Efficacy of the methods of age determination using artificial intelligence in panoramic radiographs - a systematic review.

The aim of this systematic review is to analyze the literature to determine whether the methods of artificial intelligence are effective in determining age in panoramic radiographs. Searches without language and year limits were conducted in PubMed/Medline, Embase, Web of Science, and Scopus databases. Hand searches were also performed, and unpublished manuscripts were searched in specialized journals. Thirty-six articles were included in the analysis. Significant differences in terms of root mean square error and mean absolute error were found between manual methods and artificial intelligence techniques, favoring the use of artificial intelligence (p < 0.00001). Few articles compared deep learning methods with machine learning models or manual models. Although there are advantages of machine learning in data processing and deep learning in data collection and analysis, non-comparable data was a limitation of this study. More information is needed on the comparison of these techniques, with particular emphasis on time as a variable.

TGF-ß uncouples glycolysis and inflammation in macrophages and controls survival during sepsis.

Changes in metabolism of macrophages are required to sustain macrophage activation in response to different stimuli. We showed that the cytokine TGF-ß (transforming growth factor-ß) regulates glycolysis in macrophages independently of inflammatory cytokine production and affects survival in mouse models of sepsis. During macrophage activation, TGF-ß increased the expression and activity of the glycolytic enzyme PFKL (phosphofructokinase-1 liver type) and promoted glycolysis but suppressed the production of proinflammatory cytokines. The increase in glycolysis was mediated by an mTOR-c-MYC-dependent pathway, whereas the inhibition of cytokine production was due to activation of the transcriptional coactivator SMAD3 and suppression of the activity of the proinflammatory transcription factors AP-1, NF-κB, and STAT1. In mice with LPS-induced endotoxemia and experimentally induced sepsis, the TGF-ß-induced enhancement in macrophage glycolysis led to decreased survival, which was associated with increased blood coagulation. Analysis of septic patient cohorts revealed that the expression of PFKL, TGFBRI (which encodes a TGF-ß receptor), and F13A1 (which encodes a coagulation factor) in myeloid cells positively correlated with COVID-19 disease. Thus, these results suggest that TGF-ß is a critical regulator of macrophage metabolism and could be a therapeutic target in patients with sepsis.

A prospective non-fatal injuries assessment: A multivariate analysis in medical-legal examinations.

OBJECTIVE: To prospectively determine injury recovery time in the medical-legal examinations of non-fatal injuries and their associated factors, carried out by the National Institute of Legal Medicine and Forensic Sciences of Colombia to create a multivariate analysis. METHODS: A prospective medical-legal assessment of non-fatal injuries was carried out on 281 individuals with complete follow-up, in which the observational unit of analysis was the most serious injury. Variables, such as sex, circumstances of the injury, the mechanism that caused the injury, medical certificate of incapacity to work, among others were related to the injury recovery time, measured in days. The Kruskal Wallis (K-W) ANOVA and a multivariate analysis using the ordinal regression model were applied. RESULTS: In the multivariate analysis, the factors most associated with longer recovery time were the extent of joint damage (CR95%:1.47-5.94,p = 0.0001) and bone damage (CR95%:2.92-7.42,p < 0.001). In terms of circumstances of the injury, traffic accidents (CR95%:1.03-2.96,p < 0.001), medical-legal impairments (CR95%:0.34-2.19,p = 0.007), and complications of the primary injury (CR95%: 1.18-2.57,p < 0.001) had the greatest impact on recovery time. Others factors that significantly impacted injury recovery time are surgical treatments (IC95%: 0.33-3.26,p = 0.0164) and delayed treatment (CR95%:1.41-4.72,p < 0.001). A direct correlation (significant and moderately strong) was found between the recovery time of the injury and the days of incapacity for work (r = 0.802, p < 0.001). CONCLUSION: This prospective analysis determined which variables were most strongly related to the medical-legal assessment of non-fatal injuries and the recovery time of said injuries. Further studies aimed at improving the strategies to help individuals complete the legal process are required.

BMPR1A and BMPR1B Missense Mutations Cause Primary Ovarian Insufficiency.

