number variations (CNVs). In this study, we evaluated the efficiency of this approach to detect mosaic CNVs, using seven postnatal and 19 tumor samples, previously characterized by chromosomal microarray analyses (CMA). These samples harbored a total of 28 genomic imbalances ranging in size from 0.68 to 171 Mb, and present in 10-80% of the cells. All CNV regions covered by the platform were correctly identified in postnatal samples, and only seven out of 19 CNVs from tumor samples were not identified either because of a lack of target probes in the affected genomic regions or an absence of minimum reads for an alteration call. These results demonstrate that, in a research setting, this is a robust approach for detecting mosaicism in cases of segmental and whole chromosome alterations. Although the current sequencing platform presented a resolution similar to genomic microarrays, it is still necessary to further validate this approach in a clinical setting in order to replace CMA and sequencing analyses by a single test.

Effect of surface preparation and light curing on penetration of silver particles from 38% silver diamine fluoride in dentin of primary teeth: An in vitro evaluation.

PURPOSE: To evaluate the effect of light cure, as well as various dentin surface treatment approaches, on the penetration depth of silver precipitating from 38% silver diamine fluoride into primary dentin tubules. METHODS: The occlusal dentin surfaces of 42 non-carious primary molars were exposed and then sectioned into halves bucco-lingually. The halves from each tooth pair were randomly split in two mega-groups, and each mega-group was divided randomly as follows into six experimental groups: prepared by either carbide bur (G1, G2), ceramic bur (G3, G4), or erbium laser (G5, G6). SDF was then applied to all prepared surfaces, and finally even-numbered groups (G2, G4, G6) were light cured. One mega-group was assigned to quantitative evaluation of silver penetration depth along the axial wall, and the other mega-group was reserved for qualitative observation of relative silver distribution on the occlusal surface, both via scanning electron microscope. RESULTS: No significant difference was observed in silver penetration depth between light cure and non-light cure groups (P= 0.8908). There was a statistically significant association between tooth preparation method and depth of silver penetration (P< 0.000001); laser-treated groups had significantly deeper silver penetration (1,148.9 µm G5, 1160.4 µm G6) than carbide bur (P< 0.05; 184.7 µm G1, 301.8 µm G2) or ceramic bur (P< 0.05; 184.1 µm G3, 131.0 µm G4) groups. A significant difference (P< 0.05) was noted in percentage occlusal surface coverage of particles between laser (51.4% G5, 35.8% G6) and carbide groups (21.1% G1, 19.3% G2). Light cure had no significant effect on the depth of silver penetration from 38% SDF in the dentin of primary teeth. Laser preparation resulted in deeper silver penetration than carbide or ceramic bur. CLINICAL SIGNIFICANCE: Exposure of 38% silver diamine fluoride-treated dentin to light cure did not affect the depth of penetration of silver particles into the dentin tubules of primary teeth. Rather, tooth preparation approaches that reduce the smear layer, like laser ablation, resulted in the deepest penetration of silver into the tubules. Clinical application of these findings will depend on scenario and treatment aim.

Atraumatic removal of a zirconia-framework fixed partial denture using an Er,Cr:YSGG laser: a case report.

A 3-unit fixed partial denture spanning the maxillary left central incisor, lateral incisor, and canine was fabricated with a zirconia framework and a pontic veneered with porcelain to replace the maxillary left lateral incisor. The prosthesis was attached to the abutment teeth using lingual wings that sat in preparations approximately 1.5 mm deep axially. When the veneering porcelain on the pontic developed a void 5 months after cementation, the patient was offered 3 options to eliminate the defect: (1) restoration with composite resin; (2) destructive removal of the prosthesis followed by fabrication of a replacement; or (3) attempted nondestructive removal of the prosthesis with an erbium, chromium-doped: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser followed by laboratory repair of the void with low-fusing porcelain. The patient selected the third option. The laser was set to 3 W, 20 Hz, 50% air, and 50% water using the turbo handpiece with a 700-µm-diameter tip, per the manufacturer's recommendations. The Er,Cr:YSGG laser operates in a noncontact mode at an ideal distance of 3 to 5 mm from the target tissue, in this case, the lingual surfaces of the maxillary left central incisor and canine. No anesthetic was administered to the patient. The laser beam was applied to the wings of the prosthesis intermittently for 8 cycles at a maximum of 60 seconds per cycle. During irradiation, the handpiece moved side-to-side in a scanning motion. Darkening of the cement was observed under the wing on the central incisor, indicating the initial disruption of the adhesive seal. Slight torquing pressure was applied using specially designed crown removal pliers, and the prosthesis was removed atraumatically. The recovered prosthesis was repaired with low-fusing porcelain and adhesively luted. Atraumatic debonding of a high-strength ceramic can be predictably performed with the Er,Cr:YSGG laser when appropriate settings are used.

