Condiciones de salud enfermedad bucal en adolescentes embarazadas indígenas y no indígenas de Maracaibo-Venezuela / Health conditions oral disease in indigenous and non-indigenous pregnant adolescents from Maracaibo- Venezuela

RESUMEN Objetivo : Comparar las condiciones de salud enfermedad bucal en adolescentes embarazadas indígenas y no indígenas de Maracaibo-Venezuela. Material y Métodos : Fue un estudio epidemiológico, transversal con un diseño de campo, no experimental. La muestra fue de 121 adolescentes embarazadas de la consulta prenatal del Hospital Materno Infantil "Eduardo Soto Peña" y de la Maternidad "Dr. Armando Castillo Plaza" de Maracaibo. Se realizó examen clínico según los criterios de la Organización Mundial de la Salud, se midieron los índices CPO, ISC, IHO-S e IPC. Para procesar los datos se utilizó el software SPSS® versión 25 y se aplicó tanto prueba de t para muestras independientes como Chi cuadrado. Resultados : La experiencia de caries fue 91,7%, el índice CPO fue de 6,84±4,173 y el ISC 11,42±2,374. Para la condición periodontal, el 41,3% presentó condición sana, un 53,7% tenía gingivitis y el 5% presentó periodontitis. El grupo de embarazadas no indígenas tuvo un índice CPO más alto 7,19±4,066 en comparación con un 5,52±4,398 para las indígenas. Respecto a la condición periodontal consolidada, el 52% del grupo de indígenas presentó gingivitis y el resto resultó sana y entre las no indígenas el 54% presentó gingivitis, el 6% tenía periodontitis y el 40% estaba sana. Conclusiones : En este estudio, no se observaron diferencias entre las condiciones de salud enfermedad bucal de las adolescentes embarazadas indígenas y no indígenas. Sin embargo, es necesario dar prioridad a la atención del componente bucal de las adolescentes durante el embarazo.

ABSTRACT Objective: Compare the oral disease health conditions in indigenous and non-indigenous pregnant adolescents from Maracaibo-Venezuela. Material and Methods : It was an epidemiological, cross-sectional study with a field design, not experimental. The sample was 121 pregnant adolescents from the prenatal consultation of the Maternal and Child Hospital "Eduardo Soto Peña" and the Maternity "Dr. Armando Castillo Plaza" in Maracaibo. Clinical examination was performed according to the criteria of the World Health Organization, DMFT, SCI, SOH-I and PCI indexes were measured. SPSS® version 25 software was used to process the data and both the t-test for independent samples and Chi square were applied. Results : the caries experience was 91.7%, the DMFT index was 6.84 ± 4.173 and the SCI was 11.42 ± 2.374. For the periodontal condition, 41.3% had a healthy condition, 53.7% had gingivitis and 5% had periodontitis. The group of non-indigenous pregnant women had a higher DMFT index of 7.19 ± 4.066 compared to 5.52 ± 4.398 for indigenous women. Regarding the consolidated periodontal condition, 52% of the indigenous group presented gingivitis and the rest were healthy and among the non-indigenous 54% presented gingivitis, 6% had periodontitis and 40% was healthy. Conclusion : In this study, no differences were observed between the oral disease health conditions of indigenous and non-indigenous pregnant adolescents. However, it is necessary to give priority to the attention of the oral component of adolescent girls during pregnancy.

Ceramide Imbalance and Impaired TLR4-Mediated Autophagy in BMDM of an ORMDL3-Overexpressing Mouse Model.

Increased orosomucoid-like 3 (ORMDL3) expression levels, due to single nucleotide polymorphisms (SNPs), have been associated with several inflammatory diseases, including asthma and inflammatory bowel diseases. ORMDL proteins inhibit serine palmitoyltransferase (SPT), the first rate-limiting enzyme in de novo sphingolipid synthesis and alter cellular calcium homeostasis. Both processes are essential for immune response. The present study addresses ORMDL3 protein involvement in macrophage physiology using an overexpressing knock-in mouse model. Ceramide content was notably different in the bone-marrow-derived macrophages (BMDM) from the transgenic mouse model compared with the wild type (WT) macrophages. Our data revealed an alteration of de novo production of sphinganine upon BMDM activation in the transgenic mouse. Gene-expression analysis showed that alteration in ORMDL3 expression levels did not affect activation or macrophage polarization. Nevertheless, we studied phagocytosis and autophagy-crucial processes that are dependent on lipid membrane composition. Phagocytosis in transgenic macrophages was not affected by ORMDL3 overexpression, but we did find a reduction in toll-like receptor 4 (TLR-4)-mediated autophagy. Both genetic and functional studies have pointed to autophagy as an essential pathway involved in inflammation. We believe that our work provides new insights into the functional link between ORMDL3 expression and inflammatory diseases.

Correction: Natural Killer Cell Receptors and Cytotoxic Activity in Phosphomannomutase 2 Deficiency (PMM2-CDG).

[This corrects the article DOI: 10.1371/journal.pone.0158863.].

Dihydroceramide accumulation mediates cytotoxic autophagy of cancer cells via autolysosome destabilization.

Autophagy is considered primarily a cell survival process, although it can also lead to cell death. However, the factors that dictate the shift between these 2 opposite outcomes remain largely unknown. In this work, we used Δ9-tetrahydrocannabinol (THC, the main active component of marijuana, a compound that triggers autophagy-mediated cancer cell death) and nutrient deprivation (an autophagic stimulus that triggers cytoprotective autophagy) to investigate the precise molecular mechanisms responsible for the activation of cytotoxic autophagy in cancer cells. By using a wide array of experimental approaches we show that THC (but not nutrient deprivation) increases the dihydroceramide:ceramide ratio in the endoplasmic reticulum of glioma cells, and this alteration is directed to autophagosomes and autolysosomes to promote lysosomal membrane permeabilization, cathepsin release and the subsequent activation of apoptotic cell death. These findings pave the way to clarify the regulatory mechanisms that determine the selective activation of autophagy-mediated cancer cell death.