CONTEXT: Primary ovarian insufficiency (POI) is a frequently occurring disorder affecting approximately 1% of women under 40 years of age. POI, which is characterized by the premature depletion of ovarian follicles and elevated plasma levels of follicle-stimulating hormone, leads to infertility. Although various etiological factors have been described, including chromosomal abnormalities and gene mutations, most cases remain idiopathic. OBJECTIVE: To identify and to functionally validate new sequence variants in 2 genes that play a key role in mammalian ovarian function, BMPR1A and BMPR1B (encoding for bone morphogenic protein receptor), leading to POI. METHODS: The impact on bone morphogenic protein (BMP) signaling of BMPR1A and BMPR1B variants, previously identified by whole-exome sequencing on 69 women affected by isolated POI, was established by different in vitro functional experiments. RESULTS: We demonstrate that the BMPR1A-p.Arg442His and BMPR1B-p.Phe272Leu variants are correctly expressed and located but lead to an impairment of downstream BMP signaling. CONCLUSION: In accordance with infertility observed in mice lacking Bmpr1a in the ovaries and in Bmpr1b-/- mice, our results unveil, for the first time, a link between BMPR1A and BMPR1B variants and the origin of POI. We show that BMP signaling impairment through specific BMPR1A and BMPR1B variants is a novel pathophysiological mechanism involved in human POI. We consider that BMPR1A and BMPR1B variants constitute genetic biomarkers of the origin of POI and have clinical utility.

Recomendaciones de la Academia Nacional de Medicina de Colombia para enfrentar los conflictos éticos secundarios a la crisis de COVID-19 en el inicio y mantenimiento de medidas de soporte vital avanzado / Recommendations of the National Academy of Medicine of Colombia to face the ethical conflicts secondary to the COVID-19 crisis in the initiation and maintenance of advanced life support measures

La profesión médica, fiel a su compromiso de buscar el beneficio de los pacientes y lograr los mejores resultados posibles en el proceso de atención, respetando sus derechos y voluntad, se enfrenta al reto de la pandemia de COVID-19 y la consecuente situación de emergencia sanitaria, la cual conlleva un estado de excepcionalidad que modifica de manera fundamental algunos elementos tradicionales de la práctica médica.

ATG7 and ATG9A loss-of-function variants trigger autophagy impairment and ovarian failure.

PURPOSE: Primary ovarian insufficiency (POI) is a frequent disorder that affects ~1% of women under 40 years of age. POI, which is characterized by the premature depletion of ovarian follicles and elevated plasma levels of follicle-stimulating hormone (FSH), leads to infertility. Although various etiological factors have been described, including chromosomal abnormalities and gene variants, most cases remain idiopathic. The aim of the present study was to identify and validate functionally new sequence variants in ATG (autophagy-related genes) leading to POI. METHODS: We have reanalyzed, in silico, the exome sequencing data from a previously reported work performed in 69 unrelated POI women. Functional experiments using a classical hallmark of autophagy, the microtubule-associated protein 1 light chain 3ß (LC3), were then used to link these genes to this lysosomal degradation pathway. RESULTS: We venture a functional link between ATG7 and ATG9A variants and POI. We demonstrated that variant ATG7 and ATG9A led to a decrease in autophagosome biosynthesis and consequently to an impairment of autophagy, a key biological process implicated in the preservation of the primordial follicles forming the ovarian reserve. CONCLUSION: Our results unveil that impaired autophagy is a novel pathophysiological mechanism involved in human POI.

Functional evidence implicating NOTCH2 missense mutations in primary ovarian insufficiency etiology.

Primary ovarian insufficiency (POI) is a frequently occurring disease affecting women under 40 years old. Recently, we have analyzed unrelated POI women via whole exome sequencing (WES) and identified NOTCH2 mutations underlying possible functional effects. The present study involved reanalyzing of WES assays. We used in the KGN granulosa-like cell model, a synthetic gene reporter construct driving luciferase gene expression to assess the functional effects of five NOTCH2 mutations identified in POI patients. We found that NOTCH2-p.Ser1804Leu, p.Ala2316Val, and p.Pro2359Ala mutations had a functional impact on the protein's transcriptional activity. The results have demonstrated for the first time that NOTCH2 mutations contribute to POI etiology. We therefore recommend sequencing NOTCH2's open reading frame in large panels of POI patients to establish an accurate genotype-phenotype correlation. We cannot rule out the fact that patients affected by Alagille syndrome carrying NOTCH2 mutations may suffer ovarian dysfunction.