Pharmacodynamic Attainment of the Synergism of Meropenem and Fosfomycin Combination against Pseudomonas aeruginosa Producing Metallo-ß-Lactamase.

Fosfomycin combined with other antimicrobials has shown good efficacy against multidrug-resistant (MDR) bacteria in both in vitro and clinical studies; however, the activity of fosfomycin combined with other antimicrobials against metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa strains has not been tested. The objective of this study was to determine the synergism and optimal intravenous dosing regimens of fosfomycin with meropenem against MDR and MBL-producing P. aeruginosa strains. The MICs of both antimicrobials were determined by the checkerboard method and analyzed by two synergism tests with 19 clones of P. aeruginosa isolates, 10 of which were MBL producers. A pharmacodynamic (PD) analysis was performed for meropenem (administered at 1 g every 8 h [q8h], 1.5 g every 6 h [q6h], and 2 g q8h) and fosfomycin (administered at 4 g q8h, 4 g q6h, 6 g q8h, and 8 g q8h) regimens with a dose reduction for renal impairment by determining the probability of target attainment (PTA) for target PD indices of meropenem (the percentage of the time in a 24-h duration at which the free drug concentration remains above the MIC [fT>MIC], ≥40%) and fosfomycin (the ratio of the area under the free drug concentration-versus-time curve over 24 h and the MIC [fAUC/MIC], ≥40.8). The combination reduced the MIC50 and MIC90 by 8-fold. Seven (44%) isolates with MICs in the intermediate or resistant ranges became sensitive to meropenem. For the MBL-producing isolates, the combination resulted in 40% of isolates becoming sensitive to meropenem. The meropenem regimens reached a PTA of ≥90% (MIC = 4 µg/ml) in 6 (32%) isolates when they were used as monotherapy and 13 (68%) isolates when they were combined with fosfomycin. None of the fosfomycin monotherapy regimens reached the PTA of ≥90% (MIC = 16 µg/ml). When combined with meropenem, the fosfomycin regimens reached the PTA of ≥90% in 14 (74%) isolates. The increase in pharmacodynamic activities resulting from the synergistic action of meropenem with fosfomycin demonstrates the potential relevance of this combination to fight infections caused by MDR and MBL-producing P. aeruginosa strains.

Evaluation of SHOX defects in the era of next-generation sequencing.

Short stature homeobox (SHOX) haploinsufficiency is a frequent cause of short stature. Despite advances in sequencing technologies, the identification of SHOX mutations continues to be performed using standard methods, including multiplex ligation-dependent probe amplification (MLPA) followed by Sanger sequencing. We designed a targeted panel of genes associated with growth impairment, including SHOX genomic and enhancer regions, to improve the resolution of next-generation sequencing for SHOX analysis. We used two software packages, CONTRA and Nexus Copy Number, in addition to visual analysis to investigate the presence of copy number variants (CNVs). We evaluated 15 patients with previously known SHOX defects, including point mutations, deletions and a duplication, and 77 patients with idiopathic short stature (ISS). The panel was able to confirm all known defects in the validation analysis. During the prospective evaluation, we identified two new partial SHOX deletions (one detected only by visual analysis), including an intragenic deletion not detected by MLPA. Additionally, we were able to determine the breakpoints in four cases. Our results show that the designed panel can be used for the molecular investigation of patients with ISS, and it may even detect CNVs in SHOX and its enhancers, which may be present in a significant fraction of patients.

PID temperature controller in pig nursery: spatial characterization of thermal environment.

The use of enhanced technologies of temperature control can improve the thermal conditions in environments of livestock facilities. The objective of this study was to evaluate the spatial distribution of the thermal environment variables in a pig nursery with a heating system with two temperature control technologies based on the geostatistical analysis. The following systems were evaluated: overhead electrical resistance with Proportional, Integral, and Derivative (PID) controller and overhead electrical resistance with a thermostat. We evaluated the climatic variables: dry bulb temperature (Tbs), air relative humidity (RH), temperature and humidity index (THI), and enthalpy in the winter, at 7:00, 12:00, and 18:00 h. The spatial distribution of these variables was mapped by kriging. The results showed that the resistance heating system with PID controllers improved the thermal comfort conditions in the pig nursery in the coldest hours, maintaining the spatial distribution of the air temperature more homogeneous in the pen. During the hottest weather, neither system provided comfort.

