Mechanical activation with Easy Clean device enhanced organic tissue removal from simulated internal root resorption in a laboratory evaluation.

BACKGROUND: Considering the irregular shape of the root canal, removing inflamed pulp and granulation tissue completely from internal resorption cavities during chemomechanical preparation can be challenging. This study aimed to evaluate the effectiveness of passive ultrasonic irrigation (PUI) compared to mechanical activation with Easy Clean in the removal of organic tissue from simulated areas of internal root resorption. METHODS: The root canals of 72 extracted single-rooted teeth with oval canals were instrumented with Reciproc R25 instruments. After root canal preparations, the specimens were split longitudinally, and semicircular cavities were prepared using a round bur on each half of the roots. Samples obtained from bovine muscle tissue were weighed and adapted into semicircular cavities. The roots were reassembled and joined, and the teeth were divided into six groups (n = 12) according to the irrigation protocol: Sodium hypochlorite (NaOCl) without activation; NaOCl + PUI; NaOCl + Easy Clean; distilled water without activation; distilled water + PUI; and distilled water + Easy Clean. After irrigation protocols, the teeth were disassembled, and the remaining organic tissue was weighed. Data were analyzed by two-way ANOVA and Tukey's post hoc test (p < 0.05). RESULTS: None of the experimental protocols totally removed the bovine tissue from simulated cavities. Tissue weight reduction was significantly affected by the activation method (p < 0.05) and by irrigation solution (p < 0.05). Groups with NaOCl irrigation presented higher tissue weight loss when compared to distilled water, for all irrigation methods (p < 0.05). The use of Easy Clean resulted in the greatest tissue weight loss (42.0%-Distilled water/45.5%- NaOCl) compared to those of PUI (33.3%-Distilled water/37.7%- NaOCl) and no activation (33.4%-Distilled water/38.8%- NaOCl) (p < 0.05). However, no differences were observed between PUI and no activation groups (p > 0.05). CONCLUSIONS: Mechanical activation with Easy Clean enhanced organic tissue removal from simulated internal resorption more effectively than PUI. Easy Clean for agitation of the irrigating solution is effective in removing simulated organic tissues from artificial internal resorption cavities, being an alternative to the use of PUI.

Role of Goats in the Epidemiology of Coxiella burnetii.

Since its first description in the late 1930s, Q fever has raised many questions. Coxiella burnetii, the causative agent, is a zoonotic pathogen affecting a wide range of hosts. This airborne organism leads to an obligate, intracellular lifecycle, during which it multiplies in the mononuclear cells of the immune system and in the trophoblasts of the placenta in pregnant females. Although some issues about C. burnetii and its pathogenesis in animals remain unclear, over the years, some experimental studies on Q fever have been conducted in goats given their excretion pattern. Goats play an important role in the epidemiology and economics of C. burnetii infections, also being the focus of several epidemiological studies. Additionally, variants of the agent implicated in human long-term disease have been found circulating in goats. The purpose of this review is to summarize the latest research on C. burnetii infection and the role played by goats in the transmission of the infection to humans.

The first European consensus on principles of management for achondroplasia.

Achondroplasia is the most common type of skeletal dysplasia, caused by a recurrent pathogenic variant in the fibroblast growth factor receptor 3 (FGFR3). The management of achondroplasia is multifaceted, requiring the involvement of multiple specialties across the life course. There are significant unmet needs associated with achondroplasia and substantial differences in different countries with regard to delivery of care. To address these challenges the European Achondroplasia Forum (EAF), a network of senior clinicians and orthopaedic surgeons from Europe and the Middle East representative of the achondroplasia clinical community, came together with the overall aim of improving patient outcomes. The EAF developed a consensus on guiding principles of management of achondroplasia to provide a basis for developing optimal care in Europe. All members of the EAF were invited to submit suggestions for guiding principles of management, which were consolidated and then discussed during a meeting in December 2020. The group voted anonymously on the inclusion of each principle, with the requirement of a 75% majority at the first vote to pass the principle. A vote on the level of agreement was then held. A total of six guiding principles were developed, which cover management over the lifetime of a person with achondroplasia. The principles centre on the lifelong management of achondroplasia by an experienced multidisciplinary team to anticipate and manage complications, support independence, and improve quality of life. There is focus on timely referral to a physician experienced in the management of achondroplasia on suspicion of the condition, shared decision making, the goals of management, access to adaptive measures to enable those with achondroplasia to access their environment, and the importance of ongoing monitoring throughout adolescence and adulthood. All principles achieved the 75% majority required for acceptance at the first vote (range 91-100%) and a high level of agreement (range 8.5-9.6). The guiding principles of management for achondroplasia provide all healthcare professionals, patient advocacy groups and policy makers involved in the management of achondroplasia with overarching considerations when developing health systems to support the management of achondroplasia.