New mutations in non-syndromic primary ovarian insufficiency patients identified via whole-exome sequencing.

STUDY QUESTION: Is it possible to identify new mutations potentially associated with non-syndromic primary ovarian insufficiency (POI) via whole-exome sequencing (WES)? SUMMARY ANSWER: WES is an efficient tool to study genetic causes of POI as we have identified new mutations, some of which lead to protein destablization potentially contributing to the disease etiology. WHAT IS KNOWN ALREADY: POI is a frequently occurring complex pathology leading to infertility. Mutations in only few candidate genes, mainly identified by Sanger sequencing, have been definitively related to the pathogenesis of the disease. STUDY DESIGN, SIZE, DURATION: This is a retrospective cohort study performed on 69 women affected by POI. PARTICIPANTS/MATERIALS, SETTING, METHODS: WES and an innovative bioinformatics analysis were used on non-synonymous sequence variants in a subset of 420 selected POI candidate genes. Mutations in BMPR1B and GREM1 were modeled by using fragment molecular orbital analysis. MAIN RESULTS AND THE ROLE OF CHANCE: Fifty-five coding variants in 49 genes potentially related to POI were identified in 33 out of 69 patients (48%). These genes participate in key biological processes in the ovary, such as meiosis, follicular development, granulosa cell differentiation/proliferation and ovulation. The presence of at least two mutations in distinct genes in 42% of the patients argued in favor of a polygenic nature of POI. LIMITATIONS, REASONS FOR CAUTION: It is possible that regulatory regions, not analyzed in the present study, carry further variants related to POI. WIDER IMPLICATIONS OF THE FINDINGS: WES and the in silico analyses presented here represent an efficient approach for mapping variants associated with POI etiology. Sequence variants presented here represents potential future genetic biomarkers. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by the Universidad del Rosario and Colciencias (Grants CS/CIGGUR-ABN062-2016 and 672-2014). Colciencias supported Liliana Catherine Patiño´s work (Fellowship: 617, 2013). The authors declare no conflict of interest.

A homozygous donor splice-site mutation in the meiotic gene MSH4 causes primary ovarian insufficiency.

Premature ovarian insufficiency (POI) is a frequent pathology that affects women under 40 years of age, characterized by an early cessation of menses and high FSH levels. Despite recent progresses in molecular diagnosis, the etiology of POI remains idiopathic in most cases. Whole-exome sequencing of members of a Colombian family affected by POI allowed us to identify a novel homozygous donor splice-site mutation in the meiotic gene MSH4 (MutS Homolog 4). The variant followed a strict mendelian segregation within the family and was absent in a cohort of 135 women over 50 years of age without history of infertility, from the same geographical region as the affected family. Exon trapping experiments showed that the splice-site mutation induced skipping of exon 17. At the protein level, the mutation p.Ile743_Lys785del is predicted to lead to the ablation of the highly conserved Walker B motif of the ATP-binding domain, thus inactivating MSH4. Our study describes the first MSH4 mutation associated with POI and increases the number of meiotic/DNA repair genes formally implicated as being responsible for this condition.

A potential functional association between mutant BMPR2 and primary ovarian insufficiency.

Primary ovarian insufficiency (POI) affects ~1% of women in the general population. Despite numerous attempts at identifying POI genetic aetiology, coding mutations in only a few genes have been functionally related to POI pathogenesis. It has been suggested that mutant BMPR2 might contribute towards the phenotype. Several BMP15 (a BMPR2 ligand) coding mutations in human species have been related to POI pathogenesis. The BMPR2 p.Ser987Phe mutation, previously identified in a woman with POI, might therefore lead to cellular dysfunction contributing to the phenotype. To explore such an assumption, the present study assessed potential pathogenic subcellular localization/aggregation patterns associated with the p.Ser987Phe mutant form of BMPR2 in a relevant model for studying ovarian function. A significant increase in protein-like aggregation patterns was identified at the endoplasmic reticulum (ER) which permitted us to establish, for the first time, a potential functional association between mutant BMPR2 and POI aetiology. Since BMPR2 mutant forms were previously related to idiopathic pulmonary arterial hypertension, BMPR2 mutations may be related to an as-yet-to-be described syndromic form of POI involving pulmonary dysfunction. Additional assays are necessary to confirm that BMPR2 abnormal subcellular patterns are composed by aggregates. ABBREVIATIONS: POI: primary ovarian insufficiency; ER: endoplasmic reticulum; NGS: next generation sequencing.