CTLA-4 gene polymorphisms are associated with obesity in Turner Syndrome.

Turner syndrome (TS) is characterized by a set of clinical conditions, including autoimmune/inflammatory diseases and infectious conditions, that can compromise a patient's quality of life. Here we assessed polymorphisms in CTLA-4 +49A/G (rs231775), PTPN22 +1858G/A (rs2476601), and MBL2 -550 (H/L) (rs11003125), -221(X/Y) (rs7096206) and exon 1 (A/O) in women from northeastern Brazil to determine whether polymorphisms within these key immune response genes confer differential susceptibility to clinical conditions in TS. A case-control genetic association study was performed, including 86 female TS patients and 179 healthy women. An association was observed for the A/G genotype of CTLA-4 +49A/G in TS patients (p=0.043, odds ratio [OR]=0.54). In addition, an association between the CTLA-4 G/G genotype and obesity was detected in TS patients (p=0.02, OR=6.04). Regarding, the -550(H/L) polymorphism in the MBL2 promoter, the frequency of the H/L genotype was significantly higher in the TS group than healthy controls (p=0.01, OR=1.96). The H/H genotype indicated a protective effect in TS patients (p=0.01, OR=0.23). No differences were observed in the distribution of -221(X/Y), MBL2 exon 1 variants, and PTPN22 +1858G/A in any assessed groups. CTLA-4 variants are potentially involved in obesity in this cohort of TS patients from northeastern Brazil.

PID temperature controller in pig nursery: improvements in performance, thermal comfort, and electricity use.

The use of smarter temperature control technologies in heating systems can optimize the use of electric power and performance of piglets. Two control technologies of a resistive heating system were assessed in a pig nursery: a PID (proportional, integral, and derivative) controller and a thermostat. The systems were evaluated regarding thermal environment, piglet performance, and use of electric power for 99 days. The heating system with PID controller improved the thermal environment conditions and was significantly (P < 0.001) more efficient in terms of electricity use to produce 1 kg of body weight (2.88 kWh kg(-1)), specific cost (0.75 R$ kg(-1)), weight gain (7.3 kg), daily weight gain (0.21 kg day(-1)), and feed conversion (1.71) than the system with thermostat (3.98 kWh kg(-1); 1.03 R$ kg(-1); 5.2 kg; 0.15 kg day(-1), and 2.62, respectively). The results indicate that the PID-controlled heating system is more efficient in electricity use and provides better conditions for thermal comfort and animal performance than heating with thermostat.

Polymorphisms in folate pathway genes are not associated with somatic nondisjunction in turner syndrome.

Folate metabolism dysfunction can lead to DNA hypomethylation and abnormal chromosomal segregation. Previous investigations of this association have produced controversial results. Here we performed a case-control study in patients with Turner syndrome (TS) to determine the effects of genetic polymorphisms of folate pathway genes as potential risk factors for somatic chromosomal nondisjunction. TS is a useful model for this investigation because patients with TS show a high frequency of chromosome mosaicism. Here we investigated the possible association of polymorphisms of the MTHFR gene with TS risk, which has been previously investigated with controversial results. We also examined the effects of MTR, RFC1, and TYMS gene polymorphisms in TS for the first time. The risk was evaluated according to allelic and genotype (independent and combined) frequencies among 70 patients with TS and 144 age-matched healthy control subjects. Polymorphism genotyping was performed by PCR, PCR-RFLP, and PCR-ASA. The polymorphisms MTHFR 677C>T and 1298A>C, MTR 2756A>G, RFC1 80G>A, and TYMS 2R/3R-alone or in combinations-were not associated with the risk of chromosomal aneuploidy in TS. In conclusion, our present findings did not support a link between impaired folate metabolism and abnormal chromosome segregation leading to somatic nondisjunction in TS patients.

Molecular study of Trypanosoma caninum isolates based on different genetic markers.