Natural Killer Cell Receptors and Cytotoxic Activity in Phosphomannomutase 2 Deficiency (PMM2-CDG).

BACKGROUND: PMM2-CDG is the most common N-glycosylation defect and shows an increased risk of recurrent and/or severe, sometimes fatal, infections in early life. We hypothesized that natural killer (NK) cells, as important mediators of the immune response against microbial pathogens and regulators of adaptive immunity, might be affected in this genetic disorder. OBJECTIVE: To evaluate possible defects on PMM2-CDG NK peripheral blood cell number, killing activity and expression of membrane receptors. METHODS: We studied fresh and activated NK cells from twelve PMM2-CDG cells. The number and expression of lymphoid surface receptors were studied by flow cytometry. The NK responsiveness (frequency of degranulated NK cells) and killing activity against K562 target cells was determined in the NK cytotoxicity assay. RESULTS: We found an increase of blood NK cells in three patients with a severe phenotype. Two of them, who had suffered from moderate/severe viral infections during their first year of life, also had reduced T lymphocyte numbers. Patient activated NK cells showed increased expression of CD54 adhesion molecule and NKG2D and NKp46 activating receptors. NKp46 and 2B4 expression was inversely correlated with the expression of NKG2D in activated PMM2-CDG cells. Maximal NK activity against K562 target cells was similar in control and PMM2-CDG cells. Interestingly, the NK cell responsiveness was higher in patient cells. NKG2D and specially CD54 increased surface expression significantly correlated with the increased NK cell cytolytic activity according to the modulation of the killer activity by expression of triggering receptors and adhesion molecules. CONCLUSIONS: Our results indicate that hypoglycosylation in PMM2-CDG altered NK cell reactivity against target cells and the expression of CD54 and NKG2D, NKp46 and 2B4 activating receptors during NK cell activation. This suggests a defective control of NK cell killing activity and the overall anti-viral immune response in PMM2-CDG patients. The present work improves our understanding of the immunological functions in PMM2-CDG and possibly in other CDG-I types.

Coordinated regulation of the orosomucoid-like gene family expression controls de novo ceramide synthesis in mammalian cells.

The orosomucoid-like (ORMDL) protein family is involved in the regulation of de novo sphingolipid synthesis, calcium homeostasis, and unfolded protein response. Single nucleotide polymorphisms (SNPs) that increase ORMDL3 expression have been associated with various immune/inflammatory diseases, although the pathophysiological mechanisms underlying this association are poorly understood. ORMDL proteins are claimed to be inhibitors of the serine palmitoyltransferase (SPT). However, it is not clear whether individual ORMDL expression levels have an impact on ceramide synthesis. The present study addressed the interaction with and regulation of SPT activity by ORMDLs to clarify their pathophysiological relevance. We have measured ceramide production in HEK293 cells incubated with palmitate as a direct substrate for SPT reaction. Our results showed that a coordinated overexpression of the three isoforms inhibits the enzyme completely, whereas individual ORMDLs are not as effective. Immunoprecipitation and fluorescence resonance energy transfer (FRET) studies showed that mammalian ORMDLs form oligomeric complexes that change conformation depending on cellular sphingolipid levels. Finally, using macrophages as a model, we demonstrate that mammalian cells modify ORMDL genes expression levels coordinately to regulate the de novo ceramide synthesis pathway. In conclusion, we have shown a physiological modulation of SPT activity by general ORMDL expression level regulation. Moreover, because single ORMDL3 protein alteration produces an incomplete inhibition of SPT activity, this work argues against the idea that ORMDL3 pathophysiology could be explained by a simple on/off mechanism on SPT activity.

GPI-anchor and GPI-anchored protein expression in PMM2-CDG patients.

BACKGROUND: Mutations in PMM2 impair phosphomannomutase-2 activity and cause the most frequent congenital disorder of glycosylation, PMM2-CDG. Mannose-1-phosphate, that is deficient in this disorder, is also implicated in the biosynthesis of glycosylphosphatidyl inositol (GPI) anchors. OBJECTIVE: To evaluate whether GPI-anchor and GPI-anchored proteins are defective in PMM2-CDG patients. METHODS: The expression of GPI-anchor and seven GPI-anchored proteins was evaluated by flow cytometry in different cell types from twelve PMM2-CDG patients. Additionally, neutrophil CD16 and plasma hepatic proteins were studied by Western blot. Transferrin glycoforms were evaluated by HPLC. RESULTS: Patients and controls had similar surface expression of GPI-anchor and most GPI-anchored proteins. Nevertheless, patients displayed a significantly diminished binding of two anti-CD16 antibodies (3G8 and KD1) to neutrophils and also of anti-CD14 (61D3) to monocytes. Interestingly, CD16 immunostaining and asialotransferrin levels significantly correlated with patients' age. Analysis by flow cytometry of CD14 with MΦP9, and CD16 expression in neutrophils by Western blot using H-80 ruled out deficiencies of these antigens. CONCLUSIONS: PMM2 mutations do not impair GPI-anchor or GPI-anchored protein expression. However, the glycosylation anomalies caused by PMM2 mutations might affect the immunoreactivity of monoclonal antibodies and lead to incorrect conclusions about the expression of different proteins, including GPI-anchored proteins. Neutrophils and monocytes are sensitive to PMM2 mutations, leading to abnormal glycosylation in immune receptors, which might potentially affect their affinity to their ligands, and contribute to infection. This study also confirms less severe hypoglycosylation defects in older PMM2-CDG patients.