BMP15 Mutations Associated With Primary Ovarian Insufficiency Reduce Expression, Activity, or Synergy With GDF9.

CONTEXT: Bone morphogenetic protein (BMP)15 is an oocyte-specific growth factor, which, together with growth differentiation factor (GDF) 9, regulates folliculogenesis and ovulation rate. Multiple mutations in BMP15 have been identified in women with primary ovarian insufficiency (POI), supporting a pathogenic role; however, the underlying biological mechanism of many of these mutants remains unresolved. OBJECTIVES: To determine how mutations associated with ovarian dysfunction alter the biological activity of human BMP15. DESIGN: The effects of 10 mutations in BMP15 on protein production, activation of granulosa cells, and synergy with GDF9 were assessed. RESULTS: Sequencing of 35 patients with POI identified both an unrecognized BMP15 variant (c.986G>A, R329H) and a variant (c.581T>C, F194S) previously associated with the condition. Assessing expression and activity of these and 8 other BMP15 mutants identified: (1) multiple variants, including L148P, F194S, and Y235C, with reduced mature protein production; (2) three variants (R138H, A180T, and R329H) with ∼fourfold lower activity than wild-type BMP15; and (3) 3 variants (R68W, F194S, and N196K) with a significantly reduced ability to synergize with GDF9. CONCLUSIONS: Mutations in BMP15 associated with POI reduce mature protein production, activity, or synergy with GDF9. The latter effect is perhaps most interesting given that interactions with GDF9 most likely underlie the physiology of BMP15 in the human ovary.

Next generation sequencing in women affected by nonsyndromic premature ovarian failure displays new potential causative genes and mutations.

OBJECTIVE: To identify new molecular actors involved in nonsyndromic premature ovarian failure (POF) etiology. DESIGN: This is a retrospective case-control cohort study. SETTING: University research group and IVF medical center. PATIENT(S): Twelve women affected by nonsyndromic POF. The control group included 176 women whose menopause had occurred after age 50 and had no antecedents regarding gynecological disease. A further 345 women from the same ethnic origin (general population group) were also recruited to assess allele frequency for potentially deleterious sequence variants. INTERVENTION(S): Next generation sequencing (NGS), Sanger sequencing, and bioinformatics analysis. MAIN OUTCOME MEASURE(S): The complete coding regions of 70 candidate genes were massively sequenced, via NGS, in POF patients. Bioinformatics and genetics were used to confirm NGS results and to identify potential sequence variants related to the disease pathogenesis. RESULT(S): We have identified mutations in two novel genes, ADAMTS19 and BMPR2, that are potentially related to POF origin. LHCGR mutations, which might have contributed to the phenotype, were also detected. CONCLUSION(S): We thus recommend NGS as a powerful tool for identifying new molecular actors in POF and for future diagnostic/prognostic purposes.

Medición de la citotoxicidad y producción de citocinas en fibroblastos gingivales humanos estimulados con Mercurius-Heel®S: Un estudio piloto / Cytotoxicity measurement and cytokine production in human gingival fibroblasts stimulated with Mercurius-Heel®S: Pilot Study / Medição de citotoxidade e produção de citocinas em fibrobastos gengivais humanos estimulados com Mercurius-Heel®®S: Estudo piloto