Trypanosoma caninum is a parasite recently described in dogs, whose life cycle is rather unknown. Here, we performed a genetic study with T. caninum samples obtained in different Brazilian regions. The study was based on PCR assays target to small and large subunit ribosomal DNA (rDNA) (18S rDNA and 24Sα rDNA), cytochrome B (Cyt b), and internal transcribed spacer 1 rDNA (ITS1 rDNA) following by the sequence analysis. Additionally, we used primers for the variable regions of kinetoplast DNA (kDNA) minicircles and endonucleases restriction in the ITS1 rDNA amplification product. T. caninum samples displayed the same patterns. Tree construction confirmed the close relationship between T. caninum samples, regardless of the molecular target used and endonuclease restriction digestion revealed that all samples have the same restriction profile. Therefore, T. caninum seems to be a genetically homogeneous specie. In the kDNA assay, T. caninum possessed a different molecular size profile with respect to others trypanosomes, 330 and 350 bp. This study provides nucleotide sequences from different regions of the genome of T. caninum that certainly facilitate future studies.

Targeted and Shallow Whole-Genome Sequencing Identifies Therapeutic Opportunities in p53abn Endometrial Cancers.

PURPOSE: Shallow whole-genome sequencing (sWGS) can detect copy-number (CN) aberrations. In high-grade serous ovarian cancer (HGSOC) sWGS identified CN signatures such as homologous recombination deficiency (HRD) to direct therapy. We applied sWGS with targeted sequencing to p53abn endometrial cancers to identify additional prognostic stratification and therapeutic opportunities. EXPERIMENTAL DESIGN: sWGS and targeted panel sequencing was performed on formalin-fixed, paraffin-embedded p53abn endometrial cancers. CN alterations, mutational data and CN signatures were derived, and associations to clinicopathologic and outcomes data were assessed. RESULTS: In 187 p53abn endometrial cancers, 5 distinct CN signatures were identified. Signature 5 was associated with BRCA1/2 CN loss with features similar to HGSOC HRD signature. Twenty-two percent of potential HRD cases were identified, 35 patients with signature 5, and 8 patients with BRCA1/2 somatic mutations. Signatures 3 and 4 were associated with a high ploidy state, and CCNE1, ERBB2, and MYC amplifications, with mutations in PIK3CA enriched in signature 3. We observed improved overall survival (OS) for patients with signature 2 and worse OS for signatures 1 and 3. Twenty-eight percent of patients had CCNE1 amplification and this subset was enriched with carcinosarcoma histotype. Thirty-four percent of patients, across all histotypes, had ERBB2 amplification and/or HER2 overexpression on IHC, which was associated with worse outcomes. Mutations in PPP2R1A (29%) and FBXW7 (16%) were among the top 5 most common mutations. CONCLUSIONS: sWGS and targeted sequencing identified therapeutic opportunities in 75% of patients with p53abn endometrial cancer. Further research is needed to determine the efficacy of treatments targeting these identified pathways within p53abn endometrial cancers.

Predicting Risk of Ammonia Exposure in Broiler Housing: Correlation with Incidence of Health Issues.

The study aimed to forecast ammonia exposure risk in broiler chicken production, correlating it with health injuries using machine learning. Two chicken breeds, fast-growing (Ross®) and slow-growing (Hubbard®), were compared at different densities. Slow-growing birds had a constant density of 32 kg m-2, while fast-growing birds had low (16 kg m-2) and high (32 kg m-2) densities. Initial feeding was uniform, but nutritional demands led to varied diets later. Environmental data underwent selection, pre-processing, transformation, mining, analysis, and interpretation. Classification algorithms (decision tree, SMO, Naive Bayes, and Multilayer Perceptron) were employed for predicting ammonia risk (10-14 pmm, Moderate risk). Cross-validation was used for model parameterization. The Spearman correlation coefficient assessed the link between predicted ammonia risk and health injuries, such as pododermatitis, vision/affected, and mucosal injuries. These injuries encompassed trachea, bronchi, lungs, eyes, paws, and other issues. The Multilayer Perceptron model emerged as the best predictor, exceeding 98% accuracy in forecasting injuries caused by ammonia. The correlation coefficient demonstrated a strong association between elevated ammonia risks and chicken injuries. Birds exposed to higher ammonia concentrations exhibited a more robust correlation. In conclusion, the study effectively used machine learning to predict ammonia exposure risk and correlated it with health injuries in broiler chickens. The Multilayer Perceptron model demonstrated superior accuracy in forecasting injuries related to ammonia (10-14 pmm, Moderate risk). The findings underscored the significant association between increased ammonia exposure risks and the incidence of health injuries in broiler chicken production, shedding light on the importance of managing ammonia levels for bird welfare.

Risks of soil chemical degradation from atmospheric gypsum plumes around selected extraction and processing enterprises, Northeast Brazil.