Hi-C Identifies Complex Genomic Rearrangements and TAD-Shuffling in Developmental Diseases.

Genome-wide analysis methods, such as array comparative genomic hybridization (CGH) and whole-genome sequencing (WGS), have greatly advanced the identification of structural variants (SVs) in the human genome. However, even with standard high-throughput sequencing techniques, complex rearrangements with multiple breakpoints are often difficult to resolve, and predicting their effects on gene expression and phenotype remains a challenge. Here, we address these problems by using high-throughput chromosome conformation capture (Hi-C) generated from cultured cells of nine individuals with developmental disorders (DDs). Three individuals had previously been identified as harboring duplications at the SOX9 locus and six had been identified with translocations. Hi-C resolved the positions of the duplications and was instructive in interpreting their distinct pathogenic effects, including the formation of new topologically associating domains (neo-TADs). Hi-C was very sensitive in detecting translocations, and it revealed previously unrecognized complex rearrangements at the breakpoints. In several cases, we observed the formation of fused-TADs promoting ectopic enhancer-promoter interactions that were likely to be involved in the disease pathology. In summary, we show that Hi-C is a sensible method for the detection of complex SVs in a clinical setting. The results help interpret the possible pathogenic effects of the SVs in individuals with DDs.

Autosomal-Recessive Mutations in MESD Cause Osteogenesis Imperfecta.

Osteogenesis imperfecta (OI) comprises a genetically heterogeneous group of skeletal fragility diseases. Here, we report on five independent families with a progressively deforming type of OI, in whom we identified four homozygous truncation or frameshift mutations in MESD. Affected individuals had recurrent fractures and at least one had oligodontia. MESD encodes an endoplasmic reticulum (ER) chaperone protein for the canonical Wingless-related integration site (WNT) signaling receptors LRP5 and LRP6. Because complete absence of MESD causes embryonic lethality in mice, we hypothesized that the OI-associated mutations are hypomorphic alleles since these mutations occur downstream of the chaperone activity domain but upstream of ER-retention domain. This would be consistent with the clinical phenotypes of skeletal fragility and oligodontia in persons deficient for LRP5 and LRP6, respectively. When we expressed wild-type (WT) and mutant MESD in HEK293T cells, we detected WT MESD in cell lysate but not in conditioned medium, whereas the converse was true for mutant MESD. We observed that both WT and mutant MESD retained the ability to chaperone LRP5. Thus, OI-associated MESD mutations produce hypomorphic alleles whose failure to remain within the ER significantly reduces but does not completely eliminate LRP5 and LRP6 trafficking. Since these individuals have no eye abnormalities (which occur in individuals completely lacking LRP5) and have neither limb nor brain patterning defects (both of which occur in mice completely lacking LRP6), we infer that bone mass accrual and dental patterning are more sensitive to reduced canonical WNT signaling than are other developmental processes. Biologic agents that can increase LRP5 and LRP6-mediated WNT signaling could benefit individuals with MESD-associated OI.

Author Correction: RSPO2 inhibition of RNF43 and ZNRF3 governs limb development independently of LGR4/5/6.

In this Letter, the surname of author Lena Vlaminck was misspelled 'Vlaeminck'. In addition, author Kris Vleminckx should have been associated with affiliation 16 (Center for Medical Genetics, Ghent University, Ghent, Belgium). These have been corrected online.

Phenolic compounds are highly correlated to the antioxidant capacity of genotypes of Oenocarpus distichus Mart. fruits.

This research aimed to evaluate 32 genotypes of Oenocarpus distichus fruits regarding the contents of total phenolic compounds, flavonoids, flavanols, monomeric anthocyanins, antioxidant capacity (ABTS and DPPH assays), and the phenolic compound profiles of the five genotypes that presented the highest yields of bioactive compounds. The genotypes were harvested in three different locations in Pará State, Northern Brazil, (Belém, São João do Araguaia and Marabá). Among the 32 genotypes, the highest bioactive compound contents and antioxidant capacity were found for three genotypes harvested in Belém (B-3, B-7 and B-8) and two harvested in São João do Araguaia (SJ-1 and SJ-4), and the total phenolic compounds varied from 131.97 to 363.01 mg gallic acid equivalent/100 g, total flavonoids from 24.23 to 38.19 mg quercetin equivalent/100 g, total flavanols from 72.29 to 259.18 mg catechin equivalent/100 g, and monomeric anthocyanins from 21.31 to 67.76 mg cyanidin 3-rutinoside/100 g. The main phenolic compounds tentatively identified in the five selected genotypes were cyanidin 3-O-rutinoside (48.47 to 196.51 µg/g), which could be identified and quantified as the major phenolic compound in Oenocarpus distichus fruits, for the first time, followed by chlorogenic acid (0.71 to 64.56 µg/g) and rutin (13.98 to 56.76 µg/g).