El Mercurius-Heel®S es un medicamento homeopático utilizado para tratar patologías infecciosas como la gingivitis y la periodontitis; principalmente de la cavidad oral. El componente principal de este medicamento es el Mercurius solubilis. Se estableció el efecto citotóxico a través de la viabilidad celular o antiinflamatorio por medición de citocinas del Mercurius-Heel®S sobre fibroblastos gingivales humanos (FGH). Las células fueron tratadas con Mercurius-Heel®S a concentraciones desde 300 mg/ml hasta 0.0005 mg/ml y como control, células sin tratamiento. La viabilidad celular fue medida mediante la prueba colorimétrica MTS® y la expresión de citocinas (IL1b, TNFa e IL10) presentes en sobrenadantes celulares, fue realizada con Quantiquine® ELISA. Se observó incremento de la viabilidad celular significativamente mayor con respecto al control (concentraciones bajas del medicamento) a partir de 3,7 mg/ml hasta 0.0005 mg/ml. La producción de citocinas no mostró diferencias significativas a diferentes concentraciones de Mercurius-Heel®S. No se evidenció incremento significativo de IL1b, IL10 y TNFa y el Mercurius-Heel®S no mostró efecto citotóxico sobre FGH.

Mercurius-Heel®S is a homeopathic remedy used to treat infectious diseases such as gingivitis and periodontitis, mainly in the oral cavity. The main component of this drug is the Mercurius solubilis. Its cytotoxic effect was established based on cell viability and its anti-inflammatory effect by measuring the cytokines Mercurius-Heel®S on human gingival fibroblasts (FGH). We treated cells with Mercurius-Heel®S at concentrations of 300 mg/ml to 0.0005 mg/ml and left untreated cells as a control. Cell viability was measured using the MTS® colorimetric assay and the expression of cytokines (IL1b, TNFa e IL10) present in cell supernatants was performed with Quantiquine® ELISA. From 3.7 mg/ml to 0.0005 mg/ml, we observed a significantly higher increase in cell viability in comparison to the control (lower concentrations of the drug). Cytokine production was not significantly different at different concentrations of Mercurius-Heel®S. There was no significant increase in IL1b, IL10 and TNFa and Mercurius-Heel®S showed no cytotoxic effect on FGH.

Mercurius-Heel®S é um medicamento homeopático utilizado para tratar patologías infecciosas como la gengvite e a periodontite; principalmente da cavidade oral. O componente principal deste medicamento é o Mercurius solubilis. Estabeleceu-se o efeito citotóxico através da viabilidade celular a anti-inflamatório por medição de citocinas do Mercurius-Heel®S sobre fibroblastos gengivais humanos (FGH). As células foram tratadas com Mercurius-Heel®S a concentrações desde 300 mg/ml até 0.0005 mg/ml e como controlo, células sem tratamento. A viabilidade celular foi medida a prova colorimétrica MTS® e a expressão de citocinas (IL1b, TNFae IL10) presentes em sobrenadantes celulares, foi realizada com Quantiquine® ELISA. Observou-se incremento da viabilidade celular significativamente maior com respeito ao controlo (concentrações baixas do medicamento) a partir de 3,7 mg/ml até 0.0005 mg/ml. A produção de citocinas não mostrou diferenças significativas a diferentes concentrações de Mercurius-Heel®S. Não se evidenciou incremento significativo de IL1b, IL10 e TNFa e o Mercurius-Heel®S não mostrou efeito citotóxico sobre FGH.

Exome sequencing is an efficient tool for variant late-infantile neuronal ceroid lipofuscinosis molecular diagnosis.

The neuronal ceroid-lipofuscinoses (NCL) is a group of neurodegenerative disorders characterized by epilepsy, visual failure, progressive mental and motor deterioration, myoclonus, dementia and reduced life expectancy. Classically, NCL-affected individuals have been classified into six categories, which have been mainly defined regarding the clinical onset of symptoms. However, some patients cannot be easily included in a specific group because of significant variation in the age of onset and disease progression. Molecular genetics has emerged in recent years as a useful tool for enhancing NCL subtype classification. Fourteen NCL genetic forms (CLN1 to CLN14) have been described to date. The variant late-infantile form of the disease has been linked to CLN5, CLN6, CLN7 (MFSD8) and CLN8 mutations. Despite advances in the diagnosis of neurodegenerative disorders mutations in these genes may cause similar phenotypes, which rends difficult accurate candidate gene selection for direct sequencing. Three siblings who were affected by variant late-infantile NCL are reported in the present study. We used whole-exome sequencing, direct sequencing and in silico approaches to identify the molecular basis of the disease. We identified the novel c.1219T>C (p.Trp407Arg) and c.1361T>C (p.Met454Thr) MFSD8 pathogenic mutations. Our results highlighted next generation sequencing as a novel and powerful methodological approach for the rapid determination of the molecular diagnosis of NCL. They also provide information regarding the phenotypic and molecular spectrum of CLN7 disease.