Gypsum plays a prominent role in agriculture, being considered an effective alternative to alleviate subsurface acidity due to its higher solubility and containing sulfur. However, another significant aspect is which pose long-term risks of groundwater contamination due to excessive applications of salts, pesticides, and other chemicals that will be leached, or even soil chemical depletion. So far, no study has focused on understanding the impacts of the atmospheric gypsum plume originating from gibbsite mining and processing on the leaching of soil bases and chemical degradation surrounding these sites. In this study, we evaluated the behavior of chemical characteristics in soil profiles distributed along the dispersion of the atmospheric plume and in areas without interference from the industrial sector in the state of Maranhão, Northeast Brazil. Fifty-three sampling points were collected at 7 locations based on the dispersion of the dust plume through wind drift. Each sampling point was represented by three composite soil samples at depths of 0.0-0.20, 0.40-0.60, and 1.00-1.20 m, where the chemical soil characteristics were evaluated. The average levels of Ca, Mg, and K in the studied layers are classified as low, with minimum values below the method's detection limit, and they also show imbalance due to higher concentrations of Ca in the surface layer in areas affected by atmospheric dispersion. The sum and saturation of bases at all depths are classified as low. Higher aluminum saturation values were observed in the deeper soil layers. The gypsum dust altered the soil's chemical characteristics at the evaluated depths; therefore, it is necessary to seek means to mitigate gypsum dust release during gibbsite extraction and processing and ensure that the soils in areas near these enterprises maintain their natural characteristics.

Use of Psychotropic Drugs during the COVID-19 pandemic in Minas Gerais, Brazil.

OBJECTIVE: To describe the profile of dispensation of mental health drugs by analyzing trends in use before and during the COVID-19 pandemic within the Unified Health System (Sistema Único de Saúde [SUS]). METHODS: Pharmacoepidemiological study based on the retrospective analysis of records regarding the dispensation of psychotropic medicines in the SUS database in the state of Minas Gerais between 2018 and 2021, considering the periods before (2018-2019) and during the COVID-19 pandemic (2020-2021). A database with the records of dispensation of municipalities was created, and the consistency of releases was verified using the Analysis of Variance (ANOVA) test. Medicine consumption was measured in a defined daily dose (DDD) per 1,000 inhabitants/day for SUS, and the difference between periods was evaluated using Student's t-test. RESULTS: During the COVID-19 pandemic, there was an increase in the consumption of psychotropic drugs in SUS-MG. The most consumed medicines were fluoxetine hydrochloride, diazepam and phenobarbital sodium (DDD=5.89; 3.42; 2.49) in the Basic Pharmaceutical Services Component(CBAF), and olanzapine, risperidone and quetiapine hemifumarate (DDD=0.80; 0.47; 0.38) in the Specialized Pharmaceutical Services Component (CEAF). The highest percentage increase in consumption was attributed to clonazepam (75.37%) and lithium carbonate (35.35%), in CBAF, and levetiracetam (3,000.00%) and memantine hydrochloride (340.0%) in CEAF. CONCLUSION: The change in the psychotropic drug dispensation profile during the COVID-19 pandemic highlights the need to produce more studies to complete, confirm or rule out this profile and monitor the use of psychotropic drugs by the population in the post-pandemic context.

Does functionalised nanoplastics modulate the cellular and physiological responses of aquatic fungi to metals?

Co-contamination of freshwaters by nanoplastics (NPs; ≤ 1 µm) and metals is an emerging concern. Aquatic hyphomycetes play a crucial role as primary decomposers in these ecosystems. However, concurrent impacts of NPs and metals on the cellular and physiological activities of these fungi remain poorly understood. Here, the effects of environmentally realistic concentrations of two types of polystyrene (PS) NPs (bare and -COOH; up to 25 µg L-1) and copper (Cu; up to 50 µg L-1) individually and all possible combinations (NPs types and Cu) on Articulospora tetracladia, a prevalent aquatic hyphomycete, were investigated. Endpoints measured were intracellular reactive oxygen species accumulation, plasma membrane disruption and fungal growth. The results suggest that functionalised (-COOH) NPs enhance Cu adsorption, as revealed by spectroscopic analyses. Notably, NPs, Cu and their co-exposure to A. tetracladia can lead to ROS accumulation and plasma membrane disruption. In most cases, exposure to treatments containing -COOH NPs with Cu showed greater cellular response and suppressed fungal growth. By contrast, exposure to Cu individually showed stimulatory effects on fungal growth. Overall, this study provides novel insight that functionalisation of NPs facilitates metal adsorption, thus modulating the impacts of metals on aquatic fungi.