RSPO2 inhibition of RNF43 and ZNRF3 governs limb development independently of LGR4/5/6.

The four R-spondin secreted ligands (RSPO1-RSPO4) act via their cognate LGR4, LGR5 and LGR6 receptors to amplify WNT signalling1-3. Here we report an allelic series of recessive RSPO2 mutations in humans that cause tetra-amelia syndrome, which is characterized by lung aplasia and a total absence of the four limbs. Functional studies revealed impaired binding to the LGR4/5/6 receptors and the RNF43 and ZNRF3 transmembrane ligases, and reduced WNT potentiation, which correlated with allele severity. Unexpectedly, however, the triple and ubiquitous knockout of Lgr4, Lgr5 and Lgr6 in mice did not recapitulate the known Rspo2 or Rspo3 loss-of-function phenotypes. Moreover, endogenous depletion or addition of exogenous RSPO2 or RSPO3 in triple-knockout Lgr4/5/6 cells could still affect WNT responsiveness. Instead, we found that the concurrent deletion of rnf43 and znrf3 in Xenopus embryos was sufficient to trigger the outgrowth of supernumerary limbs. Our results establish that RSPO2, without the LGR4/5/6 receptors, serves as a direct antagonistic ligand to RNF43 and ZNRF3, which together constitute a master switch that governs limb specification. These findings have direct implications for regenerative medicine and WNT-associated cancers.

Digoxin Induces Cardiac Hypertrophy Without Negative Effects on Cardiac Function and Physical Performance in Trained Normotensive Rats.

Cardiotonic drugs and exercise training promote cardiac inotropic effects, which may affect training-induced cardiac adaptations. This study investigated the effects of long-term administration of digoxin on heart structure and function, and physical performance of rats submitted to high-intensity interval training (HIIT). Male Wistar rats, 60 days old, were divided into control (C), digoxin (DIGO), trained (T), and trained with digoxin (TDIGO). Digoxin was administered by gavage (30 µg/kg/day) for 75 days. The HIIT program consisted of treadmill running 60 min/day (8 min at 80% of the maximum speed (MS) and 2 min at 20% of the MS), 5 days per week during 60 days. The main cardiac parameters were evaluated by echocardiograph and cardiomyocyte area was determined by histology. There were no group x time effects of digoxin, HIIT or interactions (digoxin and HIIT) on functional echocardiographic parameters (heart rate; ejection fraction) or in the maximum exercise test. There was a group x time interaction, as evidenced by observed cardiac hypertrophy in the TDIGO group evaluated by ratio of left ventricle weight to body weight (p<0.002) and cardiomyocyte area (p<0.000002). Long-term administration of digoxin promoted cardiac hypertrophy without affecting cardiac function and physical performance in rats submitted to HIIT.

Mutations in EXTL3 Cause Neuro-immuno-skeletal Dysplasia Syndrome.

EXTL3 regulates the biosynthesis of heparan sulfate (HS), important for both skeletal development and hematopoiesis, through the formation of HS proteoglycans (HSPGs). By whole-exome sequencing, we identified homozygous missense mutations c.1382C>T, c.1537C>T, c.1970A>G, and c.2008T>G in EXTL3 in nine affected individuals from five unrelated families. Notably, we found the identical homozygous missense mutation c.1382C>T (p.Pro461Leu) in four affected individuals from two unrelated families. Affected individuals presented with variable skeletal abnormalities and neurodevelopmental defects. Severe combined immunodeficiency (SCID) with a complete absence of T cells was observed in three families. EXTL3 was most abundant in hematopoietic stem cells and early progenitor T cells, which is in line with a SCID phenotype at the level of early T cell development in the thymus. To provide further support for the hypothesis that mutations in EXTL3 cause a neuro-immuno-skeletal dysplasia syndrome, and to gain insight into the pathogenesis of the disorder, we analyzed the localization of EXTL3 in fibroblasts derived from affected individuals and determined glycosaminoglycan concentrations in these cells as well as in urine and blood. We observed abnormal glycosaminoglycan concentrations and increased concentrations of the non-sulfated chondroitin disaccharide D0a0 and the disaccharide D0a4 in serum and urine of all analyzed affected individuals. In summary, we show that biallelic mutations in EXTL3 disturb glycosaminoglycan synthesis and thus lead to a recognizable syndrome characterized by variable expression of skeletal, neurological, and immunological abnormalities.