BMP15 c.-9C>G promoter sequence variant may contribute to the cause of non-syndromic premature ovarian failure.

BMP15 has drawn particular attention in the pathophysiology of reproduction, as its mutations in mammalian species have been related to different reproductive phenotypes. In humans, BMP15 coding regions have been sequenced in large panels of women with premature ovarian failure (POF), but only some mutations have been definitely validated as causing the phenotype. A functional association between the BMP15 c.-9C>G promoter polymorphism and cause of POF have been reported. The aim of this study was to determine the potential functional effect of this sequence variant on specific BMP15 promoter transactivation disturbances. Bioinformatics was used to identify transcription factor binding sites located on the promoter region of BMP15. Reverse transcription polymerase chain reaction was used to study specific gene expression in ovarian tissue. Luciferase reporter assays were used to establish transactivation disturbances caused by the BMP15 c.-9C>G variant. The c.-9C>G variant was found to modify the PITX1 transcription factor binding site. PITX1 and BMP15 co-expressed in human and mouse ovarian tissue, and PITX1 transactivated both BMP15 promoter versions (-9C and -9G). It was found that the BMP15 c.-9G allele was related to BMP15 increased transcription, supporting c.-9C>G as a causal agent of POF.

Plasmodium falciparum rhoptry neck protein 5 peptides bind to human red blood cells and inhibit parasite invasion.

Plasmodium falciparum malaria parasite invasion of erythrocytes is an essential step in host infection and the proteins involved in such invasion are the main target in developing an antimalarial vaccine. Secretory organelle-derived proteins (micronemal AMA1 protein and the RON2, 4, and 5 rhoptry neck proteins) have been recently described as components of moving junction complex formation allowing merozoites to move into a newly created parasitophorous vacuole. This study led to identifying RON5 regions involved in binding to human erythrocytes by using a highly robust, sensitive and specific receptor-ligand interaction assay; it is further shown that the RON5 protein remains highly conserved throughout different parasite strains. It is shown that the binding peptide-erythrocyte interaction is saturable and sensitive to chymotrypsin and trypsin. Invasion inhibition assays using erythrocyte binding peptides showed that the RON5-erythrocyte interaction could be critical for merozoite invasion of erythrocytes. This work provides evidence (for the first time) suggesting a fundamental role for RON5 in erythrocyte invasion.

Ethanol exposure disrupts extraembryonic microtubule cytoskeleton and embryonic blastomere cell adhesion, producing epiboly and gastrulation defects.

Fetal alcohol spectrum disorder (FASD) occurs when pregnant mothers consume alcohol, causing embryonic ethanol exposure and characteristic birth defects that include craniofacial, neural and cardiac defects. Gastrulation is a particularly sensitive developmental stage for teratogen exposure, and zebrafish is an outstanding model to study gastrulation and FASD. Epiboly (spreading blastomere cells over the yolk cell), prechordal plate migration and convergence/extension cell movements are sensitive to early ethanol exposure. Here, experiments are presented that characterize mechanisms of ethanol toxicity on epiboly and gastrulation. Epiboly mechanisms include blastomere radial intercalation cell movements and yolk cell microtubule cytoskeleton pulling the embryo to the vegetal pole. Both of these processes were disrupted by ethanol exposure. Ethanol effects on cell migration also indicated that cell adhesion was affected, which was confirmed by cell aggregation assays. E-cadherin cell adhesion molecule expression was not affected by ethanol exposure, but E-cadherin distribution, which controls epiboly and gastrulation, was changed. E-cadherin was redistributed into cytoplasmic aggregates in blastomeres and dramatically redistributed in the extraembryonic yolk cell. Gene expression microarray analysis was used to identify potential causative factors for early development defects, and expression of the cell adhesion molecule protocadherin-18a (pcdh18a), which controls epiboly, was significantly reduced in ethanol exposed embryos. Injecting pcdh18a synthetic mRNA in ethanol treated embryos partially rescued epiboly cell movements, including enveloping layer cell shape changes. Together, data show that epiboly and gastrulation defects induced by ethanol are multifactorial, and include yolk cell (extraembryonic tissue) microtubule cytoskeleton disruption and blastomere adhesion defects, in part caused by reduced pcdh18a expression.

Efecto del Mercurius-Heel®s sobre la citotoxicidad de fibroblastos gingivales humanos en un modelo in vitro / Effect of Mercurius-Heel®s on the cytotoxicity of human gingival fibroblasts in an in vitro model

La enfermedad periodontal es una patología de origen infeccioso, caracterizada por ocasionar secuelas destructivas al tejido de soporte del diente, y cuyo tratamiento va encaminado a la destrucción de los agentes etiológicos y a la regeneración periodontal. Una alternativa es el uso de agentes homeopáticos ya que son naturales y se administran a muy bajas concentraciones, uno de ellos es el Mercurius Heel® S como coadyuvante en enfermedades infecciosas. En este trabajo se presentan los resultados de una investigación cuyo objetivo fue evaluar el efecto del Mercurius Heel® S en la viabilidad de fibroblastos gingivales humanos, que fueron sometidos a tratamiento con Mercurius Heel® S durante 15 minutos y dos horas a concentraciones desde 300mg hasta 0.00006mg. Transcurridos esos tiempos, se retiró el tratamiento y las células fueron mantenidas durante 24, 48 y 72 horas más. Posteriormente se realizó un ensayo colorimétrico de viabilidad y proliferación celular denominado MTS de promega® Los fibroblastos gingivales humanos tratados con Mercurius Heel® S mostraron un aumento en la proliferación celular comparada con las células no tratadas. Bajas concentraciones del medicamento 0,0001mg y 0,00006mg mostraron una mayor proliferación observando diferencias estadísticamente significativas. El tratamiento a 15 minutos mostró mejores resultados con respecto al tratamiento de 2 horas con diferencias estadísticamente significativas también. Finalmente el efecto del Mercurius Heel® se mantuvo hasta las primeras 48 horas. Considerando lo anterior, el Mercurius Heel® no presentó ningún efecto citotóxico en los fibroblastos gingivales; por el contrario, las células proliferaron, lo que sugiere su utilidad como tratamiento complementario en la enfermedad periodontal.

Periodontal disease is pathology of infectious origin, characterized because it causes destructive consequences to the supporting tissue of the tooth, its treatment is aimed at the destruction of the etiologic agents and to the periodontal regeneration. An alternative is the use of homeopathic agents because they are natural and are managed to very low concentrations, one of them is the Mercurius Heel ® S as an adjunctive therapy in infectious diseases. In this work, we present the results of an investigation whose objective was to evaluate the effect of Mercurius Heel® S on the viability of human gingival fibroblasts, which were undergoing treatment with Mercurius Heel® S for 15 minutes and two hours to concentrations from 300mg to 0.00006mg. After these times, the treatment was removed and the cells were maintained for 24, 48, and 72 more hours. It was followed by a colorimetric assay for cell viability and proliferation called MTS of Promega ®. Human gingival fibroblasts treated with Mercurius Heel® S showed an increase in cell proliferation compared with the untreated cells. Low medication concentrations of 0.0001 mg and 0.00006 mg showed a greater proliferation showed greater statistically differences. Finally, the effect of Mercurius Heel® was maintained for the first 48 hours. Considering the above, the Mercurius Heel® did not provide any cytotoxic effect on human gingival fibroblasts; on the contrary, the cells proliferated, suggesting its usefulness as supplementary treatment for periodontal disease.

A novel familial case of diffuse leukodystrophy related to NDUFV1 compound heterozygous mutations.

NDUFV1 mutations have been related to encephalopathic phenotypes due to mitochondrial energy metabolism disturbances. In this study, we report two siblings affected by a diffuse leukodystrophy, who carry the NDUFV1 c.1156C>T (p.Arg386Cys) missense mutation and a novel 42-bp deletion. Bioinformatic and molecular analysis indicated that this deletion lead to the synthesis of mRNA molecules carrying a premature stop codon, which might be degraded by the nonsense-mediated decay system. Our results add information on the molecular basis and the phenotypic features of mitochondrial disease caused by NDUFV1 mutations.