Treating Nerve Injury after Endodontic Microsurgery Using Laser Photobiomodulation: A Report of 2 Cases.

The purpose of this case report was to inform dentists and dental specialists about a noninvasive, viable treatment option that could aid in the recovery of patients who have experienced iatrogenic nerve injuries. Nerve injury is an inherent risk of many dental procedures and a complication that can negatively impact a patient's quality of life and activities of daily living. Managing neural injuries presents a challenge for clinicians because there are no standard protocols reported in the literature. Although spontaneous healing of these injuries can occur, the duration and degree of healing can vary greatly between individuals. Photobiomodulation (PBM) therapy is used as an adjunct in medicine for functional nerve recovery. Once target tissues are illuminated with a low-level laser during PBM, the light energy is absorbed by the mitochondria causing adenosine triphosphate production, modulation of reactive oxygen species, and the release of nitric oxide. These cellular changes explain why PBM has been shown to aid in cell repair, vasodilation, a reduction in inflammation, accelerated healing, and an improvement in postoperative pain. This case report presents 2 patients with neurosensory alterations after endodontic microsurgery with a significant improvement in their condition after PBM treatment using a 940-nm diode laser.

Body Mass Index but not Physical Activity Level Moderates Lowered Cardiac Baroreflex Sensitivity in People Living with HIV.

Reduced cardiac baroreflex sensitivity (cBRS) is an autonomic marker associated with a worse cardiovascular prognosis. Whether cBRS is lowered in people living with HIV (PLHIV) is yet unclear, as well as potential moderator effects of body mass index (BMI) or physical activity (PA) level. The present study aims to compare the spontaneous cBRS in PLHIV vs. HIV-uninfected controls, and to determine among PLHIV the relationship between cBRS vs. body mass index (BMI) and PA level. Total, upward (cBRS+), and downward (cBRS-) cBRS gains were assessed using the sequential method from beat-to-beat blood pressure at rest in 16 PLHIV (46.5±8.4 years) under antiretroviral therapy for at least 6 months, and 16 HIV-uninfected controls (CTL; 42.1±8.0 years). PA level was assessed by the Physical Activity Questionnaire (IPAQ short version) overall score. PLHIV showed lower total cBRS (8.7±3.1 vs. 15.3±7.7 ms.mmHg-1; p < 0.01), cBRS+ (9.2±4.9 vs. 16.0±6.8 ms.mmHg-1; p < 0.01) and cBRS- (9.5±4.9 vs. 15.3±9.3 ms.mmHg-1; p < 0.01) vs. CTL. No between-group difference was found for BMI (PLHIV: 25.2±2.6 vs. CTL: 26.8±3.2 kg.m-2; p > 0.05) or IPAQ score (PLHIV: 2.4±1.0 vs. CTL: 2.0±1.4; p > 0.05). In PLHIV, total cBRS was inversely correlated vs. BMI (r = -0.44; p = 0.04), but not vs. IPAQ score (r = 0.17; p = 0.26). HIV infection may reduce spontaneous cBRS, which seemed to be moderated by higher BMI, but not PA level of PLHIV.

Novel Nonablative Radiofrequency Approach for the Treatment of Anal Incontinence: A Phase 1 Clinical Trial.

OBJECTIVE: We aimed to describe the action, impact on quality of life, and side effects of perianal nonablative radiofrequency (RF) application in the treatment of anal incontinence (AI) in women. METHODS: This was a pilot, randomized clinical trial conducted between January and October 2016. We enrolled women who consecutively attended the Attention Center of the Pelvic Floor (CAAP) with complaints of AI for more than six months. Nonablative RF was applied to the perianal region of the participants using Spectra G2 (Tonederm®, Rio Grande do Sul, Brazil). The reduced or complete elimination of the need for protective undergarments (diapers and absorbents) was considered a partial therapeutic response. RESULTS: Nine participants reported treatment satisfaction, while one reported dissatisfaction with the nonablative RF treatment of AI based on the Likert scale. No patient interrupted treatment sessions because of adverse effects, although adverse effects occurred in six participants. However, the clinical and physical examination of the participants with burning sensations showed no hyperemia or mucosal lesions. CONCLUSIONS: This study showed a promising reduction of fecal loss, participant satisfaction with treatment, and improved lifestyle, behavior, and depression symptoms with minimal adverse effects.