Individual lithium disilicate crowns in a full-arch, implant-supported rehabilitation: a clinical report.

This clinical report presents the clinical outcome of a maxillary full-arch implant-supported fixed rehabilitation with lithium disilicate reinforced glass ceramic monolithic crowns opposing a mandibular metal-acrylic implant-supported fixed rehabilitation in a 62-year-old woman. Eight implants were successfully placed (four maxillary, four mandibular), and no complications occurred in the postoperative or maintenance periods. Six months after delivery, the maxillary and mandibular prostheses were found to be clinically, biologically, and mechanically stable, and the patient was satisfied with the esthetics and her ability to function. Although the present indications for the use of lithium disilicate are still restricted to tooth-borne restorations, it is possible to successfully rehabilitate edentulous patients through implant-supported fixed prostheses using lithium disilicate reinforced glass ceramic monolithic crowns.

Loss-of-function mutations in PTPN11 cause metachondromatosis, but not Ollier disease or Maffucci syndrome.

Metachondromatosis (MC) is a rare, autosomal dominant, incompletely penetrant combined exostosis and enchondromatosis tumor syndrome. MC is clinically distinct from other multiple exostosis or multiple enchondromatosis syndromes and is unlinked to EXT1 and EXT2, the genes responsible for autosomal dominant multiple osteochondromas (MO). To identify a gene for MC, we performed linkage analysis with high-density SNP arrays in a single family, used a targeted array to capture exons and promoter sequences from the linked interval in 16 participants from 11 MC families, and sequenced the captured DNA using high-throughput parallel sequencing technologies. DNA capture and parallel sequencing identified heterozygous putative loss-of-function mutations in PTPN11 in 4 of the 11 families. Sanger sequence analysis of PTPN11 coding regions in a total of 17 MC families identified mutations in 10 of them (5 frameshift, 2 nonsense, and 3 splice-site mutations). Copy number analysis of sequencing reads from a second targeted capture that included the entire PTPN11 gene identified an additional family with a 15 kb deletion spanning exon 7 of PTPN11. Microdissected MC lesions from two patients with PTPN11 mutations demonstrated loss-of-heterozygosity for the wild-type allele. We next sequenced PTPN11 in DNA samples from 54 patients with the multiple enchondromatosis disorders Ollier disease or Maffucci syndrome, but found no coding sequence PTPN11 mutations. We conclude that heterozygous loss-of-function mutations in PTPN11 are a frequent cause of MC, that lesions in patients with MC appear to arise following a "second hit," that MC may be locus heterogeneous since 1 familial and 5 sporadically occurring cases lacked obvious disease-causing PTPN11 mutations, and that PTPN11 mutations are not a common cause of Ollier disease or Maffucci syndrome.

Consumo máximo de oxigênio e fatores de risco cardiovascular em adultos jovens / Maximum uptake oxygen consumption and cardiovascular risk factors in young adults

Objetivos: Analisar a interação do consumo máximo de oxigênio com diferentes indicadores de risco cardiovascular entre adultos jovens do sexo masculino e sem histórico prévio de doenças. Métodos: Estudo transversal realizado com universitários. A amostra foi composta por 32 adultos jovens do sexo masculino e sem histórico prévio de doenças. Foram adotados como indicadores de risco: peso corporal, circunferência de cintura, percentual de gordura, glicemia em jejum, pressão arterial sistólica e diastólica. O consumo máximo de oxigênio foi avaliado de maneira direta em teste de esteira. A análise de variância e teste qui-quadrado analisaram os dados. Resultados: Menor consumo máximo de oxigênio associou-se com maior excesso de peso (p= 0,005), mas não com hipertensão (p= 0,059). Além disso, o mesmo associou-se com a menor ocorrência de riscos cardiovasculares. Conclusão: O consumo máximo de oxigênio é um indicador de risco mesmo entre adultos jovens sem diagnóstico prévio de doenças.

Objectives: To analyze the relationship between maximum oxygen uptake and cardiovascular risk factors among young male adults with no previously detected disease. Methods: Cross-sectional study with undergraduate subjects. The sample was composed by 32 young male adults without previously detected disease. Body weight, waist circumference, percentage body fat, fasting glucose, systolic and diastolic blood pressure were utilized as cardiovascular risk factors. Maximum oxygen uptake was assessed through treadmill test. Analysis of variance and chi-square test analyzed the data. Results: Lower maximum oxygen uptake was associated with higher overweight (p= 0,005), but not with arterial hypertension (p= 0,059). Moreover, the maximum oxygen uptake was also associated with lower occurrence of cardiovascular risk. Conclusion: The maximum oxygen uptake is an indicator of higher risk even among young adults without previously detected